Traditional Chinese medicine composition for treating nasopharynx cancer and preparation method of nano freeze-dried tablet of traditional Chinese medicine composition

Abstract

The invention discloses a traditional Chinese medicine composition for treating nasopharynx cancer and a preparation method of a nano freeze-dried tablet of the traditional Chinese medicine composition. The traditional Chinese medicine composition is characterized by being prepared from the following raw materials in parts by weight: 25-35 parts of astragalus membranaceus, 25-35 parts of ganoderma lucidum spore powder, 25-35 parts of centipeda minima, 25-35 parts of cortex juglandis mandshuricae, 8-12 parts of dried toad skin, 2-4 parts of centipede, 1-3 parts of gecko, 10-20 parts of ligusticum wallichii, 10-20 parts of sculellaria barbata, 25-35 parts of oldenlandia diffusa, 10-20 parts of paris polyphylla, 5-10 parts of liquorice, 8-12 parts of magnolia flower and 5-10 parts of fresh ginger. The nasopharynx cancer can be treated through the traditional Chinese medicine, so that the defects of operative treatment, radiotherapy and chemical treatment are overcome, and the treatment effect is improved.

Description

Technical Field

[0001] This invention relates to the field of biopharmaceutical technology, and in particular to a traditional Chinese medicine composition for treating nasopharyngeal carcinoma and a method for preparing its nano-lyophilized tablets. Background Technology

[0002] Nasopharyngeal carcinoma (NPC) is one of the most common malignant tumors in my country, originating from the epithelial tissue of the nasopharynx. It accounts for 78.08% of head and neck malignancies and 92.99% of upper respiratory tract cancers. NPC originates in the nasopharyngeal mucosa and is characterized by its hidden primary site, making early detection difficult. It also exhibits poor pathological differentiation, high malignancy, and a tendency for invasive growth and early metastasis. Male patients are 2-3 times more likely to be affected than female patients, and the disease typically develops after age 40. Clinical symptoms include bloody nasal discharge, epistaxis, nasal congestion, tinnitus, hearing loss, headache, double vision, and neck masses. Early detection and diagnosis are crucial for the treatment of this disease.

[0003] Currently, radiotherapy is the preferred treatment for nasopharyngeal carcinoma, but traditional Chinese medicine is also an important means of treating cancer. It can make up for the shortcomings of surgery, radiotherapy, and chemotherapy, and can both consolidate the effects of radiotherapy and chemotherapy and eliminate the toxic side effects of radiotherapy and chemotherapy. Summary of the Invention

[0004] The purpose of this invention is to provide a traditional Chinese medicine composition for treating nasopharyngeal carcinoma and a method for preparing its nano-lyophilized tablets. This method can treat nasopharyngeal carcinoma with traditional Chinese medicine, thereby compensating for the shortcomings of surgical treatment, radiotherapy, and chemotherapy, and thus improving the treatment effect.

[0005] The above-mentioned technical objective of the present invention is achieved through the following technical solution: A traditional Chinese medicine composition for treating nasopharyngeal carcinoma is made from the following raw materials in parts by weight: Astragalus membranaceus 25-35 parts, Ganoderma lucidum spore powder 25-35 parts, Centipeda minima 25-35 parts, Evodia rutaecarpa 25-35 parts, Toad skin 8-12 parts, Scolopendra subspinipes 2-4 parts, Gastrodia elata 1-3 parts, Ligusticum chuanxiong 10-20 parts, Scutellaria barbata 10-20 parts, Hedyotis diffusa 25-35 parts, Paris polyphylla 10-20 parts, Glycyrrhiza uralensis 5-10 parts, Magnolia biondii 8-12 parts, and Zingiber officinale 5-10 parts.

[0006] This traditional Chinese medicine composition is reconstructed according to the principles of principal, adjuvant, and guide herbs: Principal drug group (antiviral core) Astragalus 30g: Polysaccharides activate the TLR4 / NF-κB pathway and enhance NK cell activity; 30g of Ganoderma lucidum spore powder: Ganoderic acid D inhibits EB virus nuclear antigen EBNA1; Drug group (tumor killing) 30g of *Centipeda minima*: Sesquiterpene lactones downregulate the LMP1 carcinogenic protein; 10g of dried toad skin: Bufotoxin selectively induces apoptosis in EBV⁺ cells; Adjuvant drug group (synergistic effect) Three centipedes + two dragonfish: Peptides inhibit VEGF-R2 phosphorylation (anti-angiogenic activity EC50=85nM); Scutellaria barbata 15g + Hedyotis diffusa 30g: Flavonoid glycosides synergistically block the STAT3 signaling pathway; Paris polyphylla 15g + 30g of gentian root: Steroidal saponins inhibit DNMT1 methyltransferase (a key target of epigenetic regulation); Drug group (attenuated targeted therapy) Chuanxiong 15g + Xinyi 10g: Increases nasal mucosal permeability; Licorice 6g + Ginger 8g: β-glucuronidase inhibition reduces toad venom hepatotoxicity.

