CN108743813B - Traditional Chinese medicine nano-emulsion for resisting nasopharyngeal carcinoma - Google Patents
Traditional Chinese medicine nano-emulsion for resisting nasopharyngeal carcinoma Download PDFInfo
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- CN108743813B CN108743813B CN201810920190.7A CN201810920190A CN108743813B CN 108743813 B CN108743813 B CN 108743813B CN 201810920190 A CN201810920190 A CN 201810920190A CN 108743813 B CN108743813 B CN 108743813B
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Abstract
The invention discloses an anti-nasopharyngeal carcinoma traditional Chinese medicine nano-emulsion. The traditional Chinese medicine nano-emulsion for treating nasopharyngeal carcinoma comprises the following traditional Chinese medicine raw materials: herba Gynostemmatis, herba Dendrobii, herba Berchemiae Lineatae, herba Pteridis Multifidae, Oletum Trogopterori, resina Toxicodendri, herba Selaginellae Doederleinii, radix Sophorae Tonkinensis, rhizoma Bolbostematis, Scolopendra, flos Magnoliae, radix Ophiopogonis, rhizoma Pinelliae Preparata, Glycyrrhrizae radix, herba Solani Lyrati, and herba Hedyotidis Diffusae. The traditional Chinese medicine nano-emulsion for resisting nasopharyngeal carcinoma is extracted after carrying out wet superfine grinding on traditional Chinese medicine raw materials, is more thoroughly extracted, has more and more complete active ingredients, is prepared by taking chitosan derivatives as carriers, directly reaches focuses, and improves curative effects. Compared with the prior art, the traditional Chinese medicine nano-emulsion for treating nasopharyngeal carcinoma can make up the defects of operative treatment, radiotherapy and chemotherapy, not only can consolidate the effects of radiotherapy and chemotherapy, but also can eliminate the toxic and side effects of radiotherapy and chemotherapy.
Description
Technical Field
The invention belongs to the field of traditional Chinese medicines, and particularly relates to a traditional Chinese medicine nanoemulsion for resisting nasopharyngeal carcinoma.
Background
Nasopharyngeal carcinoma is the most common malignant tumor of head and neck which is better to be found in the nasopharynx, the malignant degree is high, the onset is hidden, no obvious symptom is caused in the early stage, and the enlargement of cervical lymph nodes and the metastasis of organs of the whole body are caused in the late stage. The etiology of the disease includes genetic susceptibility, dietary environment, EB virus infection and the like, the canceration process is relatively long, and various oncogenes, cancer suppressor genes and interaction thereof are involved.
The basic causes of diseases mainly include the following aspects: the genetic factors comprise: 1. the disease has ethnic heterogeneity, more yellow than white, and higher incidence rate of nasopharyngeal carcinoma than local residents in Guangdong nationality or descendants and blood-mixed infants in foreign provinces and foreign countries, and the incidence rate of nasopharyngeal carcinoma in foreign countries is 40 times higher than that in local residents in men and 50 times higher than that in women. 2. There is a phenomenon of family aggregation. The nasopharyngeal carcinoma has obvious family aggregation phenomenon, more than 10 percent of patients with the nasopharyngeal carcinoma have family history, and the genetic susceptibility of the nasopharyngeal carcinoma can be an important factor of the onset of the nasopharyngeal carcinoma. 3. The immune genetic marker is found, and the HLA-A2 site of the A site in leukocyte antigen (HLA) is related to nasopharyngeal carcinogenesis.
The clinical symptoms mainly include the following aspects: 1. snivel of opium: in the early stage, blood may be carried in sputum after nose sucking, or in nasal discharge when blowing nose, or sometimes. In the late stage, there may be significant epistaxis. 2. Ear symptoms: when nasopharyngeal carcinoma occurs on the side wall of nasopharynx or the upper lip of the opening of the eustachian tube, unilateral tinnitus or hearing loss can occur when the tumor presses the eustachian tube, secretory otitis media, unilateral tinnitus or hearing loss, occlusion in the ear and the like can also occur, and early nasopharyngeal carcinoma symptoms can be possibly caused. 3. Headache: headache is a common symptom, and can be the first symptom or the only symptom, the headache part in the early stage is not fixed and is intermittent, the migraine in the later stage is persistent migraine, and the part is fixed. Early headache may be caused by neurovascular reflex or by stimulation of the first peripheral nerve of the trigeminal nerve, and late headache is often caused by tumor destruction of the cranial base or by cranial nerve involvement in the intracranial process. 4. Other symptoms: such as diplopia, facial numbness, nasal obstruction, cervical lymph node metastasis, tongue muscle atrophy and tongue extension deviation, eyelid drooping, eyeball fixation, hypopsia or disappearance, etc.
Radiation therapy has been the first choice for the treatment of nasopharyngeal carcinoma. But traditional Chinese medicine is also an important means for treating cancer. The traditional Chinese medicine in the prior art has poor treatment effect on nasopharyngeal carcinoma, cannot be well matched with radiotherapy, generates synergistic effect, and improves the effective rate of the nasopharyngeal carcinoma. The invention provides an anti-nasopharyngeal carcinoma traditional Chinese medicine nano-emulsion which can make up the defects of operative treatment, radiotherapy and chemotherapy, consolidate the effects of radiotherapy and chemotherapy and eliminate the toxic and side effects of radiotherapy and chemotherapy.
Disclosure of Invention
Aiming at the defects of the prior art. The purpose of the invention is realized by the following technology:
the invention aims to solve the technical problem of providing the traditional Chinese medicine nano-emulsion for resisting nasopharyngeal carcinoma.
An anti-nasopharyngeal carcinoma traditional Chinese medicine nano-emulsion comprises the following traditional Chinese medicine raw materials: herba Gynostemmatis, herba Dendrobii, herba Berchemiae Lineatae, herba Pteridis Multifidae, Oletum Trogopterori, resina Toxicodendri, herba Selaginellae Doederleinii, radix Sophorae Tonkinensis, rhizoma Bolbostematis, Scolopendra, flos Magnoliae, radix Ophiopogonis, rhizoma Pinelliae Preparata, Glycyrrhrizae radix, herba Solani Lyrati, and herba Hedyotidis Diffusae.
Further preferably, the traditional Chinese medicine nano-emulsion for treating nasopharyngeal carcinoma comprises the following traditional Chinese medicine raw materials in parts by weight: 8-12 parts of gynostemma pentaphylla, 4-15 parts of dendrobium, 13-17 parts of berchemia lineate, 4-15 parts of Chinese brake herb, 8-12 parts of trogopterus dung, 1-5 parts of dried lacquer, 1-6 parts of selaginella doederleinii, 1-8 parts of vietnamese sophora root, 4-15 parts of rhizoma bolbostemmae, 1-5 parts of centipede, 5-20 parts of biond magnolia flower, 10-20 parts of radix ophiopogonis, 5-20 parts of rhizoma pinellinae praeparata, 5-20 parts of liquorice, 4-15 parts of solanum lyratum and 4-16 parts of oldenlandia diffusa.
