Use of a scabioside derivative in the preparation of a drug for treating glioma

Abstract

The application discloses application of a swertianol derivative in preparation of a glioma treatment drug and belongs to the technical field of medicines. The application finds that the compound has significant inhibitory activity on SF126, T98G, A172, U-87 MG, U251 and various glioma cell lines, and the activity is better than that of temozolomide, a commonly used clinical anti-glioma drug, and the compound has lower toxicity to normal brain cells. The application further provides a preparation method of the compound and a pharmaceutical composition containing the compound, and the pharmaceutical composition can be prepared into injection, tablets and various dosage forms. The application provides a new effective drug selection for the treatment of glioma, has a mature preparation process, various drug dosage forms and a wide clinical application prospect.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically to the application of a gentamicin lactone derivative in the preparation of drugs for treating gliomas. Background Technology

[0002] Gliomas are the most common malignant tumors of the central nervous system, accounting for approximately 40%-50% of intracranial tumors. They are characterized by high invasiveness, rapid growth, high recurrence rate, and poor prognosis, seriously threatening human life and health. Currently, treatment methods for gliomas mainly include comprehensive therapies such as surgical resection, radiotherapy, and chemotherapy, but the clinical treatment outcomes remain unsatisfactory.

[0003] Surgical resection often fails to completely remove tumor tissue, and residual tumor cells easily lead to recurrence. While radiotherapy can inhibit tumor growth to some extent, it can also damage normal brain tissue, triggering a series of adverse reactions. Chemotherapy is an important means of postoperative adjuvant therapy and treatment for advanced gliomas; however, existing chemotherapy drugs generally suffer from poor targeting, significant toxic side effects, and a tendency to develop drug resistance, limiting their clinical application. Therefore, developing novel anti-glioma drugs with high efficacy, low toxicity, and strong targeting has become an important issue that urgently needs to be addressed in the current medical field. Summary of the Invention

[0004] To address the shortcomings of existing glioma treatments, such as insufficient efficacy, significant side effects, and difficulty in obtaining naturally occurring active compounds, this invention provides a glioma lactone derivative. This derivative is prepared through artificial chemical synthesis and structural modification using glioma lactone alcohol extracted from *Gynostemma pentaphyllum* (a plant in the *Gynostemma* genus of the Asteraceae family) as the core raw material, rather than directly using naturally extracted glioma lactone alcohol. This compound has a simple synthetic route, can be prepared in large quantities, and exhibits highly efficient inhibitory activity against glioma cells, making it suitable for the preparation of drugs for treating gliomas.

[0005] This invention provides the application of a gentamicin lactone derivative in the preparation of a drug for treating gliomas. The gentamicin lactone derivative is used to prepare a drug for treating gliomas, wherein the gliomas include human brain tumor cells (SF126), human brain glioma cells (T98G), human glioblastoma cells (A172), and human brain astrocytoblastoma cells (U-87 MG).

[0006] The beneficial effects of this application, based on the above technical solutions, are as follows:

[0007] This invention is the first to discover that the ring-opening product of 12-methylaminogentinol lactone derivative (compound A) has significant anti-glioma activity, inhibiting various glioma cell lines such as SF126, T98G, A172, U-87 MG, and U251, and its inhibitory activity is superior to that of temozolomide, a commonly used anti-glioma drug in clinical practice. At the same time, this compound has low toxicity to normal brain cells (such as human astrocytes) and better safety.

[0008] The preparation method of this compound is mature, the reaction parameters are well-defined, the yield is stable, and the purity is controllable, which is conducive to industrial production. Drugs prepared with this compound as the active ingredient can be formulated into various dosage forms according to clinical needs, which are convenient to use and provide a new and effective option for the treatment of gliomas, with broad clinical application prospects. Attached Figure Description

[0009] Figure 1 This is a schematic diagram of the chemical structure of the gentian lactone derivative provided in an embodiment of the present invention;

[0010] Figure 2 This is a schematic diagram of the chemical structure of the preparation process of 12-methylaminoglycinol lactone alcohol ring-opening product (compound A). Detailed Implementation

