CN108815214A - For treating drug and the application of liver cancer - Google Patents

For treating drug and the application of liver cancer Download PDF

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Publication number
CN108815214A
CN108815214A CN201811051143.XA CN201811051143A CN108815214A CN 108815214 A CN108815214 A CN 108815214A CN 201811051143 A CN201811051143 A CN 201811051143A CN 108815214 A CN108815214 A CN 108815214A
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China
Prior art keywords
parts
drug
liver cancer
mixing
extract
Prior art date
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Pending
Application number
CN201811051143.XA
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Chinese (zh)
Inventor
韦范
王雯丽
周小雷
缪剑华
欧春丽
龚小妹
王硕
莫单丹
唐炳兰
侯小利
宋志军
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Guangxi Botanical Garden of Medicinal Plants
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Guangxi Botanical Garden of Medicinal Plants
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Priority to CN201811051143.XA priority Critical patent/CN108815214A/en
Publication of CN108815214A publication Critical patent/CN108815214A/en
Pending legal-status Critical Current

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Abstract

The invention discloses a kind of for treating the drug of liver cancer, including refutes bone extract greatly, and the preparation for refuting bone extract greatly includes the following steps：S1, it will refute bone greatly and be placed in boiling water and extract, and obtain Aqueous extracts；S2, bone extract will be refuted greatly after the Aqueous extracts separating-purifying in S1 to obtain the final product.Of the invention including to refute bone extract greatly for treating the drug of liver cancer, experiment, which shows to refute bone extract greatly to have the growth of BEL-7402, Hep-B3 hepatoma cell strain, significantly inhibits effect.

Description

For treating drug and the application of liver cancer
Technical field
The present invention relates to pharmaceutical technology fields.It is more particularly related to a kind of drug for treating liver cancer and Using.
Background technique
Primary carcinoma of liver is one of China's common cancer.The death rate is high, is only second in mortality of malignant tumors cis-position Stomach, esophagus and occupy third position, account for second in the rural area of Electric Field At Ground In Partial Region, be only second to gastric cancer.China dies of liver cancer about 11 every year Ten thousand people account for the 45% of whole world PLC mortality number.Therefore, reinforce the research of primary liver cancer, extend patient survival, Curative effect is improved, the task of top priority that patients ' life quality is clinical tumor worker is improved.Liver cancer essential therapeutic arsenals are that operation is cut It removes, early stage, postoperative 1 year, 3 years and 5 years survival rate was respectively 80~92%, 61~86% and 41~75%.But 90% patient is because swollen Tumor is larger or cirrhosis and loses operative chance；Normal liver cell is sensitive to radioactive ray, and then sensibility is poor for liver cancer cells, > 4000cGy is needed to be likely to obtain certain Partial controll；Local ablation therapy can play Partial controll tumour, alleviation symptom etc. Effect, can be used as to undergo surgery and cuts off the therapeutic choice of patient, but only effective less than the liver cancer of 5cm to tumour；Chemotherapeutic Object is to the part remission rate of liver cancer 20% hereinafter, curative effect is undesirable；Trans-hepatic artery conduit chemoembolization (TACE) or TACE Add portal vein embolization (PVE) that effective percentage is made to improve, whether also there is improvement in life cycle, but complete, best in arterial embolism Medication combined and optimal embolism type etc. still need to further study；Biological target therapy is to the effective of primary carcinoma of liver Rate is below 10%.In conclusion modern medicine is to advanced primary liver cancer still without good treatment method.Traditional Chinese medicine treatment Advanced primary liver cancer has potential advantages, how to develop safe and efficient traditional Chinese medicine, has very important significance.
Summary of the invention
It is an object of the invention to solve at least the above problems, and provide the advantages of at least will be described later.
It is a further object to provide a kind of for treating the drug of liver cancer, including refutes bone extract greatly, tests Show to refute greatly bone extract and have to the growth of BEL-7402, Hep-B3 hepatoma cell strain and significantly inhibits effect.
In order to realize object of the present invention and further advantage, provide it is a kind of for treating the drug of liver cancer, including Bone extract is refuted greatly, and the preparation for refuting bone extract greatly includes the following steps：
S1, it will refute bone greatly and be placed in boiling water and extract, and obtain Aqueous extracts；
S2, bone extract will be refuted greatly after the Aqueous extracts separating-purifying in S1 to obtain the final product.
