Musa basjoo root extract and application thereof in preparation of antitumor drugs

Abstract

The invention belongs to the technical field of medicines, and particularly relates to a Japanese banana root extract and application thereof in preparation of antitumor drugs, and a preparation method of the Japanese banana root extract comprises the following steps: 1) raw material pretreatment; 2) extracting; (3) concentrating and purifying; and 4) extracting and drying. The Japanese banana root extract shows broad-spectrum activity in the anti-tumor aspect, in-vitro experimental data shows that the Japanese banana root extract has an inhibiting effect on various tumors such as cervical cancer, triple negative breast cancer, lung cancer, liver cancer and colon cancer, and in addition, the Japanese banana root extract can be used as a chemotherapy sensitizer and has multiple application values in tumor comprehensive treatment.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically to a banana root extract and its application in the preparation of antitumor drugs. Background Technology

[0002] Banana root, a traditional Chinese medicine, was first recorded in the *Compendium of Materia Medica*. It is cold in nature and sweet in taste, and enters the stomach, spleen, and liver meridians. Traditionally, it is used to clear heat and detoxify, promote urination, and quench thirst. Modern chemical analysis shows that banana root contains various bioactive substances, including polysaccharides (approximately 14.86%), flavonoids, amino acids, and cellulose. However, pharmacological studies have long focused on its anti-inflammatory and antioxidant effects, with very few reports on its anti-tumor activity.

[0003] Existing literature only mentions that banana root may exert anti-cancer effects by regulating immune function or inducing tumor cell apoptosis, but these studies remain at the preliminary screening stage, lacking systematic elucidation of the mechanism of action and in vitro and in vivo experimental verification. This research gap contrasts sharply with the widespread use of banana root in traditional medicine, suggesting that its anti-tumor potential has not yet been fully explored.

[0004] Cancer is one of the leading diseases threatening human health worldwide. Chemotherapy, as one of the three traditional treatment methods (surgery, radiotherapy, and chemotherapy), plays an important role in the control of advanced tumors and postoperative adjuvant therapy. However, the clinical application of chemotherapy has long faced two major challenges: First, the non-selective toxicity of chemotherapy drugs to normal cells leads to serious side effects, such as bone marrow suppression, gastrointestinal reactions, and cardiotoxicity; second, tumor cells are prone to developing drug resistance through various mechanisms (such as overexpression of drug efflux pumps and enhanced DNA repair), resulting in approximately 90% of chemotherapy failure cases being related to drug resistance.

[0005] Despite breakthroughs in targeted therapy and immunotherapy in recent years, natural products remain a crucial source for anti-tumor drug development. Data shows that approximately 50% of currently used anti-tumor drugs are derived directly or indirectly from natural products, such as paclitaxel and camptothecin. However, the application of natural products in anti-tumor treatment still faces significant limitations: most components target only a single site, making it difficult to address the complexity of tumors with multiple genes and pathway abnormalities; low bioavailability and poor water solubility limit their clinical translation; and traditional extraction processes are often crude, resulting in insufficient purity of active ingredients and unclear mechanisms of action.

[0006] Chemotherapy sensitizers can significantly improve treatment efficacy and reduce toxic side effects by reversing tumor cell drug resistance or enhancing the sensitivity of chemotherapeutic drugs. Currently, commonly used sensitizers in clinical practice include verapamil and cyclosporine A, but these suffer from poor selectivity and significant toxic reactions. Sensitizers derived from natural products have become a research hotspot due to their potential advantages of multi-target and low toxicity.

[0007] Recent studies have found that vitamin K2 can inhibit and The sensitization mechanism of 5-FU enhances the killing effect of 5-FU on colorectal cancer cells through multi-pathway synergistic regulation. However, such studies mostly focus on single components or single tumor types, lacking broad-spectrum sensitization strategies. How to efficiently isolate components with both anti-tumor and sensitizing activities from natural products remains a technical bottleneck. Summary of the Invention

[0008] To address the aforementioned technical problems, this invention provides a banana root extract and its application in the preparation of antitumor drugs. The banana root extract of this invention exhibits broad-spectrum antitumor activity. In vitro experimental data show that it has inhibitory effects on various tumors, including cervical cancer, triple-negative breast cancer, lung cancer, liver cancer, and colon cancer. Furthermore, the banana root extract of this invention can be used as a chemotherapy sensitizer, possessing multiple application values ​​in comprehensive tumor treatment.

