CN110201005A - The synthetic method of the visualization guidance tumour combined immunization treatment nanometer formulation of Gd:CuS mineralising sendai virus - Google Patents
The synthetic method of the visualization guidance tumour combined immunization treatment nanometer formulation of Gd:CuS mineralising sendai virus Download PDFInfo
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- 206010028980 Neoplasm Diseases 0.000 title claims abstract description 26
- 241000711408 Murine respirovirus Species 0.000 title claims abstract description 17
- 238000009472 formulation Methods 0.000 title claims abstract description 7
- 239000000203 mixture Substances 0.000 title claims abstract description 7
- 238000002649 immunization Methods 0.000 title claims description 8
- 230000003053 immunization Effects 0.000 title claims description 8
- 238000010189 synthetic method Methods 0.000 title claims description 8
- 238000012800 visualization Methods 0.000 title claims description 5
- 238000006243 chemical reaction Methods 0.000 claims abstract description 17
- 239000002105 nanoparticle Substances 0.000 claims abstract description 14
- 239000007864 aqueous solution Substances 0.000 claims abstract description 12
- 229910003317 GdCl3 Inorganic materials 0.000 claims abstract description 6
- MEANOSLIBWSCIT-UHFFFAOYSA-K gadolinium trichloride Chemical compound Cl[Gd](Cl)Cl MEANOSLIBWSCIT-UHFFFAOYSA-K 0.000 claims abstract description 6
- 238000005580 one pot reaction Methods 0.000 claims abstract description 6
- 239000000243 solution Substances 0.000 claims description 48
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 30
- 229910021591 Copper(I) chloride Inorganic materials 0.000 claims description 15
- OXBLHERUFWYNTN-UHFFFAOYSA-M copper(I) chloride Chemical compound [Cu]Cl OXBLHERUFWYNTN-UHFFFAOYSA-M 0.000 claims description 15
- XLYOFNOQVPJJNP-ZSJDYOACSA-N heavy water Substances [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 claims description 15
- 229910021642 ultra pure water Inorganic materials 0.000 claims description 15
- 239000012498 ultrapure water Substances 0.000 claims description 15
- 239000000843 powder Substances 0.000 claims description 11
- 238000004090 dissolution Methods 0.000 claims description 6
- BMPYETKQLHJVDE-UHFFFAOYSA-N O.O.O.O.O.O.[Gd] Chemical class O.O.O.O.O.O.[Gd] BMPYETKQLHJVDE-UHFFFAOYSA-N 0.000 claims description 5
- 238000013019 agitation Methods 0.000 claims description 5
- MPTQRFCYZCXJFQ-UHFFFAOYSA-L copper(II) chloride dihydrate Chemical compound O.O.[Cl-].[Cl-].[Cu+2] MPTQRFCYZCXJFQ-UHFFFAOYSA-L 0.000 claims description 5
- 150000002500 ions Chemical class 0.000 claims description 5
- 238000003760 magnetic stirring Methods 0.000 claims description 5
- 239000011734 sodium Substances 0.000 claims description 5
- 238000003756 stirring Methods 0.000 claims description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 5
- 239000003795 chemical substances by application Substances 0.000 claims description 3
- CYPRMUMKDSHJER-UHFFFAOYSA-N O.O.O.O.O.O.O.O.O.[Na] Chemical compound O.O.O.O.O.O.O.O.O.[Na] CYPRMUMKDSHJER-UHFFFAOYSA-N 0.000 claims description 2
- 238000004458 analytical method Methods 0.000 claims 1
- 238000009738 saturating Methods 0.000 claims 1
- 238000000034 method Methods 0.000 abstract description 14
- 230000000694 effects Effects 0.000 abstract description 5
- 238000003384 imaging method Methods 0.000 abstract description 5
- 238000002560 therapeutic procedure Methods 0.000 abstract description 4
- 238000002360 preparation method Methods 0.000 abstract description 3
- 238000004088 simulation Methods 0.000 abstract description 3
- 230000009885 systemic effect Effects 0.000 abstract description 3
- 230000001225 therapeutic effect Effects 0.000 abstract description 3
- 230000004913 activation Effects 0.000 abstract description 2
- 230000036039 immunity Effects 0.000 abstract description 2
- 229910021592 Copper(II) chloride Inorganic materials 0.000 abstract 1
- ORTQZVOHEJQUHG-UHFFFAOYSA-L copper(II) chloride Chemical compound Cl[Cu]Cl ORTQZVOHEJQUHG-UHFFFAOYSA-L 0.000 abstract 1
- 238000007626 photothermal therapy Methods 0.000 description 5
- 210000004027 cell Anatomy 0.000 description 4
- 238000002604 ultrasonography Methods 0.000 description 4
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 201000011510 cancer Diseases 0.000 description 3
- 229910052708 sodium Inorganic materials 0.000 description 3
- 229910052979 sodium sulfide Inorganic materials 0.000 description 3
- GRVFOGOEDUUMBP-UHFFFAOYSA-N sodium sulfide (anhydrous) Chemical compound [Na+].[Na+].[S-2] GRVFOGOEDUUMBP-UHFFFAOYSA-N 0.000 description 3
- 230000007812 deficiency Effects 0.000 description 2
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- 210000004881 tumor cell Anatomy 0.000 description 2