Gynostemma pentaphylla is sweet and bitter in taste and slightly cold in nature, has the effects of resisting and preventing cancer, inhibiting and killing cancer cells, enhancing the activity of lymphocytes in human blood, enhancing the immune function of human body, diminishing inflammation, resisting intestinal ulcer, gastric ulcer, swelling and ulcer and the like; herba Dendrobii has effects and effects of benefiting stomach, promoting fluid production, nourishing yin, clearing heat, and treating stomach heat and yin deficiency, and can be combined with rhizoma Pinelliae Preparata, radix Ophiopogonis, and Glycyrrhrizae radix to enhance therapeutic effect.
Oletum Trogopterori, resina Toxicodendri and herba Selaginellae Doederleinii have effects of promoting blood circulation, removing blood stasis, relieving swelling and pain; radix Sophorae Tonkinensis, rhizoma Bolbostematis, and Scolopendra have effects of clearing away heat and toxic materials and resisting cancer.
Radix ophiopogonis: sweet in taste, slightly bitter in flavor and slightly cold in nature. It enters heart, lung and stomach meridians. Has the functions of nourishing yin, promoting the production of body fluid, moistening lung and clearing away heart-fire; can be used for treating dry cough due to lung dryness. Cough due to consumptive disease, thirst due to body fluid consumption, insomnia due to vexation, internal heat, diabetes, constipation due to intestinal dryness.
Rhizoma pinelliae preparata: pungent flavor and warm nature. It enters spleen, stomach and lung meridians. It can eliminate dampness and phlegm. Can be used for treating cough and asthma due to excessive phlegm, dizziness and palpitation due to phlegm and fluid retention, dizziness due to wind-phlegm, phlegm syncope and headache.
Licorice root: sweet in flavor and neutral in nature. It enters heart, lung, spleen and stomach meridians. Has the functions of invigorating spleen and replenishing qi, clearing away heat and toxic material, eliminating phlegm and stopping cough, relieving spasm and pain and harmonizing the drugs. Can be used for treating weakness of spleen and stomach, asthenia, palpitation, short breath, cough, excessive phlegm, abdominal and limb spasm, pain, carbuncle, swelling, sore, and drug toxicity and strong nature.
Berchemia lineate, slightly bitter and flat, nontoxic, can remove blood stasis and blood, dispel wind-damp and eliminate swelling and toxin;
the Chinese brake herb is light and slightly bitter in taste and cold in nature, clears heat and promotes diuresis, cools blood and stops bleeding, and reduces swelling and detoxifies;
and (3) magnolia flower: pungent and slightly bitter with warm nature. It enters lung and stomach meridians. Has the functions of dispelling wind and cold and dredging nasal orifices. Can be used for treating headache due to wind-cold evil, nasal obstruction, nasosinusitis, and thick nasal discharge.
Solanum lyratum Thunb has effects of clearing heat, promoting diuresis, removing toxic substance, relieving swelling, and resisting cancer; the Chinese pulsatilla root has the effects of clearing away heat and toxic materials, cooling blood, relieving dysentery, eliminating dampness and killing parasites; the oldenlandia heat-clearing and stasis-dissipating, carbuncle-eliminating and detoxifying.
The invention is researched on the basis of comprehensively knowing the basic pathogenesis of nasopharyngeal carcinoma, the nasopharyngeal carcinoma takes bloody rhinorrhea and neck tumor as main clinical manifestations, and takes the mixing of lung and stomach channel heat toxin and dampness as main pathogenesis. Under the guidance of the theory of traditional Chinese medicine, the traditional Chinese medicine composition has the advantages of good effect, quick response, obvious effect and quick response, and all the medicines are matched and complement each other to play the roles of strengthening body resistance and banking up root, tonifying qi and nourishing blood, activating blood and dissolving stasis, and clearing heat, detoxicating and resisting cancer.
The components of the Chinese herbal medicine selected by the invention meet the regulation of the Chinese medicine management law, the comprehensive effect of the Chinese herbal medicines is utilized to treat nasopharyngeal carcinoma, no toxic or side effect is caused to human bodies, no toxic reaction is caused by acute and chronic animal toxicity tests, main organs of the Chinese herbal medicine are not obviously changed, and the medicine is safe and reliable.
The invention also discloses a preparation method of the traditional Chinese medicine nano-emulsion for resisting nasopharyngeal carcinoma, which comprises the following steps:
a preparation method of traditional Chinese medicine nano-emulsion for resisting nasopharyngeal carcinoma comprises the following steps:
s1, respectively crushing the traditional Chinese medicine raw materials to 20-100 meshes, and mixing to obtain traditional Chinese medicine coarse powder;
s2, treating the traditional Chinese medicine coarse powder, water, ethanol and sodium polyacrylate for 10-30 min under wet superfine grinding to obtain traditional Chinese medicine slurry;
s3, refluxing and extracting the traditional Chinese medicine slurry at 60-80 ℃ for 1-3 h, centrifuging, evaporating under reduced pressure, and freeze-drying in vacuum to obtain traditional Chinese medicine active powder;
s4, adding the traditional Chinese medicine active powder into a 1.5-3 mg/mL n-hexanoyl carboxymethyl chitosan aqueous solution, and dialyzing in distilled water for 16-32 hours to obtain dialysate; taking out the dialyzate, adding a fluorocarbon material accounting for 1.5-3.2% of the dialyzate in mass and a stabilizer accounting for 0.8-1.6% of the dialyzate in mass into the dialyzate, stirring, introducing oxygen to saturate, maintaining the oxygen content in the solution to be 25-35 mg/L, then carrying out ultrasonic emulsification under the conditions of an ice bath at 0-4 ℃ and introducing oxygen to maintain the oxygen content in the solution to be 25-35 mg/L, centrifuging for 5-10 min, and taking out the lower-layer emulsion for sterilization.
Preferably, the preparation method of the traditional Chinese medicine nano-emulsion for resisting nasopharyngeal carcinoma comprises the following steps:
s1, crushing the traditional Chinese medicine raw materials to 20-100 meshes, weighing the crushed traditional Chinese medicine raw materials according to the proportion, and uniformly mixing to obtain traditional Chinese medicine coarse powder;
s2, treating the traditional Chinese medicine coarse powder, water, ethanol and sodium polyacrylate for 10-30 min under wet superfine grinding to obtain traditional Chinese medicine slurry; the mass ratio of the traditional Chinese medicine coarse powder to the water to the ethanol to the sodium polyacrylate is 1 (4-6) to (12-20) to (0.01-0.02);
s3, refluxing and extracting the traditional Chinese medicine slurry at 60-80 ℃ for 1-3 h, centrifuging, concentrating the obtained supernatant to 1/10-1/6 of the original volume by reduced pressure evaporation, and stopping evaporation to obtain a concentrated solution, wherein the concentrated solution is subjected to vacuum freeze drying to obtain traditional Chinese medicine active powder;
s4, preparing a hexanoyl carboxymethyl chitosan aqueous solution with the concentration of 1.5-3 mg/mL, adding traditional Chinese medicine active powder into the hexanoyl carboxymethyl chitosan aqueous solution, stirring and mixing uniformly, then filling into a dialysis bag, dialyzing in distilled water for 16-32 hours, wherein the liquid in the dialysis bag is dialysate; after dialysis, taking out dialysate, adding a fluorocarbon material with the mass of 1.5-3.2% of that of the dialysate and a stabilizer with the mass of 0.8-1.6% of that of the dialysate into the dialysate, uniformly stirring and mixing, introducing oxygen to saturate, maintaining the oxygen content in the solution to be 25-35 mg/L, then carrying out ultrasonic emulsification under the conditions of ice bath at 0-4 ℃ and oxygen introduction to maintain the oxygen content in the solution to be 25-35 mg/L, wherein the ultrasonic power is 20-30W, the time is 5-15 min, the ultrasonic process is started for 3s, closed for 1s, and repeated circulation, centrifuging for 5-10 min under the condition of 3500-0 r/min, discarding supernatant, taking out lower-layer emulsion, and sterilizing to obtain the traditional Chinese medicine nano emulsion for resisting nasopharyngeal carcinoma; the mass-volume ratio of the traditional Chinese medicine active powder to the hexanoyl carboxymethyl chitosan aqueous solution is 1 (5-20) g/mL.
