Anti-tumor pharmaceutical composition and preparation method thereof

Abstract

The invention discloses an anti-tumor pharmaceutical composition containing an acidophilic thiobacillus ferrooxidans realgar leaching solution. The composition is prepared from the acidophilic thiobacillus ferrooxidans Dom-BY3 realgar leaching solution and paclitaxel. Compared with single use, the drug combination has the advantages of remarkable liver cancer resistance, lower dosage demand and lower toxicity.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and particularly relates to a composition of an extract of Acidithiobacillus ferrooxidans realgar domesticated strain Dom-BY3 (RTS) and paclitaxel (Taxol) that can effectively inhibit the proliferation of liver cancer. The composition can effectively kill liver cancer cells and reduce the dosage requirement when a single drug is used. Background Art

[0002] Liver cancer is a malignant tumor that seriously threatens human health. Chemotherapy plays an important role in the treatment of liver cancer. Although the commonly used liver cancer chemotherapy drugs can inhibit tumor growth to a certain extent, they have many disadvantages. Chemotherapy drugs usually lack specificity. While killing cancer cells, they also cause serious damage to normal cells, causing patients to experience a series of toxic side effects such as nausea, vomiting, hair loss, and decreased immunity. In addition, chemotherapy drugs are prone to drug resistance, which gradually reduces the therapeutic effect. Paclitaxel, as an important chemotherapy drug, plays an important role in the treatment of various tumors. In recent years, the application of paclitaxel in the treatment of liver cancer has gradually attracted attention. Paclitaxel can induce tumor cell apoptosis and inhibit tumor angiogenesis. However, as a drug widely used in the treatment of various cancers, paclitaxel also has its limitations in the treatment of liver cancer, and the effective rate of monotherapy is relatively low. In order to improve the therapeutic effect, paclitaxel is often used in combination with other chemotherapy drugs, targeted drugs or immunotherapy drugs.

[0003] As a traditional Chinese medicine, realgar has a long history of application and rich clinical experience in my country. The main ingredient, arsenic disulfide, has clear anti-tumor activity. It can induce apoptosis of liver cancer cells and inhibit the proliferation and invasion of liver cancer cells. However, realgar contains arsenic. Although it has anti-tumor effects at a certain dose, it also has a greater toxicity risk. On the one hand, arsenic poisoning will occur; on the other hand, the therapeutic window is narrow, that is, the effective dose is close to the toxic dose. In clinical applications, it is necessary to strictly control the dose and monitor the blood arsenic concentration to avoid poisoning. Through microbial hydrometallurgy technology, the metabolic activities of microorganisms can be used to dissolve arsenic and other components in realgar to form soluble substances, thereby improving its bioavailability and enabling the drug to play a better role. Acidithiobacillus ferrooxidans, a common microbial hydrometallurgical bacterium, was used to biochemically dissolve realgar to release arsenic and other active ingredients. The prepared realgar microbial extract has significant anti-tumor efficacy and good safety.

[0004] The present invention unexpectedly found that the combination of the realgar microbial extract of Acidithiobacillus ferrooxidans Dom-BY3 and paclitaxel has a significant effect of inhibiting proliferation of the liver cancer cell line HepG2 and significantly improves safety. Summary of the invention

[0005] The present invention discloses an anti-tumor drug composition. The active ingredients of the composition include acidithiobacillus ferroxidans realgar microbial extract and paclitaxel.

[0006] Another aspect of the present invention discloses the use of the above composition in preparing a drug for treating liver cancer.

[0007] Preferably, the acidithiobacillus ferroxidans is Dom-BY3 (Acidithiobacillus ferroxidans Dom-BY3), which has been deposited in the China Center for Type Culture Collection, the deposit address is Wuhan University, Wuhan, China, the deposit date is November 25, 2024, and the deposit number is CCTCC NO: M 20242585.

