CN115707459A - Synergistic composition of drug-resistant tumor treatment drugs - Google Patents
Synergistic composition of drug-resistant tumor treatment drugs Download PDFInfo
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- CN115707459A CN115707459A CN202110952808.XA CN202110952808A CN115707459A CN 115707459 A CN115707459 A CN 115707459A CN 202110952808 A CN202110952808 A CN 202110952808A CN 115707459 A CN115707459 A CN 115707459A
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Abstract
The invention belongs to the technical field of biological medicines, relates to a synergistic composition of drugs for treating drug-resistant tumors, and particularly relates to a composition for enhancing the treatment of drug-resistant tumors by chemotherapeutic drugs and inhibiting lung metastasis, a combined preparation thereof and a construction method thereof. The composition consists of ambroxol, pluronic and phospholipid materials; the composition can reverse drug resistance of drug-resistant tumor cells to chemotherapeutic drugs, inhibit growth and proliferation of tumor cells, and inhibit lung metastasis of tumor cells. The invention also provides a preparation of the composition, which comprises ambroxol, phospholipid and pluronic composition solution, liposome and micelle preparation, wherein the composition can remarkably improve the cytotoxic effect and in-vivo anti-tumor effect of chemotherapeutic drugs on drug-resistant tumors and inhibit the lung metastasis of the drug-resistant tumors.
Description
Technical Field
The invention belongs to the technical field of biological medicines, relates to a synergistic composition of drugs for treating drug-resistant tumors, and particularly relates to a composition for enhancing the treatment of drug-resistant tumors by chemotherapeutic drugs and inhibiting lung metastasis, and construction and application of a combined preparation of the composition.
Background
The prior art discloses that in cancer chemotherapy, the development of multidrug resistance (MDR) is a major cause of chemotherapy failure. MDR refers to the phenomenon of drug resistance of malignant cells to various drugs unrelated to molecular structure, the classical mechanism of MDR is the overexpression of adenosine triphosphate binding cassette (ABC) transmembrane transporter, such as P-glycoprotein (P-glycoprotein) and multidrug resistance-associated protein (MRP), and the overexpressed P-gp leads to a decrease in the accumulation of an anticancer drug in cancer cells, and thus requires an increase in dose to produce equivalent toxicity. Third-generation P-gp inhibitors, such as tariquidar, have shown superior efficacy in overcoming MDR. However, since P-gp plays an important role in the physiological regulation of both endogenous and exogenous compounds in vivo, systemic use of P-gp inhibitors has significant side effects. Therefore, it is important to limit exposure of normal cells and tissues to efflux inhibitors. Studies have reported that co-administration of an anti-cancer drug with an efflux inhibitor can achieve co-localization of the drug and inhibitor in the same tumor cell population, maximizing therapeutic efficacy and minimizing systemic toxicity.
Numerous studies have shown that autophagy plays an important role in the development, progression and resistance of tumors. Relevant clinical data suggest that autophagy can be activated as a protective mechanism mediating MDR during tumor therapy. Therefore, inhibiting autophagy of tumor cells can re-sensitize drug-resistant cancer cells, induce apoptosis of tumor cells, and enhance the cytotoxic effect of chemotherapeutic drugs. Ambroxol is used as an expectorant and a pulmonary surfactant synthesis inducer, has the effects of oxidation resistance, inflammation resistance and the like, and is mainly used for respiratory diseases. According to the preliminary research of the research team laboratory of the literature research and the application, ambroxol can regulate the autophagy of cells, for example, can improve the expression level of autophagy-related proteins LC3B-II, p62/SQSTM1 in neurons and tumor cells. It is currently believed that its mechanism of regulation of autophagy is primarily by inhibiting fusion between lysosomes and autophagosomes, increasing the pH of lysosomes, thereby blocking the autophagy flux.
