A combined drug combination for anti-tumor treatment

Abstract

The present invention discloses a combined drug for anti-tumor use, and the combined drug comprises an anti-tumor active substance and an antibacterial drug. In view of the fact that bacteria in tumors can reduce the anti-tumor treatment effect through various mechanisms such as promoting tumor growth, metastasis and inactivating therapeutic drugs, the present invention discovers that using antibacterial drugs in anti-tumor drugs can improve the cancer treatment effect of anti-tumor drugs by eliminating the adverse effects of bacteria, make up for the deficiencies of only using anti-tumor active substances, improve the anti-tumor effect, improve the prognosis or quality of life of patients, and has broad application prospects in the field of tumor treatment.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical technology, and particularly relates to a combined drug for anti-tumor use. Background Art

[0002] Globally, cancer is a major cause of death and an important obstacle to increasing life expectancy. Currently, there are various treatment options for tumors, such as chemotherapy, phototherapy, ion interference therapy, etc., which are widely used in tumor treatment. Chemotherapy refers to a treatment method that uses cytotoxic drugs to prevent the proliferation, infiltration, and metastasis of cancer cells until the cancer cells are killed. It is one of the most effective means for treating cancer at present. However, chemotherapy drugs are highly toxic and can cause a continuous weight loss in cancer patients, reducing the treatment effect. Phototherapy is a new means of cancer treatment. It irradiates the lesion area with a near-infrared light source to stimulate the photosensitizer to generate singlet oxygen with a killing effect on tumor cells or generate heat to cause local heating of the lesion site to kill tumor cells, so as to achieve the treatment purpose. Phototherapy has small side effects and can produce a synergistic treatment effect when combined with other treatment methods. Ion interference therapy is to regulate the interaction between bioactive ions and the physiological processes of tumor cells, induce metabolic disorders of tumor cells, and thus play a role in inhibiting tumor growth and metastasis. At the same time, in terms of antibacterial, metal ions are positively charged and can be adsorbed onto the cell membrane through electrostatic interaction, resulting in the rupture of the cell membrane; metal ions can also play a catalytic role in inducing oxidative stress reactions, ultimately causing bacteria to die.

[0003] In recent years, the correlation between bacteria and malignant tumors has attracted wide attention. Studies have found that bacteria are almost present in all cancer types. These bacteria, especially intracellular bacteria in tumor cells, are important factors promoting the occurrence and development of cancer. Bacteria can trigger pathogen-associated molecular patterns, thereby inducing inflammatory responses and continuously producing inflammatory mediators, such as tumor necrosis factor α, interleukin-6, etc., directly or indirectly promoting tumor progression. Bacteria may also cause chemotherapy drug resistance by metabolizing chemotherapy drugs, reducing the anti-tumor treatment effect. At the same time, the use of antibacterial drugs in the treatment of solid tumors can have a positive impact on tumor treatment, such as inhibiting tumor growth, reducing metastasis, and improving prognosis. However, limited by the complex physiological environment of tumors and the characteristics of intracellular bacteria, traditional antibacterial drugs cannot completely eliminate intracellular bacteria, resulting in the long-term survival of tumor-promoting bacteria.

[0004] Currently, anti-tumor drug treatments centered on clearing tumor cells are limited by tumor multi-drug resistance and the complex tumor microenvironment, and usually have limited efficacy. During cancer treatment, the administration of antibacterial drugs has positive significance for inhibiting tumor growth, metastasis, and improving tumor prognosis. Therefore, the combination therapy of anti-tumor active substances and antibacterial drugs has broad prospects in the field of anti-tumor treatment. Summary of the Invention

[0005] The object of the present invention is to provide a combined drug for anti-tumor use.

[0006] To achieve the above object, the present invention adopts the following technical solutions:

[0007] Use of the combination of an anti-tumor active substance and an antibacterial drug in the preparation of a tumor treatment drug;

[0008] The anti-tumor active substance is selected from at least one of chemotherapeutic drugs, photosensitizers, and inorganic metal salts with anti-cancer activity. The chemotherapeutic drugs are selected from doxorubicin, daunorubicin, idarubicin, cisplatin, oxaliplatin or mitoxantrone. The photosensitizers are selected from indocyanine green (IR-808), neo-indocyanine green (IR-820), chlorin e6 or temoporfin. The inorganic metal salts with anti-cancer activity are selected from ferrous sulfate, ferric chloride, copper sulfate, manganese chloride or magnesium chloride;

[0009] The antibacterial drug is selected from vancomycin, teicoplanin, benzylpenicillin, cefaclor, metronidazole or sulfamethoxazole.

[0010] Further, the dosage ratio w:w of the anti-tumor drug active substance to the antibacterial drug is 1 to 10:1.

[0011] Further, the active ingredient of the tumor treatment drug is a composition of an anti-tumor active substance and an antibacterial drug, or a self-assembled nanoparticle of an anti-tumor active substance and an antibacterial drug.

[0012] A tumor treatment drug, comprising an anti-tumor active substance and an antibacterial drug, as well as a pharmaceutically acceptable carrier and / or excipient;

[0013] The anti-tumor active substance is selected from at least one of chemotherapeutic drugs, photosensitizers, and inorganic metal salts with anti-cancer activity. The chemotherapeutic drugs are selected from doxorubicin, daunorubicin, idarubicin, cisplatin, oxaliplatin or mitoxantrone. The photosensitizers are selected from indocyanine green (IR-808), neo-indocyanine green (IR-820), chlorin e6 or temoporfin. The inorganic metal salts with anti-cancer activity are selected from ferrous sulfate, ferric chloride, copper sulfate, manganese chloride or magnesium chloride;

[0014] The antibacterial drug is selected from vancomycin, teicoplanin, benzylpenicillin, cefaclor, metronidazole or sulfamethoxazole.

[0015] Further, the dosage ratio w:w of the anti-tumor drug active substance to the antibacterial drug is 1 to 10:1.

[0016] Further, the active ingredient of the tumor treatment drug is a composition of an anti-tumor active substance and an antibacterial drug.

[0017] Furthermore, the active ingredient of the tumor therapeutic drug is a self-assembled nanoparticle of an anti-tumor active substance and an antibacterial drug.

[0018] Compared with the prior art, the present invention has the following advantages:

[0019] 1. Based on the fact that tumor-promoting bacteria in tumors can promote tumor growth, metastasis, and reduce the therapeutic effect of chemotherapeutic drugs through various mechanisms, the present invention discovers that introducing antibacterial drugs to eliminate tumor-promoting bacteria in tumor treatment can eliminate the adverse effects of tumor-promoting bacteria on tumor treatment and improve the tumor treatment effect of anti-tumor active substances, which is specifically reflected in the increase in tumor inhibition rate and the reduction of distant metastasis. Therefore, the combined application of anti-tumor active substances and antibacterial drugs has excellent synergistic anti-tumor effects and can provide a new combined treatment strategy for clinical anti-tumor treatment.

[0020] 2. The present invention provides various types of anti-tumor active substances for tumor treatment, including chemotherapeutic drugs, photosensitizers, and inorganic metal salts with anti-tumor effects. Using anti-tumor active substances with different mechanisms in combination during anti-tumor treatment can reduce drug resistance and toxicity, thereby improving cancer treatment.