[0008] Preferably, the Acidithiobacillus ferrooxidans realgar microbial extract is prepared by the following method:

[0009] S1: Prepare the culture medium of Acidithiobacillus ferrooxidans Dom-BY31K: weigh 3.00 g of (NH4)2SO4, 0.50 g of K2HPO4, 0.50 g of MgSO4·7H2O, 0.10 g of KCl, 0.01 g of Ca(NO3)2, and 4.478 g of FeSO4·7H2O, and dissolve them in 950 mL of distilled water with stirring. Pipette 1 mL of concentrated sulfuric acid to adjust the pH value to 1.8-2.0, and make up to 1000 mL for use.

[0010] S2: Take out the Dom-BY3 strain preserved in the laboratory, draw 10mL and inoculate it into the above-prepared 1K medium containing 0.5g / 100mL realgar and 0.4478g / 100mL FeSO4·7H2O, activate it on a shaker at 180rpm, 30℃ for 4d, observe the color of the bacterial culture solution under the 1K growth condition, it changes from light yellow to light reddish brown, indicating that the next leaching experiment can be carried out, continue to carry out the leaching experiment under the condition of 1K medium containing 0.5g / 100mL realgar, 180rpm, 30℃, add 2% of the total volume of 1K medium to the culture bottle every 2d, leaching for 68d, and prepare the acidophilic ferrothiobacillus Dom-BY3 realgar microbial leaching solution.

[0011] Preferably, the ratio of acidithiobacillus ferrooxidans Dom-BY3 realgar microbial extract to paclitaxel is 15-120:6.25-50, and the concentration of acidithiobacillus ferrooxidans Dom-BY3 realgar microbial extract is calculated as arsenic.

[0012] Preferably, the arsenic concentration in the Acidithiobacillus ferrooxidans Dom-BY3 realgar microbial extract is 0.15 μg / mL to 1.2 μg / mL.

[0013] Preferably, the concentration of paclitaxel is 0.0625 μg / mL to 0.5 μg / mL.

[0014] Preferably, the ratio of the Acidithiobacillus ferrooxidans Dom-BY3 realgar microbial extract to paclitaxel is 2.4:1.

[0015] Preferably, the composition is one of powder, granules, powder, pills, tablets, capsules, pills, and oral liquid.

[0016] Beneficial effects of the present invention:

[0017] (1) When the acidithiobacillus ferrooxidans Dom-BY3 realgar microbial extract is used in combination with paclitaxel, it has a significant inhibitory effect on the proliferation of liver cancer cells. The acidithiobacillus ferrooxidans Dom-BY3 realgar microbial extract can improve the therapeutic effect of paclitaxel on liver cancer. (2) When the acidithiobacillus ferrooxidans Dom-BY3 realgar microbial extract is used in combination with paclitaxel, the synergistic index (Cindex, CI) values ​​are all less than 1, and there is a significant synergistic effect. (3) When the acidithiobacillus ferrooxidans Dom-BY3 realgar microbial extract is used in combination with paclitaxel, the dosage of a single drug is significantly reduced, which is beneficial to reduce the toxic side effects caused by high doses of a single drug and improve the safety of treatment. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 The results of Taxol and RTS on the cell proliferation inhibition of HepG2 cell line

[0019] Figure 2 The results of the combination of Taxol and RTS on the cell proliferation inhibition of HepG2 liver cancer cell line

[0020] Figure 3 The results of the combination of 10-HCPT and RTS on the cell proliferation inhibition of liver cancer cell line HepG2

[0021] Figure 4 The results of the combination of DOX and RTS on the cell proliferation inhibition of HepG2 liver cancer cell line

[0022] Figure 5 DRI for the combined dose reduction of Taxol and RTS

[0023] Figure 6 The dose-effect curve of Taxol and RTS DETAILED DESCRIPTION

[0024] The following specific embodiments are provided to implement the technical solutions described in the present invention, but are not limited to these embodiments.

