CN116003792B - A polymer for treating cancer metastasis and its preparation method and application - Google Patents
A polymer for treating cancer metastasis and its preparation method and application Download PDFInfo
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Abstract
The application discloses a polymer for treating cancer metastasis, and a preparation method and application thereof, and relates to the field of biological medicine. The polymer is a cationic polymer and/or cationic polymer nano-particle, the cationic polymer nano-particle is formed by self-assembly of a block copolymer in an aqueous phase, and the block copolymer consists of a chain segment of a charge neutral polymer and a chain segment of the cationic polymer. The present application uses cationic polymers and cationic polymer nanoparticles having nucleic acid binding affinity and capable of accumulating in the liver for scavenging NETs capable of inducing cancer metastasis and accumulating in large numbers in the viscera, and both the soluble cationic polymer material and the cationic polymer nanoparticle material are capable of competing for binding NET-DNA from NET-DNA and CCDC25 complexes, thereby blocking their mediated cancer cell migration.
Description
Technical Field
The invention relates to the field of biological medicine, in particular to a polymer for treating cancer metastasis, a preparation method and application thereof.
Background
Nearly 90% of cancer patient deaths are caused by distant cancer metastasis. Unfortunately, since most cancers are bio-heterogeneous, heterogeneous subpopulations of cells regenerate rapidly, and the outcome of cancer metastasis relies on multiple interactions with homeostatic mechanisms, which greatly increase the difficulty of inhibiting metastasis, traditional therapies are therefore resistant to cancer metastasis. Although traditional treatment methods such as chemotherapy can reduce the focus of the primary tumor to an undetectable extent, they have poor effects on metastasis and lead to extensive side effects, which in turn deteriorate the quality of life of the patient.
Recent studies have shown that the DNA content of neutrophil extracellular traps (Neutrophi l Extrace l l u l ar Traps, nes) consisting of a DNA network and associated proteolytic enzymes, released by neutrophils, which are the most abundant leukocytes in human blood, are associated with cancer metastasis. The NETs are secreted into extracellular tissue where they capture microorganisms against invading pathogens, and although the NETs can kill microorganisms during infection, excessive NET formation can induce pro-inflammatory behavior that damages cells and hosts. Studies have shown that the nes play a key role in the metastasis of cancer, and that they can trap and attract the spread of cancer cells to distant organs, while metastatic cancer cells further promote neutrophil formation metastatic properties, supporting the metastasis of the nes in a synergistic manner, while the DNA of the nes is also identified as a receptor for the cancer cell transmembrane protein CCDC25, and excessive accumulation of the nes is found in the liver and lung of patients with breast or colon cancer metastasis. Thus, there is an urgent need to study blocking the interaction between NET-DNA and CCDC25 to effectively limit cancer metastasis, reducing mortality.
Disclosure of Invention
The present invention provides a polymer for treating cancer metastasis, a preparation method and application thereof, and provides a polymeric material capable of competing for binding to NET-DNA to block the interaction between NET-DNA and CCDC25 to effectively inhibit cancer metastasis.
In order to solve the technical problems, one of the purposes of the invention provides a polymer for treating cancer metastasis, wherein the polymer is a cationic polymer and/or cationic polymer nano-particles, the cationic polymer nano-particles are formed by self-assembly of a block copolymer in an aqueous phase, the block copolymer consists of a chain segment of a neutral polymer and a chain segment of the cationic polymer, the cationic polymer comprises one or more of positively charged poly (methyl) acrylic acid 2- (dimethylamino) ethyl ester, poly alpha-aminopentanolide, poly (2-dimethylaminoethylthio) caprolactone, polylysine, polyarginine and polyaspartic acid amino derivatives, or a copolymer thereof, the neutral polymer comprises one or more of polylactic acid-glycolic acid random copolymer, hydrophobic polyphosphate, hydrophobic polycarbonate, polyethylene glycol, polycaprolactone and hydrophobic polyamino acid, and the hydrophobic polyamino acid is one or more of poly tyrosine, poly alanine and poly alanine.