[0009] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0010] A banana root extract, the preparation method of which includes the following steps:

[0011] 1) Raw material pretreatment: Take dried banana roots, crush them, sieve them, defatt them with petroleum ether or diethyl ether, filter them, remove the solvent from the residue, and obtain defatted banana roots for later use;

[0012] 2) Extraction: Take the defatted banana root prepared in step 1), add 70% to 90% ethanol aqueous solution, adjust the pH to 5-6, extract at 60-70℃, filter to obtain extract A; add 50% to 60% ethanol aqueous solution to the filter residue, adjust the pH to 8-9, extract at 60-70℃, filter to obtain extract B;

[0013] 3) Concentration and purification: Concentrate extract A and extract B from step 2) to 1 / 10-1 / 5 of their original volumes to obtain concentrated extract A and concentrated extract B. Load concentrated extract A and concentrated extract B separately or in combination onto a macroporous adsorption resin column, elute with water until the effluent is nearly colorless, and discard the aqueous solution; then elute with 50%-60% ethanol (v / v) and collect the ethanol eluent.

[0014] 4) Extraction and drying: The ethanol eluent from step 3) is depressurized to recover ethanol and concentrated until there is no alcohol odor to obtain concentrate C; concentrate C is extracted with n-butanol, the n-butanol phase is taken, the n-butanol is recovered under reduced pressure, and the resulting extract is vacuum dried to obtain the banana root extract.

[0015] Further, step 1) specifically involves: taking dried banana root, crushing it, passing it through a No. 3 sieve, and defatting it with petroleum ether or diethyl ether, wherein the volume-to-weight ratio of petroleum ether or diethyl ether to banana root is 1 ml: 1 g, the defatting is performed twice, filtering, and drying the residue at 40°C under reduced pressure until there is no solvent odor, to obtain defatted banana root for later use.

[0016] Further, step 2) specifically involves: taking the defatted banana root prepared in step 1), adding an 80% ethanol aqueous solution at a material-to-liquid ratio of 1g:20mL, adjusting the pH to 5.5 with 0.1mol / L sodium dihydrogen phosphate buffer, refluxing at 65℃ for 2 hours, filtering to obtain extract A, and adding a 55% ethanol aqueous solution to the filter residue at a material-to-liquid ratio of 1g:15mL, adjusting the pH to 8.5 with 0.1mol / L sodium borate buffer, refluxing at 70℃ for 2 hours, filtering to obtain extract B.

[0017] Further, step 3) specifically involves: concentrating extract A and extract B from step 2) to 1 / 8 of their original volumes to obtain concentrated extract A and concentrated extract B. Combining concentrated extract A and concentrated extract B, the mixture is loaded onto a D101 macroporous resin column and eluted with water until the eluent is nearly colorless, then the aqueous solution is discarded. Next, the mixture is eluted with 3-5 BV of 55% ethanol and the ethanol eluent is collected.

[0018] Further, step 4) specifically involves: the extraction operation specifically involves mixing concentrated liquid C with n-butanol at a volume ratio of 1:1, allowing it to stand and separate into layers, separating and collecting the n-butanol phase; repeating this operation 3 times, and combining all the n-butanol liquids; the water content of the banana root extract is ≤5%.

[0019] This invention provides the application of the banana root extract in the preparation of antitumor drugs, wherein the tumors include cervical cancer, triple-negative breast cancer, lung cancer, liver cancer, and colon cancer.

[0020] The present invention also provides the application of the banana root extract in the preparation of a chemotherapy sensitizer, wherein the chemotherapy sensitizer is a sensitizer that increases the sensitivity to 5-FU or DDP chemotherapy.

[0021] The present invention provides a composition containing the above-mentioned banana root extract, the composition comprising banana root extract and 5-FU.

[0022] The present invention also provides a composition containing the above-mentioned banana root extract, the composition comprising banana root extract and DDP.

[0023] The present invention also provides the use of the above composition in the preparation of antitumor drugs.

[0024] In this invention, the banana root is the dried rhizome of the banana plant Musa basjoo Sied. et Zucc. of the Musaceae family.