[0021] 3. The inorganic metal salt with anti-tumor effects among the anti-tumor active substances in the present invention also has antibacterial effects. When used in combination with antibacterial drugs, it can enhance the killing effect on intracellular bacteria in tumors, thereby completely eliminating tumor-promoting bacteria.

[0022] 4. The drug combination using anti-tumor drugs and antibacterial drugs in the present invention can eliminate bacteria in tumor tissues, effectively improve the anti-tumor treatment effect, and improve the prognosis of cancer treatment. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 It is the lung H&E picture of mice after treatment with different groups in Example 1.

[0024] Figure 2 It is the quantitative result of the number of bacteria in tumors of mice after treatment with different groups in Example 1.

[0025] Figure 3 It is a representative picture of plating the tumor homogenate of mice after treatment with different groups in Example 1. DETAILED DESCRIPTION OF THE INVENTION

[0026] The following examples further illustrate the content of the present invention, but should not be construed as limiting the present invention. Without departing from the spirit and essence of the present invention, the modifications and substitutions made to the methods, steps, or conditions of the present invention all belong to the scope of the present invention. If not specifically indicated, the technical means used in the examples are conventional means well known to those skilled in the art.

[0027] The anti-tumor combined drug provided by the embodiment of the present invention includes the combined combination in Table 1.

[0028] Table 1 Composition of anti-tumor combined drug

[0029]

[0030] Example 1

[0031] Evaluation of anti-tumor, anti-metastatic and antibacterial properties of the combination of doxorubicin, ferrous sulfate and vancomycin against tumors with bacterial infections

[0032] Preparation of doxorubicin + ferrous sulfate + vancomycin multi-component composite nano-system: Weigh accurately 2 mg of doxorubicin hydrochloride, 3 mg of ferrous sulfate and 5 mg of vancomycin hydrochloride and dissolve them in purified water. Then mix and stir, and inject 8 times the total volume of the mixed solution of 10% ethanol water under stirring conditions. Continue to stir for 10 min at a stirring speed of 1500 r / min, stand in a water bath at 30 °C for 12 h, centrifuge, redissolve the precipitate in water, and probe sonicate for 10 min to obtain the multi-component composite nano-system.

[0033] Take 1 mL of the prepared multi-component composite nano-system and dilute it to 3 mL with water, and measure it with a particle size analyzer (Malvem Instruments, Malvern, UK). The results are shown in Table 2. It can be seen from the table that for the prepared multi-component composite nano-system, the particle size of the nano-system reaches the nanometer level and the particle size distribution is uniform.

[0034] Table 2 Particle size characterization of the nano-particles of the composition prepared from doxorubicin, ferrous sulfate and vancomycin

[0035]

[0036] Using high performance liquid chromatography, measure the mass of doxorubicin or vancomycin in the self-assembled nanoparticles. Prepare a series of standard solutions of doxorubicin or vancomycin with different concentrations, and the solvent is dimethyl sulfoxide in each case. Detect the peak areas of the respective series of standard solutions at the maximum absorption wavelength of doxorubicin or vancomycin, and draw a concentration-peak area standard curve. Take 1 mL of the self-assembled nanoparticle solution, add 10 mL of dimethyl sulfoxide, measure its peak area at the maximum absorption wavelength of doxorubicin or vancomycin, substitute it into the standard curve, and calculate the mass of doxorubicin and vancomycin in the self-assembled nanoparticles. The mass of ferrous sulfate in the self-assembled nanoparticles is measured by inductively coupled plasma mass spectrometry. Finally, the ratio (w:w:w) of doxorubicin, ferrous sulfate and vancomycin in the self-assembled nanoparticles is calculated to be 2:2:1.

[0037] Next, Balb / c mice were used to evaluate the antitumor effect of the combined antitumor drugs on tumor-bearing mice with bacterial infection. After adaptive feeding, 4T1 tumor cells (2×10 6 cells) infected with Staphylococcus xylosus (MOI 2) were inoculated in situ to establish a breast cancer tumor model with bacterial infection. When the tumor volume of the mice grew to 50 mm 3 , drug treatment was started, and the tumor volume was measured daily during the treatment process, and the mice were weighed. The tumor-bearing mice were randomly divided into groups of 3 each. The experimental animals were grouped as follows: control group (Control), doxorubicin chemotherapy drug group, ferrous sulfate inorganic metal salt group, vancomycin antibacterial drug group, physical mixture of doxorubicin + ferrous sulfate + vancomycin group (with a dosage ratio of 2:2:1), and nanoformulation of doxorubicin / ferrous sulfate / vancomycin group. On days 1, 3, and 5, the drugs were injected via the tail vein, and the dosing doses of doxorubicin, ferrous sulfate, and vancomycin were 5 mg / kg, 5 mg / kg, and 2.5 mg / kg, respectively. On day 14, the mice were sacrificed, the tumor weight was weighed, and the tumor inhibition rate was calculated. The results are shown in Table 3. The combined antitumor drugs (including the physical mixture group and the nanoformulation group) had a higher tumor inhibition rate for tumor-bearing mice with bacterial infection, indicating that the combination of doxorubicin, ferrous sulfate, and vancomycin had the effect of improving the antitumor treatment effect. At the same time, it was found that the body weight of the mice in the doxorubicin chemotherapy drug group decreased significantly, while the body weight of the doxorubicin + ferrous sulfate + vancomycin physical mixture group only decreased slightly, and the body weight of the doxorubicin / ferrous sulfate / vancomycin nanoformulation group increased slightly, indicating that the combined drug group of antitumor drugs and antibacterial drugs reduced the toxicity of chemotherapy drugs and improved cancer treatment.

[0038] Table 3 Evaluation of the antitumor performance of the combination of doxorubicin, ferrous sulfate, and vancomycin on tumors with bacterial infection

[0039]

[0040] Note: Compared with the Control group * P < 0.05, ** P < 0.01, *** P < 0.001, compared with free doxorubicin # P < 0.05, ## P < 0.01, compared with free vancomycin @ P < 0.05, @@ P < 0.01

[0041] Furthermore, the lung metastasis status was determined by H&E staining of the lungs, and software was used to calculate the lung metastasis areas of the doxorubicin chemotherapy drug group, the ferrous sulfate inorganic metal salt group, the vancomycin antibacterial drug group, the physical mixture doxorubicin + ferrous sulfate + vancomycin group, and the nanoplatform doxorubicin + ferrous sulfate + vancomycin group. The metastasis areas of each group are shown in Table 4, and the representative pictures are as shown in Figure 1 The black circles indicate the lung metastasis sites. A large area of lung metastasis occurred in the Control group. The doxorubicin chemotherapy drug group, the ferrous sulfate inorganic metal salt group, and the vancomycin antibacterial drug group significantly reduced the lung metastasis area. Almost no lung metastasis (including the physical mixture group and the nanoplatform group) occurred in the anti-tumor combination drug group, indicating that the anti-tumor combination drug group can reduce the risk of tumor metastasis and improve the tumor prognosis.

