CN110898033A - Drug delivery system with tumor microenvironment regulation and targeting functions and application of drug delivery system in pharmacy - Google Patents
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Abstract
The invention belongs to the field of screening of effective components of traditional Chinese medicines and preparation thereof, and relates to a drug delivery system with functions of adjusting tumor microenvironment and targeting and application thereof in pharmacy, in particular to application thereof in pancreatic tumor treatment. The nano delivery system is a biocompatible, biodegradable nano particle which is constructed by medicinal high-molecular polylactic acid materials and has the functions of regulating a tumor microenvironment and increasing blood perfusion, and the surface of the nano delivery system is covalently combined with a segment of functional polypeptide CGKRK which can target heparan sulfate highly expressed in the tumor microenvironment and improve the tumor targeting property; the natural drug monomers are physically entrapped in the hydrophobic core of the nano particles by an emulsified solvent evaporation method, so that the solubility of the drug and the in vivo circulation time are improved. The established drug-loaded nano system can inhibit the over-activation of CAFs, reduce the proportion of M2 type macrophages, effectively improve the tumor microenvironment and increase the accumulation of drugs in tumor parts.
Description
Technical Field
The invention belongs to the field of screening of effective components of traditional Chinese medicines and preparation thereof, and relates to a drug delivery system with functions of adjusting tumor microenvironment and targeting and application thereof in pharmacy, in particular to application thereof in pancreatic tumor treatment. The invention relates to a method for encapsulating a hydrophobic natural drug monomer with short half-life period in biodegradable polylactic acid nanoparticles, delivering the hydrophobic natural drug monomer to a tumor part in a targeted manner to improve the tumor microenvironment, increasing the penetration and accumulation of subsequent chemotherapeutic drugs at the tumor part, and investigating the action mechanism of the hydrophobic natural drug monomer.
Background
The prior art discloses digestive tract cancer with malignant pancreatic cancer, the treatment tolerance is mainly related to immunosuppressive Tumor Microenvironment (TME), and research shows that more than 80% of tumor microenvironment is mesenchymal cells, including tumor-associated fibroblasts (CAFs), endothelial cells, pericytes, tumor-associated macrophages and the like, wherein the content of CAFs is the most, a large amount of abnormal activation is performed in TME of pancreatic cancer, and besides the function of supporting tumor, a large amount of collagen fibers can be generated, extracellular matrix can be secreted to generate irregular deposition, and higher interstitial fluid pressure is formed; various immunosuppressive cytokines are secreted to interact with tumor cells, thereby hindering drug delivery at the tumor site and promoting cancer progression and metastasis. In addition, immunosuppressive M2-type macrophages also secrete immunosuppressive molecules that affect tumor vasculature and permeability blocking drug delivery. The existing chemotherapy combined scheme shows that the curative effect of resisting pancreatic cancer is improved, but the tumor response rate is still low and the prognosis is poor, so that for the tumor with an immunosuppressive tumor microenvironment, such as pancreatic cancer, a drug with low toxicity is selected to regulate the tumor microenvironment, an activated signal channel is blocked to inhibit abnormal activation of CAFs, the deposition of tumor extracellular matrix is reduced, and the polarization of macrophages to M2 type is inhibited, and the chemotherapeutic drug is combined to improve the anti-cancer curative effect, so that the chemotherapy combined scheme has a high clinical application prospect and a high conversion value.
Natural product monomers extracted from traditional Chinese medicines have proved to have great application prospects in recent years in order to improve the tumor microenvironment and improve the penetration and accumulation of chemotherapeutic drugs. However, natural products have the defects of poor solubility, short in-vivo circulation half-life period, easy removal, incapability of accumulating in tumor parts and the like, and based on the current situation of the prior art, the inventor of the application intends to provide a drug delivery system with functions of regulating tumor microenvironment and targeting and application thereof in pharmacy, particularly application thereof in pancreatic tumor treatment.