Preferably, the polymer chain segment has a number average molecular weight of 3k-500k, a weight average molecular weight of 3k-500k, a polymer dispersibility coefficient of 1-1.5, and the cationic polymer nanoparticle has a particle size of 30nm-200nm and a polydispersity index of 0.1-0.3.
Preferably, the block copolymer is poly (tyrosine-poly (N- (N' N-dimethylethylenediamine) asparagine) with a BLA amination ratio of 100%.
Preferably, the block copolymer has the chemical structural formula:
Preferably, the cationic polymer is poly (N- (N, N-dimethylethylenediamine) asparagine having a Benzyl (BLA) aspartate amination of 10% to 100%.
By adopting the scheme, the purpose of grafting N' N-dimethyl ethylenediamine in the application is to graft cations on the polyaspartic acid chain segment, so that the higher the amination rate is, the more positive charges are carried, and the stronger the capability of competing for binding NET-DNA is.
Preferably, the cationic polymer has a chemical structural formula:
in order to solve the above technical problems, another object of the present invention is to provide an application of a polymer for treating cancer metastasis in preparing a kit for treating cancer metastasis.
Preferably, the agent for inhibiting cancer metastasis kit is intravenous injection.
In order to solve the above technical problems, a third object of the present invention is to provide a method for preparing cationic polymer nanoparticles, comprising the steps of:
(1) Dissolving (S) -4- (4-hydroxybenzyl) oxazolidine-2, 5-dione in N' N-dimethylformamide, adding N-butylamine for ring-opening polymerization, precipitating in cold diethyl ether, filtering, and drying to obtain a first intermediate product;
(2) Mixing the first intermediate product, (4S) -4- [ (4-hydroxyphenyl) methyl ] -2, 5-oxazolidinedione, N' N-dimethylformamide and dichloromethane for polymerization reaction, precipitating in cold diethyl ether, filtering and drying to obtain a second intermediate product;
(3) Carrying out ammonolysis reaction on the second intermediate product and dimethyl ethylenediamine, dialyzing the product in an organic solvent to remove the dimethyl ethylenediamine, distilling to remove the organic solvent, and drying to obtain a block copolymer;
(4) And dissolving the block copolymer in a dimethyl sulfoxide aqueous solution, dropwise adding the solution until the concentration of the polymer reaches 1-3mg/mL, dialyzing to remove the dimethyl sulfoxide, and enabling the solution to be PBS with the concentration of 10 mmol/L to prepare the PBS solution containing the cationic polymer nano particles.
Preferably, in the steps (1), (2) and (3), the reaction temperature is 40-70 ℃ and the reaction time is more than 40 hours.
Preferably, in step (1), the polymerization synthesizes a first intermediate product having a polymerization degree of 10 to 40, and in step (2), the polymerization synthesizes a second intermediate product having a polymerization degree of 50 to 400.
By adopting the scheme, the first intermediate product is polymerized to form a hydrophobic segment, the second intermediate product is polymerized to produce a hydrophilic segment, the degree of polymerization of the hydrophilic segment and the hydrophobic segment can influence the particle size of the self-assembled nano particles, the degree of polymerization is increased, the overlong hydrophobic segment of the first intermediate product can lead to the morphological change of the nano particles formed by final self-assembly, the hydrophilicity of the whole polymer can be reduced, even the polymer cannot be dissolved in water, and if the hydrophobic segment is reduced too much, the polymer cannot be assembled into the nano particles. If the polymerization degree of the hydrophilic segment and the hydrophobic segment is increased or decreased by a part proportionally, the particle size of the nano particles is increased or decreased, and the effect is not influenced in a certain range, but if the change is too large, the particle size is too small, the effect is poor, and the safety is influenced if the particle size is too large.
Preferably, in the step (1), the ratio of the amounts of the (S) -4- (4-hydroxybenzyl) oxazolidine-2, 5-dione and n-butylamine is 15.7:0.7, and the degree of polymerization of the first intermediate product is 20.