[0042] Table 4 Evaluation of the anti-metastatic effect of anti-tumor combination drugs in tumor-bearing mice with bacterial infection

[0043]

[0044] Note: Compared with the Control group * P < 0.05, ** P < 0.01, compared with free doxorubicin # P < 0.05, ## P < 0.01, compared with free antibacterial drug @P < 0.05, @@P < 0.01

[0045] The number of bacteria in the tumors was counted by spreading the tumor homogenate and then spreading it on a plate to evaluate the antibacterial effects of different groups. Figure 2 This is the quantitative calculation result chart of the number of bacteria in the tumors. Figure 3 This is the representative picture of the MSA plate spread with the homogenate. Analysis Figure 2 showed that compared with the Control group, the number of bacteria in the tumors in the ferrous sulfate inorganic metal salt group and the vancomycin antibacterial drug group was significantly reduced. Doxorubicin + ferrous sulfate + vancomycin (including the physical mixture group and the nanoplatform group) further reduced the number of bacteria in the tumors. The nanoplatform group showed a more excellent bactericidal effect, and almost no bacteria survived in the tumors in the nanoplatform group.

[0046] Example 2

[0047] Evaluation of the anti-tumor, anti-metastatic and antibacterial properties of the combination of cisplatin, indocyanine green and teicoplanin against tumors with bacterial infection

[0048] Preparation of cisplatin and teicoplanin binary composite nanosystem: Weigh 2 mg of cisplatin and 3 mg of teicoplanin accurately and dissolve them in methanol. Vortex and mix them respectively, then mix and stir. Under stirring conditions, inject 10 times the total volume of the mixed solution of purified water, continue stirring for 10 min, the stirring speed is 1000 r / min, stand still in a water bath at 30 °C for 2 h, centrifuge, dissolve the precipitate in water, and probe sonicate for 10 min to obtain cisplatin and teicoplanin binary nanoparticles.

[0049] Preparation of indocyanine green and teicoplanin binary composite nanosystem: Weigh 2 mg of indocyanine green and 5 mg of teicoplanin accurately and dissolve them in water. Vortex and mix them respectively, then mix and stir. Continue stirring for 10 min, the stirring speed is 2000 r / min, and dialyze for 24 h to obtain indocyanine green and teicoplanin binary nanoparticles.

[0050] Preparation of cisplatin, indocyanine green, teicoplanin ternary composite nanosystem: Weigh 1 mg of cisplatin, 2 mg of indocyanine green, and 3 mg of teicoplanin accurately and dissolve them in methanol. Vortex and mix them respectively. Mix and stir the prepared cisplatin, indocyanine green, and teicoplanin solutions. Under stirring conditions, inject 10 times the total volume of the mixed solution of purified water, continue stirring for 10 min, the stirring speed is 1000 r / min, stand still in a water bath at 30 °C for 2 h, centrifuge, dissolve the precipitate in water, and probe sonicate for 10 min to obtain the ternary composite nanosystem.

[0051] Take 1 mL of the prepared nanosystem and dilute it to 3 mL with water, and measure it with a particle size analyzer (Malvem Instruments, Malvern, UK). The results are shown in Table 5. It can be seen from the table that for the prepared ternary composite nanosystem, the particle size of the nanosystem reaches the nanometer level and the particle size distribution is uniform.

[0052] Table 5 Particle size characterization of the composite nanoparticles of cisplatin, indocyanine green and teicoplanin

[0053]

[0054] The high performance liquid chromatography method was adopted to determine the mass of cisplatin, indocyanine green or teicoplanin in the self-assembled nanoparticles. Standard solutions of cisplatin, indocyanine green or teicoplanin with a series of concentrations were respectively prepared, and the solvent was dimethyl sulfoxide. The peak areas of the standard solutions with respective series of concentrations were detected at the maximum absorption wavelengths of cisplatin, indocyanine green or teicoplanin, and the concentration-peak area standard curves were plotted. Take 1 mL of the self-assembled nanoparticle solution, add 5 mL of dimethyl sulfoxide, determine its peak area at the maximum absorption wavelength of cisplatin, indocyanine green or teicoplanin, substitute it into the standard curve, and calculate the mass of cisplatin, indocyanine green or teicoplanin in the self-assembled nanoparticles. Finally, the ratio (w:w) of cisplatin and teicoplanin in the self-assembled nanoparticle "cisplatin + teicoplanin" was calculated to be 1:1; the ratio (w:w) of indocyanine green and teicoplanin in the self-assembled nanoparticle "indocyanine green + teicoplanin" was 2:1; the ratio (w:w:w) of cisplatin, indocyanine green and teicoplanin in the self-assembled nanoparticle "cisplatin + indocyanine green + teicoplanin" was 1:2:1.

[0055] Next, Balb / c mice were used to evaluate the antitumor effect of the combined antitumor drugs on tumor-bearing mice with bacterial infection. After adaptive feeding, 4T1 tumor cells (2×10 6 cells) after Enterococcus infection (MOI 1) were inoculated in situ to establish a breast cancer tumor model with bacterial infection. When the tumor volume of the mice grew to 50 mm 3 , drug treatment was started, and the tumor volume was measured every day during the treatment process, and the mice were weighed. The tumor-bearing mice were randomly divided into groups of 3 each. The experimental animals were grouped as follows: control group (Control), cisplatin group (chemotherapeutic drug), indocyanine green group (photosensitizer), teicoplanin group (antibacterial drug), cisplatin + teicoplanin physical mixture group (with a dosage ratio of 1:1), indocyanine green + teicoplanin physical mixture group (with a dosage ratio of 2:1), cisplatin + indocyanine green + teicoplanin physical mixture group (with a dosage ratio of 1:2:1), cisplatin + teicoplanin nanoparticle preparation group, indocyanine green + teicoplanin nanoparticle preparation group, and cisplatin + indocyanine green + teicoplanin nanoparticle preparation group. On the 1st, 3rd, and 5th days, the drugs were injected through the tail vein. The administration doses of cisplatin, indocyanine green and teicoplanin were 3 mg / kg, 2 mg / kg and 5 mg / kg respectively. For the groups containing indocyanine green, 12 h after administration, the tumors of the mice were irradiated with an 808 nm laser at 1 W / cm 2 for 5 min. The mice were sacrificed on the 14th day, the tumor weights were weighed, and the tumor inhibition rate was calculated. The results are shown in Table 6. The combined antitumor drugs (including the physical mixture group and the nanoparticle preparation group) had a higher tumor inhibition rate for tumor-bearing mice with bacterial infection, indicating that the combination of cisplatin, indocyanine green and teicoplanin had the effect of improving the antitumor treatment effect.

[0056] Table 6 Evaluation of the Antitumor Performance of the Combination of Cisplatin, Indocyanine Green, and Teicoplanin against Tumors with Bacterial Infection

[0057]

[0058] Note: Compared with the Control group * P < 0.05, ** P < 0.01, *** P < 0.01, compared with cisplatin # P < 0.05, ## P < 0.01

[0059] The lung metastasis status was further determined by H&E staining of the lungs, and the lung metastasis area of each group was calculated using software. The metastasis area of each group is shown in Table 7. A large area of lung metastasis occurred in the Control group, and the lung metastasis in the combined antitumor drug group was significantly reduced (including the physical mixture group and the nanoparticle formulation group), indicating that the combined antitumor drug group can reduce the risk of tumor metastasis and improve the tumor prognosis.

[0060] Table 7 Evaluation of the Antimetastatic Effect of the Combination of Cisplatin, Indocyanine Green, and Teicoplanin in Tumor-Bearing Mice with Bacterial Infection

[0061]

[0062] Note: Compared with the Control group * P < 0.05, ** P < 0.01, *** P < 0.01, compared with cisplatin # P < 0.05, ## P < 0.01, compared with indocyanine green @ P < 0.05, @@ P < 0.01, @@@ P < 0.001.