Preferably, the reduced pressure evaporation concentration condition is that the absolute pressure is 0.080-0.09 MPa and the temperature is 50-60 ℃.
Preferably, the vacuum freeze drying condition is to control the thickness of the material to be 6-8 mm, set the pre-freezing temperature to be-28 to-22 ℃, maintain the sample for 1-3 hours after the temperature of the sample is reduced to-28 to-22 ℃, set the sublimation temperature to be 5-15 ℃, the analysis temperature to be 30-40 ℃, and the absolute pressure to be 15-35 Pa, and dry the sample for 20-30 hours.
Preferably, the cut-off molecular weight of the dialysis bag is 3500 Da.
Preferably, the fluorocarbon material is perfluoro-n-pentane and/or decafluoropentane.
Preferably, the fluorocarbon material is formed by mixing perfluoro-n-pentane and decafluoropentane according to a mass ratio of 1 (1-3).
The liquid fluorocarbon nano-droplet has the particle size less than 1 μm, so the liquid fluorocarbon nano-droplet can penetrate through the gap between tumor endothelial cells to reach the tumor cells, can escape phagocytosis of macrophages and removal of organs such as liver, spleen and the like, has a half-life period in vivo of several hours, is longer than the circulation time of microbubbles in vivo, and has a longer residence time in vivo. Common wrapping materials currently used for preparing liquid fluorocarbon nano-droplets are lipids and high molecular polymers. When lipid and high molecular polymer are used for preparation, organic solvent is needed to be used for dissolving the lipid and high molecular polymer, and residual organic solvent is easy to accumulate in organisms to generate toxicity, so that the nano liquid drops prepared from the lipid and high molecular polymer have safety risk in practical application.
The n-hexanoyl carboxymethyl chitosan added in the invention is a water-soluble substance, does not need to use an organic solvent during dissolution, and has no biological toxicity. Moreover, the n-hexanoyl carboxymethyl chitosan also has good biocompatibility, and the amphiphilic structure of the n-hexanoyl carboxymethyl chitosan enables the n-hexanoyl carboxymethyl chitosan to be subjected to self-assembly so as to wrap hydrophobic substances such as liquid fluorocarbon and the like, and can be used as a drug delivery carrier.
Preferably, the stabilizer is dipalmitoylphosphatidylcholine and/or palmitic acid.
More preferably, the stabilizer is formed by mixing dipalmitoyl phosphatidylcholine and palmitic acid according to a mass ratio of (2-4): 1.
The added stabilizer can well maintain the stability of the nano-emulsion, prevent the nano-droplets from coagulating and ensure the storage time and the curative effect of the anti-nasopharyngeal-cancer traditional Chinese medicine nano-emulsion.
Preferably, the sterilization is performed by irradiating for 10-30 min under the ultraviolet light with the wavelength of 200-280 nm.
Preferably, the particle size of the nano liquid drops in the traditional Chinese medicine nano emulsion for resisting nasopharyngeal carcinoma is 200-700 nm.
The invention has the beneficial effects that:
(1) the invention provides an anti-nasopharyngeal carcinoma traditional Chinese medicine nano-emulsion, which is prepared by extracting active ingredients of a traditional Chinese medicine after carrying out wet superfine grinding on natural traditional Chinese medicine raw materials, and then preparing the anti-nasopharyngeal carcinoma nano-emulsion with good biocompatibility through n-hexanoyl carboxymethyl chitosan-fluorocarbon nano-liquid drops, so that the nano-emulsion can directly reach a focus and improve the curative effect of the medicine.
(2) The stabilizer is added in the process of preparing the n-hexanoyl carboxymethyl chitosan-traditional Chinese medicine active ingredient-fluorocarbon nano emulsion, so that the stability of the nano emulsion can be maintained, and the stability and the using curative effect of the anti-nasopharyngeal-cancer traditional Chinese medicine nano emulsion are improved.
(3) The traditional Chinese medicine nano-emulsion for resisting nasopharyngeal carcinoma has a remarkable effect of inhibiting tumor cells, and is remarkable in treatment of nasopharyngeal carcinoma.
Detailed Description
In the examples, the sources of the raw materials are as follows:
gynostemma pentaphylla, dendrobium, berchemia lineate, Chinese brake herb, trogopterus dung, resina toxicodendri, selaginella doederleinii, subprostrate sophora, rhizoma bolbostemmae, centipede, biond magnolia flower, dwarf lilyturf tuber, rhizoma pinellinae praeparata, liquorice, solanum lyratum and oldenlandia diffusa are all conventional traditional Chinese medicinal materials sold in the market. The herba Gynostemmatis is native Shaanxi, herba Dendrobii is native Zhejiang, herba Berchemiae Giraldianae is native Sichuan, herba Pteridis Multifidae is native Yunnan, Oletum Trogopterori is native Anhui, resina Toxicodendri is native Fujian, herba Selaginellae is native Anhui, radix Sophorae Tonkinensis is native Guangxi, rhizoma Bolbostematis is native Shanxi, Scolopendra is native Hubei, flos Magnoliae is native Guangxi, radix Ophiopogonis is native Hubei, rhizoma Pinelliae Preparata is native Sichuan, radix Glycyrrhizae is native Gansu, Solanum dulcamara is native Hubei, and herba Hedyotidis Diffusae is native Jiangxi.
Sodium polyacrylate: CAS number: 9003-04-7, molecular weight of 2000-3000, food grade, Huaxing bioengineering Co., Ltd in Hunan century.
The preparation method of the n-hexanoyl carboxymethyl chitosan comprises the following steps: dissolving 0.50g of carboxymethyl chitosan in 10g of water, stirring for 12h until the carboxymethyl chitosan is completely dissolved, adding 10mL of methanol, stirring for 30min, adding 0.33mL of n-hexanoic anhydride under a rapid stirring state, and reacting for 12 h. Dialyzing the solution for 24h by using 25% by mass of ethanol aqueous solution, dialyzing for 24h by using pure water, and carrying out vacuum freeze drying on the dialysate to finally obtain the n-hexanoyl carboxymethyl chitosan; the vacuum freeze drying condition is that the thickness of the material is controlled to be 8mm, the pre-freezing temperature is set to be-22 ℃, the sample is kept for 2 hours after the temperature is reduced to-22 ℃, the sublimation temperature is set to be 15 ℃, the analysis temperature is 35 ℃, the absolute pressure is 35Pa, and the drying is carried out for 24 hours; the cut-off molecular weight of the dialysis bag for dialysis is 3500 Da.
Carboxymethyl chitosan: CAS 83512-85-0, molecular weight 20 ten thousand, Hipporo Biotech, Hangzhou.
