Application of sanggenol L in preparation of medicine for treating chemotherapy-resistant small cell lung cancer

Abstract

The invention relates to the technical field of medicines, and discloses application of sanggenol L in preparation of a medicine for treating chemotherapy-resistant small cell lung cancer. The invention also provides a medicine containing sanggenol L and used for treating chemotherapy resistant small cell lung cancer. The invention finds that sanggenol L is targeted to Cx32 protein so as to reverse the chemotherapy drug resistance of SCLC to CDDP and VP-16 in an EP chemotherapy regimen, and provides a new safe and effective drug combination strategy.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and in particular to the use of sangenone alcohol L in preparing a drug for treating chemotherapy-resistant small cell lung cancer. Background Art

[0002] Small cell lung cancer (SCLC), the most malignant subtype of lung cancer, is characterized by rapid proliferation, early metastasis, high risk of drug resistance and recurrence, and a low survival rate. For four decades, treatment progress has been slow, and effective targets and targeted drugs are lacking. The current standard first-line chemotherapy regimen, the EP chemotherapy regimen of etoposide (VP-16) combined with cisplatin (CDDP), while initially sensitive, has a high rate of drug resistance and recurrence. Therefore, identifying key targets and innovative drug molecules that can reverse SCLC's malignant progression and drug resistance is crucial for overcoming the challenge of chemotherapy resistance in SCLC and improving patient outcomes.

[0003] The connexin (Cx) family plays an important role in cell communication and signal transduction. Abnormal Cx expression is often associated with chemoresistance in tumors. Cx32, a member of the Cx family, has recently garnered significant attention for its role in tumors. Its expression level is closely correlated with the prognosis of various cancers, but its role in regulating SCLC malignant progression and chemoresistance remains unclear.

[0004] Sanggenon L (San L) is a natural flavonoid compound isolated from the root bark of Morus alba L., a plant of the Moraceae family. It is also known as 2-(2,4-dihydroxyphenyl)-2,3-dihydro-5-hydroxy-8-methyl-8-(4-methylpent-3-enyl)-4H,8H-benzo[1,2-b:3,4-b']dipyran-4-one. It belongs to the flavonoid compound with a molecular formula of C 25 H 26 O6, molecular weight 422.47, CAS number: 329319-20-2. Its structural formula is as follows:

[0005]

[0006] Sanggenconol L has attracted increasing attention due to its multi-target anti-tumor mechanism, but there are currently no reports on the use of Sanggenconol L in the treatment of drug-resistant small cell lung cancer. Summary of the Invention

[0007] The first object of the present invention is to provide a use of sangenol L in the preparation of a drug for treating chemotherapy-resistant small cell lung cancer. The second object of the present invention is to provide a drug containing sangenol L for treating chemotherapy-resistant small cell lung cancer.

[0008] Specifically, the drug resistance includes resistance to etoposide and / or cisplatin. More specifically, the drug resistance is resistance caused by the combined use of etoposide and cisplatin in the EP chemotherapy regimen.

[0009] Specifically, the sangenone alcohol L targets the Cx32 protein to reverse chemotherapy resistance.

[0010] Specifically, the drug comprises sangenone alcohol L and pharmaceutically acceptable excipients. Pharmaceutically acceptable excipients refer to various conventional excipients required for preparing different dosage forms, such as diluents, binders, disintegrants, glidants, lubricants, flavoring agents, inclusion materials, adsorbents, etc. The drug dosage forms include tablets, powders, granules, capsules, or injections.

[0011] The beneficial effects of the present invention include:

[0012] 1. This study clarified the central role of the gap junction protein Cx32 in regulating SCLC malignant progression and chemoresistance, providing a novel mechanism of action and filling a research gap in SCLC treatment. Based on the Cx32 target structure, a high-affinity natural compound, San L, was identified through high-throughput virtual screening. San L specifically binds to Cx32 and effectively reverses SCLC chemoresistance to CDDP and VP-16.