[0025] In summary, due to the adoption of the above technical solution, the beneficial effects of the present invention are:

[0026] This invention innovatively combines acidic (pH 5-6) and alkaline (pH 8-9) ethanol extraction with a staged pH-controlled extraction technique to achieve targeted enrichment of moderately polar active ingredients. The banana root extract of this invention exhibits broad-spectrum anti-tumor activity, with in vitro experimental data showing inhibitory effects against various tumors including cervical cancer, triple-negative breast cancer, lung cancer, liver cancer, and colon cancer. Its mechanism may involve inducing tumor cell apoptosis, inhibiting proliferation, and regulating related signaling pathways, providing a potential natural drug source for the treatment of multiple types of cancer.

[0027] Furthermore, the banana root extract of this invention can serve as a chemotherapy sensitizer, significantly enhancing the antitumor effects of 5-FU (fluorouracil) and DDP (cisplatin). This helps reduce the required chemotherapy dosage, thereby minimizing toxic side effects. It has multiple applications in comprehensive cancer treatment. Detailed Implementation

[0028] Example 1

[0029] A banana root extract, the preparation method of which includes the following steps:

[0030] 1) Raw material pretreatment: Take dried banana root, crush it, pass it through No. 3 sieve, and then use petroleum ether to degrease the powder twice at a liquid-solid ratio of 1ml:1g (filter out the solvent after each treatment). Dry the degreased residue under reduced pressure at 40℃ until there is no solvent odor to obtain defatted banana root for later use.

[0031] 2) Extraction: Take the defatted banana root prepared in step 1), add 80% ethanol aqueous solution at a material-to-liquid ratio of 1g:20mL, adjust the pH to 5.5 with 0.1mol / L sodium dihydrogen phosphate buffer, reflux at 65℃ for 2 hours, filter to obtain extract A, add 55% ethanol aqueous solution to the filter residue at a material-to-liquid ratio of 1g:15mL, adjust the pH to 8.5 with 0.1mol / L sodium borate buffer, reflux at 70℃ for 2 hours, filter to obtain extract B;

[0032] 3) Concentration and purification: Extract A and extract B from step 2) are concentrated under vacuum at ≤70℃ to 1 / 8 of their original volume to obtain concentrated solution A and concentrated solution B. The concentrated solution A and concentrated solution B are combined and loaded onto a D101 macroporous resin column. The column is eluted with water until the eluent is nearly colorless and the aqueous solution is discarded. Then, the column is eluted with 4 BV of 55% ethanol and the ethanol eluent is collected.

[0033] 4) Extraction and drying: The ethanol eluent from step 3) is depressurized to recover ethanol and concentrated until no alcohol odor is detected, yielding concentrate C; concentrate C is extracted with n-butanol, the n-butanol phase is collected, the n-butanol is recovered under reduced pressure, and the resulting extract is vacuum dried at ≤70℃ to obtain the banana root extract; the extraction operation specifically involves mixing concentrate C with n-butanol at a volume ratio of 1:1, allowing it to stand and separate into layers, separating and collecting the n-butanol phase, repeating this operation 3 times, and combining all the n-butanol solutions; the water content of the banana root extract is ≤5%.

[0034] Example 2

[0035] A banana root extract, the preparation method of which includes the following steps:

[0036] 1) Raw material pretreatment: Take dried banana root, crush it, pass it through No. 3 sieve, and then use ether to degrease the powder twice at a liquid-solid ratio of 1ml:1g (filter out the solvent after each treatment). Dry the degreased residue under reduced pressure at 40℃ until there is no solvent odor to obtain defatted banana root for later use.

[0037] 2) Extraction: Take the defatted banana root prepared in step 1), add 90% ethanol aqueous solution at a material-to-liquid ratio of 1g:15mL, adjust the pH to 5 with 0.1mol / L sodium dihydrogen phosphate buffer, reflux at 65℃ for 3 hours, filter to obtain extract A, add 60% ethanol aqueous solution to the filter residue at a material-to-liquid ratio of 1g:20mL, adjust the pH to 9 with 0.1mol / L sodium borate buffer, reflux at 70℃ for 2 hours, filter to obtain extract B;

[0038] 3) Concentration and purification: Extract A and extract B from step 2) are concentrated under vacuum at ≤70℃ to 1 / 10 of their original volume to obtain concentrated solution A and concentrated solution B. The concentrated solution A and concentrated solution B are combined and loaded onto a D101 macroporous resin column. The column is eluted with water until the eluent is nearly colorless and the aqueous solution is discarded. Then, the column is eluted with 50% ethanol (5 BV) and the ethanol eluent is collected.