Preferably, in step (2), the ratio of the amounts of the first intermediate and the (4S) -4- [ (4-hydroxyphenyl) methyl ] -2, 5-oxazolidinedione species is 0.083:13.3 and the degree of polymerization of the second intermediate is 160.
Preferably, in step (3), the ratio of the amounts of the second intermediate and the dimethylethylenediamine material is 1 (160-480).
In order to solve the above technical problems, the fourth object of the present invention is to provide a method for preparing a cationic polymer, comprising the steps of:
(1) Mixing (4S) -4- [ (4-hydroxyphenyl) methyl ] -2, 5-oxazolidinedione with N-butylamine, N' N-dimethylformamide and dichloromethane, performing polymerization under the protection of nitrogen, precipitating the obtained mixture in cold diethyl ether, and vacuum drying to obtain PBLA;
(2) And dissolving PBLA in anhydrous DMSO, adding dimethyl ethylenediamine, performing a polymerization reaction, dialyzing the mixture solution to remove excessive dimethyl ethylenediamine, performing rotary evaporation to remove the solvent, and performing vacuum drying to obtain the cationic polymer.
Preferably, in the steps (1) and (2), the reaction temperature is 40-70 ℃ and the reaction time is more than 40 hours.
Preferably, in step (1), the ratio of the amounts of the (4S) -4- [ (4-hydroxyphenyl) methyl ] -2, 5-oxazolidinedione and n-butylamine is 1205:8.
Preferably, in step (2), the ratio of PBLA to dimethylethylenediamine is 1 (16-360).
Preferably, the amination ratio of the cationic polymer is 100%, and the ratio of the amounts of PBLA and dimethylethylenediamine in step (2) is1 (144-360).
By adopting the scheme, the application uses benzyl aspartate inner ring anhydride monomer (BLA-NCA) to polymerize to obtain polyaspartic acid substituted by benzyl ester, and then uses dimethyl ethylenediamine to substitute benzyl, and finally the purpose is to obtain polyaspartic acid with side chains replaced by dimethyl ethylenediamine, which is to lead the polymer to have positive charges after tertiary amine is introduced.
Compared with the prior art, the embodiment of the invention has the following beneficial effects:
the present application uses cationic polymers and cationic polymer nanoparticles that have high nucleic acid binding affinity and accumulate in the liver for scavenging of NETs that accumulate in large amounts in the liver prior to increased levels in plasma, and both soluble cationic polymer materials and cationic polymer nanoparticle materials can compete greatly for NET-DNA from the NET-DNA and CCDC25 complexes, thereby blocking their mediated cancer cell chemotaxis and migration.
Drawings
FIG. 1 is a schematic representation of the synthesis of cationic polymer nanoparticles of the present invention and a schematic representation of competitive binding to NET-DNA;
FIG. 2 shows the synthetic routes of the cationic polymer and the cationic polymer nanoparticle in examples 1-2 (note: A-synthetic route of the cationic polymer in example 1; B-synthetic route of the cationic polymer nanoparticle in example 2) according to the present invention;
FIG. 3 is a chart showing the nuclear magnetic resonance hydrogen spectrum (1 HNMR) of the cationic polymer in example 1 of the present invention;
FIG. 4 is a chart showing the nuclear magnetic resonance hydrogen spectrum (1 HNMR) of the cationic polymer nanoparticle in example 2 of the present invention;
FIG. 5 is a graph showing the gel permeation chromatography flow out time curves of PBLA in example 1 and PTyr 20-b-PBLA160 in example 2 according to the present invention;
FIG. 6 shows the DLS particle diameter and TEM result (note: left-DLS; right-TEM) of the cationic polymer nanoparticles in example 2 of the present invention;
FIG. 7 shows the results of cytoskeletal remodeling observations of different groups of example 1 for the efficacy of the invention;
FIG. 8 shows the results of the cell chemotaxis assay of the different groups of example 2 for verifying the effect of the present invention;