[0013] 2. A synergistic drug use strategy of San L and the standard first-line EP chemotherapy regimen (VP-16+CDDP) is proposed, which can significantly enhance the sensitivity of tumor cells to chemotherapy drugs and break through the problem of SCLC resistance. Compared with single chemotherapy or San L monotherapy, combined drug use can more effectively inhibit tumor growth and improve survival prognosis.

[0014] 3. The treatment regimen of San L alone and in combination with CDDP and VP-16 has no obvious toxicity to major organs such as the heart, liver, spleen, lungs, and kidneys, and has good safety for liver and kidney function. It can also reverse the liver function damage caused by EP chemotherapy, has a certain protective effect on the liver, and is highly safe. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 :(A) Schematic diagram of the construction process of drug-resistant SCLC cell lines; (B) CCK-8 assay for changes in IC50 of drug-resistant SCLC cells; (CD) Western blot assay for changes in the expression of MRP1 and P-gp proteins in drug-resistant SCLC cells. Among them, **p<0.01, ***p<0.001, ****p<0.0001; ( n=3).

[0016] Figure 2:(AB) qRT-PCR results of mRNA levels in H1688VP and H69AR cells overexpressing Cx32; (CD) Western blot results of protein levels in H1688VP and H69AR cells overexpressing Cx32. Among them, **p<0.01, ***p<0.001, ****p<0.0001, "ns" indicates no significant difference; ( n=3).

[0017] Figure 3 :(AB) Cell viability and IC of H1688VP cells treated with different concentrations of VP-16 and cisplatin for 48 h 50 ; (CD) Cell viability and IC of H69AR cells treated with different concentrations of VP-16 and cisplatin for 48 h 50 . Among them, *p<0.05, **p<0.01, ***p<0.001; ( n=3).

[0018] Figure 4 :(A) The effect of San L on Cx32 protein stability was detected by CETSA method; (B) Protein grayscale quantification. Among them, *p<0.05, **p<0.01; ( n=3).

[0019] Figure 5 : MST analysis was performed on the binding of San L and Cx32, and the fitted Kd curve was 9.48 μM.

[0020] Figure 6 : Synergistic effect score of CDDP and VP-16 combined with San L on H69AR cells; (n=3).

[0021] Figure 7 :(AB) Macroscopic observation of tumor-bearing nude mice treated in vivo and corresponding tumor photos; (C) Tumor volume growth curve. (D) Tumor weight statistics; (E) Body weight statistics. Among them, * / #p<0.05, ****p<0.0001, "ns" indicates no significant difference; ( n=7).

[0022] Figure 8 : HE staining was used to detect the pathomorphological changes of the main organs of nude mice in each group (scale bar = 200 μm).

[0023] Figure 9 : Serum biochemical analysis of liver and kidney function in nude mice in different treatment groups. *p<0.05, "ns" indicates no significant difference; ( n=7). DETAILED DESCRIPTION

[0024] The present invention discloses the use of sangenone alcohol L in the preparation of a drug for treating chemotherapy-resistant small cell lung cancer. Those skilled in the art can refer to the present disclosure and appropriately modify the process parameters to achieve the desired effect. It is particularly important to note that all similar substitutions and modifications obvious to those skilled in the art are considered encompassed by the present invention.

[0025] The present invention provides the use of sangenconol L in the preparation of a drug for treating chemotherapy-resistant small cell lung cancer. Corresponding to this use, the present invention also provides a drug for treating chemotherapy-resistant small cell lung cancer containing sangenconol L.

[0026] The drug also includes one or more inert, non-toxic, pharmacologically suitable excipients. The excipients include carriers, solvents, emulsifiers, dispersants, wetting agents, binders, stabilizers, colorants, flavorings, etc. The drug can be in the form of tablets, powders, granules, capsules, injections, etc.

[0027] The medicaments according to the invention can act systemically and / or locally and, for this purpose, can be administered in a suitable manner, for example, by the oral, parenteral, pulmonary or nasal route. The medicaments according to the invention can be administered in dosage forms suitable for these administration routes.