Clinical practice has shown that most patients may develop different degrees of tumor metastasis when hospitalized, and data shows that metastatic spread of primary tumors accounts for more than 90% of the mortality of solid tumor patients. Therefore, inhibition of tumor metastasis is an important part of cancer therapy. Matrix Metalloproteinases (MMPs) are a highly conserved group of endoproteolytic enzymes containing zinc atoms, which can be classified into collagenases, gelatinases, matrilysins, macrophage elastase, membrane-type MMPs depending on the substrates used, and these enzymes mainly degrade various types of collagens, gelatinases, fibronectin, laminin and other extracellular matrices. MMPs promote tumor invasion and metastasis by degrading the extracellular matrix (ECM) and basement membrane surrounding the tumor. Research shows that the matrix metalloproteinase inhibitor has obvious effect of resisting tumor metastasis. Triblock copolymer pluronic consisting of Polyoxyethylene (PEO) and polyoxypropylene ether (PPO) blocks is a common amphiphilic polymer material, and special block ratio types such as P123, P85 and P105 of the triblock copolymer pluronic have a biological function of reducing MMP-9 expression, and have a good prevention and treatment effect on tumor cell metastasis.
Based on the research background and the current situation of the prior art, the application aims at the characteristics of drug-resistant tumors: the composition and the preparation thereof are formed by a certain proportion, and the composition and the preparation thereof are used for enhancing the killing power of the chemotherapeutic drug to the drug-resistant tumor by multiple strategies and multiple measures, and simultaneously inhibiting the invasion and the metastasis of the drug-resistant tumor to the lung, thereby further improving the anti-tumor effect.
Disclosure of Invention
The invention aims to solve the problems of the prior art that the drug-resistant tumor has the following characteristics: the synergistic composition for treating the drug-resistant tumor medicament is provided by utilizing the inhibition effect of pluronic on drug-resistant tumor efflux protein and matrix metalloenzyme, the autophagy regulation effect of ambroxol on drug-resistant tumor cells and the affinity capacity of phospholipid cells and lung tissues, so as to enhance the lethality of chemotherapeutic medicaments on the drug-resistant tumor, inhibit the invasion and metastasis of the drug-resistant tumor to the lung and improve the anti-tumor effect.
The composition consists of ambroxol, pluronic and phospholipid materials, and is further prepared into a pluronic liposome or micelle preparation containing the ambroxol, wherein the medicinal preparation can enhance the treatment of drug-resistant tumors by chemotherapeutic drugs and inhibit the lung metastasis of the tumors.
More specifically, the present invention is directed to a method for producing,
the invention provides a synergistic composition of a drug for treating drug-resistant tumors, which consists of Ambroxol, pluronic and phospholipid materials, wherein the Ambroxol is Ambroxol hydrochloride, the English name is Ambroxol (Ax), the chemical formula of the Ambroxol is trans-4- [ (2-amino-3, 5-dibromo benzyl) amino ] cyclohexanol, the molecular weight is 378.1, the Pluronic (Pluronic) comprises Pluronic P105, pluronic P85 and Pluronic P123, the general name is poloxamer, and the Pluronic is a PEO-PPO nonionic triblock copolymer consisting of Polyoxyethylene (PEO) and polyoxypropylene (PPO), and the phospholipid materials are selected from Dipalmitoylphosphatidylcholine (DPPC), distearoylphosphatidylcholine (DSPC), lecithin and soybean lecithin.
The synergistic composition is further prepared into a solution, a liposome or a micelle preparation containing ambroxol, pluronic and a phospholipid composition;
wherein the dose concentration range of the ambroxol is 5-100mg/ml; the dosage concentration range of the pluronic is 0.005-5mg/ml; the molar ratio of phospholipid to pluronic was 100.
In the invention, the solution preparation is prepared by the following method:
ambroxol, pluronic and phospholipid material are dissolved in deionized water or PBS and then are subjected to ultrasonic dispersion to prepare a composition solution.
In the invention, the composition liposome or micelle preparation is prepared by adopting a film dispersion method:
dissolving phospholipid and Poncirus into organic solvent, removing the organic solvent by rotary evaporation, adding deionized water or PBS buffer solution containing ambroxol for hydration, performing ultrasonic treatment, and extruding through polycarbonate membrane to obtain liposome or micelle preparation.
The organic solvent in the preparation method is trichloromethane, methanol, dichloromethane and ethanol.