Disclosure of Invention
The invention aims to provide a drug delivery system with functions of regulating tumor microenvironment and targeting and application thereof in pharmacy, in particular application in pancreatic tumor treatment based on the current state of the prior art.
The invention prepares the screened natural product monomer into drug-loaded nanoparticles by high-performance biodegradable materials, modifies functional polypeptide to realize targeted delivery to tumor microenvironment, inhibits the activation of CAFs, reduces the proportion of M2 type macrophages, and increases blood perfusion to promote the penetration and accumulation of the drug at tumor sites.
Specifically, the invention provides a drug capable of regulating a tumor microenvironment and a nano delivery system with a targeting function, wherein the drug is constructed by a medical high-molecular polylactic acid material approved by FDA, has good biocompatibility, is biodegradable, has a function of regulating the tumor microenvironment and increasing blood perfusion, and is covalently combined with a segment of functional polypeptide CGKRK on the surface, and can target heparan sulfate highly expressed in the tumor microenvironment so as to improve the tumor targeting property; the natural drug monomers are physically entrapped in the hydrophobic core of the nano particles by an emulsified solvent evaporation method, so that the solubility of the drug and the in vivo circulation time are improved.
In the invention, the high molecular polylactic acid material contains a methylated polyethylene glycol-polylactic acid block copolymer (mPEG-PLA); the molecular weight of the methylated polyethylene glycol-polylactic acid block copolymer mPEG-PLA is 10000-50000 Da.
In the invention, the functional polypeptide CGKRK is modified on the surface of the nanoparticle through malondialdehyde-sulfhydryl reaction, so that the aim of targeting a tumor part is fulfilled.
The model drug adopted by the invention is a natural drug monomer fraxinellone, and is encapsulated in a biodegradable polylactic acid material in a physical encapsulation mode.
The screened acerolone natural product has low toxicity, can effectively inhibit but cannot completely kill over-activated CAFs in a tumor microenvironment, reduces the secretion of extracellular matrix, and reduces the interstitial fluid pressure of tumors; meanwhile, the polarization of macrophages to M2 type is reduced, the normalization of tumor blood vessels is promoted, and the blood flow perfusion is increased, so that the delivery efficiency and accumulation of chemotherapeutic drugs are improved, and the drug effect is better exerted.
According to the invention, an emulsion solvent evaporation method is adopted to encapsulate hydrophobic natural drug monomers in biodegradable polylactic acid nanoparticles and modify a segment of functional polypeptide, so that the stability of the drug in blood circulation can be improved, the circulation time can be prolonged, a large amount of drug can be accumulated in a tumor microenvironment, and the effect of regulating the tumor microenvironment can be exerted;
according to the invention, the hydrophobic natural drug monomer is encapsulated in the biodegradable polylactic acid nanoparticles by an emulsified solvent evaporation method, so that the defects that the natural product monomer has short half-life period in vivo, is quickly removed and cannot be accumulated in a tumor microenvironment can be effectively overcome, the circulation time of the natural product monomer in vivo is greatly prolonged, and the natural product monomer is greatly enriched in the tumor microenvironment.
The murine fibroblast NIH3T3, macrophage RAW264.7 and human pancreatic cancer cell Panc-1 used in the present invention are all recognized in the art and commercially available.
The invention provides a preparation method of a drug delivery system, which adopts an emulsified solvent evaporation method to prepare a nano drug delivery system and comprises the steps of dissolving fraxinellone powder and mPEG-PLA in dichloromethane, adding 1% sodium cholate, carrying out 220W probe ultrasonic treatment under an ice bath condition, dispersing by using 0.5% sodium cholate solution, and carrying out rotary evaporation to remove residual dichloromethane; centrifuging at 4 deg.C after rotary steaming, discarding supernatant to obtain drug-loaded nanoparticles (Frax-NP, Free drug is Frax-Free), and the obtained fraxinellone nanoparticles have uniform particle diameter, low cytotoxicity to fibroblast, and good safety.
