CN101683359A - Traditional Chinese medicine composition for treating tumours and preparation method thereof - Google Patents
Traditional Chinese medicine composition for treating tumours and preparation method thereof Download PDFInfo
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- CN101683359A CN101683359A CN200810200742A CN200810200742A CN101683359A CN 101683359 A CN101683359 A CN 101683359A CN 200810200742 A CN200810200742 A CN 200810200742A CN 200810200742 A CN200810200742 A CN 200810200742A CN 101683359 A CN101683359 A CN 101683359A
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Abstract
The invention discloses a traditional Chinese medicine composition for treating tumours and a preparation method thereof. The traditional Chinese medicine composition contains a bufogenin extract anda medicinal carrier, wherein the bufogenin extract is prepared by the purification of a silica gel column after being extracted by a toad venom medicinal material, the total content is not below 92%,and then the traditional Chinese medicine composition is prepared by dissolution, filtration, encapsulation and sterilization and can be further prepared into a liquor for injection or freeze-drying powder injection. The traditional Chinese medicine composition has a favorable anticancer action for lung cancer, liver cancer, mammary cancer, stomach cancer, colonic cancer and the like, can increaseleucocyte and has less side effect.
Description
Technical field
The present invention relates to a kind of Chinese medicine composition and preparation method thereof, relate generally to a kind of composition and method of making the same that contains the bufotalin extract that is used for the treatment of tumor.
Background technology
Medicinal Bufo siccus is Amphibia Bufonidae animal Bufo siccus (Bufo bufogargarizans Cantor) or Bufo melanostictus (Bufo melanostictus Schneider), Venenum Bufonis is the ear rear gland of Bufo siccus or Bufo melanostictus and the white serosity of skin gland secretion, makes through dry processing.The Bufo siccus of proved recipe extensive use at all times, Venenum Bufonis draw out pus by applying a plaster to the affected part detumescence, analgesic therapy parasite killing, heart tonifying diuresis.Modern study shows, the mixture of the bufogenin in the Venenum Bufonis, cinobufacin, three kinds of liposoluble constituents of Toadpoison Medicine not only has obvious antineoplastic, and can also be effects such as normal cell (inducing tumor cell differentiation), inducing apoptosis of tumour cell, the formation of inhibition tumor cell new vessels, reduction neoplasm metastasis with tumour cell transformation.
Chinese medicine is a kind of novel drug-supplying system, can make Effective Components of Chinese Herb can well bring into play its therapeutical effect.Present existing document reports that the total content of three kinds of liposoluble constituents is lower than 80%, does not meet the new drug requirements of customs declaration of existing Chinese medicine.
Summary of the invention
The technical problem to be solved in the present invention is the Chinese medicine composition that how to obtain to meet existing Chinese medicine specification requirement.
For this reason, one of purpose of the present invention has provided a kind of Chinese medicine composition that meets the Chinese medicine injection specification requirement.Concrete technical scheme has provided a kind of Chinese medicine composition, and said composition contains the bufotalin extract and the lytic agent of bufogenin, cinobufacin and Toadpoison Medicine, and the total content of three kinds of compositions is not less than 92% in the described bufotalin extract.Described compositions can further be made into the solution or the lyophilized injectable powder of injection, or additionally adds the lyophilizing caffolding agent, further is made into powder ampoule agent for injection.
Lytic agent in the described Chinese medicine composition can be medical aqueous vehicle, or the solvent that contains PEG of medicine, or moisture and PEG is by the blended medical solvent of arbitrary ratio, preferred water or PEG400 or its mixing in arbitrary ratio, preferred water or PEG400 especially.
Above-mentioned Chinese medicine composition can also add the active medicine of other treatment tumor except that bufotalin extract and lytic agent, also can contain other ancillary drug such as glucose etc.