[0028] Wherein, be suitable for oral administration and work according to the state of the art and promptly and / or discharge the administration form of medicine of the present invention in an improved manner, and comprise with crystallization and / or amorphous and / or dissolving form, for example tablet (uncoated or coated tablet, it for example has and resists gastric juice or delays dissolving or insoluble coating, release according to the present invention's medicine), rapidly broken tablet in mouth, or diaphragm, film / lyophilized product, capsule (for example hard or soft capsule), sugar-coated tablet, granule, pellet, powder, emulsion, suspension, medicine according to the present invention of aerosol or solution. Parenteral administration can avoid absorption step (for example intravenous, intra-arterial, intracardiac, intra-spinal or waist or intra-articular) or comprise simultaneously that absorption (for example intramuscular, subcutaneous, intradermal, percutaneous or intraperitoneal) is carried out.The suitable form of administration that is suitable for parenteral administration is particularly for injection and injection with the preparation of solution, suspension, emulsion, lyophilized product or sterile powder form. The dosage form suitable for the nasal route generally refers to a method of administration in which the drug is absorbed through the nasal mucosa to treat local or systemic diseases. Common dosage forms for this method of administration include nasal drops or nasal sprays.

[0029] The medicament according to the invention can be converted into the dosage form described. This can be done in a manner known per se by mixing with inert, non-toxic, pharmacologically suitable excipients. These excipients include in particular carriers (e.g. microcrystalline cellulose, lactose, mannitol, starch), solvents (e.g. liquid polyethylene glycol), emulsifiers and dispersants or wetting agents (e.g. sodium lauryl sulfate, polyoxysorbitan oleate, propylene glycol), binders (e.g. polyvinylpyrrolidone), synthetic and natural polymers (e.g. albumin), stabilizers (e.g. antioxidants such as ascorbic acid), colorants (e.g. inorganic pigments such as iron oxides) and masking flavors and odors.

[0030] In order to verify the therapeutic effect of Sanggenol L, the present invention also provides multiple experimental examples for illustration. Unless otherwise specified, the raw materials, reagents, consumables and instruments involved in the present invention are all common commercial products and can be purchased from the market.

[0031] Experimental Example 1: Construction and verification of drug-resistant SCLC cell lines.

[0032] refer to Figure 1 A. The SCLC etoposide-resistant strain NCI-H1688VP (hereinafter referred to as H1688VP) was established by using a drug concentration gradient method on the parental NCI-H1688 (hereinafter referred to as H1688) cell line: starting from 5 μM VP-16, the parental cells were intermittently stimulated for 48 hours and then switched to drug-free medium for culture until the cells recovered their proliferation activity (approximately 80-90% confluence). This cycle was repeated 3-5 times to allow the cells to adapt to the concentration; the drug concentration was then gradually increased (up to 20 μM) and the cycle was repeated for about 6 months to obtain a stable resistant strain. The SCLC multidrug-resistant cell line NCI-H69AR (hereinafter referred to as H69AR) was established from NCI-H69 cells (hereinafter referred to as H69) (ATCC HTB-119). These cells were grown for 14 months at gradually increasing concentrations of doxorubicin. The number inhibitory concentration (IC) of parental cells and resistant cells was determined by the CCK-8 method. 50 ), calculate the drug resistance index (RI) (resistant cell IC 50 / Parental cell IC 50 ), RI>5 indicates that qualified drug-resistant strains have been successfully established.

[0033] The present invention successfully obtained two pairs of SCLC chemotherapy-resistant cell models (H1688VP / H69AR). CCK-8 assays revealed that the IC values ​​of the resistant cells to VP-16 and CDDP were 50 The values ​​increased significantly, and the drug resistance index RI reached 12.60 / 10.50 (H1688VP) and 18.70 / 5.36 (H69AR) ( Figure 1B), all meet the requirement of RI>5 for drug-resistant cell lines. Western blot confirmed that the expression levels of multidrug resistance markers MRP1 and P-gp proteins in drug-resistant cells were significantly upregulated compared with those in parental cells (p<0.001) ( Figure 1 CD), which verified the reliability of the drug-resistant model at the phenotypic and molecular levels and laid the foundation for subsequent mechanism research.