The invention provides the result of the action mechanism investigation of the natural drug monomer,
the invention proves that the drug delivery system can inhibit the activation of CAFs in vitro and in vivo and reduce M2 type macrophages through Western Blot experiments and flow cytometry experiments, the action mechanism of the CAFs is mainly related to related proteins of TGF- β/smad passage, the nano delivery system of the drug for adjusting the tumor microenvironment acts on the CAFs through inhibiting the expression of psmad2 and psmad3 of the TGF- β/smad passage, and the photoacoustic oxygen saturation experiment results prove that the tumor microenvironment can be obviously improved through multiple times of administration of the fraxinellone nano preparation, the normalization of blood vessels and the increase of blood perfusion are promoted, so that the permeation and accumulation of the combined chemotherapy gemcitabine at the tumor part are obviously increased.
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FIG. 1: nano preparation characterization and cytotoxicity of the screened drug fraxinellone, wherein,
(A) particle size distribution of Frax-NP, MTT results 24h after NIH3T3 cells were administered to Frax-NP.
FIG. 2: the effect of fraxinellone on M2 typing of macrophages and on CAFs cell protein expression, wherein,
(A) effect of Fraxinellone Free drug (Frax-Free) and Frax-NP on M2 typing of activated macrophages,
(B) effects of high and low doses of Frax-NP on expression of individual proteins in NIH3T3 cells after activation.
FIG. 3: the mechanism of action of fraxinellone on CAFs, wherein,
(A) Frax-Free modulation of TGF- β/smad pathway-associated proteins in NIH3T3 cells at various time points,
(B) modulation of TGF- β/smad pathway-associated proteins by Frax-NP in NIH3T3 cells at various time points.
FIG. 4: effect of fraxinellone on blood perfusion in subcutaneous and in situ pancreatic cancer model animals, wherein,
(A) qualitative changes in tumor blood flow perfusion after subcutaneous pancreatic cancer model animals were dosed,
(B) quantitative result of tumor blood flow perfusion related parameters after subcutaneous pancreatic cancer model animal administration,
(C) qualitative and quantitative results of tumor blood oxygen saturation after administration to orthotopic pancreatic cancer model animals.
FIG. 5: the in vivo pharmacodynamic evaluation of the fraxinellone nano preparation on in-situ pancreatic cancer model animals is carried out, wherein,
(A) western Blot results of fibrinectin, α -SMA and FAP proteins in tumor tissue,
(B) western Blot results of TGF- β/smad pathway-associated proteins in tumor tissues,
(C) a change in macrophage typing in tumor tissue,
(D) immunohistochemistry results for different proteins in tumor tissues. A scale: the thickness of the film is 200 mu m,
(E) the life cycle of the tumor-bearing nude mice after multiple administration;
in the table: the analysis of the significance difference between each group and the PBS group adopts a one-factor anova method, n.s. represents that p is more than 0.05, namely, no significance difference exists, and p is less than 0.05, namely, the significance difference exists. P < 0.05, p < 0.01, p < 0.001, p < 0.0001.
Detailed Description
Example 1: preparation characterization and cytotoxicity of fraxinellone nano preparation
Preparing a nano drug delivery system by adopting an emulsified solvent evaporation method, weighing 5mg of fraxinellone powder and 250mg of mPEG-PLA, dissolving in 10mL of dichloromethane, adding 2mL of 1% sodium cholate, carrying out ultrasonic treatment for 2.4min by a 220W probe under an ice bath condition at an interval of 2s, dispersing for 10min by using 80mL of 0.5% sodium cholate solution, and carrying out rotary evaporation at 40 ℃ for 20min to remove residual dichloromethane. After rotary evaporation, centrifuging at 15000rpm/min for 1h at 4 ℃, and removing the supernatant to obtain drug-loaded nanoparticles (Frax-NP, Free drug is Frax-Free). The particle size of the nanoparticles was measured with a Malvern particle size/Zeta potential meter. After NIH3T3 cells adhere to the wall for 24h, medicines with different concentration gradients are added, and after incubation for 24h, the cytotoxicity of the fraxinellone nano preparation on fibroblast NIH3T3 is detected by using an MTT colorimetric method;
the results show that: FIG. 1A shows the Frax-NP particle size around 100nm, and the quantification of FIG. 1B shows that the IC50 value of fraxinellone nanoparticles to NIH3T3 is 61. mu.M. The results show that the fraxinellone nanoparticles have uniform particle size, low cytotoxicity to fibroblasts and high safety.