[0039] 4) Extraction and drying: The ethanol eluent from step 3) is depressurized to recover ethanol and concentrated until no alcohol odor is detected, yielding concentrate C; concentrate C is extracted with n-butanol, the n-butanol phase is collected, the n-butanol is recovered under reduced pressure, and the resulting extract is vacuum dried at ≤70℃ to obtain the banana root extract; the extraction operation specifically involves mixing concentrate C with n-butanol at a volume ratio of 1:1, allowing it to stand and separate into layers, separating and collecting the n-butanol phase, repeating this operation 3 times, and combining all the n-butanol solutions; the water content of the banana root extract is ≤5%.

[0040] Example 3

[0041] A banana root extract, the preparation method of which includes the following steps:

[0042] 1) Raw material pretreatment: Take dried banana root, crush it, pass it through No. 3 sieve, and then use petroleum ether to degrease the powder twice at a liquid-solid ratio of 1ml:1g (filter out the solvent after each treatment). Dry the degreased residue under reduced pressure at 40℃ until there is no solvent odor to obtain defatted banana root for later use.

[0043] 2) Extraction: Take the defatted banana root prepared in step 1), add 80% ethanol aqueous solution at a material-to-liquid ratio of 1g:30mL, adjust the pH to 6 with 0.1mol / L sodium dihydrogen phosphate buffer, reflux at 65℃ for 2 hours, filter to obtain extract A, add 50% ethanol aqueous solution to the filter residue at a material-to-liquid ratio of 1g:10mL, adjust the pH to 8 with 0.1mol / L sodium borate buffer, reflux at 70℃ for 2 hours, filter to obtain extract B;

[0044] 3) Concentration and purification: Extract A and extract B from step 2) are concentrated under vacuum at ≤70℃ to 1 / 5 of their original volume to obtain concentrated solution A and concentrated solution B. The concentrated solution A and concentrated solution B are combined and loaded onto a D101 macroporous resin column. The column is eluted with water until the eluent is nearly colorless and the aqueous solution is discarded. Then, the column is eluted with 3 BV of 60% ethanol and the ethanol eluent is collected.

[0045] 4) Extraction and drying: The ethanol eluent from step 3) is depressurized to recover ethanol and concentrated until no alcohol odor is detected, yielding concentrate C; concentrate C is extracted with n-butanol, the n-butanol phase is collected, the n-butanol is recovered under reduced pressure, and the resulting extract is vacuum dried at ≤70℃ to obtain the banana root extract; the extraction operation specifically involves mixing concentrate C with n-butanol at a volume ratio of 1:1, allowing it to stand and separate into layers, separating and collecting the n-butanol phase, repeating this operation 3 times, and combining all the n-butanol solutions; the water content of the banana root extract is ≤5%.

[0046] To illustrate the technical effects of the present invention, the inventors have set up the following comparative examples and comparative experiments.

[0047] Comparative Example 1

[0048] The difference from Example 1 is that in step 2) of Comparative Example 1, the staged pH-controlled extraction technique is not used; only acidic ethanol extraction is employed. The specific steps are as follows:

[0049] A banana root extract, the preparation method of which includes the following steps:

[0050] 1) Raw material pretreatment: Take dried banana root, crush it, pass it through No. 3 sieve, and then use petroleum ether to degrease the powder twice at a liquid-solid ratio of 1ml:1g (filter out the solvent after each treatment). Dry the degreased residue under reduced pressure at 40℃ until there is no solvent odor to obtain defatted banana root for later use.

