CN115724449B - Mixed salt nano aluminum adjuvant and preparation method and application thereof - Google Patents
Mixed salt nano aluminum adjuvant and preparation method and application thereof Download PDFInfo
- Publication number
- CN115724449B CN115724449B CN202211464870.5A CN202211464870A CN115724449B CN 115724449 B CN115724449 B CN 115724449B CN 202211464870 A CN202211464870 A CN 202211464870A CN 115724449 B CN115724449 B CN 115724449B
- Authority
- CN
- China
- Prior art keywords
- mixed
- salt
- aluminum
- aluminum adjuvant
- adjuvant
- 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.)
- Active
Links
- 239000002671 adjuvant Substances 0.000 title claims abstract description 133
- 229910052782 aluminium Inorganic materials 0.000 title claims abstract description 121
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 title claims abstract description 113
- 150000003839 salts Chemical class 0.000 title claims abstract description 82
- 238000002360 preparation method Methods 0.000 title claims abstract description 10
- 206010028980 Neoplasm Diseases 0.000 claims abstract description 87
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims abstract description 63
- AZDRQVAHHNSJOQ-UHFFFAOYSA-N alumane Chemical class [AlH3] AZDRQVAHHNSJOQ-UHFFFAOYSA-N 0.000 claims abstract description 26
- 239000002244 precipitate Substances 0.000 claims abstract description 26
- 159000000003 magnesium salts Chemical class 0.000 claims abstract description 23
- 239000000243 solution Substances 0.000 claims abstract description 21
- 229910021642 ultra pure water Inorganic materials 0.000 claims abstract description 17
- 239000012498 ultrapure water Substances 0.000 claims abstract description 17
- 238000003756 stirring Methods 0.000 claims abstract description 11
- WNROFYMDJYEPJX-UHFFFAOYSA-K aluminium hydroxide Chemical compound [OH-].[OH-].[OH-].[Al+3] WNROFYMDJYEPJX-UHFFFAOYSA-K 0.000 claims abstract description 9
- 239000000725 suspension Substances 0.000 claims abstract description 9
- 239000011259 mixed solution Substances 0.000 claims abstract description 8
- 239000007788 liquid Substances 0.000 claims abstract description 7
- VTHJTEIRLNZDEV-UHFFFAOYSA-L magnesium dihydroxide Chemical compound [OH-].[OH-].[Mg+2] VTHJTEIRLNZDEV-UHFFFAOYSA-L 0.000 claims abstract description 7
- 239000000347 magnesium hydroxide Substances 0.000 claims abstract description 7
- 229910001862 magnesium hydroxide Inorganic materials 0.000 claims abstract description 7
- 238000000926 separation method Methods 0.000 claims abstract description 7
- 238000010335 hydrothermal treatment Methods 0.000 claims abstract description 6
- 230000007935 neutral effect Effects 0.000 claims abstract description 6
- 238000006460 hydrolysis reaction Methods 0.000 claims abstract description 5
- 238000005406 washing Methods 0.000 claims abstract description 5
- 238000000034 method Methods 0.000 claims description 20
- 201000001441 melanoma Diseases 0.000 claims description 11
- 239000011777 magnesium Substances 0.000 claims description 8
- 230000004614 tumor growth Effects 0.000 claims description 6
- 229910002651 NO3 Inorganic materials 0.000 claims description 4
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 claims description 4
- 150000001450 anions Chemical class 0.000 claims description 4
- 239000013589 supplement Substances 0.000 claims description 4
- 238000001132 ultrasonic dispersion Methods 0.000 claims description 4
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical class [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 claims description 3
- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 claims description 3
- 229940069428 antacid Drugs 0.000 claims description 3
- 239000003159 antacid agent Substances 0.000 claims description 3
- 230000001458 anti-acid effect Effects 0.000 claims description 3
- 238000001914 filtration Methods 0.000 claims description 3
- 238000001027 hydrothermal synthesis Methods 0.000 claims description 3
- 230000035484 reaction time Effects 0.000 claims description 3
- 239000007787 solid Substances 0.000 claims description 3
- 230000003213 activating effect Effects 0.000 claims description 2
- 208000035269 cancer or benign tumor Diseases 0.000 claims 2
- 238000005119 centrifugation Methods 0.000 claims 1
- 230000002401 inhibitory effect Effects 0.000 claims 1
- 159000000000 sodium salts Chemical class 0.000 claims 1
- 230000002601 intratumoral effect Effects 0.000 abstract description 15
- 230000012010 growth Effects 0.000 abstract description 6
- 230000002378 acidificating effect Effects 0.000 abstract description 5
- 210000002865 immune cell Anatomy 0.000 abstract description 5
- 230000009286 beneficial effect Effects 0.000 abstract description 3
- 238000002347 injection Methods 0.000 description 36
- 239000007924 injection Substances 0.000 description 36
- 210000001744 T-lymphocyte Anatomy 0.000 description 16
- 229940037003 alum Drugs 0.000 description 16
- 239000007864 aqueous solution Substances 0.000 description 13
- 241000699670 Mus sp. Species 0.000 description 11
- 210000001519 tissue Anatomy 0.000 description 11
- 230000005764 inhibitory process Effects 0.000 description 10
- FBOZXECLQNJBKD-ZDUSSCGKSA-N L-methotrexate Chemical compound C=1N=C2N=C(N)N=C(N)C2=NC=1CN(C)C1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 FBOZXECLQNJBKD-ZDUSSCGKSA-N 0.000 description 9
- 230000028993 immune response Effects 0.000 description 9
- 229960000485 methotrexate Drugs 0.000 description 9
- 210000004881 tumor cell Anatomy 0.000 description 9
- 239000003814 drug Substances 0.000 description 8
- 239000002245 particle Substances 0.000 description 8
- 239000000126 substance Substances 0.000 description 8
- 239000000427 antigen Substances 0.000 description 7
- 102000036639 antigens Human genes 0.000 description 7
- 108091007433 antigens Proteins 0.000 description 7
- 210000004027 cell Anatomy 0.000 description 7
- 238000011282 treatment Methods 0.000 description 7
- 230000005975 antitumor immune response Effects 0.000 description 6
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 6
- 238000001514 detection method Methods 0.000 description 5
- 229940079593 drug Drugs 0.000 description 5
- 230000001939 inductive effect Effects 0.000 description 5
- 230000004957 immunoregulator effect Effects 0.000 description 4