[0011] The present invention is illustrated by the following examples. Example

[0012] Preparation of the ring-opening product of 12-methylaminogentinol (compound A):

[0013] The raw material used in this embodiment, *Gynostemma pentaphyllum* lactone alcohol, is a natural secondary metabolite of *Gynostemma pentaphyllum*, a plant belonging to the genus *Gynostemma* of the Asteraceae family, and is obtained by extraction and separation from the whole herb of *Gynostemma pentaphyllum*. The target derivative compound A of this invention is synthesized from this naturally extracted *Gynostemma pentaphyllum* lactone alcohol through chemical modification. Figure 2 As shown, tianmingling lactone alcohol (50 mg, 1.95 × 10) was used. -1 Dissolve 1 mmol of methylamine hydrochloride in methanol (1.5 mL), then slowly add 0.25 mL of KOH aqueous solution (0.1 mol / L) to the reaction system. Stir at room temperature for 30 min, then add methylamine hydrochloride (52.65 mg, 0.78 mmol), and stir at room temperature and 300 r / min for 2 h. The reaction endpoint is detected by TLC (developing solvent: dichloromethane-methanol = 100:3, color development under UV lamp at 254 nm, the reaction is considered to be over when the starting spot disappears completely, the product spot is clear and no new impurity spots appear).

[0014] After the reaction was complete, the reaction system was evaporated to dryness by rotary evaporation. The mixture was then extracted with EtOAc (10 mL) and water (10 mL), and the extraction was repeated three times. The organic phases were combined, dried overnight with anhydrous Na2SO4, filtered to remove the desiccant, and the product was detected by TLC (dichloromethane-methanol, 100:5). After the reaction was complete, the solvent was evaporated from the filtered organic phase under reduced pressure to obtain the residue.

[0015] The residue was purified by silica gel column chromatography with a column diameter to column length ratio of 1:20, an eluent of dichloromethane-methanol = 100:1, and an elution flow rate of 1.5 mL / min. The target eluent was collected and concentrated under reduced pressure to obtain the ring-opening product of 12-methylaminogentinol (compound A) with a yield of 50%. The purity was ≥99% as determined by HPLC (column: C18, mobile phase: methanol-water = 60:40, flow rate: 1.0 mL / min, detection wavelength: 254 nm).

[0016] Product structure confirmation: LC / MS (ESI–MS) showed [M+Na] + =281.20.

[0017] 1 H-NMR (CDCl3, 400 MHz) δ: 7.22 (t, J = 7.7 Hz, 1H,), 4.76 – 4.70 (m,1H), 3.82 – 3.80 (m, 1H), 3.36 (s, 1H), 2.49 (qd, J = 12.2, 11.1, 5.0 Hz,1H), 1.53 (dtt, J = 23.8, 9.8, 5.3 Hz, 3H), 1.25 (s, 4H), 1.20 (s, 2H), 1.05(s, 4H);

[0018] 13 C-NMR (CDCl3, 201MHz) δ: 179.6, 177.6, 129.5, 71.9, 68.0, 59.2, 47.5, 39.3, 30.5, 29.7, 25.4, 23.6, 23.5, 18.4, 18.3, 10.5.

[0019] Anti-glioma activity assay of the ring-opening product of 12-methylaminogentinol:

[0020] Experimental materials:

[0021] Glioma cell lines SF126, T98G, A172, U-87 MG, and U251 (purchased from the Shanghai Cell Bank, Chinese Academy of Sciences); normal brain cell line human astrocytes (HA) (purchased from the Shanghai Cell Bank, Chinese Academy of Sciences); 12-methylaminopheninolone ring-opening product (compound A, prepared in Example 1 of this invention, purity ≥98%); temozolomide (positive control, purchased from Sigma, purity ≥99%); DMEM culture medium and fetal bovine serum (purchased from Gibco); CCK-8 kit (purchased from Beyotime Biotechnology Co., Ltd.); CO2 incubator (Thermo Fisher Scientific); microplate reader (Bio-Rad); high-performance liquid chromatograph (Agilent Technologies).