- 102000005606 Activins Human genes 0.000 description 1
- 108010059616 Activins Proteins 0.000 description 1
- 206010050685 Cytokine storm Diseases 0.000 description 1
- 102000003886 Glycoproteins Human genes 0.000 description 1
- 108090000288 Glycoproteins Proteins 0.000 description 1
- 101710154606 Hemagglutinin Proteins 0.000 description 1
- 241000167880 Hirundinidae Species 0.000 description 1
- 102000014150 Interferons Human genes 0.000 description 1
- 108010050904 Interferons Proteins 0.000 description 1
- 102000015696 Interleukins Human genes 0.000 description 1
- 108010063738 Interleukins Proteins 0.000 description 1
- 244000134336 Malus baccata Species 0.000 description 1
- 235000005079 Malus baccata Nutrition 0.000 description 1
- 102000005348 Neuraminidase Human genes 0.000 description 1
- 108010006232 Neuraminidase Proteins 0.000 description 1
- 108090001074 Nucleocapsid Proteins Proteins 0.000 description 1
- 101710093908 Outer capsid protein VP4 Proteins 0.000 description 1
- 101710135467 Outer capsid protein sigma-1 Proteins 0.000 description 1
- 102000007982 Phosphoproteins Human genes 0.000 description 1
- 108010089430 Phosphoproteins Proteins 0.000 description 1
- 101710176177 Protein A56 Proteins 0.000 description 1
- 210000001744 T-lymphocyte Anatomy 0.000 description 1
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 1
- 239000000488 activin Substances 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 230000005784 autoimmunity Effects 0.000 description 1
- 230000003592 biomimetic effect Effects 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
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- 238000009169 immunotherapy Methods 0.000 description 1
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- 229940047122 interleukins Drugs 0.000 description 1
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- 102000003390 tumor necrosis factor Human genes 0.000 description 1
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/76—Viruses; Subviral particles; Bacteriophages
- A61K35/768—Oncolytic viruses not provided for in groups A61K35/761 - A61K35/766
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
- A61K41/0052—Thermotherapy; Hyperthermia; Magnetic induction; Induction heating therapy
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
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Abstract
The present invention discloses a kind of method that the tactful human simulation by Gd:CuS mineralising sendai virus (SeV@Gd:CuS) successfully synthesizes tumor combined therapeutic nanometer formulation.Including 1) CuCl2·2H2The preparation of O aqueous solution；2)GdCl3·6H2The preparation of O aqueous solution；3) one pot process SeV@Gd:CuS nano particle.Structure is complicated for sendai virus, can systemic activation body's immunity, comprehensive killing and removing are caused to tumour.SeV@Gd:CuS nano particle has significant T1 weighted magnetic resonance/optoacoustic Double-mode imaging ability, it is higher that near-infrared laser irradiates lower photothermal conversion efficiency, subsequent tumor thermal therapy can be efficiently guided, and there is excellent biocompatibility, generation without side-effects.
Description
Technical field
It is successfully closed the present invention relates to a kind of by the tactful human simulation of Gd:CuS mineralising sendai virus (SeV@Gd:CuS)
At visualization guidance
Background technique
Currently, tumour is to threaten one of maximum disease to the mankind in the world.Immunization therapy passes through human activin siberian crabapple
System kills cancer cell and tumor tissues by autoimmunity function, to have the function that treating cancer.Therefore, immunization therapy
It is one for the treatment of tumour most efficient method.In view of sendai virus, structure is complicated, contains nucleocapsid protein, phosphoprotein, matrix egg
The ingredients such as white, glycoprotein, fusion protein, coating, hemagglutinin, neuraminidase and ss-RNA, thus its as foreign matter in vivo
Systemic immune system can be excited to generate cytokine storm (such as interferon, tumor necrosis factor and various interleukins
Deng), and make the Dendritic Cells (DC cell offers tumour antigen) played a significant role during antineoplaston, macrophage
Cell (Cell swallows tumour cell) and natural killer cells (NK cell, killing tumor cell) etc. largely activated, then
Greatly improve the increment efficiency of T cell.This composite Nano preparation based on sendai virus can systemic activation immunity of organism function
Can, comprehensive killing and removing are caused to tumour, relative to traditional tumour treatment method, which increase oncotherapy more
Effect.