The invention further provides the application of the synergistic composition for treating drug-resistant tumor in preparing a medicinal preparation for enhancing the treatment of drug-resistant tumor by chemotherapeutic drugs and inhibiting lung metastasis,
the pharmaceutical preparation inhibits the growth and proliferation of tumor cells and inhibits the pulmonary metastasis of tumor cells;
the chemotherapy drugs are paclitaxel, docetaxel, cabazitaxel taxane drugs or doxorubicin, aclarubicin and epirubicin anthracycline chemotherapy drugs.
The composition preparation of the invention is administrated by intravenous injection or inhalation.
Table 1 shows the IC of A549/T cells incubated with different concentrations of ambroxol, pluronic and phospholipid material composition and paclitaxel for 24h, 48h and 72h 50 The value is obtained.
Table 2 shows the IC of A549/T cells incubated with Pluronic P105, pluronic P85, pluronic P123, ambroxol, phospholipid material composition and paclitaxel for 24h 50 The value is obtained.
Table 3 shows the IC of the composition of pluronic, ambroxol, DPPC, DSPC, lecithin and soybean lecithin with different concentrations after incubating A549/T cells with paclitaxel for 24h 50 The value is obtained.
Table 4 shows the combination of Pluronic, ambroxol and phospholipid with docetaxel, cabazitaxel, doxorubicin, aclarubicin, epiIC 24h after Genobicin co-incubation of A549/T cells 50 The value is obtained.
TABLE 1
TABLE 2
TABLE 3
TABLE 4
The invention provides a composition which can be used for preparing a medicinal preparation for enhancing the treatment of drug-resistant tumors by chemotherapeutic drugs and inhibiting lung metastasis.
The invention achieves the following beneficial results:
(1) The invention combines the ambroxol, the pluronic and the phospholipid materials, enhances the treatment of the chemotherapy drugs to the drug-resistant tumor and inhibits the lung metastasis of the chemotherapy drugs.
(2) The composition preparation of the ambroxol-containing pluronic liposome or micelle, which is prepared by the invention, is used for enhancing the treatment of chemotherapy drugs on drug-resistant tumors and inhibiting the lung invasion and metastasis, is easy to prepare and can be produced in large quantities.
Drawings
FIG. 1 different compositions inhibit tumor cell invasion and metastasis
(A) Results of A549/T cell scratch assay. a. Scratching cells for 0h in the control group; b. the cell scratch of the control group for 24h is completely healed; c. scratching cells after adding phospholipid liposome (BL) for 24 hours; d. adding Pluronic-Phospholipid Liposome (PL) for 24h, and scratching; e. adding Pluronic-Phospholipid Micelle (PM) for 24h, and scratching the cells;
(B) Results of Transwell experiments. Control group (Control), phospholipid liposome (BL), pluronic-Phospholipid Liposome (PL);
(C) The effect of the composition preparation on the expression of MMP-9 in tumor cells. Phospholipid liposomes (BL), pluronic-Phospholipid Liposomes (PL) and pluronic-Phospholipid Micelles (PM).
FIG. 2 physicochemical Properties of composition formulations
(A) Transmission electron micrograph of composition preparation. Phospholipid liposome (a) and composition liposome (b). A scale:
100nm；
(B) And (3) characterization results of infrared spectroscopy on the pluronic and phospholipid components in the composition preparation. The combination of Pluronic (a),
BL(b)and PL(c)。
FIG. 3 Effect of different composition formulations on cellular uptake Capacity and SP-A expression level
(A) The A549/T cells take up drug-resistant tumor cells of a phospholipid liposome preparation (BRL) loaded with Rh123, a pluronic-phospholipid liposome Preparation (PRL) without ambroxol and a composition liposome preparation (PRL + Ax);
(B) A549/T cell uptake Rh 123-loaded BRL and PRL, PRL + Ax or free Rh123 flow quantification results;
(C) Ambroxol, pluronic, phospholipid material composition and effect on SP-a protein expression by a549/T cells at different concentrations;
(D) Effect of different composition formulations on the expression of SP-a protein by a549/T cells.
FIG. 4 cellular level reversal of resistance mechanism for different composition formulations
(A) The effect of different composition preparations on the expression of P-gp proteins by A549 and A549/T cells;
(B) Effect of different composition formulations on a549 and a549/T cell autophagy flux.