Preferably, described for treating the drug of liver cancer, it is used in S1 extracting in boiling water 2 times, 2~3h, merges 2 every time Secondary extracting solution is up to Aqueous extracts；Aqueous extracts separating-purifying specifically includes in S2：By Aqueous extracts 80 DEG C at a temperature of be concentrated into Specific gravity is 1.05, is then eluted using D101 macroreticular resin, and eluant, eluent is followed successively by the second that volume fraction is 20%, 50%, 95% Alcohol collects 95% ethanol eluate, removes ethyl alcohol, is concentrated into specific gravity as 1.08 and refutes bone extract to get big.
Preferably, the drug for being used to treat liver cancer, further includes Chinese medical extract, the system of the Chinese medical extract Preparation Method includes the following steps：
A1, by 2~5 parts of umbellate pore furgus of parts by weight, 3~6 parts of dandelion, 2~6 parts of subprostrate sophora, 1~3 part of akebiae, 3~6 parts of stringy stonecrop, 2~5 parts of polygonum cuspidate, 1~3 part of Chinese small iris, 2~5 parts of catharanthus roseus, 1~4 part of Radix Glycyrrhizae, 2~5 parts Radix gentianae, 2~6 parts of gynostemma pentaphylla, 1~3 part of Rabdosia rubescens, 5~8 parts of paris polyphylla, 2~6 parts of Asian puccoon, 3~6 parts Bittersweet, 3~6 parts of mock-strawberry, 2~5 parts of clearvers, 3~6 parts of gelsemium elegan, 1~3 part of Patrinia heterophylla, 2~4 parts of careless coral Coral, 2~5 parts of lobelia chinensis, 4~7 parts of radices trichosanthis, 2~5 parts of Prunella vulgaris, 3~8 parts of airpotato yam, 2~5 parts of curcuma zedoary, 1 It is mixed that first is obtained after~3 parts of the seed of cowherb, 2~6 parts of folium cycas, 3~6 parts of wartwort, 2~6 parts of Rhizoma Atractylodis Macrocephalae co-grinding Material；It is 10~15 parts by cellulase, 2~6 parts of protease, 1~3 part of pectase and parts by weight that parts by weight are 3~5 parts Water mixing after obtain water enzyme solution, by water enzyme solution spray in the first mixture after mixing evenly, and at 35 DEG C stand 15~ 20min obtains the second mixing；Second mixture is placed in closed container, saturated steam is then passed through into closed container So that pressure in closed container is 1.1Mpa, after maintaining 20~30min, then in making pressure drop in closed container in 4~6s Down to 5~10pa, after keeping 3~6min, taking-up obtains third mixing；Then into third mixing be added parts by weight be 150~ 160 parts of water is that 2~3h is decocted at 90~95 DEG C in temperature, and filtering obtains filtrate, obtains after filter vacuum is concentrated and dried First powder；
A2, by parts by weight be 2~5 parts gecko, after 3~5 parts of leech crushes, being placed in parts by weight is 15~20 parts of leaching The 4th mixing is obtained by filtration after impregnating 30~40min in bubble liquid, the preparation method of the soak is：By tomato, sansevieria trifasciata, often Spring rattan, wormwood, lemon grass (Cymbopogon citratus), cassia bark, which are dissolved in the water that temperature is 80~90 DEG C, impregnates 1~2h, filters up to soak；Wherein, it soaks Steep liquid in tomato, sansevieria trifasciata, ivy, wormwood, lemon grass (Cymbopogon citratus), cassia bark, water mass ratio be 3~5:0.5~1:0.8~1.2:1 ~1.5:0.5~1:0.5~1:50~60；
A3, the 4th mixing obtained in A2 is added in the water that parts by weight are 80~90 parts, is heated to reflux 3~4h, filtered, The first filtrate and the first filter residue are obtained, it is spare；Then, the first filter residue is placed in the ethanol solution that volume fraction is 85~90% 5~10h is impregnated, filtering obtains the second filtrate, obtains second after vacuum freeze drying after the first filtrate, the second filtrate are mixed Powder；
A4, the first powder, the second powder are mixed up to Chinese medical extract；
The bone extract and the mass ratio of Chinese medical extract of refuting greatly is 1:0.2.
Preferably, described for treating the drug of liver cancer, vacuum freeze drying is specially in A3：In -30~-25 DEG C 2~3h of lower freezing, then in vacuum degree is 5~10Pa, temperature is dry 20~30h at 60 DEG C.
Preferably, described to use power for treating the drug of liver cancer before the 4th mixing is placed in water reflux in A3 For ultraviolet 5~10min of light irradiation of 300~500W.
The drug that the present invention also provides a kind of for treating liver cancer is as the application in the drug of preparation treatment liver cancer.
