Patent application of the present invention is application number " 02117219.6 ", the applying date " on April 17th, 2002 ", the dividing an application of title " Elemene Injection and Preparation Method and Use " patent application
Summary of the invention
The purpose of this invention is to provide elemene vein emulsion injection.
Another object of the present invention provides the preparation method of elemene vein emulsion injection.
The invention provides the novel form of these medicines.With elemene, beta-elemene is that mean diameter that raw material is prepared into this dosage form of vein emulsion type respectively is generally less than or equals 1 micron, even is less than or equal to 100 nanometers.
The accompanying drawing summary
Fig. 1 is the transmission electron microscope photo of the 1A preparation of embodiment 1.
Detailed description of the present invention
Known in the prior art field that some isomer of elemene, Rhizoma Curcumae Longae extract, common turmeric extract possess the effect that suppresses tumor, some documents disclose the various dosage forms (or dosage form) of these natural drugs, for example preparations such as liposome, Emulsion, fat milk.But, because the mean diameter of these dosage forms is generally bigger, can cause the intravital various reactions of human or animal after being expelled in the human or animal body, for example zest is big, and haemolysis sometimes takes place.
The inventor is through years of researches work, discovery with sesquiterpene compounds or the extract that contains sesquiterpene that from plant, extracts make mean diameter≤1 micron or≤the various injection types of 100 nanometers, comprising the liposome dosage form, the nanometer liposome dosage form, the pro-liposome dosage form, the long circulating liposomes dosage form, the long-circulating nanoliposome dosage form, nanometer microemulsion type, vein emulsion type, lipid nanometer microsphere dosage form, fat milk dosage form etc., can overcome the problems in the ejection preparation of prior art, these dosage forms especially of the present invention have significantly reduced zest in intravenous injection is used, and their stability is further enhanced.Novel form of the present invention has significantly improved the drug effect of pharmaceutical preparation, but the mechanism of the anticancer of these dosage forms also is not fully aware of at present.The inventor makes following supposition according to a large amount of result of the tests: because preparation of the present invention has little mean diameter and has special surface nature (hydrophile/lipophile balance for example, surface charge etc.), make the medicine (or active component) in these preparations stronger affinity be arranged to cancerous cell, can fully be attached on the cancerous cell, allow active component directly act on cancerous cell, reach the effect that suppresses or kill cancerous cell.In addition, find that the preparation of nano particle size is rapid-action, prevents that from there is certain effect the cancer cell metastasis aspect.
Simultaneously, although preparation of the present invention has little mean diameter, the preparation of the various dosage forms that obtained has high stability simultaneously, and they coagulation problems can not take place in storing for a long time, makes can store the long period and do not reduce stability.
The invention provides elemene vein emulsion injection, it contains: the extract with total elemene content of isomer 〉=70wt% that extracts through the rectification of material filling type vacuum rectifying apparatus or molecular distillation instrument from plant, refining soybean phospholipid, cholesterol and water for injection, described elemene vein emulsion injection is a kind of aqueous dispersion, the mean diameter of decentralized photo≤1 micron, the weight ratio of extract and refining soybean phospholipid, cholesterol is 0.5-1.5: 2.5-4.0: 1.0-2.0, and the ultimate density of total elemene isomer is 1-10mg/ml in the elemene vein emulsion injection.
The invention provides the preparation method of elemene vein emulsion injection, it comprises the steps:
(1), prepares the extract that contains total elemene isomer by rectification of material filling type vacuum rectifying apparatus or molecular distillation instrument, total elemene content of isomer 〉=70wt% in the described extract;
(2), the extract that step (1) is obtained is that 0.5-1.5: 2.5-4.0: 1.0-2.0 utilizes Rotary Evaporators film forming or CO according to the weight ratio of extract and refining soybean phospholipid, cholesterol
2Critical or supercritical process mixes, and adds water for injection, and high pressure homogenizing is handled, and till the stabilized aqueous dispersion that obtains particle diameter of dispersing phase≤1 micron, the ultimate density that obtains total elemene isomer is the elemene vein emulsion injection of 1-10mg/ml.
The sesquiterpene that extracts from plant (sometimes being called active component here) that can be used for making novel form in the present invention consists essentially of whole sesquiterpene compounds (C15H24), the mixture of sesquiterpene compounds or the sesquiterpene extract that from plant, extracts, extracting product for these requires the purity of sesquiterpene compounds more than 70%, preferably more than 75%, more preferably more than 85%, particularly preferably in more than 90%, even particularly preferably in more than 95%.
It is to be noted, the raw material that is used to prepare the preparation of various dosage forms in the present invention can use product such as elemene product (extract), beta-elemene product (extract), Rhizoma Curcumae Longae extract or the common turmeric extract that is purchased, and the extract product of SHULANHUA, Fructus Tonnae Sinensis, Caulis Piperis Kadsurae, Flos Caryophylli, Myrrha, Hemerocallis citrina Baroni Cuculus polioephalus, caraway, Herba thymi vulgaris, parsley, Herba Pogostemonis, Atractylodes lancea (Thunb.) DC., Fructus Cnidii, Flos lupuli (Flos Humuli Lupuli), sandalwood, bergamot, opoponax, Cupressus funebris Endl., Oleum Terebinthinae, Herba Cymbopogonis Citrari, Radix Ginseng etc.If need oneself these extract of preparation in this application, the various volatile oil that are used to prepare these extracts can use and be purchased product, also can use the volatile oil that obtains from plant by common steam distillation or molecularly distilled.
Sesquiterpene comprises elemene (elemene), curcumene (curcumene), zingiberene (zingiberene), bourbonene (bourbonene), humulene (humulene), Acacia farnesiana Willd. ellagic acid (farnesene), cadinene (cadinene), selinene (selinene), maaliene (maaliene), santalene (santalene), patchoulene (patchonlene), caryophyllene (be also referred to as Flos Caryophylli alkene, caryophyllene), gima ethylenic (big myrcene, germacrene), Cubeb oil alkene (cubebene), cedrene (cedrene), longifolene (longifolene), bisabolene (or bisabolene, bisabolene), bergaptene (bergamotene), gurjunene (gurjunene), lignum-vitae alkene (guaiene), ylangene (ylangene), isocaryophyllene (isocaryophyllene), long pinene (longipinene), Rhizoma Acori Calami alkene (calarene), wood sieve alkene (muurolene), acoradiene (acoradiene), sesquiphellandrene (β-sesquiphellandrene) etc.