[0025] Example 1 Preparation of Realgar Microbial Extract (RTS) of Acidithiobacillus ferrooxidans Dom-BY3

[0026] 1. Activation of Acidithiobacillus ferrooxidans Dom-BY3:

[0027] Prepare Acidithiobacillus ferrooxidans Dom-BY31K culture medium: weigh 3.00 g of (NH4)2SO4, 0.50 g of K2HPO4, 0.50 g of MgSO4·7H2O, 0.10 g of KCl, 0.01 g of Ca(NO3)2, and 4.478 g of FeSO4·7H2O, stir and dissolve in 950 mL of distilled water, draw 1 mL of concentrated sulfuric acid to adjust the pH to 1.8-2.0, make up to 1000 mL, and set aside. The Dom-BY3 strain preserved in the laboratory was taken out, and 10 mL was pipetted with a pipette to inoculate into the above-prepared 1K medium containing 0.5 g / 100 mL of realgar and 0.4478 g / 100 mL of FeSO4·7H2O. The culture medium was activated at 180 rpm and 30°C for 4 days. The color of the bacterial culture medium under the 1K growth condition was observed, which changed from light yellow to light reddish brown, indicating that the next leaching experiment could be carried out.

[0028] 2. Preparation of Acidithiobacillus ferrooxidans Dom-BY3 Realgar Microbial Extract

[0029] The leaching experiment was continued under the conditions of 1K culture medium containing 0.5 g / 100 mL of realgar, 180 rpm, 30°C, 2% of the total volume of 1K culture medium was added to the culture bottle every 2 days, and leaching was carried out for 68 days to finally obtain the realgar microbial leaching solution of Acidithiobacillus ferrooxidans Dom-BY3.

[0030] Example 2 Determination of the proliferation of HepG2 liver cancer cell line by Taxol, RTS and the synergistic administration of Taxol and RTS (MTT experiment)

[0031] HepG2 tumor cells in the logarithmic growth phase were cultured at about 10 5The density of arsenic per well was inoculated on a 96-well plate at 100 μL per well and cultured overnight. Two separate dosing groups were set up: Taxol group and RTS group. The drugs were administered according to the preset arsenic concentration gradient, with the maximum concentration of 16 μg / mL. The experiment was repeated at least three times using a half-dilution dosing method. After 24 hours of continued culture, 5 mg / mL of MTT was added to each well and incubated at 37°C for 4 hours. The supernatant was discarded by the rapid plate flipping method, 150 μL of DMSO was added to each well, and the cells were detected on an enzyme reader. The OD values ​​were measured. 570 IC50 50 Determination of IC of Taxol and RTS 50 After the value, the drug combination regimen was standardized according to the IC 50 For quasi-gradient synergistic drug administration, add the drug according to the preset concentration gradient, and repeat at least 3 times for each gradient. After 24 hours of continuous culture, add 5 mg / mL MTT to each well, incubate at 37°C for 4 hours, quickly flip the plate to discard the supernatant, add 150 μL DMSO to each well, and detect on the microplate reader. 570 IC50 50 .

[0032] Table 1 The inhibitory effects of Taxol and RTS on the proliferation of HepG2 liver cancer cell line

[0033]

[0034] Determination of IC of Taxol and RTS 50 After the value, the drug combination regimen was standardized according to the IC 50 For quasi-gradient synergistic drug delivery, HepG2 tumor cells in the logarithmic growth phase were cultured at a rate of about 10 5 The cell suspension was plated in 96-well plates at a density of 100 μL per well and cultured overnight. 50 The concentration was set as the synergistic scheme, with RTS and Taxol concentrations of 1.2 μg / mL and 3.97 μg / mL as the maximum combined dose concentrations, and the drugs were administered at half dilution. After 24 hours of continuous culture, 5 mg / mL of MTT was added to each well, and the cells were incubated at 37°C for another 4 hours. The supernatant was discarded by the rapid plate flipping method, and 150 μL of DMSO was added to each well. The cells were tested on an ELISA instrument, and the OD values ​​were measured. 570 IC50 50 .