The present invention is include at least the following beneficial effects：
1, of the invention for treating the drug of liver cancer, including bone extract is refuted greatly, experiment shows to refute bone extract pair greatly The growth of BEL-7402, Hep-B3 hepatoma cell strain, which has, significantly inhibits effect.
2, the drug for being used to treat liver cancer of the invention, further includes Chinese medical extract, umbellate pore furgus etc. is a variety of in Chinese medical extract Vegetable Chinese herbal medicine further includes gecko, leech drug animal simultaneously, and synergistic function can be generated between animal, plant and not only may be used To further increase the inhibiting effect to the growth of BEL-7402, Hep-B3 hepatoma cell strain, while research shows that Chinese medical extract Addition also have the function of material for anti parotitis, experiment shows that it may be up to 85.9% to the swelling inhibiting rate of mouse.
3, the drug for being used to treat liver cancer of the invention, in preparing the first powder process, first choice passes through cellulase, egg After white enzyme, pectase digest various Chinese herbal medicines, this destroys the cell wall of Chinese herbal medicine, then again will be in after enzymatic hydrolysis Herbal medicine is placed in closed container, passes first into high-pressure steam, then moment pressure release, during being passed through high-pressure steam, High-pressure steam infiltrates into inside Chinese herbal medicine after the cell wall of destruction, then moment pressure release, in the process, infiltrates into High-pressure steam inside herbal medicine can be from " ejection " inside Chinese herbal medicine, so that reaching the effect out of explosion, so that Chinese herbal medicine It is burst from inside, effective medicinal ingredient in Chinese herbal medicine can be dissolved out by high-pressure steam, this compares traditional extracting method Apparent recovery rate greatly improves.
4, the drug for being used to treat liver cancer of the invention will be impregnated after gecko, leech crushing using soak, due to wall Tiger, leech are animal, inevitably have fishy smell, can remove the fishy smell of gecko, leech after impregnating by soak, in soak kind Eggplant, which contains the organic acids such as citric acid, malic acid, to be had the function of neutralizing deodorization, sansevieria trifasciata, ivy, wormwood, lemon in soak Grass, cassia bark cover the fishy smell of animal with the smell of itself.
Further advantage, target and feature of the invention will be partially reflected by the following instructions, and part will also be by this The research and practice of invention and be understood by the person skilled in the art.
Specific embodiment
The present invention is described in further detail below, to enable those skilled in the art's refer to the instruction text being capable of evidence To implement.It should be noted that experimental method described in following embodiments is unless otherwise specified conventional method, it is described Reagent and material commercially obtain unless otherwise specified.
Embodiment 1
It is a kind of including to refute bone extract greatly for treating the drug of liver cancer, the preparation for refuting bone extract greatly include with Lower step：
S1, it will refute bone greatly and be placed in boiling water and extract, and obtain Aqueous extracts；
S2, bone extract will be refuted greatly after the Aqueous extracts separating-purifying in S1 to obtain the final product.
Described to be used extracting in boiling water 2 times, each 2h in S1 for treating the drug of liver cancer, the extracting solution for merging 2 times is Obtain Aqueous extracts；Aqueous extracts separating-purifying specifically includes in S2：By Aqueous extracts 80 DEG C at a temperature of be concentrated into specific gravity be 1.05, so It is eluted afterwards using D101 macroreticular resin, eluant, eluent is followed successively by the ethyl alcohol that volume fraction is 20%, 50%, 95%, collects 95% Ethanol eluate removes ethyl alcohol, is concentrated into specific gravity as 1.08 and refutes bone extract to get big.
Embodiment 2
It is a kind of including to refute bone extract greatly for treating the drug of liver cancer, the preparation for refuting bone extract greatly include with Lower step：
S1, it will refute bone greatly and be placed in boiling water and extract, and obtain Aqueous extracts；
S2, bone extract will be refuted greatly after the Aqueous extracts separating-purifying in S1 to obtain the final product.
Described to be used extracting in boiling water 2 times, each 2h in S1 for treating the drug of liver cancer, the extracting solution for merging 2 times is Obtain Aqueous extracts；Aqueous extracts separating-purifying specifically includes in S2：By Aqueous extracts 80 DEG C at a temperature of be concentrated into specific gravity be 1.05, so It is eluted afterwards using D101 macroreticular resin, eluant, eluent is followed successively by the ethyl alcohol that volume fraction is 20%, 50%, 95%, collects 95% Ethanol eluate removes ethyl alcohol, is concentrated into specific gravity as 1.08 and refutes bone extract to get big.