[0034] Experimental Example 2: Stable overexpression of Cx32 in SCLC by lentiviral transfection.

[0035] The role of Cx32 in SCLC chemotherapy resistance is still unclear. In this study, a third-generation lentiviral vector overexpressing Cx32 was used to transfect H1688VP and H69AR to construct H1688VP OE and H69AR OE stably overexpressing Cx32. The lentiviral vector was constructed by Heyuan Biotechnology (Shanghai) Co., Ltd. H1688VP (1.5×10 4 / hole) and H69AR (5×10 4 / well) were inoculated into 24-well plates. The next day, lentivirus and culture medium containing 5μg / mL Polybrene were added according to the optimal multiplicity of infection (MOI) value in the pre-experimental study. The peak of GFP expression was observed 72 hours after transfection. When the infection efficiency reached 80% and the cell confluence was 60-70%, the culture medium containing the minimum lethal concentration of Puromycin (H1688VP: 5μg / mL, H69AR: 2μg / mL) determined in the pre-experiment was switched to screening for 2 weeks. During this period, empty cell controls were set up to monitor the toxicity of Puromycin, and finally stable transfected cells were obtained. The expression levels of Cx32 in the negative control (NC) and overexpression (OE) cell lines were detected by qPCR and Western blot to verify the efficiency.

[0036] After continuous screening with the third generation of puromycin (5 μg / mL), RT-qPCR detection showed that the relative expression of Cx32 mRNA ( Figure 2 AB) and Cx32 protein expression levels ( Figure 2 The above results indicate that we have successfully constructed H1688VP OE and H69AR OE cell models that stably overexpress Cx32, which can be used for subsequent functional experimental studies.

[0037] Experimental Example 3: CCK-8 assay to detect the IC values ​​of SCLC cells overexpressing Cx32 to VP-16 and CDDP 50 Influence.

[0038] H1688VP (NC / OE) and H69AR (NC / OE) cells were cultured at the indicated density (5×10 3 or 1×10 4cells / well, 200 μL) were seeded in a 96-well plate (3 replicates / group) and administered according to H1688VP: VP-16 (0-1600 μM), CDDP (0-400 μM); H69AR: VP-16 (0-1600 μM), CDDP (0-160 μM). After culturing for 48 h, the culture supernatant was removed and CCK-8 working solution (100 μL / well) was added at a ratio of 10% of the final volume. The cells were incubated at 37°C in the dark for 2 h. The absorbance at a wavelength of 450 nm (OD value) was measured by a microplate reader, and the cell viability was calculated according to the formula: Viability (%) = (OD of the drug group - OD adjusted to zero) / (OD of the control group - OD adjusted to zero) × 100%, and the IC was calculated. 50 .

[0039] The results showed that in the cisplatin-treated group, H1688VP OE cells IC 50 The value decreased by 43.3% compared with the control group (118.80±6.42vs 67.33±3.66μM, p<0.05) ( Figure 3 A); H69AR OE group IC 50 33.8% reduction (33.23±3.59 vs 22.01±1.70 μM, p<0.05) ( Figure 3 B) The VP-16 treated group also showed a significant reversal effect, and the H1688VP OE group IC 50 Reduced by 66.2% (294.47±38.91 vs 99.44±22.31 μM) Figure 3 C), H69AR OE group decreased by 33.8% (317.70±40.10 vs 210.43±31.01 μM) ( Figure 3 The above results indicate that overexpression of Cx32 can reduce the IC value of drug-resistant SCLC to CDDP / VP-16. 50 , thereby reversing chemotherapy resistance.

[0040] Experimental Example 4: Screening of monomeric compounds targeting Cx32.

[0041] By consulting the National Administration of Traditional Chinese Medicine's series of literature on ethnic medicine, including "Color Atlas of Anti-tumor Zhuang Medicines", "List of Chinese Zhuang Medicine Resources", "100 Commonly Used Zhuang Medicines", "Collection of Zhuang Medicine Folk Prescriptions", and "Complete Record of Chemical Components of Gui Medicines", and conducting literature surveys, we collected and compiled 189 anti-lung cancer Zhuang medicines and 294 heat-clearing, detoxifying, and strengthening the body's resistance to expel evil Gui medicines, totaling 483 anti-lung cancer ethnic medicines. Using the online databases Herb (http: / / herb.ac.cn / ) and TCMSP (https: / / old.tcmsp-e.com / tcmsp.php), we screened and removed duplicates, and constructed a database of monomeric compounds of anti-lung cancer ethnic medicines with thousands of monomeric compounds.