[0063] The number of bacteria in the tumor was counted by smearing the tumor homogenate and then plating, so as to evaluate the antibacterial effect of different groups. Table 8 shows the quantitative calculation results of the number of bacteria in the tumor. The number of bacteria in the tumor in the photosensitizer indocyanine green group and the antibacterial drug teicoplanin group was significantly reduced. The combined cisplatin + indocyanine green + teicoplanin group (including the physical mixture group and the nanoparticle formulation group) further reduced the number of bacteria in the tumor, and the nanoparticle formulation group showed a more excellent bactericidal effect.

[0064] Table 8 Evaluation of the Antibacterial Effect of the Combination of Cisplatin, Indocyanine Green, and Teicoplanin

[0065]

[0066]

[0067] Note: Compared with the Control group * P < 0.05, ** P < 0.01, *** P < 0.01, compared with teicoplanin @ P < 0.05, @@ P < 0.01, @@@ P < 0.01 Example 3

[0068] Evaluation of the anti - tumor, anti - metastasis and antibacterial properties of the combination of daunorubicin, chlorin e6, manganese chloride and benzylpenicillin against tumors with bacterial infections

[0069] Preparation of the daunorubicin and benzylpenicillin binary composite nanosystem: Weigh accurately 2 mg of daunorubicin and 3 mg of benzylpenicillin and dissolve them in purified water. Vortex and mix them separately, then mix and stir. Under stirring conditions, inject 2 times the total volume of the mixed solution of 10% ethanol - water. Continue to stir for 10 min at a stirring speed of 1000 r / min, centrifuge, redissolve the precipitate in water, and probe - sonicate for 10 min to obtain the daunorubicin and benzylpenicillin binary nanoparticles.

[0070] Preparation of the chlorin e6 and benzylpenicillin binary composite nanosystem: Weigh accurately 1 mg of chlorin e6 and 2 mg of benzylpenicillin and dissolve them in water. Vortex and mix them separately, then mix and stir. Continue to stir for 10 min at a stirring speed of 2000 r / min, and dialyze for 24 h to obtain the chlorin e6 and benzylpenicillin binary nanoparticles.

[0071] Preparation of the manganese chloride and benzylpenicillin binary composite nanosystem: Weigh accurately 3 mg of manganese chloride and 2 mg of benzylpenicillin and dissolve them in water. Vortex and mix them separately, then mix and stir. Continue to stir for 10 min at a stirring speed of 2000 r / min, and dialyze for 24 h to obtain the manganese chloride and benzylpenicillin binary nanoparticles.

[0072] Preparation of the daunorubicin, chlorin e6, manganese chloride and benzylpenicillin multi - component composite nanosystem: Weigh accurately 1 mg of daunorubicin, 2 mg of chlorin e6, 3 mg of manganese chloride and 2 mg of benzylpenicillin and dissolve them in purified water, ethanol, purified water and purified water respectively. Under stirring conditions, inject 4 times the total volume of the mixed solution of 20% ethanol - water. Continue to stir for 20 min at a stirring speed of 1200 r / min, centrifuge, redissolve the precipitate in water, and probe - sonicate for 10 min to obtain the multi - component composite nanosystem.

[0073] Take 1 mL of the prepared nanosystem and dilute it to 3 mL with water, and measure it with a particle size analyzer (Malvem Instruments, Malvern, UK). The results are shown in Table 9. It can be seen from the table that for the prepared multi - component composite nanosystem, the particle size of the nanosystem reaches the nanoscale and the particle size distribution is uniform.

[0074] Table 9 Particle size characterization of the composition nanoparticles prepared from daunorubicin, chlorin e6, manganese chloride and benzylpenicillin

[0075]

[0076] The high performance liquid chromatography method was used to determine the mass of daunorubicin, chlorin e6 or benzylpenicillin in the self-assembled nanoparticles. Standard solutions of daunorubicin, chlorin e6 or benzylpenicillin with a series of concentrations were respectively prepared, and the solvent was dimethyl sulfoxide. The peak areas of the standard solutions with respective series of concentrations were detected at the maximum absorption wavelengths of daunorubicin, chlorin e6 or benzylpenicillin, and the concentration-peak area standard curves were plotted. Take 1 mL of the self-assembled nanoparticle solution, add 8 mL of dimethyl sulfoxide, measure its peak area at the maximum absorption wavelength of daunorubicin, chlorin e6 or benzylpenicillin, substitute it into the standard curve, and calculate the mass of daunorubicin, chlorin e6 or benzylpenicillin in the self-assembled nanoparticles. The mass of manganese chloride in the self-assembled nanoparticles was determined by inductively coupled plasma mass spectrometry. Finally, the ratio (w:w) of daunorubicin to benzylpenicillin in the self-assembled nanoparticles "daunorubicin + benzylpenicillin" was calculated to be 2:1; the ratio (w:w) of manganese chloride to benzylpenicillin in the self-assembled nanoparticles "manganese chloride + benzylpenicillin" was 1:1; the ratio (w:w:w:w) of daunorubicin, chlorin e6, manganese chloride and benzylpenicillin in the self-assembled nanoparticles "daunorubicin + chlorin e6 + manganese chloride + benzylpenicillin" was 2:2:1:1.

[0077] Next, Balb / c mice were used to evaluate the antitumor effect of the combined antitumor drugs on tumor-bearing mice with bacterial infection. After adaptive feeding, 4T1 tumor cells (2×10 6 cells) after streptococcus infection (MOI 1) were inoculated in situ to establish a breast cancer tumor model with bacterial infection. When the tumor volume of the mice grew to 50 mm 3When it was time, drug treatment was initiated. During the treatment process, the tumor volume was measured daily, and the mice were weighed. The tumor-bearing mice were randomly divided into groups of 3 each. The experimental animals were grouped as follows: Control group, daunorubicin chemotherapy drug group, chlorin e6 photosensitizer group, benzylpenicillin antibacterial drug group, physical mixture of daunorubicin + benzylpenicillin group (with a dosage ratio of 2:1), physical mixture of chlorin e6 + benzylpenicillin group (with a dosage ratio of 2:1), physical mixture of manganese chloride + benzylpenicillin group (with a dosage ratio of 1:1), physical mixture of daunorubicin + chlorin e6 + manganese chloride + benzylpenicillin group (with a dosage ratio of 2:2:1:1), nanocomposite of daunorubicin + benzylpenicillin group, nanocomposite of chlorin e6 + benzylpenicillin group, nanocomposite of manganese chloride + benzylpenicillin group, and nanocomposite of daunorubicin + chlorin e6 + manganese chloride + benzylpenicillin group. On days 1, 3, and 5, the drugs were injected via the tail vein. The administration doses of daunorubicin, chlorin e6, manganese chloride, and benzylpenicillin were 5 mg / kg, 3 mg / kg, 5 mg / kg, and 10 mg / kg, respectively. For the groups containing chlorin e6, 8 h after drug administration, the tumors of the mice were irradiated with an 808 nm laser at 0.8 W / cm 2 for 5 min. On day 14, the mice were sacrificed, the tumor weights were measured, and the tumor inhibition rate was calculated. The results are shown in Table 10. The antitumor combination drugs (including the physical mixture groups and the nanocomposite groups) had a higher tumor inhibition rate for the tumor-bearing mice with bacterial infection, indicating that the combination of daunorubicin, chlorin e6, manganese chloride, and benzylpenicillin had the effect of improving the antitumor treatment effect.

