Application of 8-O-acetyl shanzhiside methyl ester in preparation of medicine for inhibiting development of nasopharyngeal carcinoma
By using 8-O-acetylganoside methyl ester (Barlerin) to prepare various drug formulations, the problem of the lack of drugs to inhibit the occurrence and development of nasopharyngeal carcinoma in the existing technology has been solved, and an effective inhibitory effect on nasopharyngeal carcinoma cells has been achieved.
Patent Information
- Application Number
- CN202511546101.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-10-28
- Publication Date
- 2026-01-13
AI Technical Summary
There is a lack of effective drugs to inhibit the development and progression of nasopharyngeal carcinoma in the current technology, especially research on Barlerin has not yet been carried out.
Using 8-O-acetylganoside methyl ester (Barlerin) as the main active ingredient, a pharmaceutical composition is prepared to inhibit the cell viability, proliferation and migration of nasopharyngeal carcinoma cells. The pharmaceutical composition can be formulated into various dosage forms such as oral and injectable preparations, with routes of administration including enteric and non-enteric, and dosage forms including liquid, solid and semi-solid. It can be further formulated into sustained-release, controlled-release and targeted preparations.
Barlerin effectively inhibits the viability, proliferation, and migration of nasopharyngeal carcinoma cells. Studies have shown that a concentration of 50 μM significantly reduces cell viability and proliferation rate, and significantly reduces the number of migrating cells, suggesting that it is an effective drug for inhibiting the development of nasopharyngeal carcinoma.
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Abstract
Description
Technical Field
[0001] This invention belongs to the field of drug technology for preparing drugs that inhibit the development of nasopharyngeal carcinoma, and relates to the application of 8-O-acetylganoside methyl ester in the preparation of drugs that inhibit the development of nasopharyngeal carcinoma. Background Technology
[0002] Nasopharyngeal carcinoma (NPC) is a malignant tumor of the head and neck, originating from the outermost layer of epithelial tissue in the nasopharynx, located above the oropharynx and hypopharynx, near the base of the skull. Early-stage NPC is often asymptomatic, mainly presenting with headache, neuropathic facial pain, neck mass, nosebleeds, or nasal congestion. Limited diagnostic methods for the nasopharynx lead to misdiagnosis, often resulting in advanced stages at the time of diagnosis, causing delayed treatment and poor prognosis. For a long time, the main treatment for NPC has been radiotherapy, chemotherapy, or a combination of both. Early-stage patients receive only radiotherapy, while advanced-stage patients receive radiotherapy combined with chemotherapy. However, a significant proportion of patients have poor prognoses, with low 5-year survival rates and distant metastases in 6-11% of cases. Therefore, developing new, better-tolerated, lower-cost, and more effective therapies is crucial to prolonging the survival of NPC patients and improving their quality of life.
[0003] Traditional Chinese medicine (TCM), as an important component of complementary and alternative medicine, has high clinical value in the treatment of tumors and related diseases, and shows good results in reducing tumor recurrence and metastasis, as well as the side effects caused by radiotherapy and chemotherapy. For example, berberine, an alkaloid isolated from Coptis chinensis, can not only control cancer development but also relieve paclitaxel-induced neuropathic pain or pain caused by tumor invasion. Furthermore, the traditional Chinese medicine formula NPC01 has also shown some efficacy in improving the symptoms and signs of head and neck tumors.
[0004] 8-O-Acetyl shanzhiside methylester (Barlerin) is a natural iridoid glycoside and the main active ingredient of *Lamiophlomis rotata*, a traditional Tibetan medicinal plant. Current research on Barlerin primarily focuses on its ability to inhibit neuroinflammatory pain, such as regulating signal transduction in spinal cord injury pathways (inhibiting the NMDAR / PKC and NO / cGMP / PKG pathways) and spinal cord neuroinflammatory responses (reducing TNF-α and IL-1β production and promoting IL-10 expression), binding to GLP-1 receptors, promoting microglia secretion of β-endorphin, and inhibiting the p38 MAPK signaling pathway. To date, there are no reported studies on Barlerin's role in inhibiting the development and progression of nasopharyngeal carcinoma. Summary of the Invention
[0005] To address the lack of existing research on the inhibition of nasopharyngeal carcinoma by Barlerin, this invention provides the application of 8-O-acetylganoside methyl ester in the preparation of drugs that inhibit the development of nasopharyngeal carcinoma. The research of this invention shows that Barlerin can effectively inhibit the cell viability, proliferation and migration of nasopharyngeal carcinoma cells, suggesting that it can be used to prepare effective drugs that inhibit the development of nasopharyngeal carcinoma.