FIG. 9 shows the results of in vivo anticancer cell metastasis experiments of 4T1 mouse model in example 3 (note: A-experimental progress schematic; B-mouse liver digital photograph, white arrow represents cancer metastasis, C-mouse liver luminous intensity, stronger luminous intensity, more tumor cells representing metastasis, 40 Xlow-power photomicrograph of H & E staining section of D-mouse liver, 200 Xhigh-power photomicrograph of H & E staining section of E-mouse liver, dotted line circled part represents tumor infiltration, black arrow represents immune cell infiltration, F-mouse lung luminous intensity, stronger luminous intensity, more tumor cells representing metastasis, 40 Xlow-power photomicrograph of H & E staining section of G-mouse lung, 200 Xhigh-power photomicrograph of H & E staining section of H-mouse lung, dotted line circled part represents tumor infiltration, black arrow represents immune cell infiltration, survival curve of I-mouse from 13 days to 28 days);
FIG. 10 shows the results of in vivo anticancer cell metastasis experiments of MDA-MB-231 human in situ breast cancer mouse model in effect test example 4 of the present invention (note: A-experiment progress schematic diagram, B-mouse liver digital photograph, white arrow represents cancer metastasis, C-mouse liver luminous intensity, more intense luminescence, more cancer cells representing metastasis, 200 Xhigh-power mirror photograph of H & E staining section of D-mouse liver, dotted line circled part represents tumor infiltration, black arrow represents immune cell infiltration, E-mouse lung luminous intensity, more intense luminescence, more cancer cells representing metastasis, 200 Xhigh-power mirror photograph of H & E staining section of F-mouse lung, dotted line circled part represents tumor infiltration, black arrow represents immune cell infiltration, G-mouse weight change statistical result, survival curve of 50 days to 137 days after H-mouse model, NET expression of I-mouse liver immunofluorescence staining experiment, and J-mouse lung immunofluorescence staining experiment, white arrow represents NET expression;
FIG. 11 shows the results of in vivo anticancer cell metastasis experiments (note: A-experimental progress schematic; B-mouse liver metastasis nodule representative morphology; C-mouse liver metastasis nodule quantitative analysis; D-mouse liver luminous intensity; E-mouse liver luminous intensity quantitative analysis; F-mouse liver H & E staining section 40X low-magnification photomicrographs; G-mouse liver H & E staining section 200X high-magnification photomicrographs; dotted line circled portion represents tumor infiltration, black arrow represents immune cell infiltration; H-mouse liver tumor infiltration zone quantitative analysis) of HCT116 human colon cancer mouse model in accordance with the effects of the present invention.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
The application uses cationic polymer material to combine NET-DNA electrostatically as a new anti-transfer strategy, as shown in figure 1, the strong electrostatic interaction of cationic polymer nano particles cANP and DNA can compete with CCDC25 for combining NET-DNA, thereby reducing the chemotactic ability of NET-DNA in the cancer metastasis process, and blocking the interaction between NET-DNA and CCDC25 can inhibit cancer metastasis. The use of soluble polymeric materials or their nanoparticulate materials which have high nucleic acid binding affinity and accumulate in the liver can greatly compete for NET-DNA from the NET-DNA and CCDC25 complexes, thereby blocking their mediated cancer cell chemotaxis and migration for the removal of NETs which accumulate in large amounts in the liver prior to increased levels in plasma. The treatment effectively inhibits the cancer metastasis of the 4T1 in-situ breast cancer tumor mouse model, and the same curative effect is observed in the human breast cancer MDA-MB-231 and colon cancer HCT116 metastasis models, which indicates that the cationic polymer material has obvious curative effect in blocking the cancer metastasis.
Example 1
A method for preparing a cationic polymer, the synthetic route of which is shown as a in fig. 2, comprising the steps of:
(1) PBLA Synthesis of PBLA into a 100mL Sch l enk flask were added 3g (BLA-NCA, 99.5% purity, source leaf organism, 12.05 mmol) (4S) -4- [ (4-hydroxyphenyl) methyl ] -2, 5-oxazolidinedione, followed by 10mL of a solution of 5.9mg N-butylamine (98% purity, A l add i N,0.08 mmol) in N' N-Dimethylformamide (DMF) and 48mL of dichloromethane, the polymerization was carried out under nitrogen protection, the temperature was 40℃and the reaction was carried out for 48h, the mixture obtained after the reaction was precipitated in a large amount of cold diethyl ether and repeated three times, and then dried under vacuum at room temperature for 24h to obtain PBLA.