[0042] Combined with the three-dimensional structure of human Cx32 protein (PDB ID: 7zxm), the active pocket was predicted and determined by computer technology. The Chinese medicine monomers in the above compound library were cleaned, conformationally searched, and energy minimized, and the compound library after docking was processed with the active pocket as the target. The molecular docking results were scored, and the five most highly rated ethnic medicine monomers were obtained: Sanggenol L, Dihydrocurcumin, Cycloolivil, Corynoxeine, and Alisol A. The CCK-8 experiment was used to detect its anticancer activity against drug-resistant SCLC H69AR cells. The results showed that among the five monomers, San L had the highest IC value for H69AR cells. 50 Molecular docking revealed that San L exhibited optimal binding properties. 2D and 3D docking models revealed multiple interactions between San L and the Cx32 active pocket. The optimal binding free energy of San L strongly aligned with in vitro efficacy data, leading to its selection for further investigation.

[0043] Experimental Example 5: Verification by Cellular Thermal Shift Assay (CETSA) SanL interacts directly with Cx32.

[0044] H69AR cells were treated with San L (final concentration 200 μM) for 2 h, then trypsinized, centrifuged (1000 rpm, 5 min) and collected. The cells were resuspended in pre-chilled PBS and evenly divided into 6 PCR tubes. After heating at 37°C, 42°C, 47°C, 52°C, 57°C and 62°C for 3 min, NP40 lysis buffer was added, and then quick-frozen in liquid nitrogen for 1 min, thawed at room temperature, and frozen and thawed three times before centrifugation (4°C, 12000g, 30 min). Subsequent operations were performed according to the Western blotting method.

[0045] In the H69AR OE cell model, it was observed that under the temperature gradient treatment conditions of 42-62°C, the soluble Cx32 protein in the San L treatment group (200 μM) showed a significant enhancement in thermal stability compared with the DMSO control group (p < 0.05) ( Figure 4 ). This suggests that San L and Cx32 have a direct interaction.

[0046] Experimental Example 6: Quantitative Analysis of SanL and Cx32 interaction relationship.

[0047] Purified Cx32 protein was provided by MedChemExpress (Monmouth Junction, NJ, USA). The protein was fluorescently labeled with NHS using the Monolith Protein Labeling Kit RED-NHS manufacturer's instructions (MO-L011, NanoTemper Technologies). Labeled Cx32 (final concentration 20 nM) was used as the target. Sixteen sample tubes were also prepared, and San L was serially diluted in Assay Buffer (final concentration range 0.00305–100 μM) as the ligand. The two were mixed and equilibrated at room temperature for 15 minutes. The samples were then loaded using capillaries and detected using a Monolith NT.115 instrument (Nanotemper, Germany). The excitation power was set to 20%, and the MST power was set to 40%. The equilibrium dissociation constant (Kd) was calculated.

[0048] Fnorm form Kd fitting diagram ( Figure 6 ) uses the molar concentration of San L as the abscissa and the normalized fluorescence intensity Fnorm as the ordinate. The Fnorm value reflects the change in Cx32 protein fluorescence intensity per unit space. The protein migration rate increases with increasing ligand (San L) concentration. The fitted curve exhibits an inverted S-shaped pattern with distinct upper and lower plateaus and a strong concentration dependence, indicating specific binding. The affinity Kd value is 9.48 μM, confirming a moderately strong specific interaction between the two.

[0049] Experimental Example 7: CCK-8 assay to verify the effect of San L on the sensitivity of H69AR cells to VP-16 and CDDP chemotherapy ring.