[0006] The objective of this invention is achieved through the following technical solution:
[0007] Application of 8-O-acetylganoside methyl ester in the preparation of drugs that inhibit the development of nasopharyngeal carcinoma.
[0008] Furthermore, the application of 8-O-acetylganoside methyl ester in the preparation of drugs that inhibit the development of nasopharyngeal carcinoma specifically involves Barlerin effectively inhibiting the cell viability, proliferation, and migration of nasopharyngeal carcinoma cells.
[0009] Furthermore, the Barlerin effectively inhibits the cell viability of nasopharyngeal carcinoma cells. Specifically, different concentrations of Barlerin effectively inhibit the cell viability of highly metastatic human nasopharyngeal carcinoma cell lines 5-8F and CNE2 cells. Among them, 50 μM Barlerin effectively reduces the cell viability, proliferation ability, and migration ability of 5-8F cells and CNE2 cells.
[0010] Furthermore, the application of 8-O-acetylganoside methyl ester in the preparation of drugs for inhibiting the development of nasopharyngeal carcinoma, wherein the drug is a pharmaceutical composition, which is prepared into a clinically acceptable pharmaceutical formulation with 8-O-acetylganoside methyl ester as the main active ingredient and pharmaceutically acceptable excipients or auxiliary ingredients, wherein the content of 8-O-acetylganoside methyl ester in the pharmaceutical composition is typically 0.01% (w / w) to 95.0% (w / w).
[0011] Generally, pharmaceuticals are used clinically only after being formulated into a pharmaceutical composition. The pharmaceutical composition described in this invention can be prepared according to methods known in the art, and can be formulated into any dosage form suitable for human or animal use by combining the pharmaceutical composition of this invention with one or more pharmaceutically acceptable solid or liquid excipients and / or adjuvants.
[0012] Furthermore, the application of the 8-O-acetylganoside methyl ester in the preparation of drugs that inhibit the development of nasopharyngeal carcinoma, wherein the drug formulation includes both oral and injectable dosage forms.
[0013] Furthermore, in the application of the 8-O-acetylganoside methyl ester in the preparation of drugs that inhibit the development of nasopharyngeal carcinoma, the oral formulation is an oral capsule, and the injectable formulation is an intravenous injection.
[0014] The drug or drug composition described in this invention can be administered in unit dose form, and the route of administration can be enteric or non-enteric, such as oral, intravenous, intramuscular, subcutaneous, nasal, oral mucosa, eye, lung and respiratory tract, skin, vagina, rectum, etc.
[0015] Dosage forms can be liquid, solid, or semi-solid. Liquid dosage forms can include solutions (including true solutions and colloidal solutions), emulsions (including o / w, w / o, and double emulsions), suspensions, injections (including aqueous injections, powder injections, and infusions), eye drops, nasal drops, lotions, and liniments, etc.; solid dosage forms can include tablets (including regular tablets, enteric-coated tablets, lozenges, dispersible tablets, chewable tablets, effervescent tablets, and orally disintegrating tablets), capsules (including hard capsules, soft capsules, and enteric-coated capsules), granules, powders, microcapsules, pellets, suppositories, films, patches, aerosols, and sprays, etc.; semi-solid dosage forms can include ointments, gels, and pastes, etc.
[0016] The drug of this invention can be formulated into conventional formulations, sustained-release formulations, controlled-release formulations, targeted formulations, and various microparticle delivery systems. To formulate the drug of this invention into tablets, a wide range of excipients known in the art can be used, including diluents, binders, wetting agents, disintegrants, lubricants, and flow aids. Diluents can be starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, dicalcium phosphate, calcium carbonate, etc.; wetting agents can be water, ethanol, isopropanol, etc.; binders can be starch paste, dextrin, syrup, honey, glucose solution, microcrystalline cellulose, gum arabic paste, gelatin paste, sodium carboxymethyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, ethyl cellulose, acrylic resin, carbomer, polyvinylpyrrolidone, polyethylene glycol, etc.; disintegrants can be dry starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, croscarmellose, croscarmellose sodium carboxymethyl cellulose, sodium carboxymethyl starch, sodium bicarbonate and citric acid, polyoxyethylene sorbitol fatty acid ester, sodium dodecyl sulfonate, etc.; lubricants and flow aids can be talc, silica, stearate, tartaric acid, liquid paraffin, polyethylene glycol, etc.
[0017] Tablets can also be further processed into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or bilayer and multilayer tablets.