Photo-thermal therapy (photothermal therapy, PTT) is a kind of novel tumor treatment method, in external near-infrared
Under the irradiation of light, the light thermit powder of tumor locus absorbs near infrared light and is translated into heat, increases tumor locus temperature rapidly
To 48 DEG C or more, cancer cell can be killed in a few minutes.The side effect that photo-thermal therapy process generates is smaller, and whole body system toxicity is low,
Will not normal tissue cause to damage, have very big clinical application potentiality.If by photo-thermal therapy and immunotherapy of tumors phase
In conjunction with, will greatly enhance oncotherapy effect, keep oncotherapy more thorough.
In view of deficiency existing for single mode imaging technique, rising in recent years multi-modality imaging technology can be by concentration side
Formula combines, and realization has complementary functions, thus accurate diagnosis comprehensive to tumour.And at present about the nanometer of multi-modal imaging
Particle is also constantly being found, is preparing and is applying.
As a kind of available strategy for preparing nano particle risen in recent years, this method has following several albumen bio-mimetic syntheses
Big advantage: 1) reaction condition is mild, and whole organic solvent-free participates in, and is properly termed as green syt 2) product characteristics can adjust precisely
Control.3) biological phasic property is good.4) raw material sources are extensive, advantage of lower cost, reproducible.Solves prior synthesizing method process mistake
In cumbersome, environmental requirement harshness, and there is the problem of certain pollution.
Summary of the invention
The present invention in order to overcome the deficiencies of the prior art, provides a kind of by Gd:CuS mineralising sendai virus (SeV@Gd:CuS)
The tactful human simulation method that successfully synthesizes tumor combined therapeutic nanometer formulation.
Technical scheme is as follows:
The synthetic method of the tumor combined therapeutic nanometer formulation of Gd:CuS mineralising sendai virus (SeV@Gd:CuS), step is such as
Under:
1) Copper dichloride dihydrate (CuCl of 8.5mg is accurately weighed2·2H2O) powder, ultrasound is molten after 5mL ultrapure water is added
Solution obtains the CuCl that concentration is 0.01M2·2H2O aqueous solution.
2) the chlorinated gadolinium hexahydrate powder (GdCl of 9.3mg is accurately weighed3·6H2O), ultrasound is molten after 1mL ultrapure water is added
Solution obtains the GdCl that concentration is 0.025M3·6H2O aqueous solution.
3) as follows using the method for one pot process SeV@Gd:CuS nano particle:
(1) single port bottle that 5ml concentration is 0.01mg/ml sendai virus solution is placed in 37 DEG C of water-bath magnetic stirring apparatus
On, then under agitation, the CuCl of above-mentioned configuration is added dropwise with disposable dropper2·2H2O and GdCl3·6H2O solution, drop
Bi Jixu is added to stir three minutes；
(2) sodium hydroxide (NaOH) solution that concentration is 1M has been configured, 0.5mL is taken to be added in above-mentioned reaction system, has been adjusted
The final ph of solution is 8-10, and solution becomes the navy blue of clear at this time；
(3) configuration concentration is the vulcanized sodium nonahydrate (Na of 242.16mg/mL2S·9H2O) solution, and 0.4mL is taken to be added
Into reaction system, solution becomes brown at this time；
(4) under 37 DEG C of water baths, after being stirred to react 4h, taking out reaction solution and pouring into molecular cut off is 8000-14000
Bag filter in, dialyse in ultrapure water for 24 hours, to remove unreacted ion；
(5) it after by the solution after having dialysed as -80 DEG C of refrigerator overnights, is transferred in freeze drier and is lyophilized, obtained
To the SeV@Gd:CuS nano particle of light green color flocculence.
Present invention has an advantage that 1) product has significant T1 weighted magnetic resonance/optoacoustic Double-mode imaging ability.2) it produces
Product photothermal conversion efficiency under near-infrared laser irradiation is higher.3) product can efficiently guide subsequent tumor thermal therapy.
4) product has excellent biocompatibility, generation without side-effects.
Detailed description of the invention
Sendai virus nano particle (SeV@Gd:CuS) the stimulation NF- κ B of Fig. 1: Gd:CuS mineralising is generated.