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 description 3
- 206010009944 Colon cancer Diseases 0.000 description 3
- 101001109501 Homo sapiens NKG2-D type II integral membrane protein Proteins 0.000 description 3
- 102100022680 NKG2-D type II integral membrane protein Human genes 0.000 description 3
- 229910019142 PO4 Inorganic materials 0.000 description 3
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 3
- 230000004900 autophagic degradation Effects 0.000 description 3
- 208000029742 colonic neoplasm Diseases 0.000 description 3
- 239000013078 crystal Substances 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 230000002519 immonomodulatory effect Effects 0.000 description 3
- 238000001727 in vivo Methods 0.000 description 3
- 239000004310 lactic acid Substances 0.000 description 3
- 235000014655 lactic acid Nutrition 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 238000006386 neutralization reaction Methods 0.000 description 3
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 3
- 239000010452 phosphate Substances 0.000 description 3
- 238000001556 precipitation Methods 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 231100000331 toxic Toxicity 0.000 description 3
- 230000002588 toxic effect Effects 0.000 description 3
- 239000012646 vaccine adjuvant Substances 0.000 description 3
- 229940124931 vaccine adjuvant Drugs 0.000 description 3
- LKKMLIBUAXYLOY-UHFFFAOYSA-N 3-Amino-1-methyl-5H-pyrido[4,3-b]indole Chemical compound N1C2=CC=CC=C2C2=C1C=C(N)N=C2C LKKMLIBUAXYLOY-UHFFFAOYSA-N 0.000 description 2
- 206010006187 Breast cancer Diseases 0.000 description 2
- 208000026310 Breast neoplasm Diseases 0.000 description 2
- GHASVSINZRGABV-UHFFFAOYSA-N Fluorouracil Chemical compound FC1=CNC(=O)NC1=O GHASVSINZRGABV-UHFFFAOYSA-N 0.000 description 2
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 2
- JLVVSXFLKOJNIY-UHFFFAOYSA-N Magnesium ion Chemical compound [Mg+2] JLVVSXFLKOJNIY-UHFFFAOYSA-N 0.000 description 2
- 230000006044 T cell activation Effects 0.000 description 2
- 238000002441 X-ray diffraction Methods 0.000 description 2
- 230000000259 anti-tumor effect Effects 0.000 description 2
- 230000005540 biological transmission Effects 0.000 description 2
- 230000000903 blocking effect Effects 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000013043 chemical agent Substances 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 231100000135 cytotoxicity Toxicity 0.000 description 2
- 230000003013 cytotoxicity Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 239000003937 drug carrier Substances 0.000 description 2
- 229960002949 fluorouracil Drugs 0.000 description 2
- 230000003832 immune regulation Effects 0.000 description 2
- 238000009169 immunotherapy Methods 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 230000008595 infiltration Effects 0.000 description 2
- 238000001764 infiltration Methods 0.000 description 2
- 230000002147 killing effect Effects 0.000 description 2
- 201000007270 liver cancer Diseases 0.000 description 2
- 208000014018 liver neoplasm Diseases 0.000 description 2
- 229910052749 magnesium Inorganic materials 0.000 description 2
- 229910001425 magnesium ion Inorganic materials 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 101150079396 trpC2 gene Proteins 0.000 description 2
- 102000003954 Autophagy-Related Proteins Human genes 0.000 description 1
- 108010082399 Autophagy-Related Proteins Proteins 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- ILRRQNADMUWWFW-UHFFFAOYSA-K aluminium phosphate Chemical compound O1[Al]2OP1(=O)O2 ILRRQNADMUWWFW-UHFFFAOYSA-K 0.000 description 1
- DIZPMCHEQGEION-UHFFFAOYSA-H aluminium sulfate (anhydrous) Chemical compound [Al+3].[Al+3].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O DIZPMCHEQGEION-UHFFFAOYSA-H 0.000 description 1
- -1 aluminum ions Chemical class 0.000 description 1
- 125000000129 anionic group Chemical group 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 150000003841 chloride salts Chemical class 0.000 description 1
- 231100000263 cytotoxicity test Toxicity 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 210000004443 dendritic cell Anatomy 0.000 description 1
- 239000006185 dispersion Substances 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 210000002889 endothelial cell Anatomy 0.000 description 1
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 238000003018 immunoassay Methods 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 210000002540 macrophage Anatomy 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 229910021645 metal ion Inorganic materials 0.000 description 1
- 238000001000 micrograph Methods 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000002105 nanoparticle Substances 0.000 description 1
- 230000017066 negative regulation of growth Effects 0.000 description 1
- 210000003463 organelle Anatomy 0.000 description 1
- 230000002688 persistence Effects 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 238000002407 reforming Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000005909 tumor killing Effects 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Abstract
The invention relates to a mixed salt nano aluminum adjuvant and a preparation method and application thereof, comprising the following steps: adding the mixed solution of magnesium salt and aluminum salt with the molar ratio of 2-4:1 into the sodium hydroxide solution in a violent stirring state, and continuously stirring for 5-10 minutes at room temperature to obtain a mixed precipitate of magnesium hydroxide and aluminum hydroxide; collecting the mixed precipitate by adopting a solid-liquid separation method, washing with ultrapure water and re-suspending until the pH of the suspension is neutral or slightly alkaline; and (3) carrying out self-hydrolysis on the mixed precipitate at room temperature to obtain the mixed salt nano aluminum adjuvant, or carrying out hydrothermal treatment by using a hydrothermal kettle to obtain the mixed salt nano aluminum adjuvant. The beneficial effects of the invention are as follows: the mixed salt nano aluminum adjuvant can effectively neutralize acidic tumor tissues and activate intratumoral immune cells after being injected around tumor, and can directly inhibit the growth of 70-95% of solid tumors.