[0018] To formulate the drug delivery unit into capsules, the active ingredient of the present invention can be mixed with a diluent and a disintegrant, and the mixture can be placed directly into hard or soft capsules. Alternatively, the active ingredient of the present invention can be first formed into granules or microspheres with a diluent, binder, and disintegrant, and then placed into hard or soft capsules. The diluents, binders, wetting agents, disintegrants, and disintegrants used to prepare tablets of the present invention can also be used to prepare capsules of the present invention.
[0019] To prepare the drug of this invention into an injectable formulation, water, ethanol, isopropanol, propylene glycol, or mixtures thereof can be used as solvents, and appropriate amounts of commonly used solubilizers, co-solvents, pH adjusters, and osmotic pressure regulators can be added. Solubilizers or co-solvents can be poloxamer, lecithin, hydroxypropyl-β-cyclodextrin, etc.; pH adjusters can be phosphates, acetates, hydrochloric acid, sodium hydroxide, etc.; osmotic pressure regulators can be sodium chloride, mannitol, glucose, phosphates, acetates, etc. If preparing a lyophilized powder for injection, mannitol, glucose, etc., can also be added as a support agent.
[0020] In addition, colorants, preservatives, flavorings, tasters or other additives may be added to pharmaceutical preparations if necessary.
[0021] Compared with the prior art, the present invention has the following beneficial effects:
[0022] 1. The application of 8-O-acetylganoside methyl ester described in this invention in the preparation of drugs that inhibit the development of nasopharyngeal carcinoma. The research results of this invention show that different concentrations of Barlerin can effectively inhibit the cell viability of highly metastatic human nasopharyngeal carcinoma cell lines 5-8F and CNE2. Among them, 50 μM Barlerin can effectively reduce the cell viability of 5-8F cells (down to 83.57±7.44%, P<0.001) and the cell viability of CNE2 cells (down to 78.83±5.26%, P<0.001).
[0023] 2. The application of 8-O-acetylganoside methyl ester described in this invention in the preparation of drugs that inhibit the development of nasopharyngeal carcinoma. The EdU proliferation experiment results of this invention show that after intervention with 50 μM Barlerin, the proportion of EdU-positive 5-8F cells decreased from 26.16±1.76% to 18.72±1.25% (P<0.0001), and the proportion of EdU-positive CNE2 cells decreased from 46.54±1.71% to 29.96±2.53% (P<0.0001), indicating that Barlerin effectively inhibits the proliferation of human nasopharyngeal carcinoma cell lines.
[0024] 3. The application of 8-O-acetylganoside methyl ester described in this invention in the preparation of drugs that inhibit the development of nasopharyngeal carcinoma. The Transwell migration experiment results of this invention show that after treatment with different concentrations of Barlerin, the number of 5-8F cells that can cross the filter membrane is significantly reduced, and the number of 5-8F and CNE2 cells that can cross the filter membrane is significantly reduced, especially at 50 μM. The number of 5-8F cells decreased from 1064.67±57.50 cells to 787.33±69.29 cells (P<0.001), and the number of CNE2 cells decreased from 1091.67±69.47 cells to 575.67±44.06 cells (P<0.0001), indicating that Barlerin significantly inhibits the migration ability of nasopharyngeal carcinoma cells.
[0025] The above results indicate that Barlerin can effectively inhibit the cell viability, proliferation, and migration of nasopharyngeal carcinoma cells, suggesting that it can be an effective drug for inhibiting the occurrence and development of nasopharyngeal carcinoma. Attached Figure Description
[0026] Figure 1 This is a graph showing the inhibition of nasopharyngeal carcinoma cell activity by Barlerin in an experimental example of this invention;
[0027] Figure 2 This is a diagram showing that Barlerin inhibits the proliferation of nasopharyngeal carcinoma cells in an experimental example of this invention;
[0028] Figure 3 This is a diagram showing that Barlerin inhibits the migration of nasopharyngeal carcinoma cells in an experimental example of this invention. Detailed Implementation
[0029] The specific embodiments of the present invention will be further described below with reference to examples.
[0030] Example 1:
[0031] Application of 8-O-acetylganoside methyl ester in the preparation of drugs that inhibit the development of nasopharyngeal carcinoma:
[0032] According to existing process requirements, it is prepared into hard capsules.
[0033] Example 2:
[0034] Application of 8-O-acetylganoside methyl ester in the preparation of drugs that inhibit the development of nasopharyngeal carcinoma:
[0035] It is prepared into an injectable form according to existing process requirements.