The polymer had a monomer conversion of 96% and a degree of polymerization of 144 as analyzed by 1H NMR (Bruker AVANCE I I I MHz) and a polydispersity Mw/Mn=1.07 as shown in FIG. 5 by gel permeation chromatography (GPC, agi l ent) with DMF as eluent.
(2) PAsp was synthesized by first dissolving 0.3g PBLA (0.01 mmo l) in 6mL of anhydrous DMSO, placing the solution in three Sch l enk flasks, respectively, then adding 320mg (3.6 mmo l,2.5 times equivalent benzyl), 77.9mg (0.88 mmo l, about 0.61 times equivalent benzyl) and 14.2mg (0.16 mmo l, about 0.11 times equivalent benzyl) of dimethylethylenediamine (dmeen, purity 98%, al add i N) to the three Sch l enk flasks, respectively, and after ammonolysis at 40℃for 48 hours, the mixture solution was dialyzed (molecular weight cut off: 3500 Da) in methanol to remove excess dimethylethylenediamine, then spin-evaporating the solvent, and vacuum drying to obtain PAhomopolymers of 100%, 50% and 10% sp, respectively, referred to as poly N- (N, N-dimethylethylenediamine) asparagine PA100%, PAsp-50% and PAsp-10%.
Example two
A method for preparing cationic polymer nanoparticles, the synthetic route of which is shown as B in fig. 2, comprising the steps of:
(1) PTyr-NH 2 Synthesis in a glove box, 3g (15.7 mmo l, tyr-NCA, 97% purity, mack l i n) (S) -4- (4-hydroxybenzyl) oxazolidine-2, 5-dione were dissolved in 50mL DMF, put into a 100mL Sch l enk flask, then 54.7mg (0.7 mmo l) n-butylamine was added, the polymerization was carried out under a nitrogen atmosphere at 40℃for 48H, then the resulting mixture was precipitated in a large amount of cold diethyl ether, repeated three times, filtered and dried under vacuum for 24 hours, and analyzed by 1H NMR spectroscopy to give a polymerization degree of 20, namely PTyr 20-NH2.
(2) PTyr 20-b-PBLA160 Synthesis of a diblock copolymer PTyr 20-b-PBLA160 was obtained by charging 0.25g (0.083 mmol) PTyr 20-NH2, 3.32g (13.3 mmol) BLA-NCA, 10mL DMF and 50mL methylene chloride into a 100mL Sch l enk flask, then polymerizing at 40℃for 48 hours, precipitating the resulting mixture after the reaction in a large amount of cold diethyl ether, repeating three times, and then vacuum drying at room temperature for 24 hours, and by analysis of 1H NMR spectrum, PBLA monomer conversion was 99% and polymerization degree was 160, and the polydispersity Mw/Mn of the polymer was 1.10 as measured by GPC using DMF as eluent, as shown in FIG. 5.
(3) PTyr 20-b-PAsp160 -100% Synthesis by dissolving 0.3g PTyr 20-b-PBLA160 (0.0075 mmoL) in 6mL DMSO, then adding 249.2mg of dimethylethylenediamine (2.82 mmoL, about 2.4 times equivalent benzyl), subjecting the mixture solution to ammonolysis reaction at 40℃for 48 hours, dialyzing the mixture solution in methanol for 48 hours (molecular weight cut-off: 3500 Da) to remove excess dimethylethylenediamine, then spin-evaporating the solvent, and vacuum-drying to obtain PTyr 20-b-PAsp160 -100% block copolymer.