Description
Technical Field
The invention relates to the field of vaccine adjuvant preparation, in particular to a mixed salt nano aluminum adjuvant and a preparation method and application thereof.
Background
Aluminum adjuvants have been widely used as human or animal vaccine adjuvants, which enhance the specific immune response of the body to antigens, thereby inducing a durable and strong antibody immune response. The aluminum adjuvant mainly consists of aluminum salt and mainly comprises: aluminum hydroxide, aluminum phosphate, aluminum sulfate and mixed salt aluminum adjuvants, with aluminum hydroxide being the most widely used.
An effective T cell immune response is an important precondition for achieving efficient tumor immunotherapy. Although aluminum adjuvants are effective in assisting antigens in inducing antibody immune responses, they are not able to induce T cell immune responses, which results in very limited application in tumor immunotherapy. Moreover, the existing aluminum adjuvant is only used for antigen delivery, and no precedent for directly controlling tumor immunity microenvironment exists. It is found that part of metal ions such as Mg2+ and the like can play a certain adjuvant role and play an important role in activating intratumoral immune cells. At the same time, neutralization of excessive accumulation of lactic acid in the tumor can also activate intratumoral immune cells. Because of the variety of aluminum adjuvants, particularly mixed salt aluminum adjuvants, there is no clear definition, the development and application of this novel aluminum adjuvant provides a great opportunity.
In recent years, the development of nano technology and materials provides an important new idea for upgrading and reforming aluminum adjuvants. Current commercial aluminum adjuvants mainly include two classes, namely: alhydrogel (major component aluminum hydroxide, consistent with clinical aluminum adjuvants) produced by InvivoGen corporation and Imject Alum (major component magnesium hydroxide and aluminum hydroxide) produced by ThermoFisher corporation, both of which can only induce antibody immune responses. Although studies indicate that Imject Alum does not induce an effective T cell immune response as a tumor vaccine adjuvant, imject Alum induces a T helper type 1 immune response more readily than Alhydrogel, which is very beneficial for the body to induce a T cell immune response. This might be related to the inclusion of mg2+ in the object Alum that favors T cell activation. In addition, due to the addition of magnesium hydroxide, the Imject Alum is weakly alkaline, which either will facilitate direct neutralization of lactic acid accumulated in the tumor by the Imject Alum, and release of mg2+ to activate intratumoral T cells. However, the Imject Alum is in large particle clusters (about 8 μm), which has a great impact on its tumor infiltration capacity and rapid distribution within the tumor.
Disclosure of Invention
The invention aims to overcome the defects in the prior art and provides a mixed salt nano aluminum adjuvant and a preparation method and application thereof. The method is a preparation method of the nano aluminum adjuvant which ensures that the obtained nano aluminum adjuvant has uniform elucidation structure, uniform particle size distribution, low cost, high safety and reliable quality. The mixed salt nano aluminum adjuvant prepared by the method can directly neutralize acidic tumor tissues, activate T cells in tumors and induce personalized anti-tumor immune response.
In a first aspect, a method for preparing a mixed salt nano aluminum adjuvant is provided, comprising:
s1, rapidly adding a magnesium salt and aluminum salt mixed solution with a molar ratio of 2-4:1 into a sodium hydroxide solution in a violent stirring state, and continuously stirring for 5-10 minutes at room temperature to obtain a mixed precipitate of magnesium hydroxide and aluminum hydroxide;
s2, collecting the mixed precipitate by adopting a solid-liquid separation method, washing with ultrapure water and re-suspending until the pH of the suspension is neutral or slightly alkaline;
s3, carrying out self-hydrolysis on the mixed precipitate at room temperature to obtain the mixed salt nano aluminum adjuvant, or carrying out hydrothermal treatment by using a hydrothermal kettle to obtain the mixed salt nano aluminum adjuvant.
Preferably, in S1, the anions of the magnesium and aluminum salts are the same, and the magnesium and aluminum salts include chloride, nitrate, carbonate, phosphate or sulfate.
Preferably, in S1, mg is 2+ +Al 3+ And OH (OH) - The molar ratio of the mixed solution of magnesium salt and aluminum salt to the sodium hydroxide solution is 1:1.5-2.0.
Preferably, in S1, the concentration of the magnesium salt is 0.07-2.4M, the concentration of the aluminum salt is 0.03-0.6M, the volume of the mixed solution is 10mL, and the concentration of the sodium hydroxide solution is 0.005-0.9M.
Preferably, in S2, the solid-liquid separation method includes: stationary, centrifugal and filtration; when the centrifugal method is adopted, the centrifugal force is not less than 4750g.
Preferably, in S3, the mixed precipitate is sufficiently resuspended in ultrapure water having a volume of 10 to 200mL using stirring, sloshing or ultrasonic dispersion in advance.
Preferably, in S3, the precipitate suspension is stirred for 6 to 96 hours by standing or shaking with a shaker or with a stirrer when the room temperature method is employed.
Preferably, in S3, when a hydrothermal method is adopted, the hydrothermal temperature is set to be 60-150 ℃ and the reaction time is 2-16 h.
In a second aspect, there is provided a mixed salt nanoaluminum adjuvant as described in the first aspect as antacid and Mg 2+ The supplement, after local injection, activates tumor immune microenvironment and inhibits the use of solid tumor growth.
Preferably, the solid tumor comprises melanoma, colon cancer, liver cancer and breast cancer; the local injection comprises: intratumoral injection, paraneoplastic injection, distal injection (1-2 cm from tumor margin).
In a third aspect, there is provided the use of a mixed salt nanoaluminum adjuvant according to the first aspect as a drug carrier for delivering a chemical drug or an immunomodulatory molecule, for killing a tumor and inducing induction of a personalized anti-tumor immune response after local injection.