Specific embodiment
Embodiment 1:
The synthetic method of SeV@Gd:CuS tumour combined immunization treatment nanometer formulation, the specific steps are as follows:
(1) Copper dichloride dihydrate (CuCl of 4.25mg is weighed2·2H2O ultrasonic dissolution after 5mL ultrapure water is added in) powder,
Obtain the CuCl that concentration is 0.005M2·2H2O aqueous solution.
(2) the chlorinated gadolinium hexahydrate powder (GdCl of 9.3mg is weighed3·6H2O), ultrasonic dissolution after addition 1mL ultrapure water,
Obtain the GdCl that concentration is 0.025M3·6H2O aqueous solution.
(3) utilize the method for one pot process SeV@Gd:CuS nano particle: will fill 2.5ml concentration is 0.005mg/
The single port bottle of ml sendai virus solution is placed on 37 DEG C of water-bath magnetic stirring apparatus, then under agitation, with disposable drop
The CuCl of above-mentioned configuration is added dropwise in pipe2·2H2O and GdCl3·6H2O solution is added dropwise and continues stirring three minutes；
(4) sodium hydroxide (NaOH) solution that concentration is 1M has been configured, 0.25mL is taken to be added in above-mentioned reaction system, has been adjusted
The final ph of solution is 8, and solution becomes the navy blue of clear at this time；Configuration concentration is the vulcanized sodium of 242.16mg/mL
Nonahydrate (Na2S·9H2O) solution, and 0.4mL is taken to be added in reaction system, solution becomes brown at this time；
(5) under 37 DEG C of water baths, after being stirred to react 2h, taking out reaction solution and pouring into molecular cut off is 8000-14000
Bag filter in, dialyse in ultrapure water for 24 hours, to remove unreacted ion；
(6) it after by the solution after having dialysed as -80 DEG C of refrigerator overnights, is transferred in freeze drier and is lyophilized, obtained
To the SeV@Gd:CuS nano particle of light green color flocculence.
Embodiment 2:
The synthetic method of SeV@Gd:CuS tumour combined immunization treatment nanometer formulation, the specific steps are as follows:
(1) Copper dichloride dihydrate (CuCl of 8.5mg is weighed2·2H2O) powder is added ultrasonic dissolution after 5mL ultrapure water, obtains
The CuCl for being 0.01M to concentration2·2H2O aqueous solution.
(2) the chlorinated gadolinium hexahydrate powder (GdCl of 18.6mg is weighed3·6H2O), ultrasound is molten after 1mL ultrapure water is added
Solution obtains the GdCl that concentration is 0.05M3·6H2O aqueous solution.
(3) utilize the method for one pot process SeV@Gd:CuS nano particle: will fill 5ml concentration is 0.01mg/ml
The single port bottle of sendai virus solution is placed on 37 DEG C of water-bath magnetic stirring apparatus, then under agitation, with disposable dropper
The CuCl of above-mentioned configuration is added dropwise2·2H2O and GdCl3·6H2O solution is added dropwise and continues stirring three minutes；
(4) sodium hydroxide (NaOH) solution that concentration is 1M has been configured, 0.5mL is taken to be added in above-mentioned reaction system, has been adjusted
The final ph of solution is 9, and solution becomes the navy blue of clear at this time；Configuration concentration is the vulcanized sodium of 242.16mg/mL
Nonahydrate (Na2S·9H2O) solution, and 0.4mL is taken to be added in reaction system, solution becomes brown at this time；
(5) under 37 DEG C of water baths, after being stirred to react 4h, taking out reaction solution and pouring into molecular cut off is 8000-14000
Bag filter in, dialyse in ultrapure water for 24 hours, to remove unreacted ion；
(6) it after by the solution after having dialysed as -80 DEG C of refrigerator overnights, is transferred in freeze drier and is lyophilized, obtained
To the SeV@Gd:CuS nano particle of light green color flocculence.
Embodiment 3:
The synthetic method of Gd:CuS nanometers of diagnosis and treatment agent of SeV@, the specific steps are as follows:
(1) Copper dichloride dihydrate (CuCl of 17mg is weighed2·2H2O) powder is added ultrasonic dissolution after 5mL ultrapure water, obtains
The CuCl for being 0.02M to concentration2·2H2O aqueous solution.
(2) the chlorinated gadolinium hexahydrate powder (GdCl of 37.2mg is weighed3·6H2O), ultrasound is molten after 1mL ultrapure water is added
Solution obtains the GdCl that concentration is 0.1M3·6H2O aqueous solution.