Example 2: fraxinellone examination of macrophage M2 typing and CAFs cell protein expression
To verify whether the fraxinellone nanosystem affects tumor-associated macrophages in the tumor microenvironment, the change in the proportion of M2 type macrophages after administration was examined. Inactivated RAW264.7 cells are used as a negative control group, RAW264.7 activated by 10ng/ml IL-13 is used as an M2 type positive control group, and the inhibition effect of the free drug containing 20 mu M fraxinellone and the nano preparation on macrophage M2 typing is compared. Specifically, a 6-well plate is planted by RAW264.7 cells, stimulation factor IL-1310 ng/ml is added in each well except a negative control group, Frax-Free and Frax-NP with 20 mu M fraxinellone concentration and Phosphate Buffer Solution (PBS) with the same volume are respectively added after 24 hours of adherence in an incubator, cells are collected after 12 hours, M2 type macrophage marker (CD206) marked by fluorescent dye PE is added for staining for 30 minutes, and the proportion of positive cells is scanned by flow cytometry after PBS is cleaned.
In addition, in order to examine the influence of fraxinellone on labeled protein in activated CAFs, after a 6-well plate is planted in NIH3T3 cells for 24h, a DMEM culture medium containing 10ng/mL TGF- β is added for stimulation for 24h, nano preparations carrying 5 mu M fraxinellone and 20 mu M fraxinellone are respectively used as a low-dose group and a high-dose group, no medicine is given for stimulation, and no medicine is given only for stimulation and no medicine is given as a negative control group and a positive control group respectively.
The results are shown in fig. 2A, which shows that CD206 positive cells are increased from 0.558% to 21.6% after RAW264.7 cells are incubated with IL-13 for 24h, indicating that macrophages are converted into M2 type, and the proportion of M2 type is obviously reduced after fraxinellone free drug and nano preparation is given, respectively 10.5% and 2.87%, the inhibition effect of fraxinellone nano preparation group on M2 type is stronger than that of free drug, indicating that fraxinellone-loaded nano preparation can obviously reduce the proportion of macrophage M2 type, fig. 2B shows that fibroblast activated protein actin 7 (Fraxinellone-loaded nano preparation) 5634 h is increased after NIH3T3 cells are incubated with TGF- β for 24h, indicating that fibroblasts are converted into CAFs, compared with a positive control group, the nano ketone activated protein (FAP) and Fibronectin (fibrictectin) expression quantity is increased greatly, indicating that the nano preparation has a good inhibitory effect on macrophage inhibition effect and the inhibition effect of macrophage activation of nano preparation on tumors is reduced by FAP 26 and the dose dependence of nano preparation on FAP.
Example 3: research on action mechanism of Fraxinellone on CAFs
The experiment uses a Western Blot method to investigate the expression condition of TGF- β/smad pathway related proteins in CAFs after administration of fraxinellone, uses NIH3T3 after 10ng/ml TGF- β stimulates for 24h as a positive control, respectively gives free drugs containing 20 mu M fraxinellone and nano preparations for 2h, 8h and 24h, examines the change conditions of phosphorylated and non-phosphorylated smad2 and smad3 proteins, uses Tranilast (tranit, 100 mu M) as a positive drug, takes the administration time of 24h as the positive drug, and prepares for protein quantification and sample preparation in the same way as the example 2;
the results show that, as can be seen from fig. 3A and 3B, after 24h of stimulation by TGF- β, the expression levels of phosphoproteins psmad2 and psmad3 in NIH3T3 cells are significantly increased, which indicates that NIH3T3 has been converted into an activated state, after the fraxinellone free drug is given, the expression of two phosphorylated proteins is gradually reduced, but the inhibition effect is not obvious at 24h, on the contrary, the fraxinellone nano-preparation can continuously inhibit the expression of the two phosphorylated proteins within 24h, and the inhibition degree is time-dependent, after the two dosage forms are administered, the non-phosphorylated proteins smad2 and smad3 are not obviously changed, and the results show that the fraxinellone nano-preparation has the effect of sustained-release drug delivery, and can continuously and effectively control the activation of TGF- β/smad pathways.
Example 4: effect of Fraxinellone on blood perfusion in subcutaneous and in situ pancreatic cancer model animals after multiple dosing
Establishment of subcutaneous pancreatic cancer tumor model: collecting pancreatic cancer cells Panc-1 and fibroblast NIH3T3 in logarithmic growth phase, digesting with pancreatin, centrifuging, and resuspending with appropriate amount of PBS to obtain two kinds of cells at a ratio of 1: 1 and a total cell concentration of 1 × 106cells/mL. Taking 4-6 weeks female nude mice of about 20g, injecting 100 μ L cell suspension subcutaneously into hind limb, and waiting for tumor volume to grow to about 200cm after 14 days3For subsequent experiments;
establishing an in-situ pancreatic cancer tumor model: collecting Panc-1 cells and fibroblasts NIH3T3 in logarithmic growth phase, digesting with pancreatin, centrifuging, washing the cells with PBS twice, counting, and adjusting the cell concentration to 1 × 106cells/mL, and placing in an ice box for standby. 150 mu L of 5% chloral hydrate is used for carrying out intraperitoneal injection and anesthesia on a nude mouse, four limbs are fixed by a medical adhesive tape, the skin of the abdomen is coated by iodine tincture, a 2-3cm incision is cut near the left lower abdomen spleen of the nude mouse by an ophthalmic scissors, the nude mouse is turned over to the outside of the skin, and the spleen is the pancreas. Resuspending the cells on ice with a pipette, inserting 50 μ L of the needle from the tail end of the pancreas to the right front direction, observing that the pancreas is obviously filled and semitransparent, slowly moving out the needle head, and pushing the pancreas and spleen back into the body with a cotton swab dipped with physiological saline. Dropping proper amount of antibiotic to the wound, sewing the inner muscle layer with biodegradable suture and the outer skin layer with biodegradable suture, and all the operations are performed in an ultraclean operating table. Observing the animal state and wound healing condition every day after inoculation, and performing experiments 14 days after inoculation;
tumor-bearing nude mice were randomly divided into 3 groups of 6 mice each, and the dose of administered Frax-Free and Frax-NP was 20 mg/kg. The administration was performed every two days by tail vein injection for a total of 4 times, and the control group was given the same volume of PBS. After the administration period is finished, a subcutaneous pancreatic cancer model nude mouse is fixed on a photoacoustic imaging console, 50 mu L of contrast agent is injected into a tail vein after a tumor part is coated with coupling gel, the blood perfusion condition of the tumor part is shot, the change of Echo intensity along with Time (Echo Power-Time) is recorded, and the perfusion volume (Peak Enhance, PE), the Time To Peak (TTP) and the flushing rate (Wash-inRate, WiR) of each group are read. The in-situ pancreatic cancer model nude mouse is fixed on a photoacoustic imaging console, the blood oxygen saturation condition of the tumor part is shot after the tumor part is coated with coupling glue, and the average blood oxygen saturation parameter is recorded;
the results show that: in fig. 4A and 4B subcutaneous pancreatic cancer models, the degree of blood perfusion was increased in both dosing groups compared to the PBS-treated group, and the effect of fraxinellone nano-formulation was much better than that of the free drug; in the quantification, Echo Power-Time records the change in contrast agent Echo intensity in the vessels over Time after administration of the contrast agent; on the whole, the echo intensity of the acerolone nano preparation group is stronger than that of the free medicine group and the PBS group, which indicates that the blood vessel is more unobstructed; PE represents the blood flow content, and the PE value of the acerola nano preparation group is 2.8 times that of the free medicine group and 5.6 times that of the PBS group; the shorter the time to peak TTP is, the more unblocked the blood vessel is, the quantitative result shows that the time to peak of the group of the administered fraxinellone nano preparation is only 4.6s, which is far lower than 8.5s of the free medicine group and 8s of the PBS group; WiR is related to PE and TTP, the larger WiR indicates that the contrast agent is cleared more quickly and the vascular structure is more complete, and as can be seen from the figure, the WiR value of the administered fraxinellone nano preparation is the largest and is 1.6 times and 3.5 times of that of the free medicine group respectively; quantitative data shows that compared with free drugs, the blood flow perfusion amount of the tumor part is larger after the fraxinellone nanoparticles are administered, and the chemotherapy drugs can permeate into tumor cells; in the in situ pancreatic cancer model of FIG. 4C, the blood oxygen saturation levels of the administered group were improved compared to the PBS-treated group, and the mean blood oxygen saturation level (sO)2Avr%) increased from 35% in PBS group to 43% in free drug group and 62% in nano-preparation group, which indicates that the effect of increasing blood oxygen saturation of fraxinellone nano-preparation is better than that of free drug. Two types of pancreatic cancerModel results show that compared with free medicines, the fraxinellone nano preparation can more effectively improve the tumor microenvironment, normalize intratumoral blood vessels, and increase blood perfusion, thereby weakening the physical barrier of chemotherapy drugs penetrating into tumor cells.
Example 5: in-vivo pharmacodynamic evaluation of fraxinellone nano preparation on in-situ pancreatic cancer model animals
After the administration period is finished, dissecting and taking out tumor tissues of the nude mice, cracking and extracting protein, carrying out Western Blot experiment, and investigating the expression conditions of fibroblast-related proteins α -SMA, FAP, Fibronectin and TGF- β/smad pathway-related proteins in vivo;
in addition, the nude mice bearing orthomas were randomly divided into 3 groups of 6 mice each, and every two days, were injected with Frax-Free and Frax-NP (20mg/kg) through the tail vein for a total of 4 times, and the control group was given the same volume of PBS. After the administration period is finished, dissecting and taking tumors, then digesting tissues for 4 hours by using mixed enzyme liquid of DNase I, hyaluronidase and collagenase, sieving and collecting cells, adding a fluorescein isothiocyanate labeled macrophage marker (FITC-F4/80) and an allophycocyanin labeled M1 type macrophage marker CD86(APC-CD86) and an phycoerythrin labeled M2 type macrophage marker CD206(PE-CD206) for screening macrophages and marking M1 type macrophages and M2 type macrophages, and inspecting the proportion of the two types of macrophages by flow cytometry;
in addition, the nude mice with orthotopic tumors were randomly divided into 4 groups of 12 mice each, and each two days, the mice were administered 4 times with the combination of Frax-NP (20mg/kg), gemcitabine (15mg/kg), and gemcitabine (15mg/kg) by tail vein injection, and the control group was administered with PBS of the same volume; after the administration period, 6 nude mice in each group of PBS and non-combined group were anesthetized with 150 μ L of 5% chloral hydrate, fixed with 4% paraformaldehyde after heart perfusion with physiological saline, embedded in paraffin for sectioning, and each antibody was stained to show the expression of protein in tumor tissue. Another 6 mice per group (including all nude mice in the combination group) were used to investigate survival status;