Preferably, the chemical agents include, but are not limited to, methotrexate, 5 fluorouracil, and the like, and the immunomodulatory molecules include, but are not limited to, cpG.
The beneficial effects of the invention are as follows:
1. the mixed salt nano aluminum adjuvant has simple synthesis process and definite components, and the synthesized nano aluminum adjuvant has uniform particle size and particle size distribution within the range of 50-150 nm.
2. The mixed salt nano aluminum adjuvant is slightly alkaline, and can effectively neutralize acidic tissue subcellular organelles.
3. The mixed salt nano aluminum adjuvant can supplement Mg into acidic tumor tissues 2+ To activate intratumoral immune cells.
4. The mixed salt nano aluminum adjuvant can effectively neutralize acidic tumor tissues and activate intratumoral immune cells after being injected around tumor, and can directly inhibit the growth of 70-95% of solid tumors.
5. The mixed salt nano aluminum adjuvant can be further used as a carrier to deliver chemical medicine Methotrexate (MTX) or immunoregulatory molecule CpG, and after being injected around tumor, the mixed salt nano aluminum adjuvant can synergistically inhibit the growth of solid tumor.
Drawings
FIG. 1 is a transmission electron microscope image of a mixed salt nano aluminum adjuvant and commercial aluminum adjuvants Alhydrogel and Imject Alum provided by the invention;
FIG. 2 is a schematic diagram of the X-ray diffraction scan results of the mixed salt nanoaluminum adjuvant and commercial aluminum adjuvants Alhydrogel and Imject Alum provided by the present invention;
FIG. 3 is a schematic diagram of the particle size distribution test results of the mixed salt nanoaluminum adjuvant and commercial aluminum adjuvants Alhydrogel and Imject Alum provided by the invention;
FIG. 4 is a schematic diagram of the results of surface potential testing of the mixed salt nanoaluminum adjuvants and commercial aluminum adjuvants Alhydrogel and Imject Alum provided by the present invention;
FIG. 5 is a schematic illustration of the pH of solutions of mixed salt nanoaluminum adjuvants and commercial aluminum adjuvants Alhydrogel and Imject Alum provided by the present invention;
FIG. 6 shows the cytotoxicity test results of the mixed salt nano-aluminum adjuvant provided by the invention on different tumors;
FIG. 7 shows the detection results of the mixed salt nano aluminum adjuvant provided by the invention interfering with the expression of tumor cell autophagy related proteins LC3B and P62;
FIG. 8 shows the detection results of the influence of the mixed salt nano aluminum adjuvant provided by the invention on the pH of tumors in different injection modes;
FIG. 9 shows the results of the detection of the retention capacity of the mixed salt nano-aluminum adjuvant (white fluorescence) in tumors in different injection modes;
FIG. 10 shows the detection results of the mixed salt nano aluminum adjuvant provided by the invention on tumor growth inhibition under different injection modes;
FIG. 11 shows the results of the tumor growth inhibition test of the mixed salt nanoaluminum adjuvant provided by the invention and commercial aluminum adjuvants Alhydrogel and Imject Alum;
FIG. 12 shows the change in the level of intratumoral toxic T cells (CD3+CD8+) and intratumoral T cell activation (CD3+NKG2D+) after tumor treatment using the intratumoral injection method with the mixed salt nanoaluminum adjuvant and commercial aluminum adjuvants provided by the present invention;
FIG. 13 shows the inhibition of tumor treatment by tumor side injection after the combination of the mixed salt nano aluminum adjuvant and the immunoregulatory molecule CpG or the chemical drug MTX;
fig. 14 shows the change of the level of antigen-specific anti-tumor toxic T cells induced in mice after tumor treatment by the tumor side injection method after the mixed salt nano aluminum adjuvant provided by the invention and the immune regulation molecule CpG or the chemical drug MTX are combined.
Detailed Description
The invention is further described below with reference to examples. The following examples are presented only to aid in the understanding of the invention. It should be noted that it will be apparent to those skilled in the art that modifications can be made to the present invention without departing from the principles of the invention, and such modifications and adaptations are intended to be within the scope of the invention as defined in the following claims.
Example 1:
a preparation method of a mixed salt nano aluminum adjuvant comprises the following steps:
s1, rapidly adding a magnesium salt and aluminum salt mixed solution with a molar ratio of 2-4:1 into a sodium hydroxide solution in a violent stirring state, and continuously stirring for 5-10 minutes at room temperature to obtain a mixed precipitate of magnesium hydroxide and aluminum hydroxide;
s2, collecting a mixed precipitate by adopting a solid-liquid separation method, washing with ultrapure water and re-suspending until the pH of the suspension is neutral or slightly alkaline (the pH is between 7 and 9);
s3, carrying out self-hydrolysis on the mixed precipitate at room temperature to obtain the mixed salt nano aluminum adjuvant, or carrying out hydrothermal treatment by using a hydrothermal kettle to obtain the mixed salt nano aluminum adjuvant.
In S1, anions of magnesium salt and aluminum salt are the same, and the magnesium salt and the aluminum salt comprise chloride salt, nitrate, carbonate, phosphate or sulfate. That is, the magnesium salt and the aluminum salt may be different anionic salts such as chloride, nitrate, carbonate, phosphate, sulfate, or the like, but in the same reaction, magnesium salt and aluminum salt having the same anions should be used.
In the present invention, since the aqueous solution of the magnesium salt and the aluminum salt is brought into contact with the sodium hydroxide solution to cause precipitation of the aluminum adjuvant, the amount of the aqueous solution of the magnesium salt and the aluminum salt to be used in the sodium hydroxide solution is not particularly limited, so long as the aqueous solution of the magnesium salt and the aluminum salt is brought into contact with the sodium hydroxide solution to cause precipitation of the aluminum adjuvant, and in order to fully utilize the raw materials, it is preferable that in S1, the components (Mg 2+ +Al 3+ ) And OH (OH) - The molar ratio of the aqueous solution of magnesium salt and aluminum salt to the sodium hydroxide solution is 1:1.5-2.0, such as 1:1.5.