[0035] Table 2 MTT results of the synergistic administration of the combination of Taxol and RTS in the liver cancer cell line HepG2

[0036]

[0037] As shown in Table 2 and Figure 2 As shown, the IC values ​​of Taxol and RTS were respectively 50 The results of The inhibition rate dosing scheme is the maximum concentration, followed by gradient dilution. MTT results show that the inhibitory effect of the synergistic concentration on the proliferation of HepG2 cells is more significant than that of RTS and Taxol alone at the same concentration. When the synergistic dose of RTS and Taxol is 0.15+0.0625μg / mL, the CI value is the smallest and the synergistic effect is the best. The synergistic administration at each concentration shows a synergistic effect (<1), indicating that the combination of drugs has a more significant effect in inhibiting the proliferation of liver cancer cells HepG2, and is a potential clinical treatment scheme.

[0038] Example 3 Determination of the proliferation of HepG2 liver cancer cell line by co-administering 10-hydroxycamptothecin (10-HCPT) and doxorubicin (DOX) as controls with RTS (MTT experiment)

[0039] 1. MTT results of 10-hydroxycamptothecin (10-HCPT) and RTS co-administration

[0040] HepG2 tumor cells in the logarithmic growth phase were cultured at about 10 5 The cells were inoculated at a density of 100 μL / mL on a 96-well plate, and cultured overnight. Two separate dosing groups were set up: 10-HCPT group and RTS group. The maximum concentration was 32 μg / mL, and the half-dilution method was used to add at least 3 replicates to each gradient. After 24 hours of continuous culture, 5 mg / mL of MTT was added to each well, and the cells were incubated at 37°C for 4 hours. The supernatant was discarded by the rapid plate flipping method, and 150 μL of DMSO was added to each well. The cells were detected on an enzyme reader, and the OD values ​​were measured. 570 IC50 50 Determination of IC of Taxol and RTS 50 After the value, the drug combination regimen was standardized according to the IC 50 For quasi-gradient synergistic drug administration, at least 3 replicates were added to each gradient according to the preset concentration gradient. After 24 hours of continuous culture, 5 mg / mL of MTT was added to each well and incubated at 37°C for 4 hours. The supernatant was discarded by the rapid plate flipping method, and 150 μL of DMSO was added to each well. The cells were detected on an ELISA reader. The OD values ​​were measured. 570 IC50 50 .

[0041] Table 3 The inhibitory effects of 10-HCPT and RTS on the proliferation of HepG2 cell line

[0042]

[0043] Determination of IC of 10-HCPT and RTS 50 After the value, the drug combination regimen was standardized according to the IC 50 For quasi-gradient synergistic drug delivery, HepG2 tumor cells in the logarithmic growth phase were cultured at a rate of about 10 5 The cells were inoculated at a density of 100 μL / mL in a 96-well plate, and cultured overnight. At least 3 replicates were added to each gradient according to the preset concentration gradient. After 24 hours of continuous culture, 5 mg / mL of MTT was added to each well, and the cells were incubated at 37°C for 4 hours. The supernatant was discarded by the rapid plate flipping method, and 150 μL of DMSO was added to each well. The cells were detected on an ELISA reader, and the OD values ​​were measured. 570 IC50 50 .

[0044] Table 4 The inhibitory effect of the combination of 10-HCPT and RTS on the proliferation of HepG2 liver cancer cell line

[0045]

[0046] IC50 inhibition rate of cancer cells 50 The concentration is the maximum concentration combination of RTS and 10-HCPT, and the drug is administered in gradient dilution. The synergistic inhibition rate is obtained according to the MTT experiment, such as Figure 2 As shown in Table 4, the maximum synergistic inhibition rate of 10-HCPT synergistically with RTS on liver cancer cells HepG2 is 0.79, while the maximum inhibition rate of Taxol synergistically with RTS is 0.95. The synergistic effect of 10-HCPT is much smaller than the effect of Taxol synergistically with RTS under the dosing regimen.