Described extract obtains in the following ways.With the alcohols solvent is to extract solvent, with the silicagel column is detached dowel, adopt a certain proportion of petroleum ether and ethyl acetate isocratic elution, extraction efficiency height not only, simplified operating procedure, also further improved the content of above-mentioned three kinds of bufotalin extracts, content is not less than 92%, preferably be not less than 95%, especially preferably be not less than 97%.Preferred dehydrated alcohol of said extracted solvent or 95% ethanol; Bufogenin, cinobufacin and Toadpoison Medicine three's weight ratio is that weight ratio is 2～3: 1～2 in the above-mentioned bufotalin extract: 1～2, and preferred weight ratio is 2: 1～1.5: 1～1.3.
Another purpose of the present invention provides a kind of preparation method of described Chinese medicine composition.This method may further comprise the steps:
A, to get the Venenum Bufonis medical material an amount of, adds alcohols solvent, heating and refluxing extraction, and the extracting solution filtered while hot is prepared into concentrated solution, preferred dehydrated alcohol of above-mentioned alcohols solvent or 95% ethanol after concentrating; With
B, above-mentioned concentrated solution and proper silica gel are stirred, with petroleum ether: ethyl acetate (volume ratio 4: 1) is soaked back wet method dress post, use petroleum ether: (volume ratio 2～7: 1) eluant carries out the constant speed eluting to ethyl acetate, collect eluent, evaporate to dryness, promptly get bufotalin extract of the present invention, the total content of three kinds of components is not less than 92% in the described extract, above-mentioned eluant petroleum ether: preferred 4.5: 1 of the volume ratio of ethyl acetate; With
C, to get the said extracted thing an amount of, with lytic agent it dissolved, and dissolving back concentration is 0.01～2g/L, and ultrasonic 15min places 48h.Under aseptic condition, membrane filtration with 0.45 μ m or 0.22 μ m, get transparent supernatant liquid, with this liquid embedding in ampoule, sterilization, promptly get Chinese medicine composition of the present invention, preferred 0.05～the 1g/L of described concentration, preferred especially 0.1～0.5g/L, lytic agent can be medical aqueous vehicle, or the solvent that contains PEG of medicine, or moisture and PEG is in the blended medical solvent of arbitrary ratio, preferred water or PEG400 or its mixing in arbitrary ratio, special preferred water or PEG400, this Chinese medicine composition can be made into injection solution agent or injectable powder; Or
D, to get the said extracted thing an amount of, and with the lytic agent dissolving, dissolving back concentration is 0.01～2g/L, and ultrasonic 15min places 48h.Under aseptic condition, membrane filtration with 0.45 μ m or 0.22 μ m, get transparent supernatant liquid, it is an amount of to get clear liquid, add an amount of lyophilizing caffolding agent, sterilization, mixed back lyophilizing promptly gets Chinese medicine composition of the present invention, preferred 0.05～the 1g/L of described concentration, preferred especially 0.1～0.5g/L, described lytic agent can be medical aqueous vehicle, or the solvent that contains PEG of medicine, or moisture and PEG is in the blended medical solvent of arbitrary ratio, preferred water or PEG400 or its mixing in arbitrary ratio, special preferred water or PEG400, described lyophilizing caffolding agent is a mannitol, or lactose, or trehalose, or the three is in the mixing of arbitrary ratio, and said composition can be made into freeze-dried powder injection.
Chinese medicine composition of the present invention is to be raw medicinal material with the Venenum Bufonis, and main component is the bufotalin extract of bufogenin, cinobufacin and Toadpoison Medicine, is a kind of to the medicable Chinese medicine composition of tumor.Show that through animal experiment diseases such as pulmonary carcinoma, hepatocarcinoma, breast carcinoma, colon cancer, gastric cancer are had the good curing effect, the curative effect of pulmonary carcinoma, hepatocarcinoma is better than Venenum Bufonis Injection, the cyclophosphamide identical with dosage is suitable.The toxicology test of mice shows that this injection toxicity is lower than Venenum Bufonis Injection, and has function of increasing leukocyte.