(3) utilize the method for one pot process SeV@Gd:CuS nano particle: will fill 10ml concentration is 0.02mg/ml
The single port bottle of sendai virus solution is placed on 37 DEG C of water-bath magnetic stirring apparatus, then under agitation, with disposable dropper
The CuCl of above-mentioned configuration is added dropwise2·2H2O and GdCl3·6H2O solution is added dropwise and continues stirring three minutes；
(4) sodium hydroxide (NaOH) solution that concentration is 1M has been configured, 1mL is taken to be added in above-mentioned reaction system, has been adjusted molten
The final ph of liquid is 10, and solution becomes the navy blue of clear at this time；Configuration concentration is the vulcanized sodium nine of 484.32mg/mL
Hydrate (Na2S·9H2O) solution, and 0.4mL is taken to be added in reaction system, solution becomes brown at this time；
(5) under 37 DEG C of water baths, after being stirred to react 8h, taking out reaction solution and pouring into molecular cut off is 8000-14000
Bag filter in, dialyse in ultrapure water for 24 hours, to remove unreacted ion；
(6) it after by the solution after having dialysed as -80 DEG C of refrigerator overnights, is transferred in freeze drier and is lyophilized, obtained
To the SeV@Gd:CuS nano particle of light green color flocculence.
Claims (2)
- The synthetic method of the visualization guidance tumour combined immunization treatment nanometer formulation of 1.Gd:CuS mineralising sendai virus, feature It is, the specific steps are as follows:1) Copper dichloride dihydrate (CuCl of 8.5mg is accurately weighed2·2H2O) powder is added ultrasonic dissolution after 5mL ultrapure water, obtains The CuCl for being 0.01M to concentration2·2H2O aqueous solution.2) the chlorinated gadolinium hexahydrate powder (GdCl of 9.3mg is accurately weighed3·6H2O), ultrasonic dissolution after addition 1mL ultrapure water, Obtain the GdCl that concentration is 0.025M3·6H2O aqueous solution.3) one pot process SeV@Gd:CuS nano particle is utilized.
- 2. the visualization guidance tumour combined immunization of Gd:CuS mineralising sendai virus according to claim 1 treats nanometer system The synthetic method of agent, characterized in that the step 3) is specific as follows:(1) single port bottle that 5ml concentration is 0.01mg/ml sendai virus solution is placed on 37 DEG C of water-bath magnetic stirring apparatus, with Afterwards under agitation, the CuCl of above-mentioned configuration is added dropwise with disposable dropper2·2H2O and GdCl3·6H2O solution, is added dropwise Continue stirring three minutes；(2) sodium hydroxide (NaOH) solution that concentration is 1M has been configured, 0.5mL is taken to be added in above-mentioned reaction system, has adjusted solution Final ph be 8-10, solution becomes the navy blue of clear at this time；(3) configuration concentration is the vulcanized sodium nonahydrate (Na of 242.16mg/mL2S·9H2O) solution, and 0.4mL is taken to be added to instead It answers in system, solution becomes brown at this time；(4) under 37 DEG C of water baths, after being stirred to react 4h, taking out reaction solution and pouring into molecular cut off is the saturating of 8000-14000 It analyses in bag, dialyses in ultrapure water for 24 hours, to remove unreacted ion；(5) it after by the solution after having dialysed as -80 DEG C of refrigerator overnights, is transferred in freeze drier and is lyophilized, obtain shallow The SeV Gd:CuS nano particle of green flocculence.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110833566A (en) * | 2019-10-31 | 2020-02-25 | 天津大学 | Method for synthesizing tumor combined treatment nano preparation based on mineralized Sendai virus |
| CN110859822A (en) * | 2019-11-14 | 2020-03-06 | 天津大学 | Synthesis method of nano preparation |
| CN110917346A (en) * | 2019-11-30 | 2020-03-27 | 天津大学 | Method for biomimetic simulated synthesis of photothermal tumor combined treatment nano preparation |
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| CN105999309A (en) * | 2016-05-24 | 2016-10-12 | 天津大学 | Protein biological template-based gadolinium-doped copper sulfide nano-particles and preparation method thereof |
| CN107158378A (en) * | 2017-05-11 | 2017-09-15 | 江苏大学 | A kind of photo-thermal effect copper sulfide manganese albumen composite nanometer particle and preparation method thereof |
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| CN104784691A (en) * | 2015-04-29 | 2015-07-22 | 天津医科大学 | Method for preparing CuS photothermal therapy nano material with good biocompatibility |
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