FIG. 5 in vivo distribution of different composition formulations and effects on microenvironment of lung tumor tissue
(A) Distribution of different composition preparations in lung tumor and other organs within 24 h;
(B) Quantification of the distribution of different composition formulations in lung tumors within 24 h;
(C) The expression condition of the lung surfactant-associated protein SP-A of the nude mice bearing the tumor 24 hours after the administration of different composition preparations;
(D) And (3) the expression conditions of IL-6, TNF-alpha, FN and alpha-SMA in the lung tumor microenvironment of each group of tumor-bearing nude mice after multiple administration treatments.
Figure 6 in vivo pharmacodynamic evaluation of different composition formulations.
(A) In the A549/T-luciferase lung tumor model, the sizes of lung tumors are changed after different administration groups are treated;
(B) Weight change of nude mice of different administration groups in A549/T-luciferase lung tumor model;
(C) In the A549/T-luciferase lung tumor model, the survival time of nude mice after different administration groups are compared.
Detailed Description
The present invention is further illustrated by the following specific examples, which are provided for the purpose of illustration only and are not intended to be limiting. If the experimental conditions specified in the examples are not indicated, they are generally according to the conventional conditions, or according to the conditions recommended by the reagents company; reagents, consumables and the like used in the following examples are commercially available unless otherwise specified.
Example 1 different compositions enhance the cytotoxic Effect of chemotherapeutic drugs against drug resistant tumors
The method comprises the following steps: (1) The growth inhibition effect of the combination of the pluronic, the ambroxol and the phospholipid material composition and the paclitaxel with different concentrations on A549 and a drug-resistant strain A549/T, MCF7 and a drug-resistant strain MCF7/ADR cells is detected by an MTT method.
(2) And detecting the IC50 value of the combination of the composition of the pluronic, the ambroxol and the phospholipid material, docetaxel, cabazitaxel, doxorubicin, aclarubicin and epirubicin after incubating the A549/T cells for 24h.
As a result: (1) As shown in Table 1, the addition of Ax and Plu can increase the toxicity of PTX to cells compared with the pure PTX group, but the composition of ambroxol, pluronic and phospholipid has more remarkable sensitization effect on PTX compared with two-by-two combination, and the sensitization effect is more remarkable in drug-resistant strain cells A549/T and MCF-7/ADR.
(2) As shown in tables 2-4, the composition of pluronic, ambroxol and phospholipid significantly enhances the cytotoxic effect of the therapeutic drugs such as docetaxel, cabazitaxel, doxorubicin, aclarubicin and epirubicin on drug-resistant tumors.
Example 2: different compositions inhibit invasion and metastasis of drug-resistant tumor cells
The method comprises the following steps: (1) cell scratch test: A549/T cells are inoculated in a 12-hole plate, and when the cell density in the hole is about 90 percent, a vertical scratch is scratched in the middle of a culture hole by using a gun head. The floating cell debris was then washed with PBS and incubated for 24h with the addition of serum-free medium containing 0.2mg/mL of BL, PL and PM. Photographs were taken after 0h and 24h incubation to compare the width of the scratch.
(2) Transwell experiment: A549/T cells were previously cultured in a complete medium containing 0.2mg/mL of BL and PL for 24 hours, followed by seeding on the upper part of a Transwell chamber. After 24h of inoculation, the cells in the upper part of the chamber were wiped off with a cotton swab, and the cells in the lower part were fixed after washing with PBS. The lower chamber cells were then stained with 0.5% crystal violet solution and after 30min the remaining crystal violet solution was washed and photographed.
(3) Effect of the combination preparation on MMP-9 expression in tumor cells: A549/T cells were seeded in 6-well plates and complete medium containing 0.2mg/mL of BL, PL or PM was added. After 24h incubation, cells in each well were washed with PBS and lysed with RIPA lysate containing protease inhibitors, and the amount of protein in the cell samples after lysis was determined. MMP-9 expression levels in BL, PL and PM treated A549/T cells were analyzed using Western Blot.