[0047] 2. Detection of the antiproliferative effect of doxorubicin (Dox) and RTS on hepatoma cells in vitro

[0048] HepG2 tumor cells in the logarithmic growth phase were cultured at about 10 5 / mL density was inoculated on a 96-well plate, 100μL per well, and cultured overnight. Two separate dosing groups were set up: Dox group and RTS group. According to the preset concentration gradient, the maximum concentration was 16μg / mL, and the drug was added in half dilution. Each gradient was repeated at least 3 times. After continuing to culture for 24 hours, 5mg / mL of MTT was added to each well, and incubated at 37℃ for 4 hours. The supernatant was discarded by the rapid flipping method, and 150μL of DMSO was added to each well. The test was performed on the microplate reader. According to the measured OD 570 IC50 50 Determination of IC of Dox and RTS 50After the value, the drug combination regimen was standardized according to the IC 50 For quasi-gradient synergistic drug administration, at least 3 replicates were added to each gradient according to the preset concentration gradient. After 24 hours of continuous culture, 5 mg / mL of MTT was added to each well and incubated at 37°C for 4 hours. The supernatant was discarded by the rapid plate flipping method, and 150 μL of DMSO was added to each well. The cells were detected on an ELISA reader. The OD values ​​were measured. 570 IC50 50 .

[0049] Table 5 The antiproliferative effects of Dox and RTS on HepG2 cell line

[0050]

[0051] Determination of IC of Dox and RTS 50 After the value, the drug combination regimen was standardized according to the IC 50 For quasi-gradient synergistic drug delivery, HepG2 tumor cells in the logarithmic growth phase were cultured at a rate of about 10 5 The cells were inoculated at a density of 100 μL / mL in a 96-well plate, and cultured overnight. At least 3 replicates were added to each gradient according to the preset concentration gradient. After 24 hours of continuous culture, 5 mg / mL of MTT was added to each well, and the cells were incubated at 37°C for 4 hours. The supernatant was discarded by the rapid plate flipping method, and 150 μL of DMSO was added to each well. The cells were detected on an ELISA reader, and the OD values ​​were measured. 570 IC50 50 .

[0052] Table 6 Antiproliferative effect of the combination of Dox and RTS in the HepG2 liver cancer cell line

[0053]

[0054] IC50% inhibition rate of cancer cells 50 The concentration is the maximum concentration combination of Dox and RTS, and the drug is administered in gradient dilution. The synergistic inhibition rate is obtained according to the MTT experiment, such as Figure 3 As shown in Table 6, the maximum synergistic inhibition rate of Dox synergistically with RTS on liver cancer cells HepG2 is 0.66, and the maximum inhibition rate of Taxol synergistically with RTS is 0.95. The effect of Dox synergistically is much smaller than the effect of Taxol synergistically with RTS under the same dosing regimen.

[0055] In summary, according to the cell experiment results of 10-HCPT, Dox, Taxol and RTS, the synergistic effect of Taxol combined with RTS was the best, and the synergistic inhibition rate on the proliferation of liver cancer cell HepG2 cells was the highest.

[0056] Example 4 Calculation and analysis results of the synergy index (CI) of the combined use of Taxol and RTS