In the invention, as the aqueous solution of magnesium salt and aluminum salt and sodium hydroxide solution are released to form nano aluminum adjuvant precipitation, the concentration and volume of the aqueous solution of magnesium salt and aluminum salt and sodium hydroxide solution can be arbitrary as long as the requirement of the dosage is met, and in order to fully utilize raw materials and facilitate operation, in S1, the concentration of magnesium salt can be 0.7M, the concentration of aluminum salt can be 0.3M, the total volume is 10mL, and the concentration of sodium hydroxide solution can be 0.45M.
S2, the solid-liquid separation method comprises the following steps: stationary, centrifugal and filtration; when the centrifugal method is employed, the centrifugal force is not less than 4750g, and the washing is performed with ultrapure water for 2 times.
In S3, when the precipitate obtained by contacting an aqueous solution of magnesium salt and aluminum salt with a sodium hydroxide solution is treated by a room temperature method to obtain a mixed salt nanoaluminum adjuvant, the precipitate should be sufficiently resuspended in ultrapure water by stirring, sloshing or ultrasonic dispersion in advance, wherein the volume of ultrapure water is not specifically limited, and for convenience of operation, the precipitate is sufficiently resuspended by using 40mL of ultrapure water and then left to stand or sloshed by using a shaker or the suspension of the precipitate is stirred by using a stirrer for 48 hours to obtain the mixed salt nanoaluminum adjuvant.
In the invention, when a hydrothermal method is used for treating a precipitate obtained by contacting an aqueous solution of magnesium salt and aluminum salt with a sodium hydroxide solution to obtain a mixed salt nano aluminum adjuvant, the precipitate is fully resuspended in ultrapure water by a stirring, sloshing or ultrasonic dispersion method, wherein the volume of ultrapure water is not limited specifically, the hydrothermal temperature is set between 60 and 150 ℃, the reaction time is between 2 and 16 hours, and for convenient operation, 40mL of ultrapure water is used for fully resuspending the precipitate, and then a hydrothermal kettle is used for carrying out hydrothermal treatment on the precipitate suspension at the temperature for 4 hours to obtain the nano aluminum adjuvant.
The above-described reactor may be various reactors known in the art. Such as glass instruments, reaction kettles, and the like.
Example 2:
mixed salt nano aluminum adjuvant used as antacid and Mg 2+ The supplement, after local injection, activates tumor immune microenvironment and inhibits the use of solid tumor growth.
The mixed salt nano aluminum adjuvant prepared by the invention is suitable for various solid tumors, which refer to tumor tissues with definite entities, including melanoma, colon cancer, liver cancer, breast cancer and the like, and the local injection mode comprises the following steps: intratumoral injection, paraneoplastic injection, distal injection (1-2 cm from tumor margin).
The mixed salt nano aluminum adjuvant provided by the invention is adopted to locally inject solid tumors, and compared with a commercial aluminum adjuvant Alhydrogel or an Imject Alum, the mixed salt nano aluminum adjuvant can obviously neutralize tumor acidity and inhibit tumor growth.
Example 3:
the application of the mixed salt nano aluminum adjuvant as a drug carrier for delivering chemical drugs or immune regulation molecules, killing tumors and inducing personalized anti-tumor immune response induction after local injection.
Chemical agents contemplated by the present invention include, but are not limited to, methotrexate, 5 fluorouracil, and the like, and immunomodulatory molecules include, but are not limited to, cpG.
The novel preparation of the mixed salt nano aluminum adjuvant and chemical drugs or immunoregulatory molecules is adopted to locally inject the solid tumor, so that the growth of the solid tumor can be obviously inhibited, and the personalized anti-tumor immune response can be induced.
Example 4:
weighing a certain amount of MgCl 2 ·6Η 2 O and AlCl 3 ·6Η 2 O, dissolving in ultrapure water to prepare 10mL of aqueous solution with the concentration of magnesium ions and aluminum ions of 0.7mol/L and 0.3 mol/L; a certain amount of NaOH is weighed and dissolved in distilled water to obtain 40mL of 0.45mol/L NaOH aqueous solution. 40mL of sodium hydroxide solution was preset in the flask and stirred at high speed using a magnetic stirrer, followed by rapid MgCl-addition using a syringe 2 ·6Η 2 O and AlCl 3 ·6Η 2 The O solution was injected into the sodium hydroxide solution and stirred at room temperature for 10 minutes. The suspension was collected, the aluminum adjuvant precipitate was collected using a centrifugal force of 4750g, and washed 2 times with ultrapure water. Dispersing the precipitate in 40mL of ultrapure water, and performing hydrothermal treatment for 4 hours at 100 ℃ by using a hydrothermal kettle to obtain the mixed salt nano aluminum adjuvant. The particle morphology of the sample is measured by adopting a transmission electron microscope, the result is shown in figure 1, and as can be seen from figure 1, compared with commercial aluminum adjuvants Alhydrogel and Imject Alum, the mixed salt nanometer aluminum adjuvant obtained by the invention is hexagonal particles, has uniform size and uniform dispersion; the crystal type of the mixed salt nano aluminum adjuvant is determined to be a layered crystal structure by adopting X-ray diffraction scanning, and the result is shown in figure 2; the surface charge of a sample is measured by adopting a nano-particle size and electrokinetic potential (Zeta potential) analyzer, the particle size of the mixed salt nano-aluminum adjuvant obtained by the invention is uniformly distributed at 50-150nm, and the result is shown in figure 3; the surface charge of the mixed salt nano aluminum adjuvant obtained by the invention in the aqueous solution is +33.4mV, and the result is shown in figure 4; the pH value of the aqueous solution of the sample is measured by adopting a pH meter, and when the concentration of the mixed salt nano aluminum adjuvant obtained by the invention is 0.5-1mg/mL, the pH value in the aqueous solution reaches 8.0-8.5, and the result is shown in figure 5.