[0051] 2) Extraction: Take the defatted banana root prepared in step 1), add 80% ethanol aqueous solution at a material-to-liquid ratio of 1g:20mL, adjust the pH to 5.5 with 0.1mol / L sodium dihydrogen phosphate buffer, reflux at 65℃ for 2 hours, filter, and obtain extract A;

[0052] 3) Concentration and purification: The extract A from step 2) is concentrated under vacuum at ≤70℃ to 1 / 8 of its original volume to obtain concentrated solution A. The concentrated solution A is loaded onto a D101 macroporous resin column and eluted with water until the eluent is nearly colorless. The aqueous solution is then discarded. Next, it is eluted with 4 BV of 55% ethanol and the ethanol eluent is collected.

[0053] 4) Extraction and drying: The ethanol eluent from step 3) is depressurized to recover ethanol and concentrated until no alcohol odor is detected, yielding concentrate C; concentrate C is extracted with n-butanol, the n-butanol phase is collected, the n-butanol is recovered under reduced pressure, and the resulting extract is vacuum dried at ≤70℃ to obtain the banana root extract; the extraction operation specifically involves mixing concentrate C with n-butanol at a volume ratio of 1:1, allowing it to stand and separate into layers, separating and collecting the n-butanol phase, repeating this operation 3 times, and combining all the n-butanol solutions; the water content of the banana root extract is ≤5%.

[0054] Comparative Example 2

[0055] The difference from Example 1 is that in step 2) of Comparative Example 2, the staged pH-controlled extraction technique is not used; only alkaline ethanol extraction is employed. The specific steps are as follows:

[0056] A banana root extract, the preparation method of which includes the following steps:

[0057] 1) Raw material pretreatment: Take dried banana root, crush it, pass it through No. 3 sieve, and then use petroleum ether to degrease the powder twice at a liquid-solid ratio of 1ml:1g (filter out the solvent after each treatment). Dry the degreased residue under reduced pressure at 40℃ until there is no solvent odor to obtain defatted banana root for later use.

[0058] 2) Extraction: Take the defatted banana root prepared in step 1), add 55% ethanol aqueous solution at a material-to-liquid ratio of 1g:15mL, adjust the pH to 8.5 with 0.1mol / L sodium borate buffer, reflux at 70℃ for 2 hours, filter to obtain extract B;

[0059] 3) Concentration and purification: The extract B from step 2) is concentrated under vacuum at ≤70℃ to 1 / 8 of its original volume to obtain concentrated solution B. The concentrated solution B is loaded onto a D101 macroporous resin column and eluted with water until the eluent is nearly colorless. The aqueous solution is then discarded. Next, it is eluted with 4 BV of 55% ethanol and the ethanol eluent is collected.

[0060] 4) Extraction and drying: The ethanol eluent from step 3) is depressurized to recover ethanol and concentrated until no alcohol odor is detected, yielding concentrate C; concentrate C is extracted with n-butanol, the n-butanol phase is collected, the n-butanol is recovered under reduced pressure, and the resulting extract is vacuum dried at ≤70℃ to obtain the banana root extract; the extraction operation specifically involves mixing concentrate C with n-butanol at a volume ratio of 1:1, allowing it to stand and separate into layers, separating and collecting the n-butanol phase, repeating this operation 3 times, and combining all the n-butanol solutions; the water content of the banana root extract is ≤5%.

[0061] Comparative Example 3

[0062] The difference from Example 1 is that Comparative Example 3 uses water extraction for preparation. The specific steps are as follows:

[0063] A banana root extract, the preparation method of which includes the following steps:

[0064] 1) Raw material pretreatment: Take dried banana root, crush it, pass it through No. 3 sieve, and then use petroleum ether to degrease the powder twice at a liquid-solid ratio of 1ml:1g (filter out the solvent after each treatment). Dry the degreased residue under reduced pressure at 40℃ until there is no solvent odor to obtain defatted banana root for later use.

[0065] 2) Extraction: Take the defatted banana root prepared in step 1), add it to the aqueous solution at a material-to-liquid ratio of 1g:20mL, extract at 90℃ for 2 hours, filter, and obtain extract A;

[0066] 3) Concentration and purification: The extract A from step 2) is concentrated under vacuum at ≤70℃ to 1 / 8 of its original volume to obtain concentrated solution A. The concentrated solution A is loaded onto a D101 macroporous resin column and eluted with water until the eluent is nearly colorless. The aqueous solution is then discarded. Next, it is eluted with 4 BV of 55% ethanol and the ethanol eluent is collected.