[0050] H69AR cells were seeded in 96-well plates using the same procedures as previously described. For the combined drug administration assay, the stock concentrations were: CDDP 3.33 mM; VP-16 33.98 mM; and Sanggenone L 60 mM. The drug concentration gradient was as follows: CDDP 0, 2.5, 5, 10, and 20 μM; VP-16 0, 50, 100, 200, and 400 μM; and Sanggenone L 0, 5, 10, 20, and 40 μM. Because each well originally contained 100 μL of culture medium, the drug concentration was adjusted to twice the desired concentration to prevent dilution. 100 μL of the corresponding drug-containing culture medium was added to each well and incubated for 48 hours. CCK-8 assay procedures were performed as previously described.

[0051] Table 1. Effects of different concentrations of VP-16 and San L alone or in combination on the viability of H69AR cells for 48 h. n=3)

[0052]

[0053] Table 2. Effects of different concentrations of CDDP and San L alone and in combination on the viability of H69AR cells for 48 h. n=3)

[0054]

[0055] Tables 1 and 2 show the effects of San L combined with VP-16 and CDDP on H69AR cell viability. In addition, we used the online SynergyFinder software to calculate the drug ZIP synergy score based on the concentration gradient of different drugs and the corresponding cell inhibition rate. A score > 0 indicates synergy, and a score > 10 is considered strong synergy. The results showed that CDDP and VP-16 combined with San L showed strong synergy in inhibiting H69AR cell proliferation (ZIP synergy scores of 13.507 and 20.008, respectively) ( Figure 6 A). The gradient of the red area indicates the intensity of the synergistic effect, the white rectangle indicates the concentration contained in the highest synergistic area, and the X-axis and Y-axis correspond to the two sides of the white rectangle, respectively, indicating the concentration of the drug combination with the greatest inhibitory effect on cell growth ( Figure 6 B) It can be seen from the figure that CDDP 5μM and VP-16 100μM combined with 5μM San L are the lowest concentrations that exert synergistic effects.

[0056] Experimental Example 8: Verification of San L's ability to reverse VP-16 and CDDP chemotherapy resistance using a nude mouse subcutaneous transplant tumor model.

[0057] The present invention constructs a H69AR drug-resistant cell BALB / c nude mouse transplant tumor model to explore the in vivo efficacy and resistance reversal mechanism of San L on drug-resistant SCLC. The experiment set up four groups (n=7 in each group): model control group (normal saline, intraperitoneal injection every other day), standard EP chemotherapy group (CDDP 2.5 mg / kg on the first day + VP-16 4 mg / kg on the 1st to 3rd day, intraperitoneal injection), San L single drug group (10 mg / kg, intraperitoneal injection every other day) and combined treatment group (combine: San L combined with EP regimen, the dosage is the same as before). The specific process is as follows: H69AR cells in the logarithmic growth phase are taken, digested with trypsin and mixed with Matrigel matrix gel at a ratio of 1:1 on ice (final density 5×10 6 The mixture was inoculated subcutaneously in the right axilla of 5-week-old female BALB / c nude mice until the tumor volume reached 50 mm. 3 The mice were then randomly divided into groups. Dosing was performed on a 7-day cycle for three cycles, with tumor volume (formula: 0.5 × long diameter × short diameter²) and body weight monitored every other day. All drugs were prepared immediately: CDDP was dissolved in saline; VP-16 and San L were both prepared using 5% DMSO + 30% PEG3000 + 65% saline. Specimen collection at the endpoint: After blood was drawn from the eye sockets, the nude mice were euthanized, tumor tissue was obtained and weighed, and organs such as the heart, liver, spleen, lungs, and kidneys were harvested for subsequent testing.