[0057] First, according to the experimental results of MTT in Example 2, the inhibition rate of tumor cells by using drugs alone (RTS, paclitaxel, Taxol) and using drugs RTS and paclitaxel (Taxol) in combination was obtained. Then, the synergy index (CI) was calculated using the Chou-Talalay method. This method is based on the median effect principle and assumes that there is a logarithmic linear relationship between the effect of the drug and the drug concentration. The specific calculation formula is: CI = (D) 1 / (Dx) 1 + (D) 2 / (Dx) 2 + α × (D) 1 × (D) 2 / (Dx) 1 × (Dx) 2. Among them, (D) 1 and (D) 2 are the concentrations of drug A and drug B in the combined drug use, respectively; (Dx) 1 and (Dx) 2 are the concentrations when drug A and drug B are used alone to achieve the same effect (such as 50% inhibition rate); α is a coefficient, when the drugs interact with each other, α = 0; when the drugs act synergistically, α = 1; when the drugs act antagonistically, α = -1. If the CI value is much less than 1, it indicates that the synergistic effect between the drugs is very strong, and its mechanism of action can be further studied to provide a basis for clinical drug combination. When the CI value is between 0.7 and 1.0, it can be considered that the two drugs have a weak synergistic effect; when the CI value is between 0.3 and 0.7, it can be considered that the two drugs have a synergistic effect; when the CI value is less than 0.3, it indicates that the two drugs have a strong synergistic effect.

[0058] Table 7 Synergistic index of Taxol combined with RTS in HepG2 liver cancer cell line

[0059]

[0060] According to Table 7, when Taxol and RTS were used in combination, they had a synergistic effect when the combined drug concentration was in the range of 0.15+0.06 (μg / mL) to 1.2+0.5 (μg / mL). At the low concentration of 0.15+0.06 (μg / mL), the CI value of Taxol and RTS was less than 0.3, showing a strong synergistic effect; while the medium and high inhibition rates When the CI values ​​of Taxol and RTS synergistic schemes at medium and high concentrations of 0.3+0.125 (μg / mL) to 1.2+0.5 (μg / mL) were both between 0.3 and 0.7, it indicated that the two drugs had a synergistic effect.

[0061] Example 5 Calculation and analysis of the dose reduction index (DRI) for the combined use of Taxol and RTS

[0062] The formula for the DRI dose reduction index is as follows: Wherein, (D)a and (D)b are the single drug concentrations when the two drugs act in combination, and (Dx)a and (Dx)b are the concentrations when the two drugs act alone. The calculated DRIa and DRIb are the reductions of the two drugs compared with the single drugs when the effect of the synergistic effect of the combination of drugs a and b is x. The DRI value is meaningful only when the combination index CI<1, that is, when the two drugs act in combination to produce a synergistic effect. According to Example 2, the CI of RTS synergistic with Taxol is <1, so the DRI value is further calculated and analyzed.

[0063] Table 8 Dose reduction index for combined use of Taxol and RTS

[0064]

[0065] according to Figure 4 As can be seen from the data results in Table 8, on the HepG2 liver cancer cell line, the relationship curve trend between the dosage concentration (x-axis) of the two drugs and the cell proliferation inhibition rate (y-axis) is consistent, indicating that their mechanisms of action on cell proliferation inhibition are similar. Therefore, when the combined use of drugs reaches a certain fixed cell proliferation inhibition rate (such as 50%), the DRI values ​​of the two drugs may be the same. This is also the reason why the data results of RTS and Taxol in Table 8 are the same. When the combination of drugs produces a synergistic inhibitory effect, it is 4 to 7 times less than the single drug dose; especially when RTS and Taxol are 5 times less than the single drug dose, the synergistic inhibition rate of the combined drug can reach 88%, which greatly reduces the use of chemotherapy drugs while improving effectiveness.

[0066] In summary, the combination of Taxol and RTS showed a significant synergistic effect on liver cancer cells HepG2. Among them, the results of the MTT experiment showed that Taxol and RTS could inhibit the proliferation of HepG2 cells in a dose-dependent manner when used alone. However, when the two were used in combination at different concentrations, the inhibitory effect on cell proliferation was significantly better than that of a single drug at the same concentration. This synergistic inhibition phenomenon indicates that the combination of the two can synergize.

[0067] We use calculations to determine whether there is a synergistic effect between drugs. The CI value is a key indicator for measuring the synergistic effect of drug combination. When CI < 1, it indicates that there is a synergistic effect between drugs. In the present invention, the CI values ​​of RTS and Taxol combined at multiple concentration combinations are all less than 1, indicating that the combination of the two can exert a synergistic effect.