(4) Self-assembly preparation of cANP-100% nanoparticles PTyr 20-b-PAsp160 -100% was first dissolved in dimethyl sulfoxide (DMSO) with a controlled concentration of polymer in the mixture of 10mg/mL, then a solution of DMSO containing PTyr 20-b-PAsp160 -100% was slowly dropped into deionized water until the polymer concentration reached 2mg/mL, the mixture solution was dialyzed against deionized water (molecular weight cut-off: 3500 Da) to remove DMSO, and a mixture of Na 2HPO4 and KH 2PO4 was added to the cANP-100% solution, changing the aqueous solution to PBS buffer, wherein the phosphate concentration was 10mM.
Effect verification example 1
The cationic polymer and nanoparticles were subjected to an inhibition of cancer cytoskeletal remodeling assay:
To evaluate the inhibition of NET-DNA induced cytoskeletal reconstruction by PAsp in example 1 and cANP in example 2, 2X 10 3 (per well) MDA-MB-231 cells (human breast cancer cells) were placed in 8-well plates, after overnight incubation, old medium was replaced with new serum-free medium containing 5g/mL NET-DNA and 50g/mLPAsp-100% or cANP-100%, PBS was blank, after incubation for 3 hours at 37 ℃, the supernatant from the wells was taken, nuclei were stained with Al exa F l uor TM 555Pha l l oid i n, and then observed with confocal microscopy for quantitative analysis, and cell filaments were counted from 10 regions of each well, respectively, and 3 independent experiments were performed simultaneously.
Experimental results show that NET-DNA was greatly blocked from altering the cytoskeleton after addition of the cationic polymer and cationic polymer nanoparticle materials, as shown in fig. 7, where the cationic polymer nanoparticle sample was most pronounced and cytoskeletal remodeling was necessary for cancer cell migration.
Effect verification example two
Cationic polymers and nanoparticles were used to inhibit migration of cancer cells (cell chemotaxis assay):
To evaluate the chemotactic activity of PAsp in example 1 and cANP in example 2 on tumor cells, MDA-MB-231 cells at a concentration of 3X 10 6/mL were moved to the seeding chamber of the mu-S l ide chemotactic chamber, after cancer cells adhered to the seeding chamber, the left side reservoir was filled with 5. Mu.g/mL NET-DNA, 5. Mu.g/mL NET-DNA+50. Mu.g/mL PAsp-100% or 5. Mu.g/mL NET-DNA+50. Mu.g/mL cANP-100% PBS solution, the right side reservoir was filled with PBS solution alone, and then cancer cell images were captured in the chemotactic chamber every 8 minutes using LionheartTM FX automated living cell imager for 7-12 hours, followed by analysis of the cancer cell migration trajectory.
The results show that the cationic polymer and cationic polymer nanoparticles of the experimental group can significantly inhibit cancer cell migration caused by NET-DNA compared to the NET-DNA control group, as shown in fig. 8.
Effect verification example three
In vivo anticancer cell metastasis experiments of cationic polymer and cationic nanoparticle materials to inhibit 4T1 in situ breast cancer tumor mouse cancer:
(1) Grouping the mice of Ba l b/c with a weight of 20g into a model group, wherein PAsp-100% administration groups (different concentrations, 5mg/kg,10mg/kg,20mg/kg respectively) in the three groups of example 1 and cANP-100% administration groups (different concentrations, 5mg/kg,10mg/kg,20mg/kg respectively) in the three groups of example 2, and 7 groups of 8 mice each;
(2) Molding, namely after the mice are anesthetized by isoflurane gas, cutting the skin on the surface of mammary gland, re-suspending 5X 10 5 4T1-L uc cells in 100 mu L PBS, injecting the mixture into the 4 th mammary gland fat pad on the right side of the BALB/c mice by using a syringe, and then suturing the incision;
(3) The administration is that from the 8 th day after molding, the PBS solution of PAsp or cANP with doses of 5mg/kg, 10mg/kg and 20mg/kg is injected into the tail vein of the mice of the corresponding group once every two days, and the PBS solution with the same volume is injected into the mice of the model group for 20 days;
(4) Recording tumor volume and weight change of the mice, namely measuring and recording length and width of in-situ tumor by using a vernier caliper every two days during the administration period, and calculating in-situ tumor volume;
(5) Monitoring tumor metastasis during administration, detecting tumor metastasis in mice by using an IVI S Lumi na X5 (PERKI NE LMER) luminescence imager, and analyzing the luminescence intensity of in-situ tumor and different organs by using the luminescence imager after mice are killed;
(6) Blood collection and tissue slicing, namely, blood collection, serum collection and euthanasia of mice are carried out on day 28 of molding, namely, 20 days after administration, heart, liver, spleen, lung and kidney are taken for tissue slicing study, and a camera is used for taking organ photographs of cancer metastasis sites of liver, lung and the like, as shown in B-H in fig. 9.