[0078] Table 10 Evaluation of the antitumor performance of the combination of daunorubicin, chlorin e6, manganese chloride, and benzylpenicillin against tumors with bacterial infection

[0079]

[0080]

[0081] Note: Compared with the Control group * P < 0.05, ** P < 0.01, *** P < 0.001, compared with daunorubicin # P < 0.05, ## P < 0.01, ### P < 0.001, compared with manganese chloride @ P < 0.05, @@ P < 0.01, @@@ P < 0.01.

[0082] Furthermore, the lung metastasis status was determined by H&E staining of the lungs, and the lung metastasis area of each group was calculated using software. The metastasis areas of each group are shown in Table 11 below. A large area of lung metastasis was observed in the Control group, while the lung metastasis was significantly reduced in the anti-tumor combination drug group (including the physical mixture group and the nano-formulation group), indicating that the anti-tumor combination drug group can reduce the risk of tumor metastasis and improve the tumor prognosis.

[0083] Table 11 Evaluation of the anti-metastatic effect of anti-tumor combination drugs in tumor-bearing mice with bacterial infection

[0084]

[0085] Note: Compared with the Control group * P < 0.05, ** P < 0.01, *** P < 0.001, compared with daunorubicin # P < 0.05, ## P < 0.01, compared with manganese chloride @ P < 0.05, @@ P < 0.01, @@@ P < 0.01, compared with benzylpenicillin $ P < 0.05, $$ P < 0.01, $$$ P < 0.001

[0086] The number of bacteria in the tumor was counted by spreading the tumor homogenate and then plating, so as to evaluate the antibacterial effect of different groups. Table 12 shows the quantitative calculation results of the number of bacteria in the tumor. The anti-tumor combination drug group (including the physical mixture group and the nano-formulation group) significantly reduced the number of bacteria in the tumor, and the nano-formulation group showed a more excellent bactericidal effect.

[0087] Table 12 Evaluation of the antibacterial effect of the combination of daunorubicin, chlorin e6, manganese chloride and benzylpenicillin

[0088]

[0089] Note: Compared with the Control group * P < 0.05, ** P < 0.01, *** P < 0.001, compared with manganese chloride @ P < 0.05, @@ P < 0.01, @@@ P < 0.01, compared with benzylpenicillin $ P < 0.05, $$ P < 0.01, $$$ P < 0.001

[0090] Example 4

[0091] Evaluation of the anti - tumor, anti - metastasis and antibacterial properties of the combined use of idarubicin, neo - indocyanine green, ferric chloride and cefaclor against tumors with bacterial infections

[0092] Preparation of the idarubicin and cefaclor binary composite nanosystem: Weigh 2 mg of idarubicin and 4 mg of cefaclor accurately and dissolve them in purified water. Vortex and mix them separately, then mix and stir. Inject methanol with a volume twice that of the total volume of the mixed solution under stirring, continue to stir for 10 min at a stirring speed of 2000 r / min, centrifuge, redissolve the precipitate in water, and probe - sonicate for 5 min to obtain the idarubicin and cefaclor binary nanoparticles.

[0093] Preparation of the neo - indocyanine green and cefaclor binary composite nanosystem: Weigh 3 mg of neo - indocyanine green and 6 mg of cefaclor accurately and dissolve them in water. Vortex and mix them separately, then mix and stir, continue to stir for 8 min at a stirring speed of 1000 r / min, and dialyze for 24 h to obtain the neo - indocyanine green and cefaclor binary nanoparticles.

[0094] Preparation of the ferric chloride and cefaclor binary composite nanosystem: Weigh 1 mg of ferric chloride and 3 mg of cefaclor accurately and dissolve them in water. Vortex and mix them separately, then mix and stir, continue to stir for 6 min at a stirring speed of 1500 r / min, and dialyze for 12 h to obtain the ferric chloride and cefaclor binary nanoparticles.

[0095] Preparation of the idarubicin, neo - indocyanine green, ferric chloride and cefaclor multi - component composite nanosystem: Weigh 1 mg of idarubicin, 3 mg of neo - indocyanine green, 2 mg of ferric chloride and 4 mg of cefaclor accurately and dissolve them in purified water respectively, mix them evenly. Mix the prepared idarubicin, neo - indocyanine green, ferric chloride and cefaclor, add methanol - water with a volume three times that of the total volume, continue to stir for 6 min at a stirring speed of 1500 r / min, stand in a water bath at 40 °C for 5 h, centrifuge, redissolve the precipitate in water, and probe - sonicate for 10 min to obtain the multi - component composite nanosystem.

[0096] Take 1 mL of the prepared nanosystem and dilute it to 3 mL with water, and measure it with a particle size analyzer (Malvem Instruments, Malvern, UK). The results are shown in Table 13. As can be seen from the table, for the prepared multi - component composite nanosystem, the particle size of the nanosystem reaches the nanoscale and the particle size distribution is uniform.

[0097] Table 13 Particle size characterization of the composite nanoparticles of the composition prepared from idarubicin, neo - indocyanine green, ferric chloride and cefaclor

[0098]

[0099] Using high performance liquid chromatography, the mass of idarubicin, neoindocyanine green or cefaclor in the self-assembled nanoparticles was determined. Standard solutions of idarubicin, neoindocyanine green or cefaclor with a series of concentrations were prepared respectively, and the solvent was dimethyl sulfoxide. The peak areas of the standard solutions with respective series of concentrations were detected at the maximum absorption wavelengths of idarubicin, neoindocyanine green or cefaclor, and a concentration-peak area standard curve was plotted. Take 1 mL of the self-assembled nanoparticle solution, add 6 mL of dimethyl sulfoxide, measure its peak area at the maximum absorption wavelength of idarubicin, neoindocyanine green or cefaclor, substitute it into the standard curve, and calculate the mass of idarubicin, neoindocyanine green or cefaclor in the self-assembled nanoparticles. The mass of ferric chloride in the self-assembled nanoparticles was determined by inductively coupled plasma mass spectrometry. Finally, the ratio (w:w) of idarubicin and cefaclor in the self-assembled nanoparticle "idarubicin + cefaclor" was calculated to be 2:1; the ratio (w:w) of neoindocyanine green and cefaclor in the self-assembled nanoparticle "neoindocyanine green + cefaclor" was 2:1; the ratio (w:w) of ferric chloride and cefaclor in the self-assembled nanoparticle "ferric chloride + cefaclor" was 3:1; the ratio (w:w:w:w) of idarubicin, neoindocyanine green, ferric chloride and cefaclor in the self-assembled nanoparticle "idarubicin + neoindocyanine green + ferric chloride + cefaclor" was 2:2:3:1.