[0036] Experimental example:
[0037] Experimental methods
[0038] 1. Cell culture:
[0039] Highly metastatic human nasopharyngeal carcinoma cell lines 5-8F and CNE2 were kindly provided by the research group of Academician Ma Jun at Sun Yat-sen University Cancer Center. The cells were cultured in RPMI 1640 medium (Gibco, USA) containing 10% fetal bovine serum (FBS, Gibco, USA) at 37°C in a cell culture incubator containing 5% CO2.
[0040] 2. Cell viability assay:
[0041] 5-8F and CNE2 cells were respectively fed with 2×10 3 Cells were seeded in 96-well plates at concentrations of 0, 20, 30, 40, 50, and 60 μM Barlerin (HY-N0758, MCE, China) and incubated at 37°C in a 5% CO2 incubator. After 48 hours, the culture medium was replaced with fresh medium. Cell viability was assessed using a Cell Count Kit-8 (CCK8, CK-04, Tongren, Japan). Cells were incubated with CCK8 reagent for 1 hour, and the absorbance at 450 nm was measured using a microplate reader. 450nm The experiment was repeated four times.
[0042] 3. EdU cell proliferation detection:
[0043] 5-8F and CNE2 cells were fed at a rate of 1×10⁻⁶. 5 Cells were seeded in 6-well plates, with 0, 30, 40, and 50 μM Barlerin added respectively. The plates were incubated at 37°C in a 5% CO2 incubator for 48 h. After 48 h, half of the original culture medium was discarded, and an equal volume of 2×EdU working solution (diluted with fresh culture medium) was added for 2 h. The culture medium was then removed, and the cells were fixed with 4% tissue cell fixative (Solepro, China) and permeabilized with 0.3% Triton X-100. 500 μL of Click reaction solution was added, and the cells were incubated at room temperature in the dark for 30 min. After washing, 1 mL of Hoechst 33342 staining solution was added for nuclear staining for 10 min. Cell proliferation was recorded under an inverted fluorescence microscope. EdU-positive cells were considered proliferating cells, and the ratio of EdU-positive cells to Hoechst-positive cells was the proliferation rate. The experiment was repeated four times.
[0044] 4. Cell migration experiment:
[0045] 5-8F and CNE2 cells were spaced at 5 × 10⁻⁶ cells per well. 4Cells were resuspended in 100 μL of serum-free RPMI 1640 medium and seeded into the upper layer of 8 μm pore size Transwell chambers. 600 μL of medium containing 10% FBS was added to the lower layer, and 0, 30, 40, and 50 μM Barlerin were added to the lower layer medium, respectively. After incubation in a 5% CO2 incubator at 37℃ for 18 h (5-8F cells) or 24 h (CNE2 cells), residual cells in the upper layer of the chamber were carefully wiped off with a cotton swab. The chambers were fixed in 4% tissue cell fixative for 15 min, stained with 1% crystal violet (Solepro, China) for 15 min, washed with PBS, and the number of cells after penetration was recorded under an inverted microscope. The experiment was repeated three times.
[0046] Experimental results
[0047] 1. Barlerin inhibits the activity of nasopharyngeal carcinoma cells:
[0048] The CCK8 assay results showed that different concentrations of Barlerin significantly inhibited the viability of nasopharyngeal carcinoma 5-8F and CNE2 cells. Specifically, at a Barlerin concentration of 30 μM, the viability of both 5-8F and CNE2 cells decreased significantly. When the Barlerin concentration reached 50 μM, the inhibition of cell activity was most pronounced, with the relative viability of 5-8F and CNE2 cells decreasing to 83.57 ± 7.44% (P < 0.001) and 78.83 ± 5.26% (P < 0.001), respectively. Figure 1 A, Figure 1 B) indicates that Barlerin can effectively inhibit the activity of nasopharyngeal carcinoma cells.
[0049] Figure 1 The graph shows the cell viability of Barlerin in inhibiting nasopharyngeal carcinoma cells. A: Cell viability results of 5-8F cells treated with 0 μM, 20 μM, 30 μM, 40 μM, 50 μM, and 60 μM Barlerin, n=4; B: Cell viability results of CNE2 cells treated with 0 μM, 20 μM, 30 μM, 40 μM, 50 μM, and 60 μM Barlerin, n=4. , ;
[0050] 2. Barlerin inhibits the proliferation of nasopharyngeal carcinoma cells:
[0051] Results from the EdU cell proliferation assay kit showed that different concentrations of Barlerin significantly inhibited the proliferation of nasopharyngeal carcinoma 5-8F and CNE2 cells. When the Barlerin concentration reached 30 μM, the proportion of EdU-positive 5-8F and CNE2 cells was significantly reduced. When the Barlerin concentration reached 50 μM, the proportions of EdU-positive 5-8F and CNE2 cells reached their lowest levels, with 5-8F cells decreasing from 26.16±1.76% to 18.72±1.25% (P<0.0001) and CNE2 cells decreasing from 46.54±1.71% to 29.96±2.53% (P<0.0001). Figure 2 A- Figure 2 D). This result indicates that Barlerin can significantly inhibit the proliferation of nasopharyngeal carcinoma cells.