In principle, all extracts that contain the above sesquiterpene of 60wt% that extract from plant all can be used for preparing novel form of the present invention, and sesquiterpene is the active component of all dosage forms.The extract that contains the above sesquiterpene of 70wt% is ideal, the extract that contains the above sesquiterpene of 75wt% is preferred, the extract that contains the above sesquiterpene of 85wt% is preferred, contain 90wt% above or even the extract that contains the above sesquiterpene of 95wt% be particularly preferred.
The extract that contains elemene that extracts from natural plants contains α-elemene, beta-elemene, δ-elemene, γ-elemene, bourbonene etc. usually.In this application as specific embodiment, with RADIX CURCUMAE Curcuma wenyujin Y.H.Chen et C.Ling volatile oil, citronella oil Cymbopoqon citratus (DC.) Stapt submember (the mixing grease of remainder after citral, citronellol are removed in the citronella oil fractional distillation) is raw material, makes with extra care out elemene through material filling type vacuum rectifying apparatus or molecular distillation instrument.Wherein be used to prepare this RADIX CURCUMAE volatile oil of elemene raw material of the present invention and described submember and can use and be purchased product, also can from plant, obtain by common steam distillation or molecularly distilled.
In addition, each elemene isomer such as α-elemene, beta-elemene, δ-elemene or γ-elemene are as long as purity also can be used in more than 85wt% and makes novel form of the present invention.
At present according to the domestic elemene quality standard (national drug standards WS that issues referring to National Drug Administration in 2000
1-(X-094)-2000Z), (carbowax-20M is an immobile phase with gas chromatography as if separated products (extract), white diatomite carrier 60~80 orders, column temperature is 135~140 ℃, number of theoretical plate calculates by beta-elemene and should be not less than 1000) detect total elemene isomer purity and reach 〉=85% this products known as elemene, reach as if detecting beta-elemene purity 〉=95% product is a beta-elemene, in fact this is inaccurate, because use the higher chromatographic apparatus of sensitivity for example 30 meters long capillary gas chromatogram-GC-MS detect this 〉=the beta-elemene sample of 95% purity, can find that it still contains at least 6% bourbonene (at the chromatographic chromatogram of muting sensitivity, the peak of beta-elemene becomes a peak with the peak overlapping of bourbonene), the elemene sample is the mixture of multiple sesquiterpene, promptly contain beta-elemene, δ-elemene, γ-elemene, bourbonene, Flos Caryophylli alkene, gima ethylenic etc.
Rhizoma Curcumae Longae Rhizoma Curcuma Longa L. extract is the product that extracts from the volatile oil that curcuma plant obtains.In the present invention as specific embodiment, Rhizoma Curcumae Longae extract refers to the extract that Rhizoma Curcumae Longae volatile oil is made with extra care out through material filling type vacuum rectifying apparatus or molecular distillation instrument, detect through GC-MS, Rhizoma Curcumae Longae extract contains 〉=70wt% usually, preferably 〉=75wt%, more preferably 〉=and 80wt%, preferred especially 〉=85wt%, even more preferably 〉=curcumin of the sesquiterpene of 90wt% such as curcumene, zingiberene etc. and a small amount of (be equivalent to whole sesquiterpene gross weights about 1/25~1/5).
RADIX CURCUMAE Curcuma wenyujin Y.H.Chen et C.Ling extract comprises the product that extracts in the volatile oil that obtains from the RADIX CURCUMAE plant.In the present invention as specific embodiment, common turmeric extract refers to that RADIX CURCUMAE volatile oil is through smart distilled component of vacuum material filling type rectifier unit or the product made with extra care out through the molecular distillation instrument, detect through GC-MS, Rhizoma Curcumae Longae extract contains 〉=70wt% usually, preferably 〉=75wt%, more preferably 〉=80wt%, preferred especially 〉=85wt%, even more preferably 〉=curcumin of the sesquiterpene of 90wt% such as various elemene isomer, gima ethylenic, Flos Caryophylli alkene etc. and a small amount of (be equivalent to whole sesquiterpene gross weights about 1/50~1/10).
The explanation of term:
Described in the present invention particle diameter or granularity are meant the particle diameter or the granularity of decentralized photo.
In this application, if contain 〉=70wt% elemene (comprising various elemene mixture of isomers) and containing<the 85wt% beta-elemene with 30 meters long capillary gas chromatograph-mass spectrometer (GC-MS) Detection and Extraction things, then this extract is called for short elemene, contain 〉=the 85wt% beta-elemene as berry extract, then this extract is called for short beta-elemene.
Modify if before dosage form, add " nanometer ", then refer to mean diameter≤150 nanometers of this dosage form, in ideal conditions mean diameter≤100 nanometers.Injection and ejection preparation are used interchangeably.
The liposome dosage form is meant the Lipid Bilayer Structure of being made up of phospholipid and cholesterol etc. that is dispersed or suspended in the water-bearing media in the present invention, and this Lipid Bilayer Structure is similar to membrane structure.
The nanometer liposome dosage form is meant that in the present invention mean diameter is less than or equal to the liposome of 150 nanometers.
The pro-liposome dosage form is meant in the present invention on the prescription basis of liposome and adds excipient again, by the prepared lyophilized powder of vacuum lyophilization (vacuum freeze-drying) method.This lyophilized powder by adding water for injection, through jolting or vibration, made it to revert to the liposome form before clinical practice.
The long circulating liposomes dosage form, briefly, being meant in the present invention increases the prepared liposome of Polyethylene Glycol PHOSPHATIDYL ETHANOLAMINE again on the prescription basis of liposome.The time of playing effectiveness in vivo than common liposome after it is expelled in the body is longer, or eliminates slowly in vivo.
Nanometer microemulsion type refers to mean diameter≤150 nanometers, in the ideal case≤100 the Emulsion of nanometer.
The vein emulsion type refers to the injection breast of mean diameter≤1 micron in the present invention.
Lipid nanometer microsphere dosage form is meant the fat milk of mean diameter≤150 nanometers in the present invention.
The fat milk dosage form requires its mean diameter≤1 micron in the present invention; In the China national pharmacopeia, also stipulate mean diameter≤1 micron in addition.
Nm is meant nanometer, and μ is meant micron.Purity is meant percent purity by weight in this application, except as otherwise noted.
Employed in this application functional component (or active component) is an elemene, knows that at present occurring in nature mainly contains four kinds of isomers, i.e. α-elemene, beta-elemene, δ-elemene, γ-elemene.The present invention can make the elemene isomer mixture various dosage forms, also can be with through each isomer of purify obtaining α-elemene for example, beta-elemene, δ-elemene, any two or more the mixture in γ-elemene or these isomers is made various dosage forms.For simplicity, use 30 meters long capillary gas chromatograph-mass spectrometer (GC-MS) (chromatographs: Bio-Rad FTS-65A, HP5890II, chromatographic column: BP-5) Detection and Extraction thing, contain 〉=extract (wherein generally also containing other elemene isomer) of 85wt% α-elemene is called α-elemene, similarly, contain 〉=extract of 85wt% beta-elemene is called beta-elemene, contain 〉=extract of 85wt% δ-elemene is called δ-elemene, contain 〉=extract of 85wt% γ-elemene is called γ-elemene.Therefore, the not necessarily simple a kind of isomer of elemene used in the present invention, it in addition can be the various mixture in any ratio of various isomers.