As a result: (1) As shown in fig. 1A, by comparing scratch widths of 0h and 24h, it was shown that the control group had completely healed the scratch at 24h, indicating that a549/T cells have strong healing ability. For the experimental group, after the treatment of BL with 0.2mg/mL for 24 hours, scratches of A549/T cells are almost completely healed, which shows that BL has no inhibition effect on the healing of scratches of A549/T cells; significant scratching was found in a549/T cells treated with PL and PM, indicating that the composition can inhibit a549/T cell invasion.
(2) The cells pretreated with BL and PL were seeded in a Transwell chamber, and the cells migrated to the lower side were stained with crystal violet, and the results are shown in FIG. 1B. The number of a549/T cells after BL treatment migrated to the lower chamber was not significantly different from that of the control group, whereas a549/T cells treated with PL migrated to the lower chamber in a smaller number and significantly less than that of the BL group, indicating that the composition could inhibit the migration of a549/T cells.
(3) MMP-9 expression levels in BL, PL and PM treated A549/T cells were analyzed using Western Blot. As shown in fig. 1C, both PL and PM showed significant inhibition of MMP-9 expression, indicating that the composition inhibited migration of a549/T cells by inhibiting the expression of MMP 9.
Example 3: preparation and physicochemical characterization of combined liposome (PL) and micelle (PM) preparations
The method comprises the following steps: (1) Preparing a composition liposome or micelle preparation by adopting a thin film hydration method, weighing the composition according to the molar ratio of the pluronic to the phospholipid of 1. Then, the deionized water solution containing 5-100mg/ml ambroxol is adopted for hydration, and the mixture is rapidly oscillated, ultrasonically treated and hydrated, and finally extruded through a polycarbonate membrane to obtain the liposome or micelle containing the composition. The results show that the molar ratio of pluronic to phospholipids is in the range of 1.
(2) The particle size and zeta potential of the nanoparticles are measured by a Malvern particle size analyzer, the morphology of the nanoparticles is observed by a transmission electron microscope, and the component characteristics of the phospholipid material and the pluronic P105 are proved by infrared spectroscopy. As shown in fig. 2, the particle sizes of the blank phospholipid liposome and the composition liposome are respectively: 103.3 +/-3.0 and 107.9 +/-3.4 nm, and the shape is spherical, and the corresponding particle size is basically consistent with the result measured by the dynamic light scattering method. Fourier Infrared Spectroscopy (FIR) analysis of the structural functional groups of Pluronic and phospholipid in the composition proves that the characteristic peak of C-O-C of Pluronic 1102cm-1 exists in a liposome preparation of the composition, and the characteristic peak of C = O1736 cm-1 of phospholipid material also exists, which indicates that Pluronic and phospholipid material can form stable liposome or micelle solution.
Example 4: effect of different compositions on cellular uptake Capacity and SP-A expression level
The method comprises the following steps: (1) A549/T cells are inoculated in a 24-well plate and cultured for 24h. Adding a proper amount of preparation solution BRL, PRL or PRL +100 muM Ax solution which is prepared by RPMI 1640 serum-free medium and is loaded with Rh123 to ensure that the final concentration of Rh123 in a sample hole is 5 muM, placing the sample hole in an incubator at 37 ℃ and keeping out of the light for incubation for 0.5h, 1h and 2h, and then washing, wherein 3 parts of each group are processed in parallel. The uptake of the cells was qualitatively observed by an inverted fluorescence microscope and photographed. And another group is respectively given with BRL, PRL +100 mu M Ax solution or Free Rh123 solution to ensure that the final concentration of Rh123 in the sample hole is 5 mu M, and after incubation for 2h in a dark place, the cells are washed, digested, collected and quantitatively analyzed and detected by a flow cytometer.
(2) A549/T cells were seeded in 6-well plates and after 24h of culture, different concentrations of Ax solution (50. Mu.M, 100. Mu.M, 200. Mu.M), different concentrations of Pluronic 105 solution (0.025 mg/mL,0.05mg/mL and 0.1 mg/mL), and different formulations and combination solutions of formulation and Ax with phospholipid concentration of 0.2mg/mL were incubated for 24h. After incubation, cells were washed and harvested, and the effect of different treatments on SP-A expression in cells was determined by the Western Blot method.