[0067] 4) Extraction and drying: The ethanol eluent from step 3) is depressurized to recover ethanol and concentrated until no alcohol odor is detected, yielding concentrate C; concentrate C is extracted with n-butanol, the n-butanol phase is collected, the n-butanol is recovered under reduced pressure, and the resulting extract is vacuum dried at ≤70℃ to obtain the banana root extract; the extraction operation specifically involves mixing concentrate C with n-butanol at a volume ratio of 1:1, allowing it to stand and separate into layers, separating and collecting the n-butanol phase, repeating this operation 3 times, and combining all the n-butanol solutions; the water content of the banana root extract is ≤5%.

[0068] In vitro antitumor activity test of banana root extract

[0069] 1. Experimental Materials

[0070] 1.1 Cell lines

[0071] Human cell lines: HeLa (cervical cancer), MB231 (triple-negative breast cancer), A549 (lung cancer), Hepa1-6 (liver cancer);

[0072] Mouse cell line: CT26 (colon cancer).

[0073] 1.2 Reagents and Instruments

[0074] Test samples: Banana root extract A (prepared from Example 1, hereinafter referred to as extract A), Banana root extract B (prepared from Comparative Example 1, hereinafter referred to as extract B), Banana root extract C (prepared from Comparative Example 2, hereinafter referred to as extract C), and Banana root extract D (prepared from Comparative Example 3, hereinafter referred to as extract D).

[0075] Positive controls: 5-fluorouracil (5-FU, purity >99%), cisplatin (DDP, purity >99%).

[0076] Solvent: Dimethyl sulfoxide (DMSO, cell culture grade);

[0077] Culture media: DMEM, RPMI-1640;

[0078] Detection reagents: MTT (3-(4,5-dimethylthiazol-2)-2,5-diphenyltetrazolium bromide), DMSO;

[0079] Instruments and equipment: Incubator, microplate reader, inverted microscope;

[0080] The above experimental conditions were all provided by the Scientific Experiment Center of Guangxi University of Chinese Medicine.

[0081] 2. Experimental Methods

[0082] 2.1 Sample Preparation

[0083] (1) Preparation of extract stock solution: Weigh the test sample accurately, dissolve it in DMSO to prepare a 20 mg / mL stock solution, and store it at -20℃ in the dark.

[0084] (2) Gradual dilution: Before the experiment, dilute the stock solution to the working concentration (containing 0.1% DMSO) with the corresponding culture medium and store at 4℃ for later use.

[0085] (3) Preparation of chemotherapy drugs: 5-FU and DDP are dissolved in sterile water and diluted to working concentration in the same way.

[0086] 2.2 Cell Processing Procedure

[0087] (1) Cell seeding: Logarithmic growth phase cells were seeded at a specific density in 96-well plates:

[0088] Hela: 3 × 10³ cells / well

[0089] MB231, CT26: 8 × 10³ cells / well

[0090] A549, Hepa1-6: 5 × 10³ cells / well;

[0091] (2) Incubation: Place at 37℃ and 5% Incubate in an incubator for 24 hours;

[0092] (3) Drug administration: Remove the original culture medium and add culture medium containing different concentrations of test samples or chemotherapy drugs, and continue to culture for 48 h;

[0093] (4) Detection: Add 10 μL of 5 mg / mL MTT solution to each well, continue incubation for 4 h, remove the supernatant, add 150 μL LDMSO to dissolve the formazan crystals, and measure the OD value at 570 nm using an ELISA reader.

[0094] 2.3 Experimental Design

[0095] (1) Screening of single-drug activity (Table 1)

[0096] Determine the half-maximal inhibitory concentration (IC50) of extracts A, B, C, and D against each tumor cell type. Each concentration was tested in 6 replicates, and the experiment was repeated 3 times independently.

[0097] (2) Combined drug trials (Tables 2-5)

[0098] Based on the single-drug activity results, the MB231 (human triple-negative breast cancer) and CT26 (mouse colon cancer) cell lines, which were most sensitive to the extracts, were selected as representative models. To ensure that the observed effect was "sensitization" rather than "toxicity additive," the concentration of the extracts in the combination therapy was set to a sub-toxic dose with a single-drug inhibition rate of <20%.