N-hexanoic anhydride: CAS, 2051-49-2, Hangzhou Hairui chemical Co., Ltd.
Perfluoro-n-pentane: CAS number: 678-26-2, Hangzhou Jie Heng Chemicals, Inc.
Decafluoropentane: 2, 3-dihydrodecafluoropentane, CAS No.: 138495-42-8, Henan Tianfu chemical Co., Ltd.
Dipalmitoyl phosphatidylcholine: CAS number: 2797-68-4, Harvest chemical, Hangzhou, China.
Palmitic acid: CAS number: 57-10-3, Hangzhou Hairui chemical Co.
Ethanol, unless specified to the mass fraction, is referred to as absolute ethanol.
Comparative example 1
An anti-nasopharyngeal carcinoma traditional Chinese medicine nano-emulsion comprises the following traditional Chinese medicine raw materials in parts by weight: 10 parts of gynostemma pentaphylla, 6 parts of dendrobium, 15 parts of berchemia lineate, 8 parts of Chinese brake herb, 10 parts of trogopterus dung, 3 parts of resina Toxicodendri, 3 parts of selaginella doederleinii, 5 parts of subprostrate sophora, 7 parts of rhizoma bolbostemmae, 4 parts of centipede, 8 parts of biond magnolia flower, 14 parts of radix ophiopogonis, 9 parts of rhizoma pinellinae praeparata, 11 parts of liquorice, 9 parts of solanum lyratum and 12 parts of oldenlandia diffusa.
The preparation method of the traditional Chinese medicine nano-emulsion for treating nasopharyngeal carcinoma comprises the following steps:
s1, respectively crushing the traditional Chinese medicine raw materials to 40 meshes, weighing the crushed traditional Chinese medicine raw materials according to the proportion, and uniformly mixing to obtain traditional Chinese medicine coarse powder;
s2 pulverizing the coarse powder, water, ethanol, and sodium polyacrylate in wet superfine pulverizer (SWFJ-20, popular in Shandong Shuangjia farming and pasturing machinery) at 7000 rpm for 15min to obtain Chinese medicinal slurry; the mass ratio of the traditional Chinese medicine coarse powder to the water to the ethanol to the sodium polyacrylate is 1:5:14: 0.015;
s3 extracting the Chinese medicinal slurry at 70 deg.C under reflux for 2h, centrifuging at 5000 r/min for 10min, concentrating the supernatant under reduced pressure to 1/8 of the original volume, and vacuum freeze drying to obtain Chinese medicinal active powder; the vacuum freeze drying condition is that the thickness of the material is controlled to be 8mm, the pre-freezing temperature is set to be-22 ℃, the sample is kept for 2 hours after the temperature is reduced to-22 ℃, the sublimation temperature is set to be 15 ℃, the analysis temperature is 35 ℃, the absolute pressure is 35Pa, and the drying is carried out for 24 hours;
s4, preparing a 2.3mg/mL hexanoyl carboxymethyl chitosan aqueous solution, adding the traditional Chinese medicine active powder into the hexanoyl carboxymethyl chitosan aqueous solution, stirring and mixing uniformly, then filling into a dialysis bag, dialyzing in distilled water for 24 hours, replacing the distilled water every 8 hours, and obtaining a dialysate as the liquid in the dialysis bag; after dialysis, taking out the dialysate, adding a fluorocarbon material with the mass of 2.1% of that of the dialysate into the dialysate, stirring and mixing uniformly, introducing oxygen to saturate and maintain the oxygen content in the solution to be 30mg/L, then carrying out ultrasonic emulsification under the conditions of ice bath at 0 ℃ and oxygen introduction to maintain the oxygen content in the solution to be 30mg/L, wherein the power is 20W, the time is 10min, the ultrasonic process is started for 3s and closed for 1s, the circulation is repeated, the mixture is centrifuged for 10min under the condition of 4000 revolutions per minute, supernatant liquid is discarded, lower-layer emulsion is taken out, and the irradiation sterilization is carried out for 20min under ultraviolet light with the wavelength of 220nm, so that the traditional Chinese medicine nano-emulsion for resisting nasopharyngeal carcinoma is obtained; the mass-volume ratio of the traditional Chinese medicine active powder to the hexanoyl carboxymethyl chitosan aqueous solution is 1:8 g/mL; the reduced pressure evaporation concentration condition is that the absolute pressure is 0.085MPa and the temperature is 50 ℃.
The particle size of the nano liquid drops in the traditional Chinese medicine nano emulsion for resisting nasopharyngeal carcinoma is 255.3 +/-44.3 nm measured by a Malvern particle sizer.
The cut-off molecular weight of the dialysis bag is 3500 Da.
The fluorocarbon material is decafluoropentane.
Example 1
An anti-nasopharyngeal carcinoma traditional Chinese medicine nano-emulsion comprises the following traditional Chinese medicine raw materials in parts by weight: 10 parts of gynostemma pentaphylla, 6 parts of dendrobium, 15 parts of berchemia lineate, 8 parts of Chinese brake herb, 10 parts of trogopterus dung, 3 parts of resina Toxicodendri, 3 parts of selaginella doederleinii, 5 parts of subprostrate sophora, 7 parts of rhizoma bolbostemmae, 4 parts of centipede, 8 parts of biond magnolia flower, 14 parts of radix ophiopogonis, 9 parts of rhizoma pinellinae praeparata, 11 parts of liquorice, 9 parts of solanum lyratum and 12 parts of oldenlandia diffusa.
The preparation method of the traditional Chinese medicine nano-emulsion for treating nasopharyngeal carcinoma comprises the following steps:
s1, respectively crushing the traditional Chinese medicine raw materials to 40 meshes, weighing the crushed traditional Chinese medicine raw materials according to the proportion, and uniformly mixing to obtain traditional Chinese medicine coarse powder;
s2 pulverizing the coarse powder, water, ethanol, and sodium polyacrylate in wet superfine pulverizer (SWFJ-20, popular in Shandong Shuangjia farming and pasturing machinery) at 7000 rpm for 15min to obtain Chinese medicinal slurry; the mass ratio of the traditional Chinese medicine coarse powder to the water to the ethanol to the sodium polyacrylate is 1:5:14: 0.015;
s3 extracting the Chinese medicinal slurry at 70 deg.C under reflux for 2h, centrifuging at 5000 r/min for 10min, concentrating the supernatant under reduced pressure to 1/8 of the original volume, and vacuum freeze drying to obtain Chinese medicinal active powder; the vacuum freeze drying condition is that the thickness of the material is controlled to be 8mm, the pre-freezing temperature is set to be-22 ℃, the sample is kept for 2 hours after the temperature is reduced to-22 ℃, the sublimation temperature is set to be 15 ℃, the analysis temperature is 35 ℃, the absolute pressure is 35Pa, and the drying is carried out for 24 hours;
s4, preparing a 2.3mg/mL hexanoyl carboxymethyl chitosan aqueous solution, adding the traditional Chinese medicine active powder into the hexanoyl carboxymethyl chitosan aqueous solution, stirring and mixing uniformly, then filling into a dialysis bag, dialyzing in distilled water for 24 hours, replacing the distilled water every 8 hours, and obtaining a dialysate as the liquid in the dialysis bag; after dialysis, taking out the dialysate, adding a fluorocarbon material with the mass of 2.1% of that of the dialysate and a stabilizer with the mass of 1.2% of that of the dialysate into the dialysate, stirring and mixing uniformly, introducing oxygen to saturate and maintaining the oxygen content in the solution at 30mg/L, then carrying out ultrasonic emulsification under the conditions of ice bath at 0 ℃ and oxygen introduction to maintain the oxygen content in the solution at 30mg/L, wherein the power is 20W and the time is 10min, the ultrasonic process is started for 3s and closed for 1s, the circulation is repeated, the centrifugation is carried out for 10min under the condition of 4000 revolutions per minute, supernatant is discarded, lower-layer emulsion is taken out, and the sterilization is carried out for 20min under the ultraviolet light with the wavelength of 220nm, so as to obtain the traditional Chinese medicine nano-emulsion for resisting nasopharyngeal; the mass-volume ratio of the traditional Chinese medicine active powder to the hexanoyl carboxymethyl chitosan aqueous solution is 1:8 g/mL; the reduced pressure evaporation concentration condition is that the absolute pressure is 0.085MPa and the temperature is 50 ℃.