Employed elemene can be bought with extract (various isomer mixture) or pure isomer (requiring purity to be higher than 85wt%) product form by commercial sources and obtain among the application, also can oneself prepare as required, for the preparation method of elemene referring to preparation example 1 and for the preparation method of beta-elemene referring to preparation example 2.
Employed elemene raw material, reagent have no particular limits among the application, as long as this elemene raw material is to extract from natural plants and total elemene isomer purity 〉=70wt% just can use in the present invention, better is, this purity 〉=75wt% is better 〉=85wt%.Beta-elemene, δ-elemene, γ-elemene and α-elemene raw material then is to extract from natural plants and the extract of each isomer purity 〉=85wt%, and that better is purity 〉=90wt%.
Except as otherwise noted, otherwise other composition of Shi Yonging requires to meet national medicinal standard as cholesterol, soybean phospholipid, lecithin, glycerol, linoleic acid, Polyethylene Glycol PHOSPHATIDYL ETHANOLAMINE, aliphatic amine, Semen sojae atricolor wet goods in the present invention, solvent also should meet national medicinal standard, and these are mainly for the consideration of healthy aspect; Similarly, modify with " making with extra care " for some raw materials, show that this raw material must meet national medicinal standard, for example refining soybean phospholipid is the available product of injection, and refined soybean oil is the available product of injection.
Except as otherwise noted, otherwise employed in this application gas chromatograph is: Bio-RadFTS-65A, and HP5890II, chromatographic column is: BP-5.
Various dosage form of the present invention can be used for the treatment of purpose with intravenous injection, topical mode.
The preferred embodiments of the invention
In following embodiments, mainly use extract " elemene ", extract " beta-elemene ", Rhizoma Curcumae Longae extract or common turmeric extract, can also use the extract of SHULANHUA, Fructus Tonnae Sinensis, Caulis Piperis Kadsurae, Flos Caryophylli, Myrrha, Hemerocallis citrina Baroni Cuculus polioephalus, caraway, Herba thymi vulgaris, parsley, Herba Pogostemonis, Atractylodes lancea (Thunb.) DC., Fructus Cnidii, Flos lupuli (Flos Humuli Lupuli), sandalwood, bergamot, opoponax, Cupressus funebris Endl., Oleum Terebinthinae and Herba Cymbopogonis Citrari, Radix Ginseng etc. as raw material.Obviously, higher (product for example 〉=85wt%) can both be as raw material of the present invention, and utilizes synthetic various sesquiterpenes of chemical method and derivant thereof can both be used for the present invention for the purity of wherein a certain sesquiterpene isomer.
The invention provides the preparation method of liposome or nanometer liposome, it comprises: with extract and soybean phospholipid, cholesterol with weight ratio 0.5-1.5: 2.5-4.0: 1.0-2.0, preferred 0.8-1.2: 2.8-3.6: 1.2-1.8, more preferably 0.9-1.1: 3.0-3.4: 1.4-1.6 mixed in preferred especially 1: 3.2: 1.5; Add solvent (as ether or acetone or chloroform), addition is enough to make mixture fully to dissolve, insert and remove the solvent film forming on the Rotary Evaporators, add the water for injection (calculating this surplus) of surplus, insert then in the conventional ultrasonic cell disruptor and carry out ultrasonic Treatment according to the ultimate density that will prepare; Sampling utilizes the microscope of band scale, the Coulter instrument, ultramicroscope is checked mean diameter, at the mean diameter that reaches regulation for example≤1 micron (liposome), or≤100 nanometers (nanometer liposome) after, regulate PH, making pH value is 4.5~6.5, through 0.8 μ filtering with microporous membrane and bottling then, 100 ℃ of vapor stream sterilizations, the ultimate density (weight of sesquiterpene active component (mg)/total volumes of formulation (ml)) of utilizing chromatograph (gas phase or liquid phase) to record active component is 1~10mg/ml, be preferably 2~7.5mg/ml, be more typically 5~7.5mg/ml.
The envelop rate of nanometer liposome detects: detect with sieve method, promptly, utilize the SephadexG25 tomographic system, the extract nano-liposome preparation is divided into two stream parts, be extract nano-liposome stream part and free extract flow part, detect the extractive content that contains in two stream parts through gas chromatograph.The extractive content ÷ of envelop rate computing formula=extract nano-liposome stream part (extractive content+free extractive content of extract nano-liposome stream part).Whether microscope or transmission electron microscope checking are liposome.
The invention provides the preparation method of pro-liposome: with extract and refining soybean phospholipid, cholesterol is with weight ratio 0.5-1.5: 2.5-4.0: 1.0-2.0, preferred 0.8-1.2: 2.8-3.6: 1.2--1.8, more preferably 0.9-1.1: 3.0-3.4: 1.4-1.6, mixed in preferred especially 1: 3.2: 1.5, add solvent (as ether or acetone or chloroform), addition is enough to fully dissolve this mixture, insert and remove the solvent film forming on the Rotary Evaporators, add the water for injection (calculating this surplus) of surplus according to the ultimate density that will prepare, insert ultrasonic cell disruptor then and carry out supersound process, sampling utilizes the microscope or the Coulter instrument of band scale to check mean diameter, behind mean diameter≤1 micron, regulate PH, making pH value is 4.5~6.5, through 0.8 μ filtering with microporous membrane and bottling then, the excipient (for example lactose or average molecular weight Mw are the low molecular dextran of about 4000-40000) that adds the 1-2wt% that accounts for total amount of formulation, 100 ℃ of vapor stream sterilizations, lyophilization under the aseptic condition, embedding.Adding injection water before using is 1~10mg/ml to reach final activity component concentration (weight of sesquiterpene active component (mg)/volumes of formulation (ml)), is preferably 2~8mg/ml, and more preferably 2~7.5mg/ml is more typically 5~7.5mg/ml.
The detection of envelop rate: use the sieve method detection method, that is: according to the final formulation concentrations (mg/ml) that will prepare, the water jolting of a certain amount of extract proliposome preparation being added the surplus volume earlier is even, become liposome this moment again, utilize Sephadex G25 tomographic system to be divided into two stream parts, be extract liposome stream part and free extract flow part, detect the extract amount that contains in two stream parts through gas chromatograph.The extractive content ÷ of envelop rate computing formula=extract liposome stream part (extractive content+free extract amount of extract liposome stream part).Whether microscope or transmission electron microscope confirm is pro-liposome.