The drug for being used to treat liver cancer, further includes Chinese medical extract, the preparation method packet of the Chinese medical extract Include following steps：
A1, by 2 parts of umbellate pore furgus of parts by weight, 3 parts of dandelion, 2 parts of subprostrate sophora, 1 part of akebiae, 3 parts of stringy stonecrop, 2 parts of polygonum cuspidate, 1 part of Chinese small iris, 2 parts of catharanthus roseus, 1 part of Radix Glycyrrhizae, 2 parts of radix gentianae, 2 parts of gynostemma pentaphylla, 1 part of winter insult Grass, 5 parts of paris polyphylla, 2 parts of Asian puccoon, 3 parts of bittersweet, 3 parts of mock-strawberry, 2 parts of clearvers, 3 parts of gelsemium elegan, 1 part Patrinia heterophylla, 2 parts of Chloranthus glaber, 2 parts of lobelia chinensis, 4 parts of radices trichosanthis, 2 parts of Prunella vulgaris, 3 parts of airpotato yam, 2 parts of cowherb The first mixing is obtained after art, 1 part of the seed of cowherb, 2 parts of folium cycas, 3 parts of wartwort, 2 parts of Rhizoma Atractylodis Macrocephalae co-grinding；By weight Part is 3 parts of cellulase, 2 parts of protease, 1 part of pectase obtain water enzyme solution after mixing with the water that parts by weight are 10 parts, In the first mixture after mixing evenly by water enzyme solution spray, and 15min is stood at 35 DEG C, obtains the second mixing；By second Mixture is placed in closed container, is then passed through saturated steam into closed container so that the pressure in closed container is 1.1Mpa, after maintaining 20min, then in making in closed container pressure reduction in 4s to 5pa, after keeping 3min, taking-up obtains the Three mixings；Then the water that parts by weight are 150 parts is added into third mixing, is to decoct 2h at 90 DEG C in temperature, filters, filtered Liquid obtains the first powder after being concentrated and dried filter vacuum；
A2, by parts by weight be 2 parts gecko, after 3 parts of leech crushes, being placed in parts by weight is to impregnate in 15 parts of soak The 4th mixing is obtained by filtration after 30min, the preparation method of the soak is：By tomato, sansevieria trifasciata, ivy, wormwood, lemon Grass, cassia bark are dissolved in the water that temperature is 80 DEG C and impregnate 1h, filter up to soak；Wherein, tomato in soak, sansevieria trifasciata, often Spring rattan, wormwood, lemon grass (Cymbopogon citratus), cassia bark, water mass ratio be 3:0.5:0.8:1:0.5:0.5:50；
A3, the 4th mixing obtained in A2 is added to parts by weight is to be heated to reflux 3h in 80 parts of water, is filtered, and obtains the One filtrate and the first filter residue, it is spare；Then, the first filter residue is placed in the ethanol solution that volume fraction is 85% and impregnates 5h, mistake Filter, obtains the second filtrate, obtains the second powder after vacuum freeze drying after the first filtrate, the second filtrate are mixed；
A4, the first powder, the second powder are mixed up to Chinese medical extract；
The bone extract and the mass ratio of Chinese medical extract of refuting greatly is 1:0.2.
It is described for treating the preparation method of the drug of liver cancer, vacuum freeze drying is specially in A3：It is cold at -30 DEG C Freeze 2h, then in vacuum degree is 5Pa, temperature is dry 20h at 60 DEG C.
It is described to use function for treating the preparation method of the drug of liver cancer before the 4th mixing is placed in water reflux in A3 Rate is the ultraviolet light irradiation 5min of 300W.
Embodiment 3
It is a kind of including to refute bone extract greatly for treating the drug of liver cancer, the preparation for refuting bone extract greatly include with Lower step：
S1, it will refute bone greatly and be placed in boiling water and extract, and obtain Aqueous extracts；
S2, bone extract will be refuted greatly after the Aqueous extracts separating-purifying in S1 to obtain the final product.
Described to be used extracting in boiling water 2 times, each 2h in S1 for treating the drug of liver cancer, the extracting solution for merging 2 times is Obtain Aqueous extracts；Aqueous extracts separating-purifying specifically includes in S2：By Aqueous extracts 80 DEG C at a temperature of be concentrated into specific gravity be 1.05, so It is eluted afterwards using D101 macroreticular resin, eluant, eluent is followed successively by the ethyl alcohol that volume fraction is 20%, 50%, 95%, collects 95% Ethanol eluate removes ethyl alcohol, is concentrated into specific gravity as 1.08 and refutes bone extract to get big.