The particle size of the nano liquid drop in the traditional Chinese medicine nano emulsion for resisting nasopharyngeal carcinoma is measured by a Malvern particle sizer to be 265.3 +/-53.7 nm.
The cut-off molecular weight of the dialysis bag is 3500 Da.
The fluorocarbon material is decafluoropentane.
The stabilizer is palmitic acid.
Example 2
An anti-nasopharyngeal carcinoma traditional Chinese medicine nano-emulsion comprises the following traditional Chinese medicine raw materials in parts by weight: 10 parts of gynostemma pentaphylla, 6 parts of dendrobium, 15 parts of berchemia lineate, 8 parts of Chinese brake herb, 10 parts of trogopterus dung, 3 parts of resina Toxicodendri, 3 parts of selaginella doederleinii, 5 parts of subprostrate sophora, 7 parts of rhizoma bolbostemmae, 4 parts of centipede, 8 parts of biond magnolia flower, 14 parts of radix ophiopogonis, 9 parts of rhizoma pinellinae praeparata, 11 parts of liquorice, 9 parts of solanum lyratum and 12 parts of oldenlandia diffusa.
The preparation method of the traditional Chinese medicine nano-emulsion for treating nasopharyngeal carcinoma comprises the following steps:
s1, respectively crushing the traditional Chinese medicine raw materials to 40 meshes, weighing the crushed traditional Chinese medicine raw materials according to the proportion, and uniformly mixing to obtain traditional Chinese medicine coarse powder;
s2 pulverizing the coarse powder, water, ethanol, and sodium polyacrylate in wet superfine pulverizer (SWFJ-20, popular in Shandong Shuangjia farming and pasturing machinery) at 7000 rpm for 15min to obtain Chinese medicinal slurry; the mass ratio of the traditional Chinese medicine coarse powder to the water to the ethanol to the sodium polyacrylate is 1:5:14: 0.015;
s3 extracting the Chinese medicinal slurry at 70 deg.C under reflux for 2h, centrifuging at 5000 r/min for 10min, concentrating the supernatant under reduced pressure to 1/8 of the original volume, and vacuum freeze drying to obtain Chinese medicinal active powder; the vacuum freeze drying condition is that the thickness of the material is controlled to be 8mm, the pre-freezing temperature is set to be-22 ℃, the sample is kept for 2 hours after the temperature is reduced to-22 ℃, the sublimation temperature is set to be 15 ℃, the analysis temperature is 35 ℃, the absolute pressure is 35Pa, and the drying is carried out for 24 hours;
s4, preparing a 2.3mg/mL hexanoyl carboxymethyl chitosan aqueous solution, adding the traditional Chinese medicine active powder into the hexanoyl carboxymethyl chitosan aqueous solution, stirring and mixing uniformly, then filling into a dialysis bag, dialyzing in distilled water for 24 hours, replacing the distilled water every 8 hours, and obtaining a dialysate as the liquid in the dialysis bag; after dialysis, taking out the dialysate, adding a fluorocarbon material with the mass of 2.1% of that of the dialysate and a stabilizer with the mass of 1.2% of that of the dialysate into the dialysate, stirring and mixing uniformly, introducing oxygen to saturate and maintaining the oxygen content in the solution at 30mg/L, then carrying out ultrasonic emulsification under the conditions of ice bath at 0 ℃ and oxygen introduction to maintain the oxygen content in the solution at 30mg/L, wherein the power is 20W and the time is 10min, the ultrasonic process is started for 3s and closed for 1s, the circulation is repeated, the centrifugation is carried out for 10min under the condition of 4000 revolutions per minute, supernatant is discarded, lower-layer emulsion is taken out, and the sterilization is carried out for 20min under the ultraviolet light with the wavelength of 220nm, so as to obtain the traditional Chinese medicine nano-emulsion for resisting nasopharyngeal; the mass-volume ratio of the traditional Chinese medicine active powder to the hexanoyl carboxymethyl chitosan aqueous solution is 1:8 g/mL; the reduced pressure evaporation concentration condition is that the absolute pressure is 0.085MPa and the temperature is 50 ℃.
The particle size of the nano liquid drops in the traditional Chinese medicine nano emulsion for resisting nasopharyngeal carcinoma is 258.3 +/-46.7 nm measured by a Malvern particle sizer.
The cut-off molecular weight of the dialysis bag is 3500 Da.
The fluorocarbon material is perfluoro-n-pentane.
The stabilizer is palmitic acid.
Example 3
An anti-nasopharyngeal carcinoma traditional Chinese medicine nano-emulsion comprises the following traditional Chinese medicine raw materials in parts by weight: 10 parts of gynostemma pentaphylla, 6 parts of dendrobium, 15 parts of berchemia lineate, 8 parts of Chinese brake herb, 10 parts of trogopterus dung, 3 parts of resina Toxicodendri, 3 parts of selaginella doederleinii, 5 parts of subprostrate sophora, 7 parts of rhizoma bolbostemmae, 4 parts of centipede, 8 parts of biond magnolia flower, 14 parts of radix ophiopogonis, 9 parts of rhizoma pinellinae praeparata, 11 parts of liquorice, 9 parts of solanum lyratum and 12 parts of oldenlandia diffusa.