[0007] Further: It is made from the following ingredients in parts by weight: Astragalus membranaceus 30 parts, Ganoderma lucidum spore powder 30 parts, Centipeda minima 28 parts, Eriocaulon buergerianum 25 parts, dried toad skin 10 parts, centipede 3 parts, Gynostemma pentaphyllum 2 parts, Ligusticum chuanxiong 15 parts, Scutellaria barbata 15 parts, Hedyotis diffusa 30 parts, Paris polyphylla 15 parts, Glycyrrhiza uralensis 7 parts, Magnolia biondii 10 parts, and fresh ginger 6 parts.

[0008] Furthermore, it can be formulated into sustained-release oral nano-lyophilized tablets and immediate-release nasal nano-lyophilized tablets.

[0009] This invention also provides a method for preparing lyophilized nanosheets for treating nasopharyngeal carcinoma, comprising the following steps: Step 1: Processing of plant medicinal materials: Grind the materials to 40 mesh, add 8 times the amount of citrate buffer solution with a pH of 5.0, mix, add compound enzyme for enzymatic hydrolysis, then extract at 50℃ with shaking for 2 hours, inactivate the enzyme, centrifuge and concentrate to obtain the supernatant. Step 2, Animal medicinal materials processing: Animal medicinal materials are flash-frozen with liquid nitrogen and then ultra-finely pulverized. Then, they are suspended in 0.1 mol / L Tris-HCl buffer, and trypsin with an enzyme-to-substrate ratio of 1:25 is added and hydrolyzed at 37°C for 4 hours. After the enzyme is inactivated by boiling water bath, the materials are freeze-dried. Step 3: Safeguarding toxic components: The dried toad skin extract and β-cyclodextrin in a molar ratio of 1:6 were ground together and stirred in a saturated aqueous solution at 45°C for 12 hours to form an inclusion complex, which was then crystallized and dried. Step 4, Carrier preparation: Dissolve 0.5% w / v chitosan in 1% acetic acid solution, dissolve 0.8% w / v sodium alginate in ultrapure water, and add the sodium alginate solution dropwise to the chitosan solution under magnetic stirring to form positively charged nanoparticles with a particle size of 102±15nm and a Zeta potential of +28.4mV. Step 5: Loading active ingredients: Water-soluble component loading: Astragalus polysaccharide + Centipeda lactone mixture and nanoparticles were incubated at 4°C for 12 hours; Lipid-soluble component loading: Bufotoxin + Ganoderma lucidum triterpenes are dissolved in ethanol and injected into the hydrophobic core of nanoparticles; Step Six: Lyophilized Targeted Tablet Formation Drug-liquid mixing: The nano suspension is mixed with the concentrated aqueous extract of Paris saponins and other ingredients, and a freeze-drying protectant is added; Step 7, Zoned Mold Filling: Create special zoned molds for oral and nasal drug release areas, then inject the mixed drug solution into the special molds, pre-freeze at -40℃ for 4 hours, and then vacuum freeze-dry at -25℃ and 10Pa for 24 hours to obtain nano-lyophilized tablets, packaged in aluminum-plastic blister packs, and stored in an environment with relative humidity <30%.

[0010] Furthermore: In step one, the complex enzyme consists of 2000 U / g cellulase + 1500 U / g pectinase.

[0011] Furthermore: In step two, the Tris-HCl buffer contains 5 mM CaCl2.

[0012] Furthermore: In step six, the freeze-drying protectant is composed of mannitol and trehalose in a ratio of 3:1.

[0013] Furthermore: In step seven, the oral delivery zone of the specially designed mold contains pH-sensitive hydroxypropyl methylcellulose, and the nasal delivery zone contains chitosan adhesive.

[0014] Furthermore: In step seven, each nano-lyophilized tablet contains 1.2g of raw medicinal material, the diameter of the nano-lyophilized tablet in the oral cavity is 8mm, and the diameter of the nano-lyophilized tablet in the nasal cavity is 3mm.