Example 5:
the mixed salt nano aluminum adjuvant prepared by the invention is incubated with tumor cells and normal cells together, the physiological effect of the mixed salt nano aluminum adjuvant on the cells is evaluated, the result is shown in figure 6, when different cells are incubated with the mixed salt nano aluminum adjuvant with the concentration of 5-1000 mug/mL, the mixed salt nano aluminum adjuvant has obvious cytotoxicity on colon cancer cells and melanoma cells at a lower concentration of 62.5 mug/mL; and for normal cells such as dendritic cells, macrophages and endothelial cells, the mixed salt nano aluminum adjuvant does not cause obvious cytotoxicity to the cells at the concentration of 250 mug/mL or below. Further researches of the invention find that the mixed salt nano aluminum adjuvant can effectively block autophagy paths of tumor cells and cause the level of autophagy blocking marker proteins LC3-II and P62 in the tumor cells to rise, and the result is shown in figure 7, which shows that the mixed salt nano aluminum adjuvant can induce inhibition of growth of the tumor cells and cause death of the tumor cells by specifically blocking autophagy paths of the tumor cells.
Example 6:
1mg of the mixed salt nano aluminum adjuvant prepared by the invention is taken, and the mixed salt nano aluminum adjuvant is prepared by a local injection mode, namely: intratumoral injection, intratumoral injection and distal injection (injection point is 0.5-2cm from tumor edge, 1cm in this example), and the injection volume is about 100mm 3 After melanoma mice in vivo, the SNARF-1 fluorescence quantification method is used for detecting the pH value of the tumors of the mice on days 1, 7 and 14, the result is shown in figure 8, and as can be seen from figure 8, the pH value of the tumors can be raised to be neutral within 1 day by adopting the nanometer aluminum adjuvant injected in the tumors, but the persistence is poor; the mixed salt nano aluminum adjuvant injected around tumor can continuously neutralize and maintain the pH value of tumor tissues to be close to neutral within 14 days; the nano aluminum adjuvant injected at the far end can also improve the pH of the tumor, but the neutralization capability of the tumor tissue pH is poorer than that of the tumor side injection. Further, when the mixed salt nanoaluminum adjuvant of the present invention was labeled with fluorescein and its distribution in tumor tissues was observed with paraneoplastic injection, it was found that the mixed salt nanoaluminum adjuvant of the present invention was mainly distributed around tumor at 1 day after injection and slowly infiltrated into the inside of tumor within the following 14 days, and the result was shown in fig. 9. Furthermore, 1mg of the nano aluminum adjuvant prepared by the invention is injected into tumor body by means of intratumoral injection, paraneoplastic injection and distal injection (1 cm away from the edge of tumor)The product is about 100mm 3 After the melanoma mice of (2) were in vivo, tumor sizes were measured using vernier calipers every two days for the following 10 days and tumor volumes were calculated (volume=0.5×length×width 2 ) As a result, as shown in fig. 10, it can be seen from fig. 10 that the tumor side injection mixed salt nano aluminum adjuvant most effectively inhibits the growth of solid melanoma, the inhibition rate of the tumor reaches 87% at the 18 th day, and the inhibition rates of the mixed salt nano aluminum adjuvant on the tumor are only 50% and 52% respectively in the modes of intra-tumor injection and distal injection.
Example 7:
taking 1mg of the mixed salt nano aluminum adjuvant and commercial aluminum adjuvant Alhydrogel, object Alum prepared by the invention, and injecting the mixed salt nano aluminum adjuvant and commercial aluminum adjuvant into a tumor with a tumor volume of about 100mm by a tumor side injection method 3 After the melanoma mice of (2) were in vivo, tumor sizes were measured using vernier calipers every two days for the following 10 days and tumor volumes were calculated (volume=0.5×length×width 2 ) The result is shown in fig. 11, and it can be seen from fig. 11 that the mixed salt nano aluminum adjuvant prepared by the invention most effectively inhibits the growth of solid melanoma, and the inhibition rate of the mixed salt nano aluminum adjuvant reaches 91% on the 18 th day, and the inhibition rate of the Alhydrogel and the Imject aluminum on the tumor is only 1.5% and 60%; the T cells in the tumor tissue are analyzed by adopting a flow immunoassay report, the result is shown in figure 12, and the mixed salt nano aluminum adjuvant prepared by the invention is injected to the side of the tumor as can be seen from figure 12, so that the CD3 is obviously improved + CD8 + Infiltration of T cells into tumor tissues and obvious increase of NKG2D marker expression on the surface of the T cells indicate that the mixed salt nano-aluminum adjuvant prepared by the invention can most effectively recruit the T cells to the tumor when the paraneoplastic injection is adopted and activate the intratumoral T cells by improving NKG2D expression.
Example 8:
1mg of the mixed salt nano aluminum adjuvant prepared by the invention is taken and respectively mixed with chemical medicine MTX (10-50 mug/mg nano aluminum adjuvant, 10 mug/mg nano aluminum adjuvant is selected for tumor killing in the example) and immunoregulatory molecule CpG (10-100 mug/mg nano aluminum adjuvant, 10 mug/mg nano aluminum adjuvant is selected for inducing personalized anti-tumor immune response in the example)) Mixing, adsorbing the above molecules by electrostatic action, and then injecting into tumor volume of about 100mm by remote injection 3 As a result of treatment of melanoma mice of (a) is shown in fig. 13, and as shown in fig. 13, the mice are treated beside the tumor on the 12 th and 16 th days, and each experimental group name represents the kind of medicine used for the two treatments, taking nano aluminum adjuvant/mtx+nano aluminum adjuvant/CpG as an example, indicating that nano aluminum adjuvant/MTX is injected 1cm from the tumor margin of the mice on the 12 th day and nano aluminum adjuvant/CpG is injected 1cm from the tumor margin of the mice on the 16 th day. As can be seen from fig. 13, the inhibition efficiency of the nano aluminum adjuvant/mtx+nano aluminum adjuvant/CpG combination on the tumor is highest, the inhibition rate on the tumor reaches 87% at the 18 th day, and the inhibition rates of the nano aluminum adjuvant/nano aluminum adjuvant and the nano aluminum adjuvant/CpG combination on the tumor are only 52% and 68% respectively. Further, the specific toxic T cell level against the melanoma classical antigen Trp2 in the spleen of mice was analyzed using flow detection technique, and as a result, as shown in fig. 14, nanoaluminum adjuvant/mtx+nanoaluminum adjuvant/CpG induced Trp2 specific anti-tumor T cells most efficiently in mice. These results indicate that the nano-aluminum adjuvant/MTX developed by the invention can effectively kill tumor cells and promote the tumor cells to release antigens, and the nano-aluminum adjuvant/CpG can effectively capture the antigens and induce personalized anti-tumor immune responses in situ.