The preparation method of the traditional Chinese medicine nano-emulsion for treating nasopharyngeal carcinoma comprises the following steps:
s1, respectively crushing the traditional Chinese medicine raw materials to 40 meshes, weighing the crushed traditional Chinese medicine raw materials according to the proportion, and uniformly mixing to obtain traditional Chinese medicine coarse powder;
s2 pulverizing the coarse powder, water, ethanol, and sodium polyacrylate in wet superfine pulverizer (SWFJ-20, popular in Shandong Shuangjia farming and pasturing machinery) at 7000 rpm for 15min to obtain Chinese medicinal slurry; the mass ratio of the traditional Chinese medicine coarse powder to the water to the ethanol to the sodium polyacrylate is 1:5:14: 0.015;
s3 extracting the Chinese medicinal slurry at 70 deg.C under reflux for 2h, centrifuging at 5000 r/min for 10min, concentrating the supernatant under reduced pressure to 1/8 of the original volume, and vacuum freeze drying to obtain Chinese medicinal active powder; the vacuum freeze drying condition is that the thickness of the material is controlled to be 8mm, the pre-freezing temperature is set to be-22 ℃, the sample is kept for 2 hours after the temperature is reduced to-22 ℃, the sublimation temperature is set to be 15 ℃, the analysis temperature is 35 ℃, the absolute pressure is 35Pa, and the drying is carried out for 24 hours;
s4, preparing a 2.3mg/mL hexanoyl carboxymethyl chitosan aqueous solution, adding the traditional Chinese medicine active powder into the hexanoyl carboxymethyl chitosan aqueous solution, stirring and mixing uniformly, then filling into a dialysis bag, dialyzing in distilled water for 24 hours, replacing the distilled water every 8 hours, and obtaining a dialysate as the liquid in the dialysis bag; after dialysis, taking out the dialysate, adding a fluorocarbon material with the mass of 2.1% of that of the dialysate and a stabilizer with the mass of 1.2% of that of the dialysate into the dialysate, stirring and mixing uniformly, introducing oxygen to saturate and maintaining the oxygen content in the solution at 30mg/L, then carrying out ultrasonic emulsification under the conditions of ice bath at 0 ℃ and oxygen introduction to maintain the oxygen content in the solution at 30mg/L, wherein the power is 20W and the time is 10min, the ultrasonic process is started for 3s and closed for 1s, the circulation is repeated, the centrifugation is carried out for 10min under the condition of 4000 revolutions per minute, supernatant is discarded, lower-layer emulsion is taken out, and the sterilization is carried out for 20min under the ultraviolet light with the wavelength of 220nm, so as to obtain the traditional Chinese medicine nano-emulsion for resisting nasopharyngeal; the mass-volume ratio of the traditional Chinese medicine active powder to the hexanoyl carboxymethyl chitosan aqueous solution is 1:8 g/mL; the reduced pressure evaporation concentration condition is that the absolute pressure is 0.085MPa and the temperature is 50 ℃.
The particle size of the nano liquid drop in the traditional Chinese medicine nano emulsion for resisting nasopharyngeal carcinoma is determined to be 249.1 +/-41.7 nm by a Malvern particle size analyzer.
The cut-off molecular weight of the dialysis bag is 3500 Da.
The fluorocarbon material is formed by mixing perfluoro-n-pentane and decafluoropentane according to the mass ratio of 1: 3.
The stabilizer is palmitic acid.
Example 4
An anti-nasopharyngeal carcinoma traditional Chinese medicine nano-emulsion comprises the following traditional Chinese medicine raw materials in parts by weight: 10 parts of gynostemma pentaphylla, 6 parts of dendrobium, 15 parts of berchemia lineate, 8 parts of Chinese brake herb, 10 parts of trogopterus dung, 3 parts of resina Toxicodendri, 3 parts of selaginella doederleinii, 5 parts of subprostrate sophora, 7 parts of rhizoma bolbostemmae, 4 parts of centipede, 8 parts of biond magnolia flower, 14 parts of radix ophiopogonis, 9 parts of rhizoma pinellinae praeparata, 11 parts of liquorice, 9 parts of solanum lyratum and 12 parts of oldenlandia diffusa.
The preparation method of the traditional Chinese medicine nano-emulsion for treating nasopharyngeal carcinoma comprises the following steps:
s1, respectively crushing the traditional Chinese medicine raw materials to 40 meshes, weighing the crushed traditional Chinese medicine raw materials according to the proportion, and uniformly mixing to obtain traditional Chinese medicine coarse powder;
s2 pulverizing the coarse powder, water, ethanol, and sodium polyacrylate in wet superfine pulverizer (SWFJ-20, popular in Shandong Shuangjia farming and pasturing machinery) at 7000 rpm for 15min to obtain Chinese medicinal slurry; the mass ratio of the traditional Chinese medicine coarse powder to the water to the ethanol to the sodium polyacrylate is 1:5:14: 0.015;
s3 extracting the Chinese medicinal slurry at 70 deg.C under reflux for 2h, centrifuging at 5000 r/min for 10min, concentrating the supernatant under reduced pressure to 1/8 of the original volume, and vacuum freeze drying to obtain Chinese medicinal active powder; the vacuum freeze drying condition is that the thickness of the material is controlled to be 8mm, the pre-freezing temperature is set to be-22 ℃, the sample is kept for 2 hours after the temperature is reduced to-22 ℃, the sublimation temperature is set to be 15 ℃, the analysis temperature is 35 ℃, the absolute pressure is 35Pa, and the drying is carried out for 24 hours;
s4, preparing a 2.3mg/mL hexanoyl carboxymethyl chitosan aqueous solution, adding the traditional Chinese medicine active powder into the hexanoyl carboxymethyl chitosan aqueous solution, stirring and mixing uniformly, then filling into a dialysis bag, dialyzing in distilled water for 24 hours, replacing the distilled water every 8 hours, and obtaining a dialysate as the liquid in the dialysis bag; after dialysis, taking out the dialysate, adding a fluorocarbon material with the mass of 2.1% of that of the dialysate and a stabilizer with the mass of 1.2% of that of the dialysate into the dialysate, stirring and mixing uniformly, introducing oxygen to saturate and maintaining the oxygen content in the solution at 30mg/L, then carrying out ultrasonic emulsification under the conditions of ice bath at 0 ℃ and oxygen introduction to maintain the oxygen content in the solution at 30mg/L, wherein the power is 20W and the time is 10min, the ultrasonic process is started for 3s and closed for 1s, the circulation is repeated, the centrifugation is carried out for 10min under the condition of 4000 revolutions per minute, supernatant is discarded, lower-layer emulsion is taken out, and the sterilization is carried out for 20min under the ultraviolet light with the wavelength of 220nm, so as to obtain the traditional Chinese medicine nano-emulsion for resisting nasopharyngeal; the mass-volume ratio of the traditional Chinese medicine active powder to the hexanoyl carboxymethyl chitosan aqueous solution is 1:8 g/mL; the reduced pressure evaporation concentration condition is that the absolute pressure is 0.085MPa and the temperature is 50 ℃.
The particle size of the nano liquid drop in the traditional Chinese medicine nano emulsion for resisting nasopharyngeal carcinoma is 248.3 +/-38.1 nm measured by a Malvern particle sizer.
The cut-off molecular weight of the dialysis bag is 3500 Da.
The fluorocarbon material is formed by mixing perfluoro-n-pentane and decafluoropentane according to the mass ratio of 1: 3.
The stabilizer is dipalmitoyl phosphatidylcholine.
Example 5
An anti-nasopharyngeal carcinoma traditional Chinese medicine nano-emulsion comprises the following traditional Chinese medicine raw materials in parts by weight: 10 parts of gynostemma pentaphylla, 6 parts of dendrobium, 15 parts of berchemia lineate, 8 parts of Chinese brake herb, 10 parts of trogopterus dung, 3 parts of resina Toxicodendri, 3 parts of selaginella doederleinii, 5 parts of subprostrate sophora, 7 parts of rhizoma bolbostemmae, 4 parts of centipede, 8 parts of biond magnolia flower, 14 parts of radix ophiopogonis, 9 parts of rhizoma pinellinae praeparata, 11 parts of liquorice, 9 parts of solanum lyratum and 12 parts of oldenlandia diffusa.