The invention provides the preparation method of long circulation extract nano-liposome, this method comprises: with extract and refining soybean phospholipid, Polyethylene Glycol (2000) PHOSPHATIDYL ETHANOLAMINE, cholesterol is with weight ratio 0.5-1.5: 1.5-3.5: 0.3-1.2: 0.8-2.0, preferably with weight ratio 0.8-1.2: 1.8-3.0: 0.4-1.0: 1.2-1.8, more preferably with weight ratio 0.9-1.1: 2.4-2.8: 0.6-1.0: 1.2-1.6, especially preferably with weight ratio 1: 2.6-2.7: 0.7-0.9: 1.3-1.5 mixes, add solvent (as ether or acetone or chloroform), addition is enough to fully dissolve this mixture, insert and remove the solvent film forming on the Rotary Evaporators, add the water for injection (calculating this surplus) of surplus according to the ultimate density that will prepare, insert conventional ultrasonic cell disruptor then and carry out ultrasonic Treatment, sampling utilizes the microscope of band scale, Coulter instrument or ultramicroscope are checked mean diameter, after reaching the mean diameter of regulation (for example≤100 nanometer), regulate PH, making pH value is 4.5~6.5, through 0.8 μ filtering with microporous membrane and bottling then, 100 ℃ of vapor stream sterilizations, the ultimate density (weight of sesquiterpene active component (mg)/total volumes of formulation (ml)) that (gas phase or liquid phase) chromatography records active component is 1~10mg/ml, be preferably 2~7.5mg/ml, more preferably 2~8mg/ml is more typically 5~7.5mg/ml.For Polyethylene Glycol (2000) PHOSPHATIDYL ETHANOLAMINE, wherein " 2000 " refer to the mean molecule quantity (dalton) of Polyethylene Glycol segment.
The detection of the envelop rate of this injection is identical with the envelop rate detection method of the lipidosome injection of front.
The invention provides the preparation method of electronegative and positively charged extract nano microemulsion:
Preparation method 1: the preparation of electronegative nanometer microemulsion
With extract and refining soybean phospholipid, cholesterol is with weight ratio 0.5-1.5: 2.5-4.0: 1.0-2.0, preferred 0.8-1.2: 2.8-3.6: 1.2-1.8, more preferably 0.9-1.1: 3.0-3.4: 1.4-1.6, mixed in preferred especially 1: 3.2: 1.5, add solvent (as ether or acetone or chloroform), addition is enough to fully dissolve this mixture, insert and remove the solvent film forming on the Rotary Evaporators, add the water for injection (calculating this surplus) of surplus according to the ultimate density that will prepare, insert conventional ultrasonic cell disruptor then and carry out ultrasonic Treatment, sampling utilizes the microscope of band scale, Coulter instrument or ultramicroscope are checked mean diameter, after reaching the mean diameter of regulation (for example≤100 nanometer), regulate PH, making pH value is 4.5~6.5, through 0.8 μ filtering with microporous membrane and bottling then, 100 ℃ of vapor stream sterilizations, the ultimate density of active component (weight of sesquiterpene active component (mg)/total volumes of formulation (ml)) is 1~10mg/ml, be preferably 2~7.5mg/ml, be more typically 5~7.5mg/ml.Microscope, transmission electron microscope detect, and confirm whether be the nanometer microemulsion.
Preparation method 2: the preparation of positively charged nanometer microemulsion
With extract, refined lecithin, C14-C22 straight or branched aliphatic amine, cholesterol is with weight ratio 0.5-1.5: 2.5-3.5: 0.2-0.6: 1.0-2.0, preferred 0.8-1.2: 2.5-3.0: 0.3-0.5: 1.2-1.8, more preferably 0.9-1.1: 2.7-2.9: 0.3-0.5: 1.4-1.6, preferred especially 1: 2.8: 0.4: 1.5 mixed, add solvent (as ether or acetone or chloroform), addition is enough to fully dissolve this mixture, insert and remove the solvent film forming on the Rotary Evaporators, add the water for injection (calculating this She's amount) of surplus then according to the densitometer that will prepare, insert conventional ultrasonic cell disruptor then and carry out supersound process, sampling utilizes the microscope of band scale, Coulter instrument or ultramicroscope are checked mean diameter, after reaching the mean diameter of regulation (for example≤100 nanometer), regulate PH, making pH value is 7.5~9.0, through 0.8 μ filtering with microporous membrane and bottling then, 100 ℃ of vapor stream sterilizations, the ultimate density of sesquiterpene active component is 1~10mg/ml, be preferably 2~7.5mg/ml, be more typically 5~7.5mg/ml.Microscope, transmission electron microscope detect, and confirm whether be the nanometer microemulsion.
The present invention also provides the preparation method of extract lipid nanospheres, and this method comprises: get extract and add in the soybean oil with soybean phospholipid, heat up and be controlled to oil phase (remaining oil phase), this oil phase is joined in the glycerinated aqueous solution.Its concrete part by weight is: extract accounts for 0.08~0.8wt% of final preparaton total amount, soybean phospholipid 0.8~2.5wt%, and soybean oil 8-12wt%, glycerol 1.8-2.8wt%, water for injection adds to 5000ml; Preferred part by weight is: extract accounts for the 0.1wt%~0.5wt% of final preparaton total amount, soybean phospholipid 1.2wt%~2.0wt%, and soybean oil 10wt%, glycerol 2.25~2.5wt%, water for injection adds to 5000ml.High-speed stirred then, regulating PH is 4.5~6.5, passing through common high pressure homogenizer (for example M110-E/H type (Japanese MIZUHO produces)) again handles, carry out foregoing ultrasonic Treatment then, sampling utilizes microscope, Coulter instrument or the ultramicroscope of band scale to check mean diameter, the mean diameter of its lipid ball reach setting for example≤100nm after, again through 0.8 μ filtering with microporous membrane and bottling then, 100 ℃~120 ℃ sterilizations.The ultimate density that (gas phase or liquid phase) chromatography records the sesquiterpene active component is 1~10mg/ml, is preferably 2~7.5mg/ml, is more typically 2~5mg/ml, and better is 2.0~3.0mg/ml.Can in this final preparation, add an amount of cholesterol, linoleic acid etc. as required.