In summary, the invention prepares the nano aluminum adjuvant which is completely consistent with the chemical composition of the Imject aluminum and has a definite crystal structure and a nano scale in the range of 50-200nm through the hydrolysis treatment of the mixture of the magnesium hydroxide and the aluminum hydroxide which is completely consistent with the composition of the Imject aluminum. The mixed salt nano aluminum adjuvant can be infiltrated into tumor tissues more easily, neutralize excessive accumulated lactic acid in tumors, and directly activate T cells in the tumors, thereby achieving high-efficiency new auxiliary treatment of the tumors.
Claims (5)
1. The preparation method of the mixed salt nano aluminum adjuvant is characterized by comprising the following steps:
s1, rapidly adding a mixed solution of magnesium salt and aluminum salt in a molar ratio of 2-4:1 into the mixtureStirring the solution of sodium hydroxide in a violent stirring state for 5 to 10 minutes at room temperature to obtain a mixed precipitate of magnesium hydroxide and aluminum hydroxide; in S1, anions of the magnesium salt and the aluminum salt are the same, and the magnesium salt and the aluminum salt are chloride or nitrate; the concentration of the magnesium salt is 0.07-2.4M, the concentration of the aluminum salt is 0.03-0.6M, the volume of the mixed solution is 10mL, and the concentration of the sodium hydroxide solution is 0.005-0.9M; by Mg 2+ + Al 3+ And OH (OH) - The molar ratio of the mixed solution of magnesium salt and aluminum salt to the dosage of sodium hydroxide solution is 1:1.5-2.0;
s2, collecting the mixed precipitate by adopting a solid-liquid separation method, washing with ultrapure water and re-suspending until the pH of the suspension is neutral or slightly alkaline;
s3, carrying out self-hydrolysis on the mixed precipitate at room temperature to obtain a mixed salt nano aluminum adjuvant, or carrying out hydrothermal treatment by using a hydrothermal kettle to obtain the mixed salt nano aluminum adjuvant; in S3, when a hydrothermal method is adopted, the hydrothermal temperature is set to be 60-150 ℃ and the reaction time is 2h-16 h.
2. The method for preparing a mixed salt nano aluminum adjuvant according to claim 1, wherein in S2, the solid-liquid separation method comprises: a standing method, a centrifugation method and a filtration method; when the centrifugal method is adopted, the centrifugal force is not less than 4750 and g.
3. The method for preparing a mixed salt nano aluminum adjuvant according to claim 1, wherein in S3, the mixed precipitate is sufficiently resuspended in ultrapure water by stirring, sloshing or ultrasonic dispersion, and the volume of the ultrapure water is 10-200mL.
4. The method for preparing a mixed salt nano-aluminum adjuvant according to claim 1, wherein in S3, the precipitate suspension 6-96h is left to stand or is shaken by a shaker or is stirred by a stirrer when a room temperature method is adopted.
5. A mixed sodium salt prepared by the method of claim 1Preparation of antacid and Mg for activating tumor immune microenvironment and inhibiting solid tumor growth by using Mialuminum adjuvant 2+ Use of a supplement, wherein the neoplasm and solid neoplasm are melanoma.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211464870.5A CN115724449B (en) | 2022-11-22 | 2022-11-22 | Mixed salt nano aluminum adjuvant and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211464870.5A CN115724449B (en) | 2022-11-22 | 2022-11-22 | Mixed salt nano aluminum adjuvant and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN115724449A CN115724449A (en) | 2023-03-03 |
| CN115724449B true CN115724449B (en) | 2023-12-12 |
Family
ID=85297210
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202211464870.5A Active CN115724449B (en) | 2022-11-22 | 2022-11-22 | Mixed salt nano aluminum adjuvant and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115724449B (en) |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002011760A1 (en) * | 2000-08-09 | 2002-02-14 | Alk-Abelló A/S | Parenteral vaccine formulations and uses thereof |
| CN101717649A (en) * | 2009-11-13 | 2010-06-02 | 西安电子科技大学 | Preparation method of enhanced magnesium-aluminum composite flame retardant |
| CN102583460A (en) * | 2011-01-12 | 2012-07-18 | 国家纳米科学中心 | Aluminium adjuvant as well as preparation method and application thereof |
| CN104800843A (en) * | 2015-04-02 | 2015-07-29 | 中国农业科学院兰州兽医研究所 | Agentia for preventing toxoplasma infection and application |
| CN106049161A (en) * | 2016-05-25 | 2016-10-26 | 西安电子科技大学 | Calcium carbonate and aluminum magnesium composite flame-retardant paper, and preparation method thereof |
| CN107128962A (en) * | 2017-05-04 | 2017-09-05 | 四川理工学院 | A kind of preparation method of nano-sized magnesium hydroxide |
| RU2678007C1 (en) * | 2017-12-05 | 2019-01-22 | Федеральное государственное бюджетное учреждение науки Федеральный исследовательский центр "Кольский научный центр Российской академии наук" (ФИЦ КНЦ РАН) | Method of obtaining layered hydroxide of magnesium and aluminum |
| CN110284360A (en) * | 2019-07-05 | 2019-09-27 | 西安电子科技大学 | A kind of paper products of Antithemolytic streptococcus and preparation method thereof |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DK2116257T3 (en) * | 2000-08-09 | 2013-02-04 | Alk Abello As | Parenteral vaccine formulations and applications thereof |