Description of drawings
Fig. 1 is the HPLC analysis chart of bufogenin, cinobufacin, three kinds of bufotalin extracts of Toadpoison Medicine
The specific embodiment
Further specify the present invention with embodiment below, but the present invention is not so limited.
Embodiment 1
Get Venenum Bufonis medical material 1 gram, add 95% ethanol 10mL, reflux 40min extracts 2 times under the above-mentioned condition.With the extracting solution filtered while hot, revolve the inspissation back of contracting and add 1 gram silica gel mixed sample, must mix sample silica gel.Get 20 gram silica gel with petroleum ether: ethyl acetate (volume ratio 4.5: 1) is soaked back wet method dress post, adds and mixes sample silica gel, and use petroleum ether: ethyl acetate (volume ratio 4.5: 1) eluant carries out isocratic elution.Analyze by the HPLC under the usual terms, as shown in Figure 1, retention time is the Toadpoison Medicine that is of 7.184min, 8.886min be cinobufacin, 9.549min be bufogenin, collect the eluent contain these three kinds of liposoluble constituents, evaporate to dryness, promptly get bufotalin extract of the present invention, wherein three kinds of active bufotalins percentage by weight of accounting for total extract is 97.88%.
Embodiment 2
Take by weighing three kinds of bufotalin extracts of 30mg, it is dissolved in the brown volumetric flask of 250mL ultrasonic 15min with distilled water, place 48h,, get transparent clarification medicinal liquid with the membrane filtration of 0.45 μ m, medicine liquid irrigation is encapsulated in the 2mL ampoule, flowing steam sterilization, and promptly getting loading amount is the injection solution agent of 2mL.
Embodiment 3
Take by weighing three kinds of bufotalin extracts of 30mg, it is dissolved in the brown volumetric flask of 250mL ultrasonic 15min with distilled water, place 48h,, get transparent clarification medicinal liquid with the membrane filtration of 0.22 μ m, add an amount of lyophilizing caffolding agent, sterilization, fill is in the cillin bottle of 10mL under the aseptic condition, lyophilizing, promptly get the injection lyophilized injectable powder, the lyophilizing caffolding agent is a mannitol, or lactose, or trehalose, or the three is mixed in arbitrary ratio.
Get well-grown rat liver cancer H22 ascites, be diluted to 1-2 * 10 with normal saline
7The cell suspension of individual/mL concentration, every the right axil subcutaneous vaccination of mice 0.2mL, random packet is established: 1) blank group (NS); 2) cyclophosphamide (CTX, 5mg/kg, ip * 7); 3) Venenum Bufonis Injection (5mg/kg, iv * 7); 4) bufogenin composite injection (1mg/kg, iv * 7); 5) bufogenin composite injection (5mg/kg, iv * 7).
Rise by above scheme administration next day the inoculation back, and the administration volume is the 0.5mL/20g body weight.Inoculate back 10 days and take off neck execution animal, dissect and get the tumor piece, claim tumor heavy, the result judges according to following formula:
The result: the continuous tail vein injection of bufogenin composite injection 7 days, 1mg/kg and 5mg/kg dosage group are respectively 57.36% and 65.97% to the tumour inhibiting rate of rat liver cancer H22; Positive drug CTX group lumbar injection 7 days, tumour inhibiting rate is 69.57%; Venenum Bufonis Injection lumbar injection 7 days, tumour inhibiting rate are 39.65%.The results are shown in Table 1.
Table 1. toad element is to the tumor-inhibiting action of rat liver cancer H22
Compare with the blank group:
*P＜0.05,
*P＜0.01.
Claims (10)
1, a kind of Chinese medicine composition, it is characterized in that containing bufotalin extract and lytic agent, described bufotalin extract is made up of bufogenin, cinobufacin and three kinds of active component of Toadpoison Medicine, and three kinds of active component total contents are not less than 92% in the described bufotalin extract.
2, Chinese medicine composition according to claim 1 is characterized in that three kinds of active component total contents are not less than 95% in the described bufotalin extract.