The invention provides the preparation method of extract intravenous injection breast, this method comprises: with extract, refining soybean phospholipid, cholesterol is with weight ratio 0.5-1.5: 2.5-4.0: 1.0-2.0, preferred 0.8-1.2: 2.8-3.6: 1.2-1.8, more preferably 0.9-1.1: 3.0-3.4: 1.4-1.6, preferred especially 1: 3.2: 1.5, mix with the water for injection (calculating this surplus) of surplus according to the ultimate density that will prepare, high pressure homogenizer (for example M110-E/H type (Japanese MIZUHO produces)) emulsifying with routine, sampling utilizes the microscope or the Coulter instrument of band scale to check mean diameter, the mean diameter that reaches regulation for example≤1 micron after, regulate PH, making pH value is 4.5~6.5, through 1.2 μ filtering with microporous membranes and bottling then, through 100 ℃~120 ℃ sterilizations, obtain extract vein breast, the ultimate density that (gas phase or liquid phase) chromatography records the sesquiterpene active component is 1~10mg/ml, be preferably 2~7.5mg/ml, be more typically 5~7.5mg/ml.
The invention provides the preparation method of extract fat emulsion, this method comprises: get extract and soybean phospholipid and add in the soybean oil, heat up and be controlled to oil phase (remaining oil phase), this oil phase is joined in the glycerinated aqueous solution.Its concrete weight ratio is: extract accounts for 0.08~0.8wt% of final preparaton total amount, soybean phospholipid 0.8~2.5wt%, and soybean oil 8-12wt%, glycerol 1.8-2.8wt%, water for injection adds to 5000ml; Preferred part by weight is: extract accounts for the 0.1wt%~0.5wt% of final preparaton total amount, soybean phospholipid 1.2wt%~2.0wt%, and soybean oil 10wt%, glycerol 2.25~2.5wt%, water for injection adds to 5000ml.High-speed stirred then, regulating PH is 4.5~6.5, passes through common high pressure homogenizer (for example M110-E/H type (Japanese MIZUHO produces)) again and handles, and makes its particle diameter less than 1 μ, again through 1.2 μ filtering with microporous membranes with bottle 100 ℃~120 ℃ sterilizations then.The ultimate density of sesquiterpene active component is 1~10mg/ml, is preferably 2~7.5mg/ml, the better 2.0~3.0mg/ml of being or be 5~7.5mg/ml.Can in this final preparation, add an amount of cholesterol, linoleic acid etc. as required.
As required, preparation of the present invention can with different size for example the microporous filter membrane of 1.2 μ, 1.0 μ, 0.9 μ, 0.8 μ, 0.7 μ, 0.6 μ, 0.5 μ etc. filter, can intercept mean diameter≤0.8 μ ,≤0.7 μ ,≤0.6 μ ,≤0.5 μ ,≤0.45 μ ,≤0.4 μ ,≤decentralized photo of 0.3 μ etc.
Below preparation example and embodiment be used to illustrate the present invention, but should not think and limit the scope of the invention.Protection scope of the present invention is defined by claim.
Preparation example 1: the preparation of elemene
(citronella oil is removed citral with RADIX CURCUMAE Curcuma wenyujin Y.H.Chen et C.Ling volatile oil or Herba Cymbopogonis Citrari Cymbopoqon citratus (DC.) Stapt submember, the mixing grease of remainder after the citronellol) be raw material, through the Guangzhou Han Wei MD-S80 of mechanical ﹠ electrical corporation type molecular distillation instrument fractional distillation elemene, its process conditions: 30~45 ℃ of vapo(u)rizing temperatures, distillation pressure 70~90Pa, 300~320 rev/mins of knifing speed, 20~25 ℃ of cryosurface temperature, 20~25 ℃ of system's insulations, 15~30 milliliters/hour of material flows, only collect and heat up in a steamer excess, will heat up in a steamer excess and carry out molecular distillation.Vapo(u)rizing temperature is 40~60 ℃, distillation pressure 20~70Pa, 300~320 rev/mins of knifing speed, 20~25 ℃ of condensation temperatures, 20~25 ℃ of system's insulations, 15~30 milliliters/hour of material flows, collect distillation respectively and heat up in a steamer excess, will heat up in a steamer excess and proceed molecular distillation.Vapo(u)rizing temperature is 30~70 ℃, distillation pressure 10~50Pa, 300~320 rev/mins of knifing speed, 20~25 ℃ of condensation temperatures, 20~25 ℃ of system temperatures, 15~30 milliliters/hour of material flows, collect distillation, bleed off and heat up in a steamer excess, the secondary distillation that obtains is mixed, continue distillation, until utilize 30 meters long capillary gas chromatograph-mass spectrometer (GC-MS) gas chromatograph (chromatographs: Bio-Rad FTS-65A, HP5890II, chromatographic column: BP-5) detect total elemene isomer purity to reach 〉=75% till.
Preparation example 2: the preparation of beta-elemene
(citronella oil is removed citral with RADIX CURCUMAE Curcuma wenyujin Y.H.Chen et C.Ling volatile oil or Herba Cymbopogonis Citrari Cymbopoqon citratus (DC.) Stapt submember, the mixing grease of remainder after the citronellol) be raw material, through the Guangzhou Han Wei MD-S80 of mechanical ﹠ electrical corporation type molecular distillation instrument fractional distillation beta-elemene, its process conditions: 30~45 ℃ of vapo(u)rizing temperatures, distillation pressure 70~100Pa, 300~320 rev/mins of knifing speed, 20~25 ℃ of cryosurface temperature, 20~25 ℃ of system's insulations, 7~10 milliliters/hour of material flows, only collect and heat up in a steamer excess, will heat up in a steamer excess and carry out molecular distillation.Vapo(u)rizing temperature is 40~60 ℃, distillation pressure 20~70Pa, 300~320 rev/mins of knifing speed, 20~25 ℃ of condensation temperatures, 20~25 ℃ of system's insulations, 7~10 milliliters/hour of material flows, collect distillation respectively and heat up in a steamer excess, will heat up in a steamer excess and proceed molecular distillation.Vapo(u)rizing temperature is 30~70 ℃, distillation pressure 10~50Pa, 300~320 rev/mins of knifing speed, 20~25 ℃ of condensation temperatures, 20~25 ℃ of system temperatures, 7~10 milliliters/hour of material flows are collected distillation, bleed off and heat up in a steamer excess, the secondary distillation that obtains is mixed, continue distillation, until with 30 meters long capillary gas chromatograph-mass spectrometer (GC-MS)s (chromatograph: Bio-Rad FTS-65A, HP5890II, chromatographic column: BP-5) the beta-elemene purity that detects product reach 〉=85% till.