[0099] The grouping design is as follows:

[0100] Negative control: 0.1% DMSO

[0101] Positive controls: 5-FU (4 μM), DDP (2 μM)

[0102] Extract groups: Banana root extract A (1.0 / 2.5 / 5.0 μg / mL), Banana root extract B (5 / 10 / 20 μg / mL), Banana root extract C (40 μg / mL), Banana root extract D (40 μg / mL);

[0103] Combination therapy group:

[0104] 5-FU (4 μM) + Banana root extract A (1.0 / 2.5 / 5.0 μg / mL)

[0105] 5-FU (4 μM) + Banana root extract B (5 / 10 / 20 μg / mL)

[0106] 5-FU (4 μM) + Banana root extract C (40 μg / mL)

[0107] DDP (2 μM) + Banana Root Extract A (1.0 / 2.5 / 5.0 μg / mL)

[0108] DDP (2 μM) + Banana Root Extract B (5 / 10 / 20 μg / mL)

[0109] DDP (2 μM) + Banana Root Extract C (40 μg / mL)

[0110] Note: Chemotherapy drug concentrations should be selected based on single-agent chemotherapy. Values ​​(5-FU: 4 μM; DDP: 2 μM). The working concentration of the extract was set as a non-cytotoxic dose (single-drug inhibition rate < 20%) based on preliminary experimental results. Given the significant activity of extract A, its combined concentration was appropriately lowered to more accurately assess its sensitizing effect rather than cytotoxicity.

[0111] 2.4 Data Analysis

[0112] 2.4.1 Calculation of Inhibition Rate

[0113] The formula for calculating the inhibition rate is:

[0114] Equation (1)

[0115] In formula (1): A2 is the test group with culture medium containing the test solution; A1 is the control group with culture medium without the test solution; A0 is the blank control group without any liquid or cells.

[0116] 2.4.2 Calculation of the Coordination Index (CI)

[0117] The formula for calculating the synergy index is:

[0118] Equation (2)

[0119] In formula (2): D1 and D2 are the concentrations of the two drugs when used in combination, D x1 D x2 CI refers to the concentration at which a single drug achieves the same effect. The criteria for determining this effect are: CI < 1 indicates synergistic effect, CI = 1 indicates additive effect, and CI > 1 indicates antagonistic effect.

[0120] 2.5 Experimental Results

[0121] Table 1 Results of single-drug activity screening

[0122]

[0123] Note: 100 represents >100 μg / mL (no significant activity)

[0124] Table 2 Synergistic effect of banana root extract combined with 5-FU on MB231 cells (representative concentrations)

[0125]

[0126] Table 3. Synergistic effect of banana root extract combined with 5-FU on CT26 cells (representative concentrations)

[0127]

[0128] Table 4. Synergistic effect of banana root extract combined with DDP on MB231 cells (representative concentrations)

[0129]

[0130] Table 5. Synergistic effect of banana root extract combined with DDP on CT26 cells (representative concentrations)

[0131]

[0132] 2.6 Results Analysis

[0133] (1) Single drug activity screening (Table 1) showed that banana root extract A had significant cytotoxicity against a variety of tumor cells (such as MB231 and CT26); while extracts B, C and D had weak or no activity.

[0134] (2) Combined drug experiments showed that, at subtoxic doses, extract A exhibited a strong synergistic effect when used in combination with 5-FU / DDP (CI=0.48-0.61, p<0.001). For example, in CT26 cells, extract A at 5.0 μg / mL (single-drug inhibition rate of 12.7%) combined with 5-FU showed an inhibition rate as high as 72.4% (CI=0.48). Extract B showed only weak synergy (CI≈0.85-0.92, p<0.05), while extract C showed no synergistic effect (CI>1.05, p>0.05). This result strongly suggests that the active ingredient in extract A can significantly enhance the antitumor effect of chemotherapeutic drugs, rather than simply adding toxicity. This synergistic effect may be related to the degree of glycosylation—moderate glycosylation can improve water solubility and bioavailability, but excessive glycosylation (such as extracts B and C) can block target binding sites or reduce cell membrane permeability, thereby weakening activity. This discovery provides a key basis for optimizing the separation and structural modification of extract components.