[0068] In addition, DRI is used to evaluate the possible reduction in the dosage of single-drug chemotherapy when combined with other drugs. In the present invention, by comparing the dosage required to achieve the same therapeutic effect with a single drug, it is found that after Taxol and RTS are used in combination, the respective DRI values ​​are greater than 1. This shows that the combination of the realgar microbial extract of Acidithiobacillus ferrooxidans Dom-BY3 and paclitaxel can significantly reduce the dosage of the drug, and the effect is far better than the combination effect with other chemical drugs. This is not only conducive to reducing the toxic and side effects that may be caused by high doses and improving the safety of treatment, but also a clinical treatment plan that effectively reduces treatment costs and has promotion value.

[0069] In summary, the combination strategy of Acidithiobacillus ferroxidans realgar microbial extract and paclitaxel provides a new idea for the treatment of liver cancer.

Claims

1. An anti-tumor drug composition, characterized in that: The active ingredients of the composition include Acidithiobacillus ferroxidans realgar microbial extract and paclitaxel.

2. The composition according to claim 1, characterized in that The acidithiobacillus ferroxidans is acidithiobacillus ferroxidans Dom-BY3, which has been deposited in the China Center for Type Culture Collection, with the deposit address being Wuhan University, Wuhan, China. The deposit date is November 25, 2024, and the deposit number is CCTCC NO: M 20242585.

3. The composition according to claim 1, characterized in that The acidithiobacillus ferrooxidans realgar microbial extract is prepared by the following method: S1: Prepare the culture medium of Acidithiobacillus ferrooxidans Dom-BY31K: weigh 3.00 g of (NH4)2SO4, 0.50 g of K2HPO4, 0.50 g of MgSO4·7H2O, 0.10 g of KCl, 0.01 g of Ca(NO3)2, and 4.478 g of FeSO4·7H2O, and dissolve them in 950 mL of distilled water with stirring. Pipette 1 mL of concentrated sulfuric acid to adjust the pH value to 1.8-2.0, and make up to 1000 mL for use. S2: Take out the Dom-BY3 strain preserved in the laboratory, draw 10mL and inoculate it into the above-prepared 1K medium containing 0.5g / 100mL realgar and 0.4478g / 100mL FeSO4·7H2O, activate it on a shaker at 180rpm, 30℃ for 4d, observe the color of the bacterial culture solution under the 1K growth condition, it changes from light yellow to light reddish brown, indicating that the next leaching experiment can be carried out, continue to carry out the leaching experiment under the condition of 1K medium containing 0.5g / 100mL realgar, 180rpm, 30℃, add 2% of the total volume of 1K medium to the culture bottle every 2d, leaching for 68d, and prepare the acidophilic ferrothiobacillus Dom-BY3 realgar microbial leaching solution.

4. The composition according to claim 1, characterized in that The ratio of acidithiobacillus ferrooxidans Dom-BY3 realgar microorganism extract to paclitaxel is 15-120:6.25-50, and the concentration of acidithiobacillus ferrooxidans Dom-BY3 realgar microorganism extract is calculated based on arsenic.

5. The composition according to claim 4, characterized in that The arsenic concentration in the acidithiobacillus ferrooxidans Dom-BY3 realgar microbial leaching solution is 0.15 μg / mL to 1.2 μg / mL.

6. The composition according to claim 4, characterized in that The concentration of paclitaxel is 0.0625 μg / mL to 0.5 μg / mL.

7. The composition according to claim 4, characterized in that The ratio of the acidithiobacillus ferrooxidans Dom-BY3 realgar microbial extract to paclitaxel is 2.4:

1.

8. The composition according to claim 1, characterized in that The composition is one of powder, granule, powder, pill, tablet, capsule, pill and oral liquid.

9. Use of the composition according to claims 1-8 in preparing a drug for treating liver cancer.

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