As a result: (1) As shown in fig. 3 (a, B), a549/T cells have relatively stronger agent uptake capacity for PRL and Ax and PRL combined incubation groups than BRL groups. Flow quantification gave the same results. Therefore, the composition of ambroxol, pluronic and phospholipid material can obviously increase the intake level of chemotherapy drugs by drug-resistant strain cells, and further proves the uniqueness and the necessity of the composition and the preparation thereof in enhancing the treatment of the chemotherapy drugs on drug-resistant tumors.
(2) As shown in FIG. 3 (C, D), ambroxol, pluronic and phospholipid material all promote the secretion of SP-A, and the promotion effect is proportional to the dosage. In addition, the composition of ambroxol, pluronic and phospholipid material and the preparation thereof have the best effect on promoting SP-A secretion.
Example 5: mechanism for reversing drug resistance of drug-resistant tumor to chemotherapeutic drugs by combined group
The method comprises the following steps: the cell processing method is as shown in example 4, and the influence of the drug resistance related protein p-gp and the autophagy marker LC3-II of different processing modes is determined by the Western Blot method.
The result is shown in fig. 4, the pluronic has no influence on autophagy of cells and the Ax has no influence on the expression of p-gp, but the composition of ambroxol, pluronic and phospholipid material has the capacity of inhibiting tumor autophagy and reducing the expression of drug-resistant related protein p-gp, which indicates that the composition has a unique increasing effect on reversing drug-resistant tumor resistance.
Example 6: in vivo pharmacodynamic evaluation of formulations of different compositions
The method comprises the following steps: (1) in vivo distribution experiment: the blank phospholipid liposomes (BL) labeled with DiR and the blank pluronic-phospholipid liposome Preparation (PL) were injected intravenously into tumor-bearing mice (DiR 0.2 mg/kg), and the liposomes were hydrated with 100 μ M Ax solution for the different compositions of liposome preparation (BL + Ax group, PL + Ax group). Mice were sacrificed at 6, 12 and 24h post-dose for each group, and the main organs were dissected after heart perfusion and photographed by scanning and semiquantitated for fluorescence intensity. Then the lung tumor tissue is fixed and dehydrated and then paraffin embedding is carried out. The expression of the lung surfactant SP-A in the lung tissue of mice bearing lung metastases was analyzed by immunohistochemical method.
(2) In vivo efficacy test: the growth condition of the lung A549/T-Luciferase tumor is observed by using IVIS living body imaging, when the lung tumor grows to a proper size, the tumor-bearing nude mice are randomly divided into 6 groups, and each group comprises 6 mice, namely a salt group (normal Saline group), a PTX group, a BPL +100mg/kg Ax group, a PPL group and a PPL +100mg/kg Ax group. Each formulation was given by tail vein injection on days 0, 4, 8, 12 and 16, respectively, and each group of chemotherapy drug Paclitaxel (PTX) was administered at a dose of 5mg/kg. The in vivo efficacy results were monitored after each group was dosed by the following criteria:
(1) growth of drug-resistant tumors: the growth of the lung tumor of the tumor-bearing mice is monitored by adopting an IVIS in-vivo imaging method, and small animal in-vivo imaging shooting is carried out on days 0,5, 10, 15 and 20.
(2) Tumor-bearing mice were measured for body weight every 3 days.
(3) After the experiment was completed, each group of tumor-bearing mice was sacrificed, and tumor-bearing lung tissues and other major tissues and organs were collected, fixed, dehydrated, paraffin-embedded and H & E stained.
(4) The expression levels of the immune factors IL-6, TNF-alpha and the tumor-associated fibroblast-associated proteins FN and alpha-SMA in the lung microenvironment were determined by immunohistochemistry.
As a result: (1) As shown in figure 5, in the drug-resistant lung metastasis model established by A549/T-luciferase cells, relative accumulation of ambroxol, pluronic and phospholipid material composition preparation (Ax + PL group) in lung is the most, and the tumor tissue responsiveness of the composition is proved, so that the accumulation of the chemotherapeutic drug in tumor sites can be enhanced. Immunohistochemical results of SP-A protein showed that the secretion of SP-A was significantly increased compared to other groups, ambroxol, ponnonik, phospholipid material composition preparation (Ax + PL group), in concert with in vivo distribution experiments.