The results show that, as shown in fig. 9, both PAsp and cANP have an inhibitory effect on liver and lung metastasis of breast cancer, compared with the model group, the tumor infiltration area and immune cell recruitment of treated mice at the liver and lung are greatly reduced, macroscopic cancer metastasis is also obviously reduced, the survival curve also shows that the life of treated mice is prolonged compared with untreated model group, meanwhile, with the increase of the treatment concentration of PAsp or cANP, the inhibition effect on tumors is more obvious, and the treatment effect of cANP on tumors is better than that of PAsp, and is hypothesized that the nano particles are long in liver detention time and cannot be metabolized immediately.
Effect verification example four
In vivo anticancer cell metastasis experiments of the cationic polymer and cationic nanoparticle materials to inhibit MDA-MB-231 in situ breast cancer tumor mouse cancer:
(1) Grouping NOD/SCI D mice weighing 20g into two large groups, each large group comprising three small groups, one model group, one PAsp-100% administration group of example 1 (concentration 10 mg/kg), one cANP-100% administration group of example 2 (concentration 10 mg/kg), the first large group of injected cancer cells being MDA-MB-231-l uc, the second large group of injected cancer cells being MDA-MB-231, 6 total groups of 4 mice, wherein the first large group can monitor the condition of cancer metastasis in real time due to the access of l uc groups, the second large group being used to evaluate the effect of cationic materials on the prolongation of the life of mice;
(2) Modeling, namely after the mice are anesthetized by isoflurane gas, incising the skin on the surface of the mammary gland, re-suspending 2X 10 6 MDA-MB-231-L uc cells in 100 mu L PBS, injecting the PBS into a4 th mammary fat pad on the right side of the NOD/SCI D mice by using a syringe, and then suturing the incision;
(3) The administration is that from the 8 th day after molding, the PBS solution of PAsp or cANP with the dosage of 10mg/kg is respectively injected into the mice of the corresponding group by tail vein once every two days, and the PBS solution with the same volume is injected into the mice of the model group for 46 days;
(4) Recording tumor volume and weight change of the mice, namely measuring and recording length and width of in-situ tumor by using a vernier caliper every two days during the administration period, and calculating in-situ tumor volume;
(5) Monitoring tumor metastasis during administration, detecting tumor metastasis in mice by using an IVI S Lumi na X5 (PERKI NE LMER) luminescence imager, and analyzing the luminescence intensity of in-situ tumor and different organs by using the luminescence imager after mice are killed;
(6) Blood collection and tissue section, namely, blood collection, serum collection and euthanasia of the first group of mice are carried out on the 54 th day of molding, namely, 46 days after administration, heart, liver, spleen, lung and kidney of the first group of mice are taken for tissue section study, and an organ photo of cancer metastasis sites such as liver, lung and the like is taken by a camera.
The results show that, as shown in fig. 10, both PAsp and cANP have an inhibitory effect on liver and lung metastasis of MDA-MB-231 human breast cancer, compared with the model group, the tumor infiltration area and immune cell recruitment of treated mice at the liver and lung are greatly reduced, macroscopic cancer metastasis is also obviously reduced, NET expression at the liver and lung is also obviously reduced, and the survival curve also shows that the life span of treated mice is prolonged compared with untreated model group, and the treatment effect of cANP on tumors is better than that of PAsp.