Embodiment 1: the preparation of the nano-lipid body injection of elemene or beta-elemene
Program 1A: with the elemene product and the refining soybean phospholipid of preparation example 1, cholesterol mixes with weight ratio at 1: 3.2: 1.5, add ether, addition is enough to fully dissolve this mixture, insert and remove the ether film forming on the Rotary Evaporators, add the water for injection (is that 5mg/ml calculates this surplus according to the ultimate density that will prepare) of surplus, insert ultrasonic cell disruptor (model JCS-204) supersound process that Jining, Shandong ultrasonic electronic instrument plant produces then, sampling utilizes the microscope of band scale, Coulter instrument or ultramicroscope are checked mean diameter, after mean diameter≤100 nanometers, regulate PH with 1M biphosphate sodium water solution, making pH value is 4.5~6.5, (new Asia, Shanghai purifies device factory and produces through 0.8 μ microporous filter membrane, specification 0.8 μ) filters and bottling then, 100 ℃ of vapor stream sterilizations, the injection product that is obtained is called for short 1A.Sampling is 5 ± 0.10mg/ml with the ultimate density that gas chromatography records active component (weight (the mg)/volumes of formulation (ml) of total elemene isomer).
Detect envelop rate with sieve method, promptly, utilize Sephadex G25 tomographic system that elemene (referring to extract product) nano liposome preparations is divided into two stream parts, be elemene nanometer liposome stream part and free elemene stream part, detect the extractive content that contains in two stream parts through gas chromatograph.Envelop rate surpasses 85%.The extractive content ÷ of envelop rate computing formula=elemene nanometer liposome stream part [extractive content+free extract amount of elemene nanometer liposome stream part].Microscope, transmission electron microscope detect, and confirmation is a nanometer liposome, provides the transmission electron microscope photo (scale 50nm) of this injection in accompanying drawing 1.
Program 1B: repeat above-mentioned 1A program, just replace the elemene product of preparation example 1 with the beta-elemene product of preparation example 2.Microscope, transmission electron microscope detect, and confirm that preparation is a nanometer liposome.The concentration of total elemene isomer is about 5 ± 0.10mg/ml in the preparation.Envelop rate is higher than 85%.This injection is called for short 1B.
Embodiment 2: the preparation of the pro-liposome of elemene or beta-elemene
2A: according to the 1A program of embodiment 1 in identical method and proportioning raw materials, repeat the 1A program of embodiment 1, just require particle diameter≤1 micron of decentralized photo after the ultrasonic Treatment and after filtering and before bottling, the lactose that adds the 1-2% that accounts for total weight of formulation, utilize 100 ℃ of vapor stream sterilizations then, lyophilization under the aseptic condition, embedding (packing).Detecting the confirmation preparation with the Coulter instrument is pro-liposome.This injection product is called for short 2A.Said preparation added injection water before injection is used be 5 ± 0.10mg/ml to reach final activity component concentration (weight (the mg)/volumes of formulation (ml) of total elemene isomer).Envelop rate is higher than 85%.
2B: repeating above-mentioned 2A, just replace the elemene of preparation example 1 with the beta-elemene of preparation example 2, is about 40000 low molecular dextran replacement lactose with average molecular weight Mw.This injection is called for short 2B.Detecting the confirmation preparation with the Coulter instrument is pro-liposome.Envelop rate is higher than 85%.Said preparation added injection water before injection is used be 5 ± 0.10mg/ml to reach final activity component concentration (weight (the mg)/volumes of formulation (ml) of total elemene isomer).
Embodiment 3: the preparation of the long-circulating nanoliposome of elemene or beta-elemene
3A: according to the 1A program of embodiment 1 in identical method, repeat the 1A program of embodiment 1, just raw material and their weight proportion are: elemene (preparation example 1): soybean phospholipid: Polyethylene Glycol (2000) PHOSPHATIDYL ETHANOLAMINE: cholesterol=1: 2.6: 0.8: 1.4.This injection is called for short 3A.Pick test mean diameter≤100nm.The ultimate density of total elemene isomer is 5 ± 0.10mg/ml in the preparation.Envelop rate is higher than 85%.
3B: repeat above-mentioned 3A, just replace the elemene of preparation example 1 with the beta-elemene of preparation example 2.The injection that is obtained is called for short 3B.Mean diameter≤100nm.The ultimate density of total elemene isomer is 5 ± 0.10mg/ml in the preparation.Envelop rate is higher than 85%.
Embodiment 4: the preparation of the nanometer microemulsion of electronegative and positively charged elemene or beta-elemene
4A-1: the preparation of electronegative nanometer microemulsion
Repeat the 1A program of embodiment 1, just do not measure envelop rate.The ultimate density that sampling records total elemene isomer is 5 ± 0.10mg/ml.Confirm it is the nanometer microemulsion with the Coulter instrument.This injection product is called for short 4A-1.
4A-2: the preparation of positively charged nanometer microemulsion
Repeat the 1A program of embodiment 1, just raw material and their weight proportion are: the elemene of preparation example 1: lecithin: n-octadecane base amine: cholesterol=1: 2.8: 0.4: 1.5, do not measure envelop rate.The ultimate density that sampling records total elemene isomer is 5 ± 0.10mg/ml.Confirm it is the nanometer microemulsion with the Coulter instrument.This injection is called for short 4A-2.
4B-1: the preparation of electronegative nanometer microemulsion
Repeat above-mentioned 4A-1, just replace the elemene of preparation example 1 with the beta-elemene of preparation example 2.The ultimate density that sampling records total elemene isomer is 5 ± 0.10mg/ml.Confirm it is the nanometer microemulsion with the Coulter instrument.The injection that is obtained is called for short 4B-1.
4B-2: the preparation of positively charged nanometer microemulsion
Repeat above-mentioned 4A-2, just replace the elemene of preparation example 1 with the beta-elemene of preparation example 2.The ultimate density that sampling records total elemene isomer is 5 ± 0.10mg/ml.Confirm it is the nanometer microemulsion with the Coulter instrument.The injection that is obtained is called for short 4B-2.
Embodiment 5: the preparation of the lipid nanospheres preparation of elemene or beta-elemene
5A: the elemene of getting preparation example 1 adds in the soybean oil with soybean phospholipid, heats up and is controlled to oil phase (promptly remaining oil phase), and this oil phase is joined in the glycerinated aqueous solution.Its concrete weight ratio is: elemene accounts for 0.2% of total weight of formulation, soybean phospholipid 1.6%, soybean oil 10%, glycerol 2.5%, surplus is a water for injection, high-speed stirred then, regulating PH with 1M biphosphate sodium water solution is 4.5~6.5, handle through M110-E/H type (Japanese MIZUHO produces) high pressure homogenizer again, carry out ultrasonic Treatment according to same procedure among the embodiment 3 then, make the particle diameter≤200nm and the mean diameter≤100nm of its lipid ball, again through 0.8 μ filtering with microporous membrane and bottling then, 100 ℃~120 ℃ sterilizations.The ultimate density that sampling records total elemene isomer is 2 ± 0.04mg/ml.The injection that is obtained is called for short 5A.Can in raw material, comprise an amount of cholesterol, linoleic acid etc. as required.