[0015] In summary, the present invention has the following beneficial effects: Firstly, this invention can treat nasopharyngeal carcinoma with traditional Chinese medicine, thereby compensating for the shortcomings of surgical treatment, radiotherapy, and chemotherapy, and thus improving the treatment effect; Secondly, this invention utilizes a nano-lyophilization integrated process to significantly improve the drug loading rate of fat-soluble components. Attached Figure Description

[0016] Figure 1 This is a process flow diagram of the present invention. Detailed Implementation

[0017] The present invention will be further described in detail below with reference to the accompanying drawings.

[0018] In the description of this invention, it should be understood that the terms "upper", "lower", "left", "right", "front", "rear", "inner", "outer", etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. They are only for the convenience of describing this invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on this invention.

[0019] Example, refer to Figure 1 A traditional Chinese medicine composition for treating nasopharyngeal carcinoma is made from the following raw materials in parts by weight: Astragalus membranaceus 30 parts, Ganoderma lucidum spore powder 30 parts, Centipeda minima 28 parts, Evodia rutaecarpa 25 parts, Toad skin 10 parts, Scolopendra subspinipes 3 parts, Gastrodia elata 2 parts, Ligusticum chuanxiong 15 parts, Scutellaria barbata 15 parts, Hedyotis diffusa 30 parts, Paris polyphylla 15 parts, Glycyrrhiza uralensis 7 parts, Magnolia biondii 10 parts, and Zingiber officinale 6 parts.

[0020] This traditional Chinese medicine composition is reconstructed according to the principles of principal, adjuvant, and guide herbs: Principal drug group (antiviral core) Astragalus membranaceus 30 parts: Polysaccharides activate the TLR4 / NF-κB pathway and enhance NK cell activity; 30 portions of Ganoderma lucidum spore powder: Ganoderic acid D inhibits EB virus nuclear antigen EBNA1; Drug group (tumor killing) 30 portions of *Centipeda minima*: Sesquiterpene lactones downregulated the LMP1 oncogenic protein; 10 portions of dried toad skin: Bufotoxin selectively induced apoptosis in EBV⁺ cells; Adjuvant drug group (synergistic effect) Centipede 3 parts + Tianlong 2 parts: Peptide inhibits VEGF-R2 phosphorylation (anti-angiogenic activity EC50=85nM); 15 parts of Scutellaria barbata + 30 parts of Hedyotis diffusa: Flavonoids synergistically block the STAT3 signaling pathway; 15 samples of Paris polyphylla + 30 samples of Erythrina variegata: Steroidal saponins inhibit DNMT1 methyltransferase (a key target of epigenetic regulation). Drug group (attenuated targeted therapy) Ligusticum chuanxiong 15 parts + Magnolia biondii 10 parts: Increases nasal mucosal permeability; Licorice 6 parts + ginger 8 parts: β-glucuronidase inhibition reduces toad venom hepatotoxicity.

[0021] It can be made into sustained-release oral nano-lyophilized tablets and immediate-release nasal nano-lyophilized tablets.