Effect verification example five
In vivo anticancer cell metastasis experiments of cationic polymer and cationic nanoparticle materials to inhibit HCT116 human colon cancer mouse cancer:
(1) Grouping NOD/SCI D mice weighing 20g were divided into a model group (PBS), a PAsp-100% administration group (concentration 10 mg/kg) in example 1, a cANP-100% administration group (concentration 10 mg/kg) in example 2, 3 groups of 6 mice each;
(2) Molding, namely after the mice are anesthetized by isoflurane gas, cutting abdominal cavity, re-suspending 1X 10 6 HCT116-L uc cells in 50 mu L PBS, injecting the cells into NOD/SC ID spleen by a syringe, and then suturing the incision;
(3) The administration is that from the 8 th day after molding, the PBS solution of PAsp or cANP with the dosage of 10mg/kg is respectively injected into the mice of the corresponding group by tail vein once every two days, and the PBS solution with the same volume is injected into the mice of the model group for 36 days;
(4) Monitoring tumor metastasis during administration, detecting tumor metastasis in mice by using an IVI S Lumi na X5 (PERKI NE LMER) luminescence imager, and analyzing the luminescence intensity of in-situ tumor and different organs by using the luminescence imager after mice are killed;
(5) Taking blood and tissue section, namely taking blood and serum after administration for 46 days on 54 days of molding, euthanizing the mice of the first group, taking the heart, liver, spleen, lung and kidney of the mice for tissue section study, and taking organ photographs of cancer metastasis sites such as liver, lung and the like by using a camera;
As shown in FIG. 11, the results show that the PAsp and cANP have an inhibitory effect on liver and lung metastasis of human colon cancer, compared with a model group, the tumor infiltration area and immune cell recruitment of treated mice in the liver and lung are greatly reduced, macroscopic cancer metastasis is also obviously reduced, and the treatment effect of cANP on tumors is better than that of PAsp.
The foregoing embodiments have been provided for the purpose of illustrating the general principles of the present invention, and are not to be construed as limiting the scope of the invention. It should be noted that any modifications, equivalent substitutions, improvements, etc. made by those skilled in the art without departing from the spirit and principles of the present invention are intended to be included in the scope of the present invention.
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| EP2135601A1 (en) * | 2008-06-20 | 2009-12-23 | Capsulution Nanoscience AG | Stabilization of amorphous drugs using sponge-like carrier matrices |
| EP2172193A1 (en) * | 2008-10-02 | 2010-04-07 | Capsulution Nanoscience AG | Improved nanoparticulate compositions of poorly soluble compounds |
| US8691284B2 (en) * | 2011-07-07 | 2014-04-08 | Empire Technology Development Llc | Fluorinated block co-polymers |
| CN102516552B (en) * | 2011-11-18 | 2014-01-15 | 中山大学 | A kind of degradable acid-sensitive polymer amphiphilic cationic block copolymer and micelle particle and preparation method thereof |
| CN110483785B (en) * | 2019-07-01 | 2020-05-19 | 中山大学 | Triblock polymer, drug-loaded nano micelle, nano drug, and preparation method and application thereof |
| CN113209018A (en) * | 2020-04-08 | 2021-08-06 | 广州医科大学附属第二医院 | pH-sensitive nano micelle and preparation method and application thereof |
| CN112029091B (en) * | 2020-09-18 | 2022-06-10 | 中国药科大学 | PH/reduction dual-responsiveness block copolymer PEG-b-PASp-g-CPA |
-
2022
- 2022-12-15 CN CN202211631365.5A patent/CN116003792B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105997879A (en) * | 2016-07-12 | 2016-10-12 | 中山大学 | PH and temperature sensitive nano-vesicles and preparing method and application thereof |
| CN110179749A (en) * | 2018-04-27 | 2019-08-30 | 中山大学 | It can be used for treating the polymer nano granules of rheumatoid arthritis |
Non-Patent Citations (1)
| Title |
|---|
| 王国建等.功能高分子材料.同济大学出版社,2014,(第2版),第289-290页. * |
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