[0052] Figure 2 The diagram shows the inhibition of nasopharyngeal carcinoma cell proliferation by Barlerin. A: EdU staining results of 5-8F cells treated with 0 μM, 30 μM, 40 μM, and 50 μM Barlerin; B: Statistical results of the proportion of EdU-positive cells after treatment with 5-8F cells; C: EdU staining results of CNE2 cells treated with 0 μM, 30 μM, 40 μM, and 50 μM Barlerin; D: Statistical results of the proportion of EdU-positive cells after treatment with CNE2 cells. Red fluorescence (EdU-positive) represents proliferating cells, blue fluorescence (Hoechst-positive) represents total cells, and the ratio of red fluorescent cells to blue fluorescent cells is the proliferation rate (%). ;
[0053] 3. Barlerin inhibits the migration of nasopharyngeal carcinoma cells:
[0054] Cell migration ability was detected using Transwell chambers. Results showed that after treatment of nasopharyngeal carcinoma cells with different concentrations of Barlerin, treatment with 30 μM Barlerin significantly reduced the number of 5-8F and CNE2 cells that could cross the filter membrane. When the Barlerin concentration reached 50 μM, the number of 5-8F and CNE2 cells crossing the filter membrane reached its lowest point, with the 5-8F cell number decreasing from 1064.67 ± 57.50 cells to 787.33 ± 69.29 cells (P < 0.001). Figure 3 A, Figure 3 B); The number of CNE2 cells decreased from 1091.67±69.47 to 575.67±44.06 cells, P<0.0001) Figure 3 C, Figure 3(D); This result indicates that Barlerin significantly inhibits the migration ability of nasopharyngeal carcinoma cells.
[0055] Figure 3 The diagram shows the inhibition of nasopharyngeal carcinoma cell migration by Barlerin. A: Cell migration assay results after treatment of 5-8F cells with 0 μM, 30 μM, 40 μM, and 50 μM Barlerin; B: 5-8F cell count results after migration; C: CNE2 cell migration assay results after treatment with 0 μM, 30 μM, 40 μM, and 50 μM Barlerin; D: CNE2 cell count results after migration. .
[0056] 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
Application of 1,8-O-acetylganoside methyl ester in the preparation of drugs that inhibit the development of nasopharyngeal carcinoma.
2. The use of 8-O-acetylganoside methyl ester according to claim 1 in the preparation of a drug for inhibiting the development of nasopharyngeal carcinoma, characterized in that: 8-O-acetylganoside methyl ester effectively inhibits the cell viability, proliferation, and migration of nasopharyngeal carcinoma cells.
3. The use of 8-O-acetylganoside methyl ester according to claim 2 in the preparation of a drug for inhibiting the development of nasopharyngeal carcinoma, characterized in that: The 8-O-acetylgenin methyl ester effectively inhibits the cell viability of nasopharyngeal carcinoma cells. Different concentrations of 8-O-acetylgenin methyl ester effectively inhibit the cell viability of highly metastatic human nasopharyngeal carcinoma cell lines 5-8F and CNE2. Among them, 50 μM 8-O-acetylgenin methyl ester effectively reduces the cell viability, proliferation ability and migration ability of 5-8F cells and CNE2 cells.
4. The use of 8-O-acetylganoside methyl ester according to any one of claims 1-3 in the preparation of a drug for inhibiting the development of nasopharyngeal carcinoma, characterized in that: The drug is a pharmaceutical composition, which is prepared into a clinically acceptable pharmaceutical formulation with 8-O-acetylganoside methyl ester as the main active ingredient and pharmaceutically acceptable excipients or auxiliary ingredients. The content of 8-O-acetylganoside methyl ester in the pharmaceutical composition is usually 0.01% w / w-95.0% w / w.
5. The use of 8-O-acetylganoside methyl ester according to claim 4 in the preparation of a drug for inhibiting the development of nasopharyngeal carcinoma, characterized in that: The pharmaceutical preparations include two dosage forms: oral preparations and injectable preparations.
6. The use of 8-O-acetylganoside methyl ester according to claim 5 in the preparation of a drug for inhibiting the development of nasopharyngeal carcinoma, characterized in that: The oral preparation is an oral capsule, and the injectable preparation is an intravenous injection solution.