The preparation method of the traditional Chinese medicine nano-emulsion for treating nasopharyngeal carcinoma comprises the following steps:
s1, respectively crushing the traditional Chinese medicine raw materials to 40 meshes, weighing the crushed traditional Chinese medicine raw materials according to the proportion, and uniformly mixing to obtain traditional Chinese medicine coarse powder;
s2 pulverizing the coarse powder, water, ethanol, and sodium polyacrylate in wet superfine pulverizer (SWFJ-20, popular in Shandong Shuangjia farming and pasturing machinery) at 7000 rpm for 15min to obtain Chinese medicinal slurry; the mass ratio of the traditional Chinese medicine coarse powder to the water to the ethanol to the sodium polyacrylate is 1:5:14: 0.015;
s3 extracting the Chinese medicinal slurry at 70 deg.C under reflux for 2h, centrifuging at 5000 r/min for 10min, concentrating the supernatant under reduced pressure to 1/8 of the original volume, and vacuum freeze drying to obtain Chinese medicinal active powder; the vacuum freeze drying condition is that the thickness of the material is controlled to be 8mm, the pre-freezing temperature is set to be-22 ℃, the sample is kept for 2 hours after the temperature is reduced to-22 ℃, the sublimation temperature is set to be 15 ℃, the analysis temperature is 35 ℃, the absolute pressure is 35Pa, and the drying is carried out for 24 hours;
s4, preparing a 2.3mg/mL hexanoyl carboxymethyl chitosan aqueous solution, adding the traditional Chinese medicine active powder into the hexanoyl carboxymethyl chitosan aqueous solution, stirring and mixing uniformly, then filling into a dialysis bag, dialyzing in distilled water for 24 hours, replacing the distilled water every 8 hours, and obtaining a dialysate as the liquid in the dialysis bag; after dialysis, taking out the dialysate, adding a fluorocarbon material with the mass of 2.1% of that of the dialysate and a stabilizer with the mass of 1.2% of that of the dialysate into the dialysate, stirring and mixing uniformly, introducing oxygen to saturate and maintaining the oxygen content in the solution at 30mg/L, then carrying out ultrasonic emulsification under the conditions of ice bath at 0 ℃ and oxygen introduction to maintain the oxygen content in the solution at 30mg/L, wherein the power is 20W and the time is 10min, the ultrasonic process is started for 3s and closed for 1s, the circulation is repeated, the centrifugation is carried out for 10min under the condition of 4000 revolutions per minute, supernatant is discarded, lower-layer emulsion is taken out, and the sterilization is carried out for 20min under the ultraviolet light with the wavelength of 220nm, so as to obtain the traditional Chinese medicine nano-emulsion for resisting nasopharyngeal; the mass-volume ratio of the traditional Chinese medicine active powder to the hexanoyl carboxymethyl chitosan aqueous solution is 1:8 g/mL; the reduced pressure evaporation concentration condition is that the absolute pressure is 0.085MPa and the temperature is 50 ℃.
The particle size of the nano liquid drop in the traditional Chinese medicine nano emulsion for resisting nasopharyngeal carcinoma is measured by a Malvern particle sizer and is 250.3 +/-42.1 nm.
The cut-off molecular weight of the dialysis bag is 3500 Da.
The fluorocarbon material is formed by mixing perfluoro-n-pentane and decafluoropentane according to the mass ratio of 1: 3.
The stabilizer is formed by mixing dipalmitoyl phosphatidylcholine and palmitic acid according to the mass ratio of 1.5: 1.
Comparative example 2
An anti-nasopharyngeal carcinoma traditional Chinese medicine nano-emulsion comprises the following traditional Chinese medicine raw materials in parts by weight: 10 parts of gynostemma pentaphylla, 6 parts of dendrobium, 15 parts of berchemia lineate, 8 parts of Chinese brake herb, 10 parts of trogopterus dung, 3 parts of resina Toxicodendri, 3 parts of selaginella doederleinii, 5 parts of subprostrate sophora, 7 parts of rhizoma bolbostemmae, 4 parts of centipede, 8 parts of biond magnolia flower, 14 parts of radix ophiopogonis, 9 parts of rhizoma pinellinae praeparata, 11 parts of liquorice, 9 parts of solanum lyratum and 12 parts of oldenlandia diffusa.
The preparation method of the traditional Chinese medicine nano-emulsion for treating nasopharyngeal carcinoma comprises the following steps:
s1 weighing the Chinese medicinal materials at a certain ratio, placing into a casserole, adding clear water, soaking 3cm above the medicinal surface, and soaking for 15 min; boiling the Chinese medicinal materials soaked in clear water of casserole with strong fire, decocting with slow fire for 25min, filtering with 100 mesh gauze, and collecting filtrate;
s2, sequentially treating the filter residue left after filtration in the step S1 again according to the operation method in the step S1, filtering by using 100-mesh gauze, and taking the filtrate for later use;
s3, mixing the filtrates obtained in the steps S1 and S2, performing rotary reduced pressure evaporation to 1/8 of the original volume under the conditions of absolute pressure of 0.085MPa and temperature of 50 ℃, and performing vacuum freeze drying on the obtained concentrated solution to obtain traditional Chinese medicine active powder; the vacuum freeze drying condition is that the thickness of the material is controlled to be 8mm, the pre-freezing temperature is set to be-22 ℃, the sample is kept for 2 hours after the temperature is reduced to-22 ℃, the sublimation temperature is set to be 15 ℃, the analysis temperature is 35 ℃, the absolute pressure is 35Pa, and the drying is carried out for 24 hours;
s4, preparing a 2.3mg/mL hexanoyl carboxymethyl chitosan aqueous solution, adding the traditional Chinese medicine active powder into the hexanoyl carboxymethyl chitosan aqueous solution, stirring and mixing uniformly, then filling into a dialysis bag, dialyzing in distilled water for 24 hours, replacing the distilled water every 8 hours, and obtaining a dialysate as the liquid in the dialysis bag; after dialysis, taking out the dialysate, adding a fluorocarbon material with the mass of 2.1% of that of the dialysate and a stabilizer with the mass of 1.2% of that of the dialysate into the dialysate, stirring and mixing uniformly, introducing oxygen to saturate and maintaining the oxygen content in the solution at 30mg/L, then carrying out ultrasonic emulsification under the conditions of ice bath at 0 ℃ and oxygen introduction to maintain the oxygen content in the solution at 30mg/L, wherein the power is 20W and the time is 10min, the ultrasonic process is started for 3s and closed for 1s, the circulation is repeated, the centrifugation is carried out for 10min under the condition of 4000 revolutions per minute, supernatant is discarded, lower-layer emulsion is taken out, and the sterilization is carried out for 20min under the ultraviolet light with the wavelength of 220nm, so as to obtain the traditional Chinese medicine nano-emulsion for resisting nasopharyngeal; the mass-volume ratio of the traditional Chinese medicine active powder to the hexanoyl carboxymethyl chitosan aqueous solution is 1:8 g/mL; the reduced pressure evaporation concentration condition is that the absolute pressure is 0.085MPa and the temperature is 50 ℃.
The particle size of the nano liquid drops in the traditional Chinese medicine nano emulsion for resisting nasopharyngeal carcinoma is 264.3 +/-48.1 nm measured by a Malvern particle sizer.