The drug for being used to treat liver cancer, further includes Chinese medical extract, the preparation method packet of the Chinese medical extract Include following steps：
A1, by 3 parts of umbellate pore furgus of parts by weight, 5 parts of dandelion, 4 parts of subprostrate sophora, 2 parts of akebiae, 5 parts of stringy stonecrop, 3 parts of polygonum cuspidate, 2 parts of Chinese small iris, 3 parts of catharanthus roseus, 2 parts of Radix Glycyrrhizae, 3 parts of radix gentianae, 4 parts of gynostemma pentaphylla, 2 parts of winter insult Grass, 6 parts of paris polyphylla, 4 parts of Asian puccoon, 4 parts of bittersweet, 5 parts of mock-strawberry, 3 parts of clearvers, 4 parts of gelsemium elegan, 2 parts Patrinia heterophylla, 3 parts of Chloranthus glaber, 3 parts of lobelia chinensis, 5 parts of radices trichosanthis, 3 parts of Prunella vulgaris, 5 parts of airpotato yam, 3 parts of cowherb The first mixing is obtained after art, 2 parts of the seed of cowherb, 4 parts of folium cycas, 5 parts of wartwort, 4 parts of Rhizoma Atractylodis Macrocephalae co-grinding；By weight Part is 4 parts of cellulase, 4 parts of protease, 2 parts of pectase obtain water enzyme solution after mixing with the water that parts by weight are 12 parts, In the first mixture after mixing evenly by water enzyme solution spray, and 18min is stood at 35 DEG C, obtains the second mixing；By second Mixture is placed in closed container, is then passed through saturated steam into closed container so that the pressure in closed container is 1.1Mpa, after maintaining 25min, then in making in closed container pressure reduction in 5s to 8pa, after keeping 4min, taking-up obtains the Three mixings；Then the water that parts by weight are 155 parts is added into third mixing, is to decoct 3h at 93 DEG C in temperature, filters, filtered Liquid obtains the first powder after being concentrated and dried filter vacuum；
A2, by parts by weight be 3 parts gecko, after 4 parts of leech crushes, being placed in parts by weight is to impregnate in 18 parts of soak The 4th mixing is obtained by filtration after 35min, the preparation method of the soak is：By tomato, sansevieria trifasciata, ivy, wormwood, lemon Grass, cassia bark are dissolved in the water that temperature is 85 DEG C and impregnate 1h, filter up to soak；Wherein, tomato in soak, sansevieria trifasciata, often Spring rattan, wormwood, lemon grass (Cymbopogon citratus), cassia bark, water mass ratio be 4:0.8:1:1.2:0.8:0.8:55；
A3, the 4th mixing obtained in A2 is added to parts by weight is to be heated to reflux 4h in 85 parts of water, is filtered, and obtains the One filtrate and the first filter residue, it is spare；Then, the first filter residue is placed in the ethanol solution that volume fraction is 88% and impregnates 8h, mistake Filter, obtains the second filtrate, obtains the second powder after vacuum freeze drying after the first filtrate, the second filtrate are mixed；
A4, the first powder, the second powder are mixed up to Chinese medical extract；
The bone extract and the mass ratio of Chinese medical extract of refuting greatly is 1:0.2.
It is described for treating the drug of liver cancer, vacuum freeze drying is specially in A3：3h is freezed at -28 DEG C, then In vacuum degree is 8Pa, temperature is dry 25h at 60 DEG C.
It is described for treating the drug of liver cancer, the use of power is 400W's before the 4th mixing is placed in water reflux in A3 Ultraviolet light irradiation 8min.
Embodiment 4
It is a kind of including to refute bone extract greatly for treating the drug of liver cancer, the preparation for refuting bone extract greatly include with Lower step：
S1, it will refute bone greatly and be placed in boiling water and extract, and obtain Aqueous extracts；
S2, bone extract will be refuted greatly after the Aqueous extracts separating-purifying in S1 to obtain the final product.
Described to be used extracting in boiling water 2 times, each 2h in S1 for treating the drug of liver cancer, the extracting solution for merging 2 times is Obtain Aqueous extracts；Aqueous extracts separating-purifying specifically includes in S2：By Aqueous extracts 80 DEG C at a temperature of be concentrated into specific gravity be 1.05, so It is eluted afterwards using D101 macroreticular resin, eluant, eluent is followed successively by the ethyl alcohol that volume fraction is 20%, 50%, 95%, collects 95% Ethanol eluate removes ethyl alcohol, is concentrated into specific gravity as 1.08 and refutes bone extract to get big.