| GB201205237D0 (en) * | 2012-03-26 | 2012-05-09 | Univ Edinburgh | Adjuvant |
| WO2019118393A1 (en) * | 2017-12-11 | 2019-06-20 | Board Of Regents, The University Of Texas System | Dry adjuvanted immune stimulating compositions and use thereof for mucosal administration |
-
2022
- 2022-11-22 CN CN202211464870.5A patent/CN115724449B/en active Active
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002011760A1 (en) * | 2000-08-09 | 2002-02-14 | Alk-Abelló A/S | Parenteral vaccine formulations and uses thereof |
| CN101717649A (en) * | 2009-11-13 | 2010-06-02 | 西安电子科技大学 | Preparation method of enhanced magnesium-aluminum composite flame retardant |
| CN102583460A (en) * | 2011-01-12 | 2012-07-18 | 国家纳米科学中心 | Aluminium adjuvant as well as preparation method and application thereof |
| CN104800843A (en) * | 2015-04-02 | 2015-07-29 | 中国农业科学院兰州兽医研究所 | Agentia for preventing toxoplasma infection and application |
| CN106049161A (en) * | 2016-05-25 | 2016-10-26 | 西安电子科技大学 | Calcium carbonate and aluminum magnesium composite flame-retardant paper, and preparation method thereof |
| CN107128962A (en) * | 2017-05-04 | 2017-09-05 | 四川理工学院 | A kind of preparation method of nano-sized magnesium hydroxide |
| RU2678007C1 (en) * | 2017-12-05 | 2019-01-22 | Федеральное государственное бюджетное учреждение науки Федеральный исследовательский центр "Кольский научный центр Российской академии наук" (ФИЦ КНЦ РАН) | Method of obtaining layered hydroxide of magnesium and aluminum |
| CN110284360A (en) * | 2019-07-05 | 2019-09-27 | 西安电子科技大学 | A kind of paper products of Antithemolytic streptococcus and preparation method thereof |
Non-Patent Citations (3)
| Title |
|---|
| Disparate adjuvant properties among three formulations of "alum";Derek W. Cain et al.;Vaccine;第31卷;第653– 660页 * |
| Stabilization of tetanus toxoid formulation containing aluminium hydroxide adjuvant against freeze-thawing;Vipul A. Solanki et al.;International Journal of Pharmaceutics;第414卷;第140– 147页 * |
| 铝佐剂的作用机制研究进展;闻晓波等;现代畜牧兽医(第11期);第47-52页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN115724449A (en) | 2023-03-03 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Togashi et al. | Surfactant-assisted one-pot synthesis of superparamagnetic magnetite nanoparticle clusters with tunable cluster size and magnetic field sensitivity | |
| Rowe et al. | Tuning the magnetic resonance imaging properties of positive contrast agent nanoparticles by surface modification with RAFT polymers | |
| Tegafaw et al. | Dual-mode T1 and T2 magnetic resonance imaging contrast agent based on ultrasmall mixed gadolinium-dysprosium oxide nanoparticles: Synthesis, characterization, and in vivo application | |
| KR101043251B1 (en) | Magnetic Resonance Imaging Contrast Agents Comprising Zinc Containing Magnetic Metal Oxide Nanoparticles | |
| Yusoff et al. | Synthesis and characterization of biocompatible Fe3O4 nanoparticles at different pH | |
| ES2675901T3 (en) | Process for the production of a magnetic iron oxide microparticle powder and an aqueous dispersion containing the magnetic particles | |
| Lee et al. | Synthesis of colloidal aqueous suspensions of a layered gadolinium hydroxide: a potential MRI contrast agent | |
| Cao et al. | Preparation of octahedral shaped Mn0. 8Zn0. 2Fe2O4 ferrites via co-precipitation | |
| CN102583460A (en) | Aluminium adjuvant as well as preparation method and application thereof | |
| Chockalingam et al. | Gum arabic modified Fe 3 O 4 nanoparticles cross linked with collagen for isolation of bacteria | |
| EP0645048A1 (en) | Preparation of controlled size inorganic particles for use in separations, as magnetic molecular switches, and as inorganic liposomes for medical applications | |
| WO2008096279A1 (en) | Compositions containing metal oxide particles and their uses | |
| CN102639153A (en) | Treating water insoluble nanoparticles with hydrophilic alpha-hydroxyphosphonic acid conjugates, the so modified nanoparticles and their use as contrast agents | |
| CN106913885B (en) | Magnetic nano particle and preparation method and application thereof | |
| CN112791181A (en) | Manganese nano adjuvant, preparation method and application thereof | |
| Magnabosco et al. | Effect of surface chemistry on incorporation of nanoparticles within calcite single crystals | |
| CN115724449B (en) | Mixed salt nano aluminum adjuvant and preparation method and application thereof | |
| JP5802237B2 (en) | Magnetic particles and method for producing the same | |
| KR101097907B1 (en) | Surface-modified Layered Gadolinium Hydroxide, Process of Preparing the Same and MRI Contrast Agent Including the Same | |
| Lassenberger et al. | Biocompatible glyconanoparticles by grafting sophorolipid monolayers on monodispersed iron oxide nanoparticles | |
| JP2007216134A (en) | Water-dispersible magnetic nanoparticle | |
| Li et al. | Facile synthesis of manganese silicate nanoparticles for pH/GSH-responsive T 1-weighted magnetic resonance imaging | |
| CN104083778B (en) | Paramagnetic metal complex that aspartic acid-phenylalanine copolymer is modified and its preparation method and application | |
| JP2007217331A (en) | Water-dispersible magnetic nano-particle | |
| US20090074671A1 (en) | Drug-immobilized inorganic nanoparticle |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |