CN115813884A - Macromolecule nano-drug for enhancing tumor treatment effect by regulating tumor extracellular matrix and preparation method and application thereof - Google Patents

Abstract

The invention relates to a high-molecular nano-drug for enhancing tumor treatment effect by regulating tumor extracellular matrix, and a preparation method and application thereof. The nano-drug can perform effective anti-tumor treatment by regulating and controlling tumor ECM, and improve the treatment effect of various anti-tumor therapies. The nano-drug can realize the controllable drug release and activation of tumor microenvironment response, further reduce the toxic and side effects on normal tissues, simultaneously enhance the anti-tumor combined effect of small molecular drugs, and has good water solubility and biocompatibility; in addition, the composite material prepared by the preparation method of the nano-drug has uniform size and particle size, can efficiently load various small-molecule drugs, also solves the problems of poor solubility and low bioavailability of part of drugs in the preparation of antitumor drugs, and has simple preparation process and easy realization of industrial production.

Description

A polymer nanoparticle that enhances the therapeutic effect of tumors by regulating the extracellular matrix of tumors Rice medicine and its preparation method and application

technical field

The invention relates to the technical field of medical materials, in particular to a macromolecular nanomedicine that enhances tumor treatment effect by regulating tumor extracellular matrix, its preparation method and application.

Background technique

Cancer has become one of the major killers that endanger human life and health worldwide. Although cancer diagnosis and treatment technology has made significant progress with the continuous development of modern medicine, the morbidity and mortality of cancer are still high.

A well-formed tumor is a complex tissue with a unique tumor microenvironment. It includes a variety of components, such as: tumor cells, tumor blood vessels, fibroblasts, immune cells, various signaling molecules, extracellular matrix, etc. Compared with the microenvironment of normal cells, _the tumor microenvironment has lower pH value, less oxygen content, overexpression of GSH and _H2O2 content, abnormal tumor blood vessels, and increased rigidity of tumor extracellular matrix (ECM). Among them, the increase of tumor ECM rigidity will not only hinder drug penetration, increase tumor drug resistance but also further promote tumor progression. However, the current tumor treatment strategy is still dominated by small molecule cytotoxic drugs targeting tumor cells, and there are relatively few studies on anti-tumor therapy by directly regulating tumor ECM. Therefore, the regulation of tumor ECM is of great significance for enhancing the research of anti-tumor therapy.

Major drivers of increased tumor ECM stiffness include phenotypically transformed tumor-associated fibroblasts, transforming growth factor-β (TGF-β), and matrix crosslinking. Among them, TGF-β released by tumor cells, fibroblasts and other stromal cells in the tumor microenvironment plays an important regulatory role. First, TGF-β can promote the differentiation of fibroblasts into tumor fibroblasts and up-regulate the expression and synthesis of many matrix proteins such as type I collagen, laminin and fibronectin; secondly, TGF-β can up-regulate the expression of lysyl The expression of oxidase promotes the cross-linking of collagen; at the same time, TGF-β can also down-regulate the synthesis of degradative proteins such as matrix metalloproteinases in tumor ECM. Therefore, blocking the TGF-β signaling pathway can help inhibit the transformation of fibroblasts into tumor-associated fibroblasts, and inhibit the excessive deposition of ECM components and excessive cross-linking of collagen, which is expected to be an effective method to reduce ECM rigidity. At present, a number of clinical trials based on blocking TGF-β signaling pathway to improve anti-tumor efficacy are underway. However, due to the important role of TGF-β in a variety of physiological functions, therapy to block TGF-β signaling pathway may lead to systemic inhibition of TGF-β pathway, which may lead to serious side effects after clinical treatment. Therefore, selectively blocking the TGF-β signaling pathway at the tumor site is of great significance for regulating tumor ECM and improving biological safety. However, there are few reports on the research of nanomedicine targeting to block the TGF-β signaling pathway at the tumor site to reduce the rigidity of the ECM.

In addition, studies have shown that TGF-β can also induce a pro-angiogenic microenvironment to stimulate tumor angiogenesis; and vascular endothelial growth factor (VEGF) inhibitors can promote the normalization of tumor ECM, suggesting that inhibiting tumor angiogenesis can effectively promote the normalization of ECM. change. Therefore, blocking TGF-β and VEGF signaling pathways at the same time is expected to normalize ECM, inhibit tumor growth, and enhance the efficacy of various anti-tumor treatments such as photodynamic and immunotherapy, which provides a new idea for anti-tumor therapy.

In addition, in order to speed up its clinical transformation, it is crucial to fully consider its metabolism in the body and biological safety issues. Biodegradable synthetic polymers have excellent biocompatibility and easy functional modification. Using them as nanocarriers to deliver hydrophobic small molecule drugs can effectively improve their water solubility and biocompatibility, and enhance penetration And the retention effect changes the distribution of the drug in the body, so that it can be efficiently enriched in the tumor lesion and improve its bioavailability. Therefore, it is very attractive and has great clinical significance to use biodegradable polymers as nanocarriers to construct nanomedicines for tumor ECM regulation.

Contents of the invention

In view of this, the present invention aims to solve the technical problems in the prior art and provide a polymer nanomedicine that enhances tumor therapeutic effect by regulating tumor extracellular matrix, its preparation method and application. The nano-medicine of the present invention can perform effective anti-tumor therapy by regulating tumor ECM, and simultaneously improve the therapeutic effect of various anti-tumor therapies.

In order to solve the problems of the technologies described above, the technical solution of the present invention is specifically as follows:

The present invention provides a polymer nanomedicine that enhances tumor therapeutic effects by regulating tumor extracellular matrix, which is obtained by self-assembling drug I, drug II and carrier III;

The drug I includes Linker, which is a tumor microenvironment response unit, and the two ends of the Linker are respectively connected with anti-angiogenic drugs and extracellular matrix normalizing drugs;

The drug II is a photothermal, photodynamic or near-infrared imaging molecular drug;

The carrier III is a polymer carrier.

Preferably, the Linker is a GSH response unit, a ROS response unit or an enzyme response unit, and the two ends of the Linker are respectively connected to the drug through hydroxyl, aldehyde, amino or sulfhydryl functional groups.

Preferably, the anti-angiogenic drug is one or more of bevacizumab, conbercept, dexamethasone, ranibizumab and Avastin, and the extracellular matrix normalizing drug It is one or more of verteporfin, papain, tranilast and hyaluronidase, and the drug II is chlorin, indocyanine green, near-infrared dye IR780 and polydopamine. One or more, the carrier III is mPEG-DSPE or mPEG-b-PLA.

Preferably, the particle diameter of the polymer nanomedicine is 50-300nm.

Further preferably, the anti-angiogenic drug is dexamethasone, the extracellular matrix normalizing drug is tranilast, the drug II is indocyanine green, and the carrier III is mPEG-DSPE, The corresponding structural formulas are as follows:

The structural formula of the drug I is shown in formula (I):

The structural formula of the drug II is shown in formula (II):

The structure of the carrier III is shown in formula (III):

The present invention also provides a method for preparing a polymer nanomedicine that enhances tumor therapeutic effects by regulating tumor extracellular matrix, comprising the following steps:

1) Connecting anti-angiogenic drugs and extracellular matrix normalizing drugs through a microenvironment-responsive linker to obtain drug I;

2) Self-assembling the drug I, the drug II and the carrier III to obtain a polymer nanomedicine containing multiple drugs.

Preferably, a specific implementation of step 1) in the preparation method is:

1-1) Mixing and reacting tranilast, microenvironment responsive linker and anhydrous solvent to obtain intermediate IV, intermediate IV has the structure shown in formula (IV);

1-2) Mixing and reacting intermediate IV, dexamethasone and anhydrous solvent to obtain drug I.

Preferably, the reaction temperature in step 1) is 10-150°C, the reaction temperature in step 2) is 10-50°C, and the mass ratio of drug I: drug II: carrier III is ( 1~100): (1~5): (100~500).

Preferably, the reaction temperature in the step 1-1) is 120-130°C, and the reaction temperature in the step 1-2) is 20-40°C.

The present invention also provides an antitumor drug comprising the polymeric nanomedicine described in the present invention, a contrast agent comprising the polymeric nanomedicine described in the present invention for near-infrared fluorescence imaging, or comprising the polymeric nanomedicine described in the present invention Nanomedicines for anti-inflammatory drugs.

The beneficial effects of the present invention are:

The macromolecule nanomedicine provided by the invention can perform effective anti-tumor treatment by regulating tumor ECM, and simultaneously improve the therapeutic effects of various anti-tumor therapies.

The small molecule drugs used in the polymer nano-medicine of the present invention are all FDA-approved drugs. It is found through experiments that the polymer nanomedicine of the present invention can realize the controllable drug release and activation in response to the tumor microenvironment, further reduce the toxic and side effects on normal tissues, and at the same time enhance the anti-tumor joint effect of small molecule drugs, and has Good water solubility and biocompatibility.

The preparation method of the polymer nano-medicine of the present invention has a uniform size and particle size of the prepared material, can efficiently load a variety of small-molecule drugs, and also solves the problems of poor solubility and low bioavailability of some small-molecule drugs in the preparation of anti-tumor drugs , and the preparation process is simple, and it is easy to realize industrial production.

The polymer nanomedicine of the present invention can be used for antitumor drugs, contrast agents for near-infrared fluorescence imaging, or anti-inflammatory drugs.

Description of drawings

The present invention will be described in further detail below in conjunction with the accompanying drawings and specific embodiments.

Fig. 1 is the TEM picture of the sample DEX/TRN/ICG NP obtained in Example 1 of the present invention;

Fig. 2 is the dynamic light scattering diagram of the sample DEX/TRN/ICG NP obtained in Example 1 of the present invention;

Fig. 3 is the mass spectrogram of DEX, TRN and TRN-SS-OH, TRN-SS-DEX prepared in Example 1 of the present invention;

Fig. 4 is the proton nuclear magnetic resonance spectra of DEX, TRN and TRN-SS-OH, TRN-SS-DEX prepared in Example 1 of the present invention;

Fig. 5 is a histogram comparing the cytotoxicity of nanomedicines in Example 4 of the present invention to tumor cells and normal cells; wherein the upper figure is 4T1 tumor cells, and the lower figure is 3T3 normal cells;

Fig. 6 is a graph showing the content change of GSH in cancer cells in the control group and the DEX/TRN/ICG NP group in Example 5 of the present invention;

Figure 7 is a diagram of the body weight changes of mice during the in vivo treatment of the control group, DEX, TRN, ICG+L, DEX/TRN/ICG NP and DEX/TRN/ICGNP+L groups in Example 5 of the present invention;

Fig. 8 is a graph showing the changes in tumor volume during the in vivo treatment of the control group, DEX, TRN, ICG+L, DEX/TRN/ICG NP and DEX/TRN/ICGNP+L groups in Example 5 of the present invention.

Detailed ways

The present invention provides a polymer nanomedicine that enhances tumor therapeutic effects by regulating tumor extracellular matrix, which is obtained by self-assembling drug I, drug II and carrier III;

The drug I includes Linker, which is a tumor microenvironment response unit, including but not limited to GSH response, ROS response or enzyme response; the two ends of the Linker are respectively connected with anti-angiogenesis drugs and extracellular matrix normalization drugs, The functional groups at both ends of the Linker can be selected arbitrarily, including but not limited to one or more of hydroxyl, aldehyde, amino and mercapto groups, through which the functional groups are respectively connected to drugs; the anti-angiogenic drugs are bevacizumab, Kang One or more of Percept, Dexamethasone, Ranibizumab and Avastin, preferably Dexamethasone; the extracellular matrix normalizing drug is Verteporfin, Papain, Tranis One or more of special and hyaluronidase, preferably tranilast.

The drug II is a photothermal, photodynamic or near-infrared imaging molecular drug;

The drug II preferably includes but is not limited to one or more of chlorin CE6, indocyanine green (ICG), polydopamine and near-infrared dye IR780, more preferably indocyanine green;

The carrier III is a polymer carrier, which can be selected as required, including but not limited to mPEG-DSPE or mPEG-b-PLA.

The above-mentioned self-assembly process is specifically as follows: the solution of drug I, the solution of drug II and the solution of carrier III are mixed according to the required ratio, and then added dropwise to the aqueous solution and vigorously stirred overnight.

The drug I mentioned above is a microenvironment responsive structure, which can respond to the tumor microenvironment to break and release two small molecule drugs. The particle size of the polymer nanomedicine in the present invention is preferably 50-300 nm, more preferably 100-150 nm. The present invention has no special requirements on the amount of small molecule drug loaded on the polymer nanomaterial, and those skilled in the art can select an appropriate loading amount according to actual needs.

Further preferably, the anti-angiogenic drug is dexamethasone, the extracellular matrix normalizing drug is tranilast, the drug II is indocyanine green, and the carrier III is mPEG-DSPE, The corresponding structural formulas are as follows:

The structural formula of the drug I is shown in formula (I):

The structural formula of the drug II is shown in formula (II):

The structure of the carrier III is shown in formula (III):

The present invention also provides a method for preparing a polymer nanomedicine that enhances tumor therapeutic effects by regulating tumor extracellular matrix, comprising the following steps:

1) Connecting anti-angiogenic drugs and extracellular matrix normalizing drugs through a microenvironment-responsive linker to obtain drug I;

2) Self-assembling the drug I, the drug II and the carrier III to obtain a polymer nanomedicine containing multiple drugs.

Preferably, a specific implementation of step 1) in the preparation method is:

1-1) Mixing and reacting tranilast, microenvironment responsive linker and anhydrous solvent to obtain intermediate IV, intermediate IV has the structure shown in formula (IV);

1-2) Mixing and reacting intermediate IV, dexamethasone and anhydrous solvent to obtain drug I.

Preferably, the reaction temperature in step 1) is 10-150°C, the reaction temperature in step 2) is 10-50°C, and the mass ratio of drug I: drug II: carrier III is ( 1~100): (1~5): (100~500).

Preferably, the reaction temperature in the step 1-1) is 120-130°C, and the reaction temperature in the step 1-2) is 20-40°C.

In the polymer nanomedicine of the present invention, the anti-angiogenic drug is dexamethasone, the extracellular matrix normalizing drug is tranilast, the drug II is indocyanine green, and the carrier III is mPEG-DSPE , when, the preparation method of the macromolecule nano-medicine that enhances tumor therapeutic effect by regulating tumor extracellular matrix comprises the following steps:

1) Connecting dexamethasone and tranilast through a microenvironment-responsive linker to obtain TRN-linker-DEX;

2) Self-assemble TRN-linker-DEX, ICG and mPEG-DSPE to obtain polymer nanomedicine containing multiple drugs.

According to the present invention, the present invention self-assembles TRN-linker-DEX, ICG and mPEG-DSPE to obtain a polymer nanomedicine containing multiple drugs; wherein, the loading is specifically the tetrahydrofuran solution of TRN-linker-DEX, The aqueous solution of ICG and the tetrahydrofuran solution of mPEG-DSPE are added dropwise into the aqueous solution at a ratio of 1:1:1 and stirred vigorously overnight to obtain a polymer nanomedicine that enhances the tumor therapeutic effect by regulating the extracellular matrix of the tumor; among them, tetrahydrofuran represents most of the volatile Solvent, but not all, those skilled in the art can select suitable solvent and volatile amount according to actual experimental situation; Described volatile solvent is preferably one or more in tetrahydrofuran, chloroform and ethanol, is preferably tetrahydrofuran; Angiogenic drugs are preferably one or more of bevacizumab, conbercept, dexamethasone, ranibizumab and Avastin, more preferably dexamethasone; the normalization of the extracellular matrix The drug is preferably one or more of verteporfin, papain, tranilast and hyaluronidase, more preferably tranilast; the photodynamic and photothermal therapy drugs are preferably two One or more in hydroporphine, indocyanine green, near-infrared dye IR780 and polydopamine, more preferably indocyanine green; The mass ratio of described TRN-linker-DEX:ICG:mPEG-DSPE is preferably ( 1~100): (1~5): (100~500).

In the present invention, the TRN-SS-DEX is prepared according to the following method:

1-1) Mix and react tranilast, HO-linker-OH, EDC·HCl and solvent to obtain TRN-linker-OH; wherein, the solvent is anhydrous acetonitrile; specifically, in order to make the reaction more smoothly To carry out, the present invention preferably dissolves tranilast in 10-100mL acetonitrile, stirs for 5-10 minutes to make the mixture uniform; adds EDC·HCl, and stirs for 1-4 hours at 20-50°C to activate; then add to the obtained solution 10-50mL HO-linker-OH solution in acetonitrile was added dropwise, and stirred at 60-120°C for 15-30 hours. Use 0.01-1mol/L hydrochloric acid for precipitation. Centrifuge at 9000-12000rpm for 8-12min to collect and wash the precipitate, remove the supernatant, freeze-dry to obtain TRN-linker-OH.

1-2) TRN-linker-OH, triphosgene, DEX and solvent are mixed and reacted to obtain TRN-linker-DEX; wherein, the solvent is anhydrous tetrahydrofuran; the reaction is a room temperature reaction, and the reaction time is 12-48 hours; more specifically, the reaction is: redisperse 50-400 mg of the obtained TRN-linker-OH in 10-30 mL of anhydrous tetrahydrofuran, add triphosgene, and set the temperature of the condenser to 1-5 °C. Then, 5-10 mL of DMAP in tetrahydrofuran solution was added dropwise to the obtained solution to form a white precipitate, and the reaction was continuously stirred for 10-50 min. Then, 5-10 mL of a tetrahydrofuran solution of DEX was added dropwise to the obtained solution, and reacted at room temperature for 15-30 hours. Use 0.01-1mol/L hydrochloric acid for precipitation. Centrifuge at 9000-12000 rpm for 8-12 minutes to collect and wash the precipitate, remove the supernatant, and freeze-dry to obtain TRN-linker-DEX.

The present invention provides an anti-tumor drug, including the polymer nano-medicine for enhancing tumor therapeutic effect through regulation of tumor extracellular matrix described in the present invention.

The invention provides a contrast agent for near-infrared fluorescence imaging, including the polymer nano-medicine that enhances tumor treatment effect by regulating tumor extracellular matrix.

The present invention provides an anti-inflammatory drug, including the polymer nano-medicine that enhances tumor treatment effect by regulating tumor extracellular matrix described in the present invention.

To sum up, the present invention provides a polymer nanomedicine that enhances the therapeutic effect of tumors by regulating the extracellular matrix of tumors and its preparation method and application. Drug resistance is a serious problem, and the loading ratio of drugs can be adjusted according to the needs, so as to truly realize one disease, one policy. And using the above method can also solve the problem of poor solubility and low bioavailability of some small molecule drugs, and can also realize the controllable drug release and activation in response to the tumor microenvironment, reduce the toxic and side effects on normal tissues, and at the same time regulate the tumor microenvironment To significantly enhance the therapeutic effect, it can be applied to the preparation of antitumor drugs with high efficiency and low toxicity. In addition, the method of the invention makes the obtained composite material uniform in size, good in water solubility and biocompatibility, simple in preparation process, low in cost, and suitable for industrial production.

The technical solution will be clearly and completely described below in conjunction with the embodiments of the present invention. Apparently, the described embodiments are only some of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without making creative efforts belong to the protection scope of the present invention.

Example 1:

The preparation of loaded dexamethasone/tranilast/indocyanine green macromolecule nanomedicine comprises the following steps:

)溶于20mL无水乙腈中，滴加至所得溶液中，在85℃条件下搅拌24小时。反应完成后用0.1mol/L的盐酸进行沉降。用10000rpm离心10min收集并洗涤沉淀，去掉上清，冷冻干燥得到TRN-SS-OH。1) Preparation of TRN-SS-OH: Dissolve 500mg tranilast in 30mL anhydrous acetonitrile, stir for 5-10 minutes to mix well; add 293mg EDC·HCl, stir at 35°C for 2 hours to activate; then 471mg HO-SS-OH (structural formula is

) was dissolved in 20 mL of anhydrous acetonitrile, added dropwise to the resulting solution, and stirred at 85° C. for 24 hours. After the reaction is completed, settle with 0.1mol/L hydrochloric acid. Centrifuge at 10,000 rpm for 10 min to collect and wash the precipitate, remove the supernatant, and freeze-dry to obtain TRN-SS-OH.

2) Preparation of TRN-SS-DEX: Take 236mg of TRN-SS-OH prepared in step (1), and evenly disperse it in 20mL of anhydrous tetrahydrofuran, stir for 5-10 minutes to mix evenly, add 55.4mg of triphosgene, The temperature of the condenser tube was set at 3°C. Next, 187 mg of DMAP was dissolved in 20 mL of anhydrous tetrahydrofuran, and added dropwise to the resulting solution to produce a white precipitate, and the reaction was continuously stirred for 30 min. Next, 200 mg of DEX was dissolved in 10 mL of anhydrous THF, added dropwise to the resulting solution, and stirred at 25°C for 24 hours. After the reaction is completed, settle with 0.1mol/L hydrochloric acid. Centrifuge at 10,000rpm for 10min to collect and wash the precipitate, remove the supernatant, and freeze-dry to obtain TRN-SS-DEX;

3) DEX/TRN/ICG NP: Dissolve 22.5 mg of TRN-SS-DEX prepared in step (2) in 4 mL of THF, 2 mg of ICG in 4 mL of ultrapure water, and 75.5 mg of mPEG-DSPE in 4 mL of THF. Mix, after ultrasonic homogeneity, slowly add 120mL ultrapure water drop by drop, continue to stir overnight. Use a 0.22 μm filter head to remove the unencapsulated drug precipitate, freeze-dry to obtain DEX/TRN/ICG NP, and measure the drug loading rate with a UV spectrophotometer after dissolving in methanol;

The results are shown in Figures 1 to 4:

Fig. 1 is the TEM image of the sample obtained in each step of Example 1 of the present invention. It can be seen from the figure that the nanomedicine DEX/TRN/ICG NP is a solid spherical structure with a uniform size and a particle size of about 60nm;

Figure 2 is the dynamic light scattering diagram of the sample obtained in each step of Example 1 of the present invention. It can be seen from the figure that the nanomedicine DEX/TRN/ICG NP has a uniform size, the hydrated particle size is about 110.5nm, and the potential is about -30.7mV .

Fig. 3 is the mass spectrogram of TRN, DEX and the TRN-SS-OH, TRN-SS-DEX prepared in the embodiment 1 of the present invention, proves that the medicine is successfully synthesized, and after loading is completed, the burden of DEX is measured by an ultraviolet spectrophotometer. The loading rate of TRN was 10%, that of TRN was 8.34%, and that of ICG was 2%. The loading efficiency of each drug was close to 100%.

Fig. 4 is the proton nuclear magnetic resonance spectrum of TRN, DEX and TRN-SS-OH, TRN-SS-DEX prepared in Example 1 of the present invention, according to the change of peak characteristic peak and the peak position of each group of peaks, it can be proved that the drug is synthesized success.

Example 2:

In order to prove that the DEX/TRN/ICG NP obtained in Example 1 of the present invention has good biocompatibility and the effect of promoting the growth of fibroblasts, we use the MTT experiment to test the proliferation of the cells after the material and the cells are co-cultured for a period of time, The specific test method is as follows:

1) Add the 4T1 and 3T3 cells (1×10 ⁵ /mL) in the logarithmic growth phase to a 96-well cell culture plate (200 μL/well), and place the cells in the 96-well plate in 5% CO ₂ at 37°C Cultivate in a carbon dioxide incubator for 24 hours until the cell monolayer covers the bottom of the well (96-well flat bottom plate).

2) Then disperse DEX, TRN, ICG (+L), DEX/TRN/ICG NP (+L) with 1640 cell culture medium, and make dispersions with different concentrations respectively. The concentrations of DEX are: 0, 15, 20, 25, 30 mg/mL, then cells and materials were incubated at 5% CO ₂ , 37°C for 24 hours, the control group did not add samples, and the light group was cultured for 12 hours, respectively, 808nm, 5min, 1.0W/cm ² near-infrared light irradiation. Observe under an inverted microscope.