[0022] Experimental methods:

[0023] Cell culture: SF126, T98G, A172, U-87 MG, U251 glioma cells and HA normal brain cells were seeded in DMEM medium containing 10% fetal bovine serum, 100 U / mL penicillin and 100 μg / mL streptomycin, respectively, and cultured in an incubator at 37℃, 5% CO2 and saturated humidity. When the cells grew to 80%-90% confluence, they were passaged at a ratio of 1:3, and cells in the logarithmic growth phase were selected for experiments.

[0024] Drug treatment: Glioma cells in the logarithmic growth phase and normal brain cells in HA were seeded into 96-well plates at a density of 5 × 10³ cells per well. 100 μL of culture medium was added to each well. After incubation for 24 h, the old culture medium was discarded and fresh culture medium containing different concentrations (0, 1, 5, 10, 20, 50 μmol / L) of the ring-opening product of 12-methylaminotetracycline lactone alcohol (compound A) was added. At the same time, a temozolomide positive control group (concentration as above) and a blank control group (containing only culture medium and cells) were set up. Each concentration was set up in 6 replicates. The cells were cultured for another 48 h.

[0025] Cell viability assay: Add 10 μL of CCK-8 reagent to each well and incubate for 2 h. Then, measure the absorbance (OD value) of each well at 450 nm using a microplate reader and calculate the cell viability. Cell viability (%) = (OD value of experimental group - OD value of blank well) / (OD value of control group - OD value of blank well) × 100%; Cell inhibition rate (%) = 1 - Cell viability (%).

[0026] Experimental results:

[0027] Experimental results showed that the ring-opening product of 12-methylaminogentinol (compound A) significantly inhibited the growth of various glioma cell types, including SF126, T98G, A172, U-87 MG, and U251, and the inhibitory effect was concentration-dependent; meanwhile, it showed low toxicity to normal HA brain cells. Specific experimental data are shown in Table 1.

[0028] Table 1. Inhibition rates (%) and IC50 values ​​of the ring-opening product of 12-methylaminogentinol (compound A) and temozolomide on various cell lines. 50 Value (μmol / L)

[0029] cell lines drug 1 μmol / L inhibition rate 5 μmol / L inhibition rate 10 μmol / L inhibition rate 20 μmol / L inhibition rate 50 μmol / L inhibition rate <![CDATA[IC 50 Value SF126 Compound A 12.3±1.2 28.5±2.1 45.7±2.5 68.2±3.1 82.5±3.3 15.3±1.1 Temozolomide 8.1±0.9 19.3±1.5 32.6±2.2 51.4±2.8 70.2±3.0 22.5±1.3 T98G Compound A 10.5±1.0 25.3±1.8 42.1±2.3 65.4±2.9 80.1±3.2 16.7±1.2 Temozolomide 7.2±0.8 17.5±1.3 29.8±2.0 48.6±2.6 67.3±2.9 24.8±1.4 A172 Compound A 13.1±1.3 30.2±2.2 47.5±2.6 70.3±3.2 83.6±3.4 14.5±1.0 Temozolomide 8.5±0.9 20.1±1.6 34.2±2.3 53.5±2.9 72.1±3.1 21.7±1.2 U-87 MG Compound A 14.2±1.4 32.1±2.3 49.8±2.7 72.5±3.3 85.3±3.5 13.8±0.9 Temozolomide 9.3±1.0 22.4±1.7 36.5±2.4 55.8±3.0 74.6±3.2 20.3±1.1 U251 Compound A 11.8±1.1 27.6±2.0 44.3±2.4 66.7±3.0 81.2±3.3 15.9±1.1 Temozolomide 7.8±0.8 18.7±1.4 31.2±2.1 50.2±2.7 68.5±2.9 23.1±1.3 HA Compound A 3.2±0.5 5.8±0.7 8.5±0.9 12.3±1.1 18.6±1.5 89.2±3.6 Temozolomide 4.5±0.6 7.2±0.8 10.3±1.0 15.6±1.2 22.4±1.6 75.8±3.2

[0030] Note: The data in the table are the mean ± standard deviation of three independent experiments (n=6).

Claims

1. The application of a gentamicin lactone derivative in the preparation of a drug for treating glioma, characterized in that, The structural formula of the tianminglin lactone derivative is shown in Formula 1: Formula 1.

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