[0135] The above description is a detailed description of the preferred embodiments of the present invention. However, the embodiments are not intended to limit the scope of the patent application of the present invention. All equivalent changes or modifications made under the technical spirit of the present invention should fall within the patent scope covered by the present invention.

Claims

1. A banana root extract, characterized in that, The preparation method of the banana root extract includes the following steps: 1) Raw material pretreatment: Take dried banana roots, crush them, sieve them, defatt them with petroleum ether or diethyl ether, filter them, remove the solvent from the residue, and obtain defatted banana roots for later use; 2) Extraction: Take the defatted banana root prepared in step 1), add 70% to 90% ethanol aqueous solution, adjust the pH to 5-6, extract at 60-70℃, filter to obtain extract A; add 50% to 60% ethanol aqueous solution to the filter residue, adjust the pH to 8-9, extract at 60-70℃, filter to obtain extract B; 3) Concentration and purification: Concentrate extract A and extract B from step 2) to 1 / 10-1 / 5 of their original volumes to obtain concentrated extract A and concentrated extract B. Load concentrated extract A and concentrated extract B separately or in combination onto a macroporous adsorption resin column, elute with water until the effluent is nearly colorless, and discard the aqueous solution; then elute with 50%-60% ethanol (v / v) and collect the ethanol eluent. 4) Extraction and drying: The ethanol eluent from step 3) is depressurized to recover ethanol and concentrated until there is no alcohol odor to obtain concentrate C; concentrate C is extracted with n-butanol, the n-butanol phase is taken, the n-butanol is recovered under reduced pressure, and the resulting extract is vacuum dried to obtain the banana root extract.

2. The banana root extract according to claim 1, characterized in that, Step 1) Specifically: Take the dried banana root, crush it, pass it through a No. 3 sieve, and defatt it with petroleum ether or ethyl ether. The volume weight ratio of petroleum ether or ethyl ether to banana root is 1 ml: 1 g. The defatting is performed twice. After filtration, the residue is dried under reduced pressure at 40°C until there is no solvent odor, and defatted banana root is obtained for later use.

3. The banana root extract according to claim 1, characterized in that, Step 2) Specifically: Take the defatted banana root prepared in Step 1), add 80% ethanol aqueous solution at a material-to-liquid ratio of 1g:20mL, adjust the pH to 5.5 with 0.1mol / L sodium dihydrogen phosphate buffer, reflux at 65℃ for 2 hours, filter to obtain extract A, add 55% ethanol aqueous solution to the filter residue at a material-to-liquid ratio of 1g:15mL, adjust the pH to 8.5 with 0.1mol / L sodium borate buffer, reflux at 70℃ for 2 hours, filter to obtain extract B.

4. The banana root extract according to claim 1, characterized in that, Step 3) Specifically, extracts A and B from step 2) are concentrated to 1 / 8 of their original volume to obtain concentrate A and concentrate B. The concentrates A and B are combined and loaded onto a D101 macroporous resin column. The column is eluted with water until the eluent is nearly colorless and the aqueous solution is discarded. Then, the column is eluted with 3-5 BV of 55% ethanol and the ethanol eluent is collected.

5. The banana root extract according to claim 1, characterized in that, Step 4) Specifically, the extraction operation involves mixing concentrated solution C with n-butanol at a volume ratio of 1:1, allowing the mixture to stand and separate into layers, then separating and collecting the n-butanol phase; repeating this operation 3 times and combining all the n-butanol solutions; the water content of the banana root extract is ≤5%.

6. The use of the banana root extract according to claim 1 in the preparation of antitumor drugs, characterized in that: The tumors include cervical cancer, triple-negative breast cancer, lung cancer, liver cancer, and colon cancer.

7. The use of the banana root extract according to claim 1 in the preparation of chemotherapy sensitizers, characterized in that: The chemotherapy sensitizer is a sensitizer that increases the sensitivity to 5-FU or DDP chemotherapy.

8. A composition containing the banana root extract of claim 1, characterized in that: The composition includes banana root extract and 5-FU.

9. A composition containing the banana root extract of claim 1, characterized in that: The composition includes banana root extract and DDP.

10. Use of the composition of claim 8 or 9 in the preparation of an antitumor drug.

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