5B: repeat above-mentioned 5A program, just replace the elemene of preparation example 1 with the beta-elemene of preparation example 2.Particle diameter≤the 200nm of lipid ball and mean diameter≤100nm.The ultimate density that chromatography records total elemene isomer is 2 ± 0.04mg/ml.The injection that is obtained is called for short 5B.
Embodiment 6: the preparation of the intravenous injection breast of elemene or beta-elemene
6A: the elemene of preparation example 1, refining soybean phospholipid, cholesterol are mixed with the water for injection (is that 5mg/ml calculates this surplus according to required final concentration) of surplus with weight ratio 1: 3.2: 1.5, with M110-E/H type (Japanese MIZUHO produces) high pressure homogenizer emulsifying, till usefulness Coulter instrument checks particle diameter less than 1 μ always.Then, regulate PH with 1M biphosphate sodium water solution, making pH value is 4.5~6.5, through 1.2 μ filtering with microporous membranes and bottling then, through 100 ℃~120 ℃ sterilizations, obtain elemene vein breast, the ultimate density that sampling records total elemene isomer is 5 ± 0.10mg/ml.This injection is called for short 6A.
6B: repeat above 6A, just replace the elemene of preparation example 1 with the beta-elemene of preparation example 2.Check particle diameter less than 1 micron with the Coulter instrument.The ultimate density of total elemene isomer is 5 ± 0.10mg/ml in the preparation.The injection that is obtained is called for short 6B.
Embodiment 7: the preparation of elemene fat emulsion
7A: elemene and the soybean phospholipid of getting preparation example 1 add in the soybean oil, heat up and are controlled to oil phase, and this oil phase is joined in the glycerinated aqueous solution.Its concrete weight ratio is: elemene accounts for 0.2% of total weight of formulation, soybean phospholipid 1.2%, and soybean oil 10%, glycerol 2.5%, surplus is a water for injection.High-speed stirred then, regulating PH with 1M biphosphate sodium water solution is 4.5~6.5, handle through M110-E/H type (Japanese MIZUHO produces) high pressure homogenizer again, until with till Coulter instrument check particle diameter≤1 μ, again through 1.2 μ filtering with microporous membranes and bottling then, 100 ℃~120 ℃ sterilizations.The ultimate density that records total elemene isomer is 2 ± 0.04mg/ml.The injection that is obtained is called for short 7A.Can in raw material, comprise an amount of cholesterol, linoleic acid etc. as required.
7B: repeat above-mentioned 7A, just replace the elemene of preparation example 1 with the beta-elemene of preparation example 2.The ultimate density that records total elemene isomer in preparation is 2 ± 0.04mg/ml.The injection that is obtained is called for short 7B.
Embodiment 8:CO
2Supercritical methanol technology prepares liposome
8A: the chloroform that will account for the 10wt% of cholesterol weight adds to be made it in the cholesterol to soak into, the elemene product adding of soybean phospholipid and preparation example 1 is wherein mixed (elemene: soybean phospholipid: cholesterol=weight ratio 1.2: 3.2: 1.5), the mixture that is obtained is put into CO
2In the extractor of supercritical extraction instrument (Han Wei mechanical ﹠ electrical corporation in Guangzhou produces, model SFE100ml), feed CO
2(pressure 50-80kg), 50 ℃ of system temperatures carry out critical or supercritical dispersion, make the mixture uniform mixing, and slowly decompression discharges CO then
2, allow CO
2Chloroform is taken out of, take out extractor, the water for injection (calculating this surplus) that in extractor, adds surplus according to the ultimate density of injection, stir with agitator, with formed dispersion ultrasonic Treatment, until till decentralized photo particle diameter≤100 nanometers of liposome (being called nanometer liposome).Then nano-lipid body and function 1M biphosphate sodium water solution is regulated PH, making pH value is 4.5~6.5, filters and bottling then through 0.8 μ microporous filter membrane (new Asia, Shanghai purifies device factory and produces, specification 0.8 μ), 100 ℃ of vapor stream sterilizations, the injection product that is obtained is called for short 8A.Sampling is 5 ± 0.10mg/ml with the ultimate density that gas chromatography records total elemene isomer.Envelop rate is higher than 88%.
8B: repeat above-mentioned 8A, just replace the elemene of preparation example 1, make liposome with the beta-elemene of preparation example 2.The ultimate density that records total elemene isomer in preparation is 5 ± 0.10mg/ml.The injection that is obtained is called for short 8B.Envelop rate is higher than 88%.
Pharmacodynamics test
The anti-tumor in vivo Evaluation on effect method of pharmaceutical preparation has had description (for example referring to CN1153168A) in above some patent documentation of enumerating.
In the present invention, the preparation of various dosage forms is to human tumor heteroplastic transplantation model QGY hepatocarcinoma, the MKN-45 efficacy in treating gastric carcinoma of subcutaneous vaccination, can be by dosage with 80mg (elemene or beta-elemene)/kg (body weight), 40mg/kg and 20mg/kg, every day, the tail intravenously administrable was 1 time, 10 days therapeutic scheme of successive administration confirms wherein there is not the group in contrast of administration.The relevant regulations " the evaluating drug effect way of antitumor drug " of the National Drug Administration that the calculating of concrete operating process and tumor control rate (tumour inhibiting rate) (referred to before the application's the applying date) according to is at present implemented.