[0058] Pharmacodynamic evaluation showed that ( Figure 7 AB): Compared with the model control group (Vehicle), the EP chemotherapy group (CDDP+VP-16) showed no significant changes in tumor volume growth times and tumor weight (p>0.05), while the San L single-drug group and the combined drug group (Combine) showed significant anti-tumor activity, and the tumor volume growth times and tumor weight were significantly reduced compared with the Vehicle group, and the differences were statistically significant (p<0.05) ( Figure 7 CD). At the same time, compared with the San L single-drug group, the Combine group showed a stronger tumor inhibitory effect (p < 0.05). The above results show that San L has a good anti-tumor effect on drug-resistant SCLC, and can synergize with chemotherapy drugs to exert a stronger anti-tumor effect, thereby reversing SCLC chemotherapy resistance. Toxicity parameters showed that there was no statistical difference in the body weight changes of nude mice among all groups (p > 0.05) ( Figure 7 E), suggesting that San L has good drug safety.

[0059] Experimental Example 9: Hematoxylin-eosin (HE) staining was used to investigate the safety of San L combined with chemotherapy on multiple organs. estimate.

[0060] Nude mouse organ slices were baked in a 60°C forced air drying oven for 30 minutes, followed by four xylene dewaxing cycles (15 minutes each, completely immersed). Rehydration was performed with graded ethanol: anhydrous ethanol, 95% ethanol, 75% ethanol, and 50% ethanol for 15 minutes each, followed by 10 minutes in pure water. The slices were then washed three times in a shaking PBS oven (5 minutes each). Hematoxylin staining was performed for 5 minutes, followed by 5-minute running water rinses, followed by pure water rinses. Differentiation solution was added dropwise for 2-3 seconds, followed by 5-minute running water rinses. Eosin staining was performed in the same manner. Finally, the slices were baked at 60°C for 1 hour and sealed with neutral resin.

[0061] The results showed that compared with the model control group (Vehicle), the standard chemotherapy group (CDDP+VP-16), San L single drug group (10 mg / kg) and the combined treatment group (Combine) did not show obvious and characteristic pathological changes in various organs ( Figure 8 ), indicating that the EP chemotherapy group, San L or combined treatment group had no obvious toxicity to various organs of nude mice.

[0062] Experimental Example 10: Blood biochemical analysis was used to evaluate the effects of San L combined with chemotherapy on liver and kidney function in nude mice.

[0063] Blood was collected from the nude mice's orbits and allowed to rest for 2 hours. The blood was then centrifuged at 3000 rpm for 15 minutes at 4°C. The supernatant was aspirated and the serum of the four groups of nude mice was used for liver and kidney function tests. Liver function alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels, as well as renal function creatinine (CREA) and creatinine (BUN) levels, were analyzed using an automated biochemical analyzer.

[0064] The results showed that compared with the Vehicle model control group, the liver function ALT and AST in the EP chemotherapy group were significantly increased, indicating that the chemotherapy drug CDDP may cause a certain degree of liver function damage (p < 0.5) ( Figure 9 AB). However, there was no significant difference in liver and kidney function between the San L monotherapy group and the Combined group (p>0.5) ( Figure 9 CD), indicating that San L has good safety for liver and kidney function. At the same time, San L not only did not increase the liver function damage in the Combine group, but also reversed the increase of ALT and AST in liver function caused by the EP chemotherapy group, indicating that San L has a certain degree of protective effect on the liver.

Claims

1. Application of sangenone alcohol L in the preparation of drugs for treating chemotherapy-resistant small cell lung cancer.

2. The use according to claim 1, characterized in that: The drug resistance includes etoposide and / or cisplatin resistance.

3. The use according to claim 1, characterized in that: The sangenone alcohol L targets the Cx32 protein.

4. The use according to claim 1, characterized in that: The medicine contains mulberry ketone L and pharmaceutically acceptable excipients.

5. The use according to claim 4, characterized in that: The dosage form of the drug includes tablets, powders, granules, capsules or injections.

6. A drug for treating chemotherapy-resistant small cell lung cancer, characterized in that: Contains Morangone Alcohol L.

7. The drug according to claim 6, characterized in that: The drug resistance includes etoposide and / or cisplatin resistance.

8. The drug according to claim 6, characterized in that: The sangenone alcohol L targets the Cx32 protein.

9. The drug according to claim 6, characterized in that: The medicine also contains pharmaceutically acceptable excipients.

10. The drug according to claim 9, characterized in that: The dosage form of the drug includes tablets, powders, granules, capsules or injections.

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