The drug for being used to treat liver cancer, further includes Chinese medical extract, the preparation method packet of the Chinese medical extract Include following steps：
A1, by 5 parts of umbellate pore furgus of parts by weight, 6 parts of dandelion, 6 parts of subprostrate sophora, 3 parts of akebiae, 6 parts of stringy stonecrop, 5 parts of polygonum cuspidate, 3 parts of Chinese small iris, 5 parts of catharanthus roseus, 4 parts of Radix Glycyrrhizae, 5 parts of radix gentianae, 6 parts of gynostemma pentaphylla, 3 parts of winter insult Grass, 8 parts of paris polyphylla, 6 parts of Asian puccoon, 6 parts of bittersweet, 6 parts of mock-strawberry, 5 parts of clearvers, 6 parts of gelsemium elegan, 3 parts Patrinia heterophylla, 4 parts of Chloranthus glaber, 5 parts of lobelia chinensis, 7 parts of radices trichosanthis, 5 parts of Prunella vulgaris, 8 parts of airpotato yam, 5 parts of cowherb The first mixing is obtained after art, 3 parts of the seed of cowherb, 6 parts of folium cycas, 6 parts of wartwort, 6 parts of Rhizoma Atractylodis Macrocephalae co-grinding；By weight Part is 5 parts of cellulase, 6 parts of protease, 3 parts of pectase obtain water enzyme solution after mixing with the water that parts by weight are 5 parts, will Water enzyme solution sprays in the first mixture after mixing evenly, and stands 20min at 35 DEG C, obtains the second mixing；Second is mixed It closes object to be placed in closed container, is then passed through saturated steam into closed container so that the pressure in closed container is 1.1Mpa, after maintaining 30min, then in making in closed container pressure reduction in 6s to 10pa, after keeping 6min, taking-up obtains the Three mixings；Then the water that parts by weight are 160 parts is added into third mixing, is to decoct 3h at 95 DEG C in temperature, filters, filtered Liquid obtains the first powder after being concentrated and dried filter vacuum；
A2, by parts by weight be 5 parts gecko, after 5 parts of leech crushes, being placed in parts by weight is to impregnate in 20 parts of soak The 4th mixing is obtained by filtration after 40min, the preparation method of the soak is：By tomato, sansevieria trifasciata, ivy, wormwood, lemon Grass, cassia bark are dissolved in water at a temperature of 90 °C and impregnate 2h, filter up to soak；Wherein, tomato in soak, sansevieria trifasciata, often Spring rattan, wormwood, lemon grass (Cymbopogon citratus), cassia bark, water mass ratio be 5:1:1.2:1.5:1:1:60；
A3, the 4th mixing obtained in A2 is added to parts by weight is to be heated to reflux 4h in 90 parts of water, is filtered, and obtains the One filtrate and the first filter residue, it is spare；Then, the first filter residue is placed in the ethanol solution that volume fraction is 90% and impregnates 10h, mistake Filter, obtains the second filtrate, obtains the second powder after vacuum freeze drying after the first filtrate, the second filtrate are mixed；
A4, the first powder, the second powder are mixed up to Chinese medical extract；
The bone extract and the mass ratio of Chinese medical extract of refuting greatly is 1:0.2.
It is described for treating the drug of liver cancer, vacuum freeze drying is specially in A3：3h is freezed at -30 DEG C, then In vacuum degree is 10Pa, temperature is dry 30h at 60 DEG C.
It is described for treating the drug of liver cancer, the use of power is 500W's before the 4th mixing is placed in water reflux in A3 Ultraviolet light irradiation 10min.
Comparative example 1
With embodiment 2, difference is, the second powder is not contained in Chinese medical extract.
Anti-tumor activity test method
Anti tumor activity in vitro test
1, cell strain and culture
BEL-7402 hepatoma cell strain is incubated at containing 12% inactivated fetal bovine serum, 90U/mL penicillin, 90 μ g/mL chains It in the RPMn640 culture medium of mycin, is cultivated under the conditions of 37 DEG C, 5%CO2 incubator and saturated humidity, passage in 3~4 days is primary.
Hep-B3 hepatoma cell strain is incubated at containing 12% inactivated fetal bovine serum, 90U/mL penicillin, 90 μ g/mL strepto-s It in the DMEM culture medium of element, is cultivated under the conditions of 37 DEG C, 5%CO2 incubator and saturated humidity, passage in 3~4 days is primary.