Tumour inhibiting rate %=[(does not have the tumor weight of model of tumor weight-administration of the model of administration)/do not have a tumor weight of the model of administration] * 100%
The tumour inhibiting rate of table 1. human tumor heteroplastic transplantation model QGY hepatocarcinoma
Injection |
Dosage (mg/kg) |
Tumour inhibiting rate |
Dosage (mg/kg) |
Tumour inhibiting rate |
Dosage (mg/kg) |
Tumour inhibiting rate |
1A |
20 |
30.2% |
40 |
39.3% |
80 |
45.2% |
1B |
20 |
30.0% |
40 |
39.1% |
80 |
45.0% |
2A |
20 |
30.1% |
40 |
37.5% |
80 |
44.0% |
2B |
20 |
30.0% |
40 |
38.5% |
80 |
43.6% |
3A |
20 |
29.6% |
40 |
39.6% |
80 |
44.5% |
3B |
20 |
29.8% |
40 |
38.6% |
80 |
44.4% |
4A-1 |
20 |
30.4% |
40 |
38.5% |
80 |
46.0% |
4A-2 |
20 |
31.6% |
40 |
37.9% |
80 |
46.2% |
4B-1 |
20 |
30.2% |
40 |
37.4% |
80 |
45.3% |
4B-2 |
20 |
29.4% |
40 |
38.6% |
80 |
44.9% |
5A |
20 |
29.3% |
40 |
38.7% |
80 |
44.3% |
5B |
20 |
29.5% |
40 |
38.6% |
80 |
44.5% |
6A |
20 |
29.8% |
40 |
37.8% |
80 |
43.2% |
6B |
20 |
29.4% |
40 |
37.9% |
80 |
43.5% |
7A |
20 |
29.2% |
40 |
37.4% |
80 |
44.0% |
7B |
20 |
29.6% |
40 |
37.2% |
80 |
43.2% |
8A |
20 |
29.4% |
40 |
37.4% |
80 |
44.0% |
8B |
20 |
29.2% |
40 |
37.2% |
80 |
43.2% |
The tumour inhibiting rate of table 2. human tumor heteroplastic transplantation model MKN-45 gastric cancer
Injection |
Dosage (mg/kg) |
Tumour inhibiting rate |
Dosage (mg/kg) |
Tumour inhibiting rate |
Dosage (mg/kg) |
Tumour inhibiting rate |
1A |
20 |
28.9% |
40 |
39.3% |
80 |
45.2% |
1B |
20 |
29.1% |
40 |
39.1% |
80 |
45.0% |
2A |
20 |
29.3% |
40 |
37.5% |
80 |
44.0% |
2B |
20 |
28.4% |
40 |
38.5% |
80 |
43.6% |
3A |
20 |
29.1% |
40 |
39.6% |
80 |
44.5% |
3B |
20 |
29.0% |
40 |
38.6% |
80 |
44.4% |
4A-1 |
20 |
29.4% |
40 |
38.5% |
80 |
46.0% |
4A-2 |
20 |
29.5% |
40 |
37.9% |
80 |
46.2% |
4B-1 |
20 |
28.8% |
40 |
37.4% |
80 |
45.3% |
4B-2 |
20 |
28.3% |
40 |
38.6% |
80 |
44.9% |
Table 3: to the curative effect (tumour inhibiting rate) of intracranial in-situ inoculating mice cerebroma G422 model.
Injection |
Dosage (mg/kg) |
Tumour inhibiting rate |
Dosage (mg/kg) |
Tumour inhibiting rate |
Dosage (mg/kg) |
Tumour inhibiting rate |
1A |
20 |
47.6% |
40 |
59.5% |
80 |
68.2% |
1B |
20 |
46.8% |
40 |
59.2% |
80 |
68.5% |
2A |
20 |
47.2% |
40 |
57.6% |
80 |
64.0% |
2B |
20 |
48.2% |
40 |
56.3% |
80 |
63.6% |
3A |
20 |
49.5% |
40 |
59.6% |
80 |
67.5% |
3B |
20 |
48.4% |
40 |
58.3% |
80 |
66.4% |
4A-1 |
20 |
49.3% |
40 |
58.4% |
80 |
67.0% |
4A-2 |
20 |
49.4% |
40 |
57.7% |
80 |
67.2% |
4B-1 |
20 |
48.6% |
40 |
57.2% |
80 |
66.3% |
4B-2 |
20 |
49.3% |
40 |
58.4% |
80 |
67.9% |
5A |
20 |
49.8% |
40 |
58.7% |
80 |
68.3% |
5B |
20 |
47.8% |
40 |
58.3% |
80 |
65.5% |
6A |
20 |
47.6% |
40 |
56.8% |
80 |
63.2% |
6B |
20 |
46.9% |
40 |
56.9% |
80 |
63.5% |
7A |
20 |
48.2% |
40 |
56.4% |
80 |
66.0% |
7B |
20 |
46.8% |
40 |
56.2% |
80 |
63.2% |
Table 4: to the curative effect (tumour inhibiting rate) of subcutaneous vaccination mice cerebroma G422 model.
Injection |
Dosage (mg/kg) |
Tumour inhibiting rate |
Dosage (mg/kg) |
Tumour inhibiting rate |
Dosage (mg/kg) |
Tumour inhibiting rate |
1A |
20 |
23.1% |
40 |
29.8% |
80 |
35.4% |
1B |
20 |
23.0% |
40 |
29.6% |
80 |
35.2% |
2A |
20 |
22.8% |
40 |
27.8% |
80 |
34.0% |
2B |
20 |
22.8% |
40 |
28.5% |
80 |
33.3% |
3A |
20 |
22.9% |
40 |
29.8% |
80 |
34.2% |
3B |
20 |
23.0% |
40 |
28.7% |
80 |
34.8% |
4A-1 |
20 |
24.2% |
40 |
28.1% |
80 |
36.2% |
4A-2 |
20 |
24.4% |
40 |
27.9% |
80 |
36.2% |
4B-1 |
20 |
23.6% |
40 |
27.8% |
80 |
35.2% |
4B-2 |
20 |
23.2% |
40 |
28.7% |
80 |
34.4% |
5A |
20 |
23.4% |
40 |
28.7% |
80 |
34.8% |
5B |
20 |
24.1% |
40 |
28.6% |
80 |
34.5% |
6A |
20 |
22.4% |
40 |
27.9% |
80 |
33.2% |
6B |
20 |
22.6% |
40 |
27.8% |
80 |
33.4% |
7A |
20 |
22.4% |
40 |
27.5% |
80 |
34.1% |
7B |
?20 |
?22.2% |
40 |
?27.3% |
80 |
?33.6% |
In addition, the injection of various dosage forms improves 21%~23% to the NK cells in mice and the matched group specific activity of lotus Lewis lung cancer.
The injection of various dosage forms has improved 1.4~1.55 to the influence of mouse lymphocyte proliferation activity, the stimulation index of lotus Lewis lung cancer.
In a word, from above-mentioned table data as can be seen, various preparations are effective through QGY hepatocarcinoma, MKN-45 gastric cancer, mice cerebroma G422 model pharmacology test.To the Immune Function test, the NK cells in mice of lotus Lewis lung cancer improves 21%~23% with the matched group ratio.Lymphocyte increment activity influence, stimulation index have improved 1.4~1.55.
So elemene injection can be used in and suppresses and/or the treatment tumor.In addition, through overtesting, said preparation can also be used for anticancer to be shifted, and is used for tumor remission pain, is used for prevention or treatment cerebral infarction.