[0100] Next, Balb / c mice were used to evaluate the antitumor effect of the combined antitumor drugs on tumor-bearing mice with bacterial infection. After adaptive feeding, 4T1 tumor cells (2×10 6 cells) infected with Fusobacterium nucleatum (MOI 2) were inoculated in situ to establish a breast cancer tumor model with bacterial infection. When the tumor volume of the mice grew to 50 mm 3 , drug treatment was started, and the tumor volume was measured every day during the treatment process, and the mice were weighed. The tumor-bearing mice were randomly divided into groups of 3 each. The experimental animals were grouped as follows: control group (Control), chemotherapy drug idarubicin group, photosensitizer neoindocyanine green group, antibacterial drug cefaclor group, physical mixture idarubicin + cefaclor group (the dosage ratio was 2:1), physical mixture neoindocyanine green + cefaclor group (the dosage ratio was 2:1), physical mixture ferric chloride + cefaclor group (the dosage ratio was 3:1), physical mixture idarubicin + neoindocyanine green + ferric chloride + cefaclor group (the dosage ratio was 2:2:3:1), nanoformulation idarubicin + cefaclor group, nanoformulation neoindocyanine green + cefaclor group, nanoformulation ferric chloride + cefaclor group, nanoformulation idarubicin + neoindocyanine green + ferric chloride + cefaclor group. On days 1, 3, and 5, the drugs were injected via the tail vein. The dosing doses of idarubicin, neoindocyanine green, ferric chloride and cefaclor were 4 mg / kg, 3 mg / kg, 5 mg / kg and 15 mg / kg respectively. For the groups containing neoindocyanine green, 8 h after dosing, an 808 nm laser was used at 1 W / cm2 The tumor of the mouse was irradiated with laser for 5 min. The mouse was sacrificed on the 14th day, the tumor weight was measured, and the tumor inhibition rate was calculated. The results are shown in Table 14. The anti-tumor combination drugs (including the physical mixture group and the nano-formulation group) had a higher tumor inhibition rate for tumor-bearing mice with bacterial infection.

[0101] Table 14 Evaluation of the anti-tumor performance of the combination of idarubicin, neo-indocyanine green, iron chloride, and cefaclor against tumors with bacterial infection

[0102]

[0103] Note: Compared with the Control group * P < 0.05, ** P < 0.01, *** P < 0.001, compared with idarubicin # P < 0.05, ## P < 0.01, ### P < 0.001, compared with iron chloride @ P < 0.05, @@ P < 0.01, @@@ P < 0.01, compared with neo-indocyanine green $ P < 0.05, $$ P < 0.01, $$$ P < 0.001

[0104] Furthermore, the lung metastasis status was determined by H&E staining of the lung, and the software was used to calculate the lung metastasis areas of the idarubicin treatment group, the photosensitizer neo-indocyanine green group, the antibacterial drug cefaclor group, the physical mixture idarubicin + cefaclor group, the physical mixture neo-indocyanine green + cefaclor group, the physical mixture iron chloride + cefaclor group, the physical mixture idarubicin + neo-indocyanine green + iron chloride + cefaclor group, the nano-formulation idarubicin + cefaclor group, the nano-formulation neo-indocyanine green + cefaclor group, the nano-formulation iron chloride + cefaclor group, and the nano-formulation idarubicin + neo-indocyanine green + iron chloride + cefaclor group. The metastasis areas of each group are shown in Table 15. A large area of lung metastasis occurred in the Control group, and the lung metastasis decreased in the anti-tumor combination drug group (including the physical mixture group and the nano-formulation group), indicating that the anti-tumor combination drug group could reduce the risk of tumor metastasis and improve the tumor prognosis.

[0105] Table 15 Evaluation of the anti-metastatic effect of the anti-tumor combination drugs in tumor-bearing mice with bacterial infection

[0106]

[0107] Note: Compared with the Control group * P < 0.05,** P < 0.01, *** P < 0.001, compared with idarubicin # P < 0.05, ## P < 0.01, compared with ferric chloride @ P < 0.05, @@ P < 0.01, @@@ P < 0.01, compared with neo - indocyanine green $ P < 0.05, $$ P < 0.01, $$$ P < 0.001. By smearing the tumor homogenate and then plating, the number of bacteria in the tumor was counted to evaluate the antibacterial effects of different groups. Table 16 shows the quantitative calculation results of the number of bacteria in the tumor. The anti - tumor combined drug groups (including the physical mixture group and the nanoparticle formulation group) significantly reduced the number of bacteria in the tumor, and the nanoparticle formulation group showed more excellent bactericidal effects.

[0108] Table 16 Evaluation of antibacterial effects of the combination of idarubicin, neo - indocyanine green, ferric chloride and cefaclor

[0109]

[0110]

[0111] Note: Compared with the Control group * P < 0.05, ** P < 0.01, *** P < 0.001, compared with ferric chloride @ P < 0.05, @@ P < 0.01, @@@ P < 0.01, compared with cefaclor $ P < 0.05, $$ P < 0.01.

[0112] Example 5

[0113] Evaluation of anti - tumor, anti - metastasis and antibacterial properties of the combination of oxaliplatin, temoporfin, copper sulfate and metronidazole against bacteria - infected tumors

[0114] Preparation of the binary composite nanosystem of oxaliplatin and metronidazole: Weigh 2 mg of oxaliplatin and 6 mg of metronidazole accurately and dissolve them in purified water. Vortex and mix them separately, then mix and stir. Inject 5 times the total volume of the mixed solution of methanol under stirring conditions, continue to stir for 10 min at a stirring speed of 2000 r / min, centrifuge, redissolve the precipitate in water, and probe - sonicate for 5 min to obtain the binary nanoparticles of oxaliplatin and metronidazole.

[0115] Preparation of temoporfin and metronidazole binary composite nanosystem: Weigh 3 mg of temoporfin and 4 mg of metronidazole precisely, dissolve them in water, vortex and mix them respectively, then mix and stir, continue to stir for 30 min at a stirring speed of 1000 r / min, and dialyze for 8 h to obtain temoporfin and metronidazole binary nanoparticles.

[0116] Preparation of copper sulfate and metronidazole binary composite nanosystem: Weigh 4 mg of copper sulfate and 2 mg of metronidazole precisely, dissolve them in water, vortex and mix them respectively, then mix and stir, continue to stir for 20 min at a stirring speed of 20000 r / min, and dialyze for 12 h to obtain copper sulfate and metronidazole binary nanoparticles.

[0117] Preparation of oxaliplatin, temoporfin, copper sulfate and metronidazole multi - component composite nanosystem: Weigh 1 mg of oxaliplatin, 5 mg of temoporfin, 4 mg of copper sulfate and 3 mg of metronidazole precisely, dissolve them in purified water respectively, mix them evenly, mix the prepared oxaliplatin, temoporfin, copper sulfate and metronidazole, continue to stir for 15 min at a stirring speed of 1000 r / min, centrifuge, dissolve the precipitate by adding water, and probe - sonicate for 10 min to obtain the multi - component composite nanosystem.

[0118] Take 1 mL of the prepared nanosystem, dilute it to 3 mL with water, and measure it with a particle size analyzer (Malvem Instruments, Malvern, UK). The results are shown in Table 17. As can be seen from the table, for the prepared multi - component composite nanosystem, the particle size of the nanosystem reaches the nanoscale and the particle size distribution is uniform.