[0022] A method for preparing lyophilized nanoparticles for treating nasopharyngeal carcinoma includes the following steps: Step 1: Processing of medicinal plants: Grind Astragalus membranaceus, Centipeda minima, Scutellaria barbata, Hedyotis diffusa, Paris polyphylla, and Glycyrrhiza uralensis into powder to 40 mesh. Add 8 times the amount of citrate buffer with a pH of 4.8-5.2. After mixing, add a compound enzyme consisting of 2000 U / g cellulase and 1500 U / g pectinase for enzymatic hydrolysis. The endpoint of the compound enzymatic hydrolysis is controlled to have a reducing sugar content ≥16 mg glucose equivalent / g of medicinal material. Then, extract at a constant temperature of 48-52℃ with shaking for 1.5-2.5 hours. After enzyme inactivation, centrifuge and concentrate to obtain the supernatant. Step 2, Animal medicinal material processing: Centipede, dragon, and dried toad skin are flash-frozen with liquid nitrogen and then ultra-finely pulverized to a particle size ≤50μm. Then, they are suspended in Tris-HCl buffer containing 0.1mol / L and 5mM CaCl2, and trypsin with an enzyme-to-substrate ratio of 1:25 is added and hydrolyzed at 36-38℃ for 3.5-4.5 hours. After inactivating the enzyme in a boiling water bath, they are freeze-dried. Step 3: Safeguarding toxic components: The dried toad skin extract and β-cyclodextrin in a molar ratio of 1:6 are ground together and stirred in a saturated aqueous solution at 42-48℃ for 10-14 hours to form an inclusion complex, which is then crystallized and dried. Step 4, Carrier preparation: Dissolve 0.5% w / v chitosan in 1% acetic acid solution, dissolve 0.8% w / v sodium alginate in ultrapure water, and add the sodium alginate solution dropwise to the chitosan solution under magnetic stirring to form positively charged nanoparticles with a particle size of 102±15nm and a Zeta potential of +28.4mV. Step 5: Loading active ingredients: Water-soluble component loading: Astragalus polysaccharide + Centipeda lactone mixture and nanoparticles were incubated at 4°C for 12 hours; Lipid-soluble component loading: Bufotoxin + Ganoderma lucidum triterpenes are dissolved in ethanol and injected into the hydrophobic core of nanoparticles; Step Six: Lyophilized Targeted Tablet Formation Drug-liquid mixing: The nano suspension is mixed with the concentrated aqueous extract of Paris saponins and other ingredients, and a freeze-drying protectant composed of mannitol and trehalose in a ratio of 3:1 is added. Step 7, Zoned Mold Injection: Create special zoned molds for oral and nasal drug release areas. The oral drug release area of ​​the special mold contains pH-sensitive hydroxypropyl methylcellulose, and the nasal drug release area contains chitosan adhesive. Next, the mixed medicine solution is injected into a special mold, and then pre-frozen at -40℃ for 4 hours. Then, it is vacuum freeze-dried at -25℃ and 10Pa for 24 hours to obtain nano-freeze-dried tablets. Each nano-freeze-dried tablet contains 1.2g of raw medicine and is packaged in aluminum-plastic blister packs and stored in an environment with relative humidity <30%. Among them, the oral drug release area has a diameter of 7-9 mm and contains a pH-sensitive polymer, which can be released slowly in an environment with pH ≥ 6.5; Nasal drug release zone: 2.5-3.5 mm in diameter, containing mucosal adhesive, with a drug release rate of ≥35% in nasal fluid within 30 minutes.

[0023] Key data 1. Quality Standards: Astragaloside A content: 1.62 mg / g (UPLC-PDA detection); Limit of bufotalin: 0.95±0.12 mg / g (determined by HPLC-MS / MS); In vitro dissolution (simulated nasal fluid): 43.2% ± 2.1% drug release in 30 minutes.

[0024] 2. Efficacy verification: In vitro: Downregulates EBNA1 expression in CNE-2 cells by 72.3% (qPCR) and inhibits migration rate by >80%.

[0025] 3. Quality controllability Establish process quality control points: The amount of reducing sugar at the end of enzymatic hydrolysis (to control extraction efficiency); PDI of nanoparticles ≤ 0.25 (to ensure uniform dispersion); The residual oxygen content after freeze-drying is ≤0.5% (to prevent oxidation of components); This process has been verified through three batches of pilot tests: The astragaloside A content RSD was 2.1%. Tablet hardness: 4.5 ± 0.3 kp; Dissolution factor f2 in the nasal cavity region > 85 (batch consistency).

[0026] 4. Finished Product Specifications: Appearance: White porous tablets (honeycomb pattern in the oral cavity area / dense pattern in the nasal cavity area); Moisture content ≤3% (determined by Karl Fischer method); In vitro drug release characteristics (flow cell method): Nasal area: ≥40% drug release in 30 minutes (simulated nasal fluid); Oral region: ≥80% cumulative drug release over 8 hours (pH 6.8 phosphate buffer).

[0027] This specific embodiment is merely an explanation of the present invention and is not intended to limit the invention. After reading this specification, those skilled in the art can make inventive modifications to this embodiment as needed, but as long as they are within the scope of the claims of the present invention, they are protected by patent law.

Claims

1. A traditional Chinese medicine composition for treating nasopharyngeal carcinoma, characterized in that, It is made from the following raw materials by weight: 25-35 parts of Huangqi, 25-35 parts of Lingzhi Spore Powder, 25-35 parts of Enotus, 25-35 parts of Qinglongyi, 8-12 parts of dry toad skin, 2-4 parts of centipede, 1-3 parts of Tianlong, 10-20 parts of Chuanxiong, 10-20 parts of Banzhi Lian, 25-35 parts of Baihua Sheche, 10-20 parts of Chonglou, 5-10 parts of Gancao, 8-12 parts of Xinyi, and 5-10 parts of Shengjiang.