2, Antitumor Activity of Drugs measures
Mtt assay：It is 3 × 10 that concentration is configured to after logarithmic growth phase cell is digested with pancreatin4The cell suspension of a/mL, 96 hole elisa Plates are inoculated in, every hole adds 200 μ L.Changing not serum-containing medium afterwards for 24 hours grows cell synchronization, every 200 μ L of hole. Then test drug is added into hole, that is, is separately added into the drug of embodiment 1, embodiment 2, comparative example 1；Separately set control group：0.1% The every hole dimethyl sulfoxide (DMSO) adds 200 μ L.Every group of test drug to set 0.01,0.1,1,10,100,1000 μ g/mL six respectively dense Degree, each concentration sets 8 parallel holes, and after cultivating for 24 hours at 37 DEG C, every hole adds serum-free without phenol red culture solution Fresh The MTT100 μ L of 0.5mg/mL continues to cultivate 4h, and then, every hole adds the DMSO of 100 μ L to dissolve MTT formazan particle, with miniature vibration After swinging device oscillation mixing, in measuring OD value (OD) in microplate reader, experiment in triplicate, is averaged.At solvent control Reason tumour cell is control group, calculates inhibiting rate by OD value, formula is：Inhibitory rate of cell growth=(control group OD value-test drug Group OD value)/control group OD × 100%, and using drug concentration as abscissa, OD value is ordinate, cell growth curve is drawn, and Acquire IC50(half inhibiting rate), IC50The drug concentration that=inhibitory rate of cell growth is 50%, experimental result are as shown in table 1.
The anti-tumor activity experimental result of table 1- difference embodiment
Embodiment BEL-7402IC50(μg/mL) Hep-B3IC50(μg/mL)
Embodiment 1 38.42 35.63
Embodiment 2 16.12 17.36
Comparative example 1 21.89 22.52
The drug for the treatment of liver cancer of the invention as can be known from Table 1, testing through external MTT proves, this pharmaceutical composition can be shown It writes and inhibits the growth of BEL-7402, Hep-B3 hepatoma cell strain, and 2 effect of embodiment is best, illustrates the Chinese medicine being added in embodiment 2 Extract is best to the inhibitory effect of cancer cell.
Mouse 120 are taken, is randomly divided into four groups, one group is control group, in addition uses Indomethacin and the present invention respectively for three groups The drug of the treatment liver cancer of embodiment 2 and the drug of comparative example 1 cause percutaneous drug delivery before inflammation, once a day, continuous three days.In most Dimethylbenzene is applied to two sides inside and outside mouse right ear by single administration half an hour after afterwards, puts to death animal after every mouse about 0.05ml, 42Min, The punch for being 6mm with diameter punches materials weighing along left and right auricle same area, and the weight differential with every group of two auricles is swollen Swollen level index compares the difference between administration group and control group, and finds out the swollen rate of suppression.Swelling inhibiting rate=(control group auricle Weight difference-administration group auricle weight difference)/control group auricle weight difference × 100%.Experimental result is as shown in table 2.
Influence of the drug of table 2- difference embodiment to the ear swelling of mouse
Embodiment Two auricle weight differences Inhibit swelling rate (%)
Control group 9.38 -
Indomethacin group 1.79 80.9
2 medicine group of embodiment 1.32 85.9
1 medicine group of comparative example 1.68 82.1
The drug of the treatment liver cancer of the embodiment of the present invention 2 as can be known from Table 2, also with the work of good material for anti parotitis With.And the effect of the material for anti parotitis of embodiment 2 is better than the drug of comparative example 1.
Although the embodiments of the present invention have been disclosed as above, but its is not only in the description and the implementation listed With it can be fully applied to various fields suitable for the present invention, for those skilled in the art, can be easily Realize other modification, therefore without departing from the general concept defined in the claims and the equivalent scope, the present invention is simultaneously unlimited In specific details.

Claims (6)

1. the drug for treating liver cancer, which is characterized in that including refuting bone extract, the preparation packet for refuting bone extract greatly greatly Include following steps：
S1, it will refute bone greatly and be placed in boiling water and extract, and obtain Aqueous extracts；
S2, bone extract will be refuted greatly after the Aqueous extracts separating-purifying in S1 to obtain the final product.
2. as described in claim 1 for treating the drug of liver cancer, which is characterized in that using extracting in boiling water 2 times in S1, every time 2~3h merges 2 extracting solutions up to Aqueous extracts；Aqueous extracts separating-purifying specifically includes in S2：Temperature by Aqueous extracts at 80 DEG C Be concentrated under degree specific gravity be 1.05, then using D101 macroreticular resin elute, eluant, eluent be followed successively by volume fraction be 20%, 50%, 95% ethyl alcohol collects 95% ethanol eluate, removes ethyl alcohol, is concentrated into specific gravity and extracts for 1.08 to get big bone of refuting Object.