3) After the culture is over, suck off the culture medium in the wells, carefully wash with PBS 2-3 times to wash away the material, then add 100 μL of MTT-containing PBS solution (0.5 mg/mL) to each well to terminate the culture, MTT and living cells Under the action of succinate dehydrogenase, formazan, an insoluble blue-purple substance, is deposited in cells.

4) After 3-5 hours, add 100 μL dimethyl sulfoxide (DMSO) solution to dissolve the formazan, and shake it on a shaker at low speed for 10 minutes to fully dissolve the crystalline substance. The absorbance value (OD value) at 570 nm was detected on a microplate reader, and the survival rate of the cells was calculated.

The results are shown in Figure 5. Figure 5 is a graph showing the selective growth promotion of 4T1 and 3T3 cells by DEX/TRN/ICG NP (+L) prepared in Example 1 of TRN, DEX, and ICG (+L). It can be seen from the figure that even if the concentration of DEX in the material reaches 30mg/mL, the activity of 3T3 cells can still be basically maintained at 100%. It can be seen that the material has good biocompatibility and can be used in living organisms. With the increase of DEX concentration, the growth of 4T1 cells in DEX/TRN/ICG NP and DEX/TRN/ICG NP+L groups was significantly inhibited. It shows that the nanoparticles have good anti-tumor effect.

Example 3:

Polymer nano-drugs that can carry multiple drugs are used in the preparation of anti-tumor drugs with high efficiency and low toxicity. Its depletion of intracellular glutathione was determined by the following steps:

1) Add 4T1 cells (1×10 ⁵ /mL) in the logarithmic growth phase to culture flasks (200 μL/well), and place the cells in the 96-well plate in a 5% CO ₂ , 37°C carbon dioxide incubator for 24 hours , until the cell monolayer was confluent.

2) Then disperse DEX/TRN/ICG NP with 1640 cell culture medium, and make dispersions with different concentrations respectively, in which the concentrations of DEX/TRN/ICG NP are: 0, 150, 300 mg/mL, and then the cells and materials are mixed in 5% CO _2. Incubate at 37°C for 24 hours, and no samples are added to the control group. Observe under an inverted microscope. GSH and GSSH detection kits were used to detect the consumption of glutathione in cells.

As shown in Fig. 6, Fig. 6 is the graph of intracellular glutathione consumed by macromolecule nanomedicine in Example 3 of the present invention, and it is found that nanomedicine can effectively consume glutathione; it can be proved that nanomedicine of the present invention can Improve the high reduction of the tumor microenvironment by effectively consuming glutathione, and reduce the consumption of singlet oxygen generated by the ICG photodynamic process due to the high reduction of the tumor, thereby effectively improving the therapeutic effect;

Example 4:

Polymer nano-drugs, which can enhance the therapeutic effect of tumors by regulating the extracellular matrix of tumors, are being applied to the preparation of anti-tumor drugs with high efficiency and low toxicity. Detecting the anti-tumor effect of the nano-medicine at the living level comprises the following steps:

Female BALB/C mice weighing 15-25 g were selected as model mice; tumor models were established by subcutaneously injecting ¹⁰⁶ mouse-derived breast cancer cells (4T1) into the forelimbs of the mice, and when the tumor volume reached ^120mm3 , the mice were randomly Divided into four groups, 10 rats in each group, intravenous injection of normal saline, TRN, DEX and ICG (+L), DEX/TRN/ICG NP (+L), injected every two days, ICG and DEX/TRN/ICG The NP light group was lighted for 5 minutes 4 hours and 24 hours after administration, respectively. Tumor diameter and mouse body weight were measured every other day. Tumor volume was calculated by the following formula: volume=tumor length×tumor ^width2 /2. Mice were sacrificed on day 15, and tumors were dissected and weighed to assess tumor treatment efficiency.

As shown in Figure 7, Figure 7 is the control group, TRN, DEX and ICG (+L), DEX/TRN/ICG NP (+L) group living body treatment course mouse body weight change figure in the embodiment 5 of the present invention, can It can be seen that the body weight of the mice remains stable, indicating that the polymer nanomedicine of the present invention has good biocompatibility.