3, Chinese medicine composition according to claim 2 is characterized in that three kinds of active component total contents are not less than 97% in the described bufotalin extract.
4, Chinese medicine composition according to claim 1, the weight ratio that it is characterized in that stating bufogenin, cinobufacin and Toadpoison Medicine in the bufotalin extract is 2: 1～1.5: 1～1.3.
5, Chinese medicine composition according to claim 1 is characterized in that described lytic agent is selected from water, PEG400 and its mixing in arbitrary ratio.
6, Chinese medicine composition according to claim 1 is characterized in that also containing the lyophilizing caffolding agent.
7, Chinese medicine composition according to claim 6 is characterized in that described lyophilizing caffolding agent is a mannitol, or lactose, or trehalose, or the three is in the mixing of arbitrary ratio.
8,, it is characterized in that described compositions can be made into injection solution agent or lyophilized injectable powder according to claim 5 or 6 described Chinese medicine compositions.
9, according to the purposes of the described Chinese medicine composition of claim 1 to 7 in preparation medicine for treating tumor thing, it is characterized in that described tumor comprises pulmonary carcinoma, hepatocarcinoma, breast carcinoma, gastric cancer or colon cancer.
10, the preparation method of the described Chinese medicine composition of a kind of claim 1 to 7 is characterized in that preparing according to the following steps:
A, to get the Venenum Bufonis medical material an amount of, adds 95% ethanol, heating and refluxing extraction, and the extracting solution filtered while hot is prepared into concentrated solution after concentrating; With
B, above-mentioned concentrated solution and proper silica gel are stirred, with petroleum ether: ethyl acetate (volume ratio 4: 1) is soaked back wet method dress post, use petroleum ether: ethyl acetate (volume ratio 4.5: 1) eluant carries out the constant speed eluting, collect eluent, evaporate to dryness, promptly get bufotalin extract of the present invention, the total content of three kinds of components is not less than 92% in the described extract; With
C, to get the said extracted thing an amount of, and with an amount of lytic agent dissolving, dissolving back concentration is 0.01～2g/L, and ultrasonic 15min places 48h.Under aseptic condition, with the membrane filtration of 0.45 μ m or 0.22 μ m, get transparent supernatant liquid, in ampoule, sterilization promptly gets Chinese medicine composition of the present invention with this liquid embedding; Or
D, to get the said extracted thing an amount of, and with an amount of lytic agent dissolving, dissolving back concentration is 0.01～2g/L, and ultrasonic 15min places 48h.Under aseptic condition, with the membrane filtration of 0.45 μ m or 0.22 μ m, get transparent supernatant liquid, it is an amount of to get clear liquid, add an amount of lyophilizing caffolding agent, sterilization, lyophilizing promptly gets described Chinese medicine composition of the present invention, described lyophilizing caffolding agent is a mannitol, or lactose, or trehalose, or the three is in the mixing of arbitrary ratio.