(2) The growth of the tumor was observed and the change of median survival was used to examine the in vivo efficacy of different experimental groups, as shown in fig. 6, the composition of ambroxol, pluronic and phospholipid material had the best effect of enhancing the inhibition of chemotherapeutic drugs on the growth of drug-resistant tumors (Ax + PPL group), and the survival could be extended to 63.5 days. By adopting an HE staining and immunohistochemical method, the examination result of the expression levels of the inflammatory factors related to lung tumor microenvironment and the marker proteins related to CAFs of each experimental group shows that Ax can obviously reduce the secretion of the inflammatory factors IL-6, TNF-alpha, FN and alpha-SMA by lung tissues and lung tumor tissues, and the down-regulation effect of the inflammatory factors is more obvious under the synergistic effect of ambroxol, pluronic and a phospholipid composition.
In conclusion, the ambroxol, pluronic and phospholipid materials are adopted to prepare the composition preparation, and the composition or the preparation thereof is combined with chemotherapeutic drugs, so that the killing effect of the chemotherapeutic drugs on drug-resistant tumor cells can be enhanced, and the lung metastasis of the drug-resistant tumor cells can be inhibited while the drug resistance of the drug-resistant tumor cells is reversed.
Claims (7)
1. A synergistic composition of a drug for treating drug-resistant tumors is composed of Ambroxol, pluronic and phospholipid materials, wherein the Ambroxol is Ambroxol hydrochloride (Ax), the chemical formula of the Ambroxol is trans-4- [ (2-amino-3, 5-dibromobenzyl) amino ] cyclohexanol, the molecular weight of the Ambroxol is 378.1, the Pluronic (Pluronic) comprises Pluronic P105, pluronic P85 and Pluronic P123, the general formula of the poloxamer is PEO-PPO nonionic triblock copolymer composed of Polyoxyethylene (PEO) and polyoxypropylene (PPO), and the phospholipid materials are selected from Dipalmitoylphosphatidylcholine (DPPC), distearoylphosphatidylcholine (DSPC), lecithin and soybean phospholipid.
2. The synergistic combination of claim 1 for the treatment of drug resistant tumors wherein said combination is formulated as a solution, liposome or micelle containing ambroxol, pluronic, phospholipid composition;
wherein the dose concentration range of the ambroxol is 5-100mg/ml; the dosage concentration range of the pluronic is 0.005-5mg/ml; the molar ratio of phospholipid to pluronic is 100.
3. The use of the synergistic combination of drugs for the treatment of drug-resistant tumors of claim 2 for the preparation of a pharmaceutical formulation for enhancing the treatment of drug-resistant tumors with chemotherapeutic drugs and inhibiting lung metastases,
the pharmaceutical preparation inhibits the growth and proliferation of tumor cells and inhibits the pulmonary metastasis of tumor cells;
the chemotherapy drugs are paclitaxel, docetaxel, cabazitaxel taxane drugs or doxorubicin, aclarubicin and epirubicin anthracycline chemotherapy drugs.
4. The synergistic combination of drugs for the treatment of drug-resistant tumors as claimed in claim 2, wherein said solution formulation is prepared by the following method:
ambroxol, pluronic and phospholipid material are dissolved in deionized water or PBS and then are subjected to ultrasonic dispersion to prepare a composition solution.
5. The synergistic composition for the treatment of drug-resistant tumor drugs according to claim 2, wherein the composition liposome or micelle preparation is prepared by a thin film dispersion method:
dissolving phospholipid and Poncirus into organic solvent, removing the organic solvent by rotary evaporation, adding deionized water or PBS buffer solution containing ambroxol for hydration, performing ultrasonic treatment, and extruding through polycarbonate membrane to obtain liposome or micelle preparation.
6. The synergistic composition for the treatment of drug-resistant neoplastic disease of claim 5 wherein the organic solvent used in the process is chloroform, methanol, methylene chloride, or ethanol.
7. The synergistic combination of claim 2, wherein the formulation is administered by intravenous injection or inhalation.
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