The cut-off molecular weight of the dialysis bag is 3500 Da.
The fluorocarbon material is formed by mixing perfluoro-n-pentane and decafluoropentane according to the mass ratio of 1: 3.
The stabilizer is formed by mixing dipalmitoyl phosphatidylcholine and palmitic acid according to the mass ratio of 1.5: 1.
Test example 1
Tumor inhibition effect experiment:
test subjects: 160 ICR mice, weight 19-24 g, male.
Test inoculation content: s180 solid tumor.
ICR mice were inoculated with 0.2mL (about 5X 10) of S180 cancer cell suspension subcutaneously in the right axilla6Individual cells/mL), the next day was randomly divided into 8 groups. The treatment components are 7 groups, and the traditional Chinese medicine nano-emulsion for treating nasopharyngeal carcinoma disclosed in the invention in the embodiments 1-5 and the comparative examples 1-2 is respectively administrated by intragastric administration for 0.2mL/10g every day, 2 times every day and 10 days continuously; the control group is a normal saline group, and the normal saline group is fed with 0.4mL/10g normal saline every day for 10 days continuously; the 8 groups of animals are sacrificed the next day after the medicine is stopped, tumor stripping and weighing are carried out, and the tumor inhibition rate is calculated according to the formula: the tumor inhibition rate (%) is (average tumor weight of normal saline control group-average tumor weight of drug administration group)/average tumor weight of normal saline control group x 100%, and the result shows that the drug of the invention has inhibition effect on S180 growth, and has significant difference P compared with the control group<0.05. The inhibitory effect and tumor-inhibiting effect on S180 are shown in table 1.
Table 1: inhibiting effect on S180 and tumor inhibiting effect
As can be seen from table 1: compared with a normal saline group, the treatment group adopting the traditional Chinese medicine composition can inhibit the growth of tumor bodies, and has a significant difference P of less than 0.05.
Test example 2
Clinical experiments are as follows:
140 patients with middle-and late-stage nasopharyngeal carcinoma diagnosed in Linchuan were selected as study subjects, 93 men and 57 women, with the average age of 53 years and the age range of 41-76 years.
Inclusion criteria were: the patient with nasopharyngeal carcinoma in middle and late stages is confirmed to be diagnosed (by pathological or cytological examination), and no serious cardiovascular, liver, kidney and other diseases exist without operation, radiotherapy and chemotherapy.
The treatment method comprises the following steps: the patients were randomized into 7 groups of 20 patients each. Each group received radiation therapy. 5 times per week, and 6 weeks as one treatment course. The traditional Chinese medicine nano-emulsion for treating nasopharyngeal carcinoma disclosed by the invention is taken 50mL every day in the radiotherapy period, and the traditional Chinese medicine nano-emulsion for treating nasopharyngeal carcinoma disclosed by the invention in the embodiment 1-5 and the comparison examples 1-2 is taken.
The effect indexes are as follows: complete remission was achieved according to the RECIST (criteria for treating malignancies) assessment method for the efficacy assessment of solid tumors: all target lesions disappear; partial mitigation: the sum of the length and diameter of the baseline lesion is reduced by more than 30%. No obvious change: the sum of the length and diameter of the baseline lesion is reduced by less than 30% or increased by less than 20%; and (3) lesion expansion: the sum of the length and diameter of the baseline lesion increased by more than 20% or new lesions appeared.
The treatment effect of two groups of patients is compared after 1 treatment course.
TABLE 2 comparison of therapeutic effects of the anti-nasopharyngeal carcinoma Chinese medicinal nanoemulsion of the present invention
Test example 3
Long-term toxicity test:
the anti-nasopharyngeal carcinoma traditional Chinese medicine nanoemulsion prepared in the embodiment 5 of the invention is respectively used for gastric irrigation on a mouse according to the dose of 0.2mL/10g, and is continuously used for 16 weeks (2 times per day) and observed and detected after stopping the administration for 4 weeks, and the result shows that: the traditional Chinese medicine preparation has no obvious influence on indexes such as hair, behaviors, excrement and urine, body weight, weight of organs, hemogram, liver and kidney functions, blood sugar, blood fat and the like of a mouse, and the naked eyes of the organs find no abnormal change and histological examination results show that after the traditional Chinese medicine preparation is used for 16 weeks and is stopped for 4 weeks, all organs of the mouse have no obvious change. The nano-emulsion of the traditional Chinese medicine for resisting nasopharyngeal carcinoma has low toxicity after long-term administration to mice, has no abnormal reaction after stopping administration, and is safe to apply.
The present invention has been described in detail, and the principle and embodiments of the present invention are explained herein by using specific examples, which are only used to help understand the method and the core idea of the present invention; meanwhile, for a person skilled in the art, according to the idea of the present invention, there may be variations in the specific embodiments and the application scope, and in summary, the content of the present specification should not be construed as a limitation to the present invention.
Claims (2)
1. The traditional Chinese medicine nano-emulsion for treating nasopharyngeal carcinoma is characterized by being prepared from the following traditional Chinese medicine raw materials in parts by weight: 8-12 parts of gynostemma pentaphylla, 4-15 parts of dendrobium, 13-17 parts of berchemia lineate, 4-15 parts of Chinese brake herb, 8-12 parts of trogopterus dung, 1-5 parts of dried lacquer, 1-6 parts of selaginella tamariscina, 1-8 parts of subprostrate sophora, 4-15 parts of rhizoma bolbostemmae, 1-5 parts of centipede, 5-20 parts of biond magnolia flower, 10-20 parts of radix ophiopogonis, 5-20 parts of rhizoma pinellinae praeparata, 5-20 parts of liquorice, 4-15 parts of solanum lyratum and 4-16 parts of oldenlandia diffusa,
the preparation method comprises the following steps:
s1, respectively crushing the traditional Chinese medicine raw materials to 20-100 meshes, and mixing to obtain traditional Chinese medicine coarse powder;
s2, treating the traditional Chinese medicine coarse powder, water, ethanol and sodium polyacrylate for 10-30 min under wet superfine grinding to obtain traditional Chinese medicine slurry;
s3, refluxing and extracting the traditional Chinese medicine slurry at 60-80 ℃ for 1-3 h, centrifuging, evaporating under reduced pressure, and freeze-drying in vacuum to obtain traditional Chinese medicine active powder;
s4, adding the traditional Chinese medicine active powder into a 1.5-3 mg/mL n-hexanoyl carboxymethyl chitosan aqueous solution, and dialyzing in distilled water for 16-32 hours to obtain dialysate; taking out the dialyzate, adding a fluorocarbon material accounting for 1.5-3.2% of the dialyzate in mass and a stabilizer accounting for 0.8-1.6% of the dialyzate in mass into the dialyzate, stirring, introducing oxygen to saturate, maintaining the oxygen content in the solution to be 25-35 mg/L, then carrying out ultrasonic emulsification under the conditions of an ice bath at 0-4 ℃ and introducing oxygen to maintain the oxygen content in the solution to be 25-35 mg/L, centrifuging for 5-10 min, and taking out the lower-layer emulsion for sterilization.
2. The anti-nasopharyngeal-cancer traditional Chinese medicine nanoemulsion according to claim 1, wherein the fluorocarbon material is formed by mixing perfluoro-n-pentane and decafluoropentane according to a mass ratio of 1 (1-3).
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