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| CN102247335A (en) * | 2011-06-02 | 2011-11-23 | 江苏省中医药研究院 | Bufalin dry powder inhalant as well as preparation method and application thereof |
| CN102302475A (en) * | 2011-06-02 | 2012-01-04 | 江苏省中医药研究院 | Resibufogenin dry powder inhalant and preparation method and application thereof |
| CN102302477A (en) * | 2011-06-02 | 2012-01-04 | 张振海 | Cinobufagin dry power inhaler and preparation method and application thereof |
| CN102533895A (en) * | 2010-12-10 | 2012-07-04 | 中国科学院长春应用化学研究所 | Preparation method for 3-surface ester bufotalin |
| CN103191133A (en) * | 2013-04-23 | 2013-07-10 | 山东中医药大学 | Preparation method for extracting cinobufagin and bufalin mixture from venenum bufonis |
| CN103191132A (en) * | 2013-04-23 | 2013-07-10 | 山东中医药大学 | Preparation method for extracting high-purity resibufogenin from venenum bufonis |
| CN104736157A (en) * | 2012-05-09 | 2015-06-24 | 香港科技大学 | Method and compounds for inhibiting the MCM complex and their application in cancer treatment |
| CN105769927A (en) * | 2016-03-17 | 2016-07-20 | 南京多宝生物科技有限公司 | Venenum Bufonis extraction method and Venenum Bufonis product |
| CN107233349A (en) * | 2017-07-07 | 2017-10-10 | 黄娇艳 | Application of the drug combination in the medicine for the treatment of leukaemia or lymthoma is prepared |
| CN108836982A (en) * | 2018-07-06 | 2018-11-20 | 合肥华方医药科技有限公司 | A kind of toad skin active component and preparation method thereof |
| CN113813284A (en) * | 2020-06-19 | 2021-12-21 | 再少年(上海)细胞技术有限公司 | Materials for inhibiting tumor formation |
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| CN102533895A (en) * | 2010-12-10 | 2012-07-04 | 中国科学院长春应用化学研究所 | Preparation method for 3-surface ester bufotalin |
| CN102533895B (en) * | 2010-12-10 | 2014-05-21 | 中国科学院长春应用化学研究所 | Preparation method for 3-surface ester bufotalin |
| CN102302475A (en) * | 2011-06-02 | 2012-01-04 | 江苏省中医药研究院 | Resibufogenin dry powder inhalant and preparation method and application thereof |
| CN102302477A (en) * | 2011-06-02 | 2012-01-04 | 张振海 | Cinobufagin dry power inhaler and preparation method and application thereof |
| CN102302477B (en) * | 2011-06-02 | 2013-08-28 | 张振海 | Cinobufagin dry power inhaler and preparation method and application thereof |
| CN102247335A (en) * | 2011-06-02 | 2011-11-23 | 江苏省中医药研究院 | Bufalin dry powder inhalant as well as preparation method and application thereof |
| CN104736157A (en) * | 2012-05-09 | 2015-06-24 | 香港科技大学 | Method and compounds for inhibiting the MCM complex and their application in cancer treatment |
| US11648258B2 (en) | 2012-05-09 | 2023-05-16 | The Hong Kong University Of Science And Technology | Method and compounds for inhibiting the MCM complex and their application in cancer treatment |
| CN110412285B (en) * | 2012-05-09 | 2022-11-15 | 香港科技大学 | Methods and compounds for inhibiting MCM protein complexes and their use in treating cancer |
| CN110412285A (en) * | 2012-05-09 | 2019-11-05 | 香港科技大学 | The method and compound of inhibition MCM albumen composition and its application in treating cancer |
| CN103191133B (en) * | 2013-04-23 | 2014-12-24 | 山东中医药大学 | Preparation method for extracting cinobufagin and bufalin mixture from venenum bufonis |
| CN103191132B (en) * | 2013-04-23 | 2014-12-24 | 山东中医药大学 | Preparation method for extracting high-purity resibufogenin from venenum bufonis |
| CN103191132A (en) * | 2013-04-23 | 2013-07-10 | 山东中医药大学 | Preparation method for extracting high-purity resibufogenin from venenum bufonis |
| CN103191133A (en) * | 2013-04-23 | 2013-07-10 | 山东中医药大学 | Preparation method for extracting cinobufagin and bufalin mixture from venenum bufonis |
| CN105769927A (en) * | 2016-03-17 | 2016-07-20 | 南京多宝生物科技有限公司 | Venenum Bufonis extraction method and Venenum Bufonis product |
| CN107233349A (en) * | 2017-07-07 | 2017-10-10 | 黄娇艳 | Application of the drug combination in the medicine for the treatment of leukaemia or lymthoma is prepared |
| CN108836982A (en) * | 2018-07-06 | 2018-11-20 | 合肥华方医药科技有限公司 | A kind of toad skin active component and preparation method thereof |
| CN113813284A (en) * | 2020-06-19 | 2021-12-21 | 再少年(上海)细胞技术有限公司 | Materials for inhibiting tumor formation |
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