As shown in Fig. 8, Fig. 8 is the tumor volume change diagram and after treatment of the control group, TRN, DEX and ICG (+L), DEX/TRN/ICG NP (+L) group in vivo treatment process in Example 5 of the present invention It can be seen that compared with the control group, the tumor suppression efficiency of the small molecule group is limited, while the growth of the tumor in the DEX/TRN/ICG NP plus light group is almost completely inhibited; Modulation of the tumor extracellular matrix combined with anti-angiogenic therapy and phototherapy has effectively improved the therapeutic effect of tumors.

The descriptions of the above embodiments are only used to help understand the method and core idea of the present invention. It should be pointed out that for those skilled in the art, without departing from the principle of the present invention, some improvements and modifications can be made to the present invention, and these improvements and modifications also fall within the protection scope of the claims of the present invention.

Claims (10)

1. A macromolecule nanometer medicine for enhancing the tumor treatment effect by regulating and controlling tumor extracellular matrix is characterized in that the macromolecule nanometer medicine is obtained by self-assembling a medicine I, a medicine II and a carrier III;

the drug I comprises a Linker which is a tumor microenvironment response unit, and two ends of the Linker are respectively connected with an anti-angiogenesis drug and an extracellular matrix normalization drug;

the medicine II is a photo-thermal imaging molecular medicine, a photodynamic imaging molecular medicine or a near-infrared imaging molecular medicine;

the carrier III is a high molecular carrier.

2. The macromolecular nano-drug according to claim 1, wherein the Linker is a GSH response unit, a ROS response unit or an enzyme response unit, and both ends of the Linker are respectively connected with the drug through hydroxyl, aldehyde, amino or sulfhydryl functional groups.

3. The polymeric nano-drug according to claim 1, wherein the anti-angiogenic drug is one or more of bevacizumab, combretacept, dexamethasone, ranibizumab, and avastin, the extracellular matrix normalization drug is one or more of verteporfin, papain, tranilast, and hyaluronidase, the drug II is one or more of dihydroporphin, indocyanine green, near infrared dye IR780, and polydopamine, and the carrier III is mPEG-DSPE or mPEG-b-PLA.

4. The polymeric nano-drug according to claim 1, wherein the polymeric nano-drug has a particle size of 50 to 300nm.

5. The polymeric nano-drug according to claim 1, wherein the anti-angiogenic drug is dexamethasone, the extracellular matrix normalization drug is tranilast, the drug II is indocyanine green, the carrier III is mPEG-DSPE, and the respective corresponding structural formulas are as follows:

the structural formula of the medicine I is shown as the formula (I):

the structural formula of the medicine II is shown as the formula (II):

the structure of the carrier III is shown as the formula (III):

6. the method for preparing a polymeric nano-drug for enhancing the tumor treatment effect by regulating the tumor extracellular matrix according to any one of claims 1 to 5, comprising the following steps:

1) Connecting the anti-angiogenesis medicine and the extracellular matrix normalization medicine through a linker responding to a microenvironment to obtain a medicine I;

2) Self-assembling the medicine I, the medicine II and the carrier III to obtain the macromolecular nano-medicine containing various medicines.

7. The method according to claim 6, wherein one embodiment of step 1) in the method is as follows:

1-1) mixing tranilast, a micro-environment response linker and an anhydrous solvent for reaction to obtain an intermediate IV, wherein the intermediate IV has a structure shown in a formula (IV);

1-2) mixing the intermediate IV, dexamethasone and an anhydrous solvent for reaction to obtain the drug I.

8. The method according to claim 6, wherein the reaction temperature in the step 1) is 10 to 150 ℃, the reaction temperature in the step 2) is 10 to 50 ℃, and the ratio of the drug I: and (2) medicine II: the mass ratio of the carrier III is (1-100): (1-5): (100-500).

9. The method according to claim 7, wherein the temperature of the reaction in the step 1-1) is 120 to 130 ℃ and the temperature of the reaction in the step 1-2) is 20 to 40 ℃.

10. An antitumor drug comprising the polymeric nano-drug according to any one of claims 1 to 5, a contrast agent for near-infrared fluorescence imaging comprising the polymeric nano-drug according to any one of claims 1 to 5, or an anti-inflammatory drug comprising the polymeric nano-drug according to any one of claims 1 to 5.

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