3. as described in claim 1 for treating the drug of liver cancer, which is characterized in that further include Chinese medical extract, it is described in The preparation method of medicament extract includes the following steps：
A1, by 2~5 parts of umbellate pore furgus of parts by weight, 3~6 parts of dandelion, 2~6 parts of subprostrate sophora, 1~3 part of akebiae, 3~6 The stringy stonecrop of part, 2~5 parts of polygonum cuspidate, 1~3 part of Chinese small iris, 2~5 parts of catharanthus roseus, 1~4 part of Radix Glycyrrhizae, 2~5 parts of rough gentian Grass, 2~6 parts of gynostemma pentaphylla, 1~3 part of Rabdosia rubescens, 5~8 parts of paris polyphylla, 2~6 parts of Asian puccoon, 3~6 parts white English, 3~6 parts of mock-strawberry, 2~5 parts of clearvers, 3~6 parts of gelsemium elegan, 1~3 part of Patrinia heterophylla, 2~4 parts of Chloranthus glaber, 2 ~5 parts of lobelia chinensis, 4~7 parts of radices trichosanthis, 2~5 parts of Prunella vulgaris, 3~8 parts of airpotato yam, 2~5 parts of curcuma zedoary, 1~3 Part the seed of cowherb, 2~6 parts of folium cycas, 3~6 parts of wartwort, obtain the first mixing after 2~6 parts of Rhizoma Atractylodis Macrocephalae co-grinding； It is 10~15 parts by cellulase, 2~6 parts of protease, 1~3 part of pectase and parts by weight that parts by weight are 3~5 parts Water mixing after obtain water enzyme solution, by water enzyme solution spray in the first mixture after mixing evenly, and at 35 DEG C stand 15~ 20min obtains the second mixing；Second mixture is placed in closed container, saturated steam is then passed through into closed container So that pressure in closed container is 1.1Mpa, after maintaining 20~30min, then in making pressure drop in closed container in 4~6s Down to 5~10pa, after keeping 3~6min, taking-up obtains third mixing；Then into third mixing be added parts by weight be 150~ 160 parts of water is that 2~3h is decocted at 90~95 DEG C in temperature, and filtering obtains filtrate, obtains after filter vacuum is concentrated and dried First powder；
A2, by parts by weight be 2~5 parts gecko, after 3~5 parts of leech crushes, being placed in parts by weight is 15~20 parts of soak The 4th mixing is obtained by filtration after 30~40min of middle immersion, the preparation method of the soak is：By tomato, sansevieria trifasciata, Chang Chun Rattan, wormwood, lemon grass (Cymbopogon citratus), cassia bark, which are dissolved in the water that temperature is 80~90 DEG C, impregnates 1~2h, filters up to soak；Wherein, it impregnates Tomato in liquid, sansevieria trifasciata, ivy, wormwood, lemon grass (Cymbopogon citratus), cassia bark, water mass ratio be 3~5:0.5~1:0.8~1.2:1~ 1.5:0.5~1:0.5~1:50~60；
A3, the 4th mixing obtained in A2 is added in the water that parts by weight are 80~90 parts, is heated to reflux 3~4h, filtered, obtain First filtrate and the first filter residue, it is spare；Then, the first filter residue is placed in the ethanol solution that volume fraction is 85~90% and is impregnated 5~10h, filtering, obtains the second filtrate, obtains the second powder after vacuum freeze drying after the first filtrate, the second filtrate are mixed；
A4, the first powder, the second powder are mixed up to Chinese medical extract；
The bone extract and the mass ratio of Chinese medical extract of refuting greatly is 1:0.2.
4. as claimed in claim 3 for treating the preparation method of the drug of liver cancer, which is characterized in that vacuum refrigeration is dry in A3 It is dry to be specially：2~3h is freezed at -30~-25 DEG C, then in vacuum degree be 5~10Pa, temperature be dry 20 at 60 DEG C~ 30h。
5. as claimed in claim 3 for treating the preparation method of the drug of liver cancer, which is characterized in that the 4th mixing is set in A3 Before Yu Shuizhong reflux, the ultraviolet 5~10min of light irradiation for the use of power being 300~500W.
6. the drug for treating liver cancer as described in Claims 1 to 5 is any is as answering in the drug of preparation treatment liver cancer With.
CN201811051143.XA 2018-09-10 2018-09-10 For treating drug and the application of liver cancer Pending CN108815214A (en)

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Application publication date: 20181116