[0119] Table 17 Particle size characterization of the nanoparticles of the composition prepared from oxaliplatin, temoporfin, copper sulfate and metronidazole

[0120]

[0121]

[0122] The high performance liquid chromatography was used to determine the mass of oxaliplatin, temoporfin or metronidazole in the self-assembled nanoparticles. Standard solutions of oxaliplatin, temoporfin or metronidazole with a series of concentrations were respectively prepared, and the solvent was dimethyl sulfoxide. The peak areas of the standard solutions with respective series of concentrations were detected at the maximum absorption wavelengths of oxaliplatin, temoporfin or metronidazole, and the concentration-peak area standard curves were plotted. Take 1 mL of the self-assembled nanoparticle solution, add 10 mL of dimethyl sulfoxide, determine its peak area at the maximum absorption wavelength of oxaliplatin, temoporfin or metronidazole, substitute it into the standard curve, and calculate the mass of oxaliplatin, temoporfin or metronidazole in the self-assembled nanoparticles. The mass of copper sulfate in the self-assembled nanoparticles was determined by inductively coupled plasma mass spectrometry. Finally, the ratio (w:w) of oxaliplatin to metronidazole in the self-assembled nanoparticle "oxaliplatin + metronidazole" was calculated to be 1:2; the ratio (w:w) of temoporfin to metronidazole in the self-assembled nanoparticle "temoporfin + metronidazole" was 1:4; the ratio (w:w) of copper sulfate to metronidazole in the self-assembled nanoparticle "copper sulfate + metronidazole" was 1:2; the ratio (w:w:w:w) of oxaliplatin, temoporfin, copper sulfate and metronidazole in the self-assembled nanoparticle "oxaliplatin + temoporfin + copper sulfate + metronidazole" was 2:1:2:4.

[0123] Next, Balb / c mice were used to evaluate the antitumor effect of the combined anti-tumor drugs on tumor-bearing mice with bacterial infection. After adaptive feeding, 4T1 tumor cells (2×10 6 cells) infected with Fusobacterium nucleatum (MOI 5) were inoculated in situ to establish a breast cancer tumor model with bacterial infection. When the tumor volume of the mice grew to 50 mm 3 , drug treatment was started, and the tumor volume was measured every day during the treatment process, and the mice were weighed. The tumor-bearing mice were randomly divided into groups of 3 each. The experimental animals were grouped as follows: control group (Control), oxaliplatin group (chemotherapeutic drug), temoporfin group (photosensitizer), metronidazole group (antibacterial drug), physical mixture oxaliplatin + metronidazole group (the dosage ratio is 1:2), physical mixture temoporfin + metronidazole group (the dosage ratio is 1:4), physical mixture copper sulfate + metronidazole group (the dosage ratio is 1:2), physical oxaliplatin + temoporfin + copper sulfate + metronidazole group (the dosage ratio is 2:1:2:4), nanoformulation oxaliplatin + metronidazole group, nanoformulation temoporfin + metronidazole group, nanoformulation copper sulfate + metronidazole group, nanoformulation oxaliplatin + temoporfin + copper sulfate + metronidazole group. On the 1st, 3rd and 5th days, the drugs were injected through the tail vein. The administration doses of oxaliplatin, temoporfin, copper sulfate and metronidazole were 10 mg / kg, 2 mg / kg, 5 mg / kg and 5 mg / kg respectively. For the groups containing temoporfin, 8 h after administration, an 808 nm laser was used at 1.5 W / cm 2The tumor of the mouse was irradiated with laser for 5 min. The mouse was sacrificed on the 14th day, the tumor weight was measured, and the tumor inhibition rate was calculated. The results are shown in Table 18. The anti-tumor combination drugs (including the physical mixture group and the nano-formulation group) had a higher tumor inhibition rate for tumor-bearing mice with bacterial infection.

[0124] Table 18 Evaluation of the anti-tumor performance of the combination of oxaliplatin, temoporfin, copper sulfate and metronidazole against tumors with bacterial infection

[0125]

[0126]

[0127] Note: Compared with the Control group * P < 0.05, ** P < 0.01, *** P < 0.001, compared with oxaliplatin # P < 0.05, ## P < 0.01, ### P < 0.001, compared with copper sulfate @ P < 0.05, @@ P < 0.01, @@@ P < 0.01.

[0128] Furthermore, the lung metastasis status was judged by H&E staining of the lung, and the lung metastasis area of different groups was calculated using software. The metastasis area of each group is shown in Table 19. A large area of lung metastasis occurred in the Control group, and the lung metastasis in the anti-tumor combination drug group decreased (including the physical mixture group and the nano-formulation group), indicating that the anti-tumor combination drug group could reduce the risk of tumor metastasis and improve the tumor prognosis.

[0129] Table 19 Evaluation of the anti-metastatic effect of the combination of oxaliplatin, temoporfin, copper sulfate and metronidazole in tumor-bearing mice with bacterial infection

[0130]

[0131]

[0132] Note: Compared with the Control group * P < 0.05, ** P < 0.01, *** P < 0.001, compared with oxaliplatin # P < 0.05, ## P < 0.01, ### P < 0.001, compared with copper sulfate @ P < 0.05, @@ P < 0.01, @@@P < 0.01

[0133] The number of bacteria in tumors was counted by coating tumor homogenate and then plating, so as to evaluate the antibacterial effects of different groups. Table 20 below shows the quantitative calculation results of the number of bacteria in tumors. The anti-tumor combined drug group (including the physical mixture group and the nano-formulation group) significantly reduced the number of bacteria in tumors, and the nano-formulation group showed more excellent bactericidal effects.

[0134] Table 20 Evaluation of the antibacterial effects of the combination of oxaliplatin, temoporfin, copper sulfate and metronidazole

[0135]

[0136] Note: Compared with the Control group * P < 0.05, ** P < 0.01, *** P < 0.001, compared with copper sulfate @ P < 0.05, @@ P < 0.01, @@@ P < 0.01 compared with metronidazole $ P < 0.05, $$ P < 0.01, $$$ P < 0.001

[0137] Example 6

[0138] Evaluation of the anti-tumor, anti-metastatic and antibacterial properties of the combination of mitoxantrone, magnesium chloride and sulfamethoxazole against bacteria-infected tumors

[0139] Preparation of the mitoxantrone + sulfamethoxazole binary composite nano-system: Weigh 3 mg of mitoxantrone and 4 mg of sulfamethoxazole accurately and dissolve them in water. Vortex and mix them separately, then mix and stir. Continue stirring for 30 min at a stirring speed of 1500 r / min and dialyze for 6 h to prepare the mitoxantrone and sulfamethoxazole binary nanoparticles.

[0140] Preparation of the magnesium chloride + sulfamethoxazole binary composite nano-system: Weigh 2 mg of magnesium chloride and 1 mg of sulfamethoxazole accurately and dissolve them in purified water. Inject 5 times the volume of water of the mixed solution under stirring conditions and continue stirring for 1 h at a stirring speed of 2000 r / min. Centrifuge, redissolve the precipitate in water, and probe sonicate for 10 min to prepare the magnesium chloride and sulfamethoxazole binary nanoparticles.

[0141] Preparation of mitoxantrone + magnesium chloride + sulfamethoxazole multi-component composite nanosystem: Accurately weigh 2 mg of mitoxantrone, 3 mg of magnesium chloride and 1 mg of sulfamethoxazole, dissolve them in purified water, then mix and stir. Under stirring conditions, inject 4 times the total volume of the mixed solution with water, continue to stir for 1 h at a stirring speed of 2000 r / min, centrifuge, redissolve the precipitate with water, and probe sonicate for 10 min to obtain the multi-component composite nanosystem.

[0142] Take 1 mL of the prepared nanosystem and dilute it to 3 mL with water, and measure it with a particle size analyzer (Malvem Instruments, Malvern, UK). The results are shown in Table 21. As can be seen from the table, for the prepared multi-component composite nanosystem, the particle size of the nanosystem reaches the nanometer level and the particle size distribution is uniform.