the results are shown in fig. 5A and 5B, which show that after administering fraxinellone nano-formulation, the expression of fibrinectin, FAP, &ttttransformation = α "&tttα &ttt/t &ttt-SMA in the tumor tissue was significantly reduced and the two phosphorylated proteins psmad2 and psmad3 associated with TGF- β/smad pathway were also well controlled, and fig. 5C, which shows that after administration, the proportion of M2-type macrophages in the tumor tissue was reduced from 34.0% to 24.4% (free drug group) and 15.3% (formulation group), respectively, which shows that the fraxinellone nano-formulation is more effective than the free drug in reducing the proportion of M2-type macrophages in the tumor tissue, while the free drug and nano-formulation have no significant effect on the M1-type of macrophages (2.25% for PBS group, 4.23% for masculin group, 2.29 for mason) and that the free drug and nano-formulation have no significant effect on the M1-type change in the tumor tissue when administered as compared to PBS group, and the collagen-PBS group, and showing no significant change in the collagen-binding effect in the tumor tissue when administered as PBS group, and the collagen-receptor group was significantly reduced;
the survival time experiment shows that the median survival time of the tumor-bearing nude mice of the gemcitabine group which is singly administered is 38.5 days, and the survival time of the tumor-bearing nude mice is remarkably prolonged to 47 days by combining the fraxinellone nano preparation and the gemcitabine group.
The results show that the prepared fraxinellone nanoparticles can obviously improve the tumor microenvironment, reduce the deposition of collagen and the expression of related proteins, reduce the proportion of macrophage M2 parting, relax blood vessels, weaken the physical barrier of chemotherapy drugs penetrating into tumor cells, and promote the penetration and accumulation of gemcitabine in tumor parts by combining with the chemotherapy drugs gemcitabine, thereby improving the curative effect.
Claims (8)
1. A drug delivery system with functions of adjusting tumor microenvironment and targeting is characterized in that a section of functional polypeptide CGKRK is covalently bonded on the surface of a nanoparticle constructed by a medicinal high-molecular polylactic acid material, and a natural drug monomer is physically entrapped in a hydrophobic core of the nanoparticle by an emulsified solvent evaporation method.
2. The drug delivery system with the functions of regulating the microenvironment and targeting tumor according to claim 1, wherein the polymeric polylactic acid material comprises methylated polyethylene glycol-polylactic acid block copolymer mPEG-PLA.
3. The drug delivery system with the functions of adjusting the microenvironment and targeting the tumor as claimed in claim 2, wherein the molecular weight of the methylated polyethylene glycol-polylactic acid block copolymer mPEG-PLA is 10000-50000 Da.
4. The drug delivery system with the functions of adjusting the microenvironment of tumors and targeting according to claim 2, wherein the functional polypeptide CGKRK is modified on the surface of the nanoparticle through malondialdehyde-sulfhydryl reaction to achieve the purpose of targeting the tumor site.
5. The drug delivery system with tumor microenvironment and targeting function of claim 1, wherein the encapsulated hydrophobic natural drug monomer is fraxinellone.
6. The drug delivery system with the functions of adjusting the microenvironment and targeting the tumor according to claim 1, wherein the method for encapsulating the hydrophobic drug monomer is an emulsion solvent evaporation method.
7. The drug delivery system of claim 1, which inhibits the activation of CAFs, reduces the conversion of macrophages to M2 type, improves the tumor microenvironment, and increases blood perfusion.
8. The drug delivery system with tumor microenvironment and targeting function of claim 1, wherein the drug delivery system inhibits the expression of psmad2 and psmad3 of TGF- β/smad pathway from acting on the CAFs.
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| CN112603995A (en) * | 2020-12-30 | 2021-04-06 | 四川省肿瘤医院 | CAFs-targeted tumor cell vaccine, preparation method and application thereof |
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| CN107875140A (en) * | 2016-09-30 | 2018-04-06 | 复旦大学 | A kind of double targeted drug delivery systems and its application in oncotherapy preparation is prepared |
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