2. The Chinese medicine composition for treating nasopharyngeal carcinoma according to claim 1, characterized in that, It is made from the following raw materials by weight: 30 parts of Huangqi, 30 parts of Lingzhi Spore Powder, 28 parts of Enotus, 25 parts of Qinglongyi, 10 parts of dry toad skin, 3 parts of centipede, 2 parts of Tianlong, 15 parts of Chuanxiong, 15 parts of Banzhi Lian, 30 parts of Baihua Sheche, 15 parts of Chonglou, 7 parts of Gancao, 10 parts of Xinyi, and 6 parts of Shengjiang.

3. The Chinese medicine composition for treating nasopharyngeal carcinoma according to any one of claims 1-2, characterized in that: It can be prepared as a sustained-release oral nanometer freeze-dried tablet and a quick-release nasal nanometer freeze-dried tablet.

4. The method of claim 1, wherein the method is a method of preparing a nano-lyophilized tablet for treating nasopharyngeal carcinoma. It comprises the following steps: Step one, processing of plant medicines: grinding to 40 mesh, adding 8 times the amount of citric acid buffer solution with a pH value of 5.0, mixing, then adding compound enzymes for enzymatic hydrolysis, followed by constant temperature oscillation extraction at 50°C for 2 hours, enzyme inactivation, centrifugal concentration, and taking the supernatant; Step two, processing of animal medicines: using liquid nitrogen to quickly freeze animal medicines, then performing ultrafine grinding, then suspending in 0.1 mol / L Tris-HCl buffer solution, adding trypsin with an enzyme substrate ratio of 1:25, hydrolyzing at 37°C for 4 hours, boiling water bath enzyme inactivation, and freeze-drying; Step three, safety of toxic ingredients: co-grinding the dry toad skin extract and β-cyclodextrin with a molar ratio of 1:6, stirring in a 45°C saturated aqueous solution for 12 hours to form an inclusion compound, and drying after crystallization; Step four, preparation of carriers: dissolving 0.5% w / v chitosan in 1% acetic acid solution, dissolving 0.8% w / v sodium alginate in ultrapure water, and under magnetic stirring, dropping the sodium alginate solution into the chitosan solution to form positively charged nanoparticles with a particle size of 102±15 nm and a Zeta potential of +28.4 mV; Step five, loading of active ingredients: Water-soluble ingredient loading: incubating the Huangqi polysaccharide + Enotus lactone mixed solution with nanoparticles at 4°C for 12 hours; Fat-soluble ingredient loading: dissolving bufalin + Lingzhi triterpenes in ethanol and injecting into the hydrophobic core of nanoparticles; Step six, formation of freeze-dried targeted tablets Mixing of medicinal liquid: mixing the nanometer suspension with the concentrated water extract of Chonglou saponin and adding a freeze-drying protectant; Step seven, zoned mold filling: making special zoned molds for oral and nasal drug release zones, then injecting the mixed medicinal liquid into the special molds, pre-freezing at -40°C for 4 hours, vacuum freeze-drying at -25°C and 10 Pa for 24 hours to obtain nanometer freeze-dried tablets, aluminum plastic blister packaging, and storage in an environment with a relative humidity of <30%.

5. The method of claim 4, wherein the method is for preparing a nano lyophilized tablet for treating nasopharyngeal carcinoma. In step one, the composition of the compound enzymes is cellulase 2000 U / g + pectinase 1500 U / g.

6. The method of claim 4, wherein the method is for preparing a nano lyophilized tablet for treating nasopharyngeal carcinoma. In step two, the Tris-HCl buffer solution contains 5 mM CaCl2.

7. The method of claim 4, wherein the method is for preparing a nano lyophilized tablet for treating nasopharyngeal carcinoma. In step six, the freeze-drying protectant is composed of mannitol: trehalose = 3:

1.

8. The method of claim 4, wherein the method is for preparing a nano lyophilized tablet for treating nasopharyngeal carcinoma. In step seven, the mouth drug release area of the special mold contains pH-sensitive hydroxypropyl methyl cellulose, and the nasal drug release area contains chitosan adhesive.

9. The method of claim 4, wherein the method is for preparing a nano lyophilized tablet for treating nasopharyngeal carcinoma. In step seven, the nano lyophilized tablet contains 1.2g of crude drug per tablet, the diameter of the nano lyophilized tablet in the oral cavity area is 7-9mm, and the diameter of the nano lyophilized tablet in the nasal cavity area is 2.5-3.5mm.

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