[0143] Table 21 Particle size characterization of the nanoparticles of the composition prepared from mitoxantrone, magnesium chloride and sulfamethoxazole

[0144]

[0145] The high performance liquid chromatography method was used to determine the mass of mitoxantrone or sulfamethoxazole in the self-assembled nanoparticles. Standard solutions of a series of concentrations of mitoxantrone or sulfamethoxazole were respectively prepared, and the solvent was dimethyl sulfoxide. The peak areas of the respective series of concentration standard solutions were detected at the maximum absorption wavelengths of mitoxantrone or sulfamethoxazole, and a concentration-peak area standard curve was plotted. Take 1 mL of the self-assembled nanoparticle solution, add 4 mL of dimethyl sulfoxide, measure its peak area at the maximum absorption wavelength of mitoxantrone or sulfamethoxazole, substitute it into the standard curve, and calculate the mass of mitoxantrone or sulfamethoxazole in the self-assembled nanoparticles. The mass of magnesium chloride in the self-assembled nanoparticles was determined by inductively coupled plasma mass spectrometry. Finally, the ratio (w:w) of mitoxantrone and sulfamethoxazole in the self-assembled nanoparticle "mitoxantrone + sulfamethoxazole" was calculated to be 3:2; the ratio (w:w) of magnesium chloride and sulfamethoxazole in the self-assembled nanoparticle "magnesium chloride + sulfamethoxazole" was 1:2; the ratio (w:w:w) of mitoxantrone, magnesium chloride and sulfamethoxazole in the self-assembled nanoparticle "mitoxantrone + magnesium chloride + sulfamethoxazole" was 3:1:2.

[0146] Next, Balb / c mice were used to evaluate the antitumor effect of the combined drug on tumor-bearing mice infected with bacteria. After the mice were adaptively fed, 4T1 tumor cells (2×10 6 cells) infected with Staphylococcus xylosus (MOI 2) were inoculated in situ to establish a breast cancer tumor model with bacterial infection. When the tumor volume of the mice grew to 50 mm 3When it was time, drug treatment was initiated. During the treatment process, the tumor volume was measured daily, and the weight of the mice was weighed. The tumor-bearing mice were randomly divided into groups of 3 each. The experimental animals were grouped as follows: Control group, mitoxantrone (a chemotherapy drug) group, magnesium chloride (an inorganic metal salt) group, sulfamethoxazole (an antibacterial drug) group, physical mixture of mitoxantrone + sulfamethoxazole group (with a dosage ratio of 3:2), physical mixture of magnesium chloride + sulfamethoxazole group (with a dosage ratio of 1:2), physical mixture of mitoxantrone + magnesium chloride + sulfamethoxazole (with a dosage ratio of 3:1:2), and nanoformulation of mitoxantrone + sulfamethoxazole group, nanoformulation of magnesium chloride + sulfamethoxazole group, nanoformulation of mitoxantrone + magnesium chloride + sulfamethoxazole group. On the 1st, 3rd, and 5th days, the drugs were injected via the tail vein. The administration doses of mitoxantrone, magnesium chloride, and sulfamethoxazole were 5 mg / kg, 5 mg / kg, and 5 mg / kg, respectively. On the 14th day, the mice were sacrificed, the tumor weight was weighed, and the tumor inhibition rate was calculated. The results are shown in Table 22. The anti-tumor combination drugs (including the physical mixture group and the nanoformulation group) had a higher tumor inhibition rate for tumor-bearing mice with bacterial infection.

[0147] Table 22 Evaluation of the anti-tumor performance of the combination of mitoxantrone, magnesium chloride, and sulfamethoxazole against tumors with bacterial infection

[0148]

[0149] Note: Compared with the Control group * P < 0.05, ** P < 0.01, *** P < 0.001, compared with free mitoxantrone # P < 0.05, ## P < 0.01, ### P < 0.01, compared with magnesium chloride @ P < 0.05, @@ P < 0.01

[0150] Furthermore, the lung metastasis status was judged by H&E staining of the lung, and the lung metastasis area of each group was calculated using software. The metastasis areas of each group are shown in Table 23 below, and the representative pictures are as Figure 1 shown. The black circles indicate the lung metastasis sites. A large area of lung metastasis occurred in the Control group. The doxorubicin (a chemotherapy drug) group, ferrous sulfate (an inorganic metal salt) group, and vancomycin (an antibacterial drug) group significantly reduced the lung metastasis area. Almost no lung metastasis (including the physical mixture group and the nanoformulation group) occurred in the anti-tumor combination drug group, indicating that the anti-tumor combination drug group can reduce the risk of tumor metastasis and improve the tumor prognosis.

[0151] Table 23 Evaluation of the anti-metastatic effect of anti-tumor combination drugs in tumor-bearing mice with bacterial infection

[0152]

[0153]

[0154] Note: Compared with the Control group * P < 0.05, ** P < 0.01, *** P < 0.001, compared with free mitoxantrone # P < 0.05, ## P < 0.01, ### P < 0.01, compared with magnesium chloride @ P < 0.05, @@ P < 0.01

[0155] The number of bacteria in tumors was counted by coating tumor homogenate and then plating, so as to evaluate the antibacterial effects of different groups. Table 24 below shows the quantitative calculation results of the number of bacteria in tumors. The anti-tumor combined drug groups (including the physical mixture group and the nano-formulation group) significantly reduced the number of bacteria in tumors, and the nano-formulation group showed more excellent bactericidal effects.

[0156] Table 24 Evaluation of the antibacterial effects of the combination of mitoxantrone, magnesium chloride and sulfamethoxazole

[0157]

[0158] Note: Compared with the Control group * P < 0.05, ** P < 0.01, *** P < 0.001, compared with magnesium chloride @ P < 0.05, @@ P < 0.01, @@@ P < 0.01 compared with sulfamethoxazole $ P < 0.05, $$ P < 0.01.

Claims

1. Use of the combination of an antitumor active substance and an antibacterial drug in the preparation of a tumor therapeutic drug, characterized in that, The anti-tumor active substances are doxorubicin and ferrous sulfate, and the antibacterial drug is vancomycin; the tumor is breast cancer with bacterial infection.

2. The application according to claim 1, wherein The dosage ratio w:w of the anti-tumor active substances to the antibacterial drug is 1 to 10:

1.

3. The application according to claim 1, wherein The active ingredient of the tumor therapeutic drug is a composition of anti-tumor active substances and antibacterial drugs, or self-assembled nanoparticles of anti-tumor active substances and antibacterial drugs.

4. A therapeutic drug for breast cancer with bacterial infection, characterized in that, It includes anti-tumor active substances and antibacterial drugs, as well as a pharmaceutically acceptable carrier; the anti-tumor active substances are doxorubicin and ferrous sulfate, and the antibacterial drug is vancomycin.

5. The therapeutic drug according to claim 4, characterized in that, The dosage ratio w:w of the anti-tumor active substances to the antibacterial drug is 1 to 10:

1.

6. The therapeutic drug according to claim 4, characterized in that, The active ingredient of the therapeutic drug is a composition of anti-tumor active substances and antibacterial drugs.

7. The therapeutic drug according to claim 4, characterized in that, The active ingredient of the therapeutic drug is self-assembled nanoparticles of anti-tumor active substances and antibacterial drugs.

Images