CN101306184B - Common turmeric extract vein emulsion injection and its production and use - Google Patents
Common turmeric extract vein emulsion injection and its production and use Download PDFInfo
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- CN101306184B CN101306184B CN200810126418.1A CN200810126418A CN101306184B CN 101306184 B CN101306184 B CN 101306184B CN 200810126418 A CN200810126418 A CN 200810126418A CN 101306184 B CN101306184 B CN 101306184B
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Abstract
Common turmeric extract vein emulsion injection of the present invention and its production and use relates to the new injection type of common turmeric extract, more specifically to vein emulsion type, and the preparation method of this dosage form and purposes.Common turmeric extract vein emulsion injection of the present invention contains from RADIX CURCUMAE plant, extract total sesquiterpene alkene class content>=70wt% extract, Refined Soybean phospholipid, cholesterol, adds water for injection, utilizes Rotary Evaporators film forming or CO
2critical or supercritical process mix homogeneously, carries out high pressure homogenizing process, obtain the stabilized aqueous dispersion of particle diameter of dispersing phase≤1 micron common turmeric extract vein emulsion injection.Common turmeric extract vein emulsion injection of the present invention significantly improves the drug effect of pharmaceutical preparation, and decreases the zest in intravenous injection application, improves stability.
Description
Patent application of the present invention is application number " 02117218.8 ", the applying date " on April 17th, 2002 ", the divisional application of title " Curcuma wenyujin extract Injection And Preparation Method And Use " patent application
Technical field
The present invention relates to the new injection type of common turmeric extract, more specifically to vein emulsion type.The invention still further relates to preparation method and the purposes of this dosage form.
Prior art
In recent years, the many research worker in anticancer natural drug research field are all devoted to study the active anticancer of paclitaxel, the various isomer of elemene, Rhizoma Curcumae Longae extract, common turmeric extract etc. and the various dosage forms of these medicines.
Elemene is a kind of sesquiterpenoids be present in certain plants, and Latin is called Elemenum, and English Elemene by name, chemical name is Elemene, molecular formula C
15h
24.Elemene has multiple isomer, beta-elemene in these isomers, δ-elemene, and γ-elemene has been proved active anticancer.Elemene is the compound that can extract from the volatile oil of the rhizome (being commonly called as Curcuma wenyujin) of plant RADIX CURCUMAE (Curcuma Wenyujin Y.H.Chen et C.ling), or the compound extracted from the oil of plant Herba Cymbopogonis Citrari (Cymbopoqoncitratus (DC.) Stapt), such as, after removing citral, citronellol from citronella oil remaining mixing grease, the compound of extraction is actually the elemene isomer mixture comprising beta-elemene.
Above-mentioned plant RADIX CURCUMAE and other two kinds can be used as plant Rhizoma Curcumae that natural drug uses and Rhizoma Curcumae Longae belongs to zingiberaceous plant monoid, originate in the province such as Guangdong, Sichuan, Fujian, Guangxi, Zhejiang of China.The plant comprising elemene has the kinds more than 50 such as RADIX CURCUMAE, Herba Cymbopogonis Citrari, Rhizoma Acori Calami, Radix Inulae, Radix Ginseng, and wherein Herba Cymbopogonis Citrari is wide at distribution in China, that resource is sufficient plant.At present, Elemene vinyl compound mainly extracts from RADIX CURCUMAE oil and citronella oil.
Having many documents to relate to directly adopts natural drug to prepare the herbal medicinal product of anti-curing cancers, the open CN1216256A of such as Chinese patent discloses the herbal medicinal product of Hepatoma therapy, it comprises containing Herba Hedyotidis Diffusae, Rhizoma Curcumae Longae, and the extract of Rhizoma Polygoni Cuspidati and Radix Sophorae Tonkinensis is as the Injectable composition (A) of main natural drug and the extract containing Herba Hedyotidis Diffusae, Rhizoma Paridis, Rhizoma Polygoni Cuspidati, Radix Sophorae Tonkinensis, Radix Gentianae, Radix Et Rhizoma Rhei, Fructus Forsythiae, Radix Paeoniae Rubra, Rhizoma Curcumae Longae and the Rhizoma Acori Graminei Orally administered composition (B) as main natural drug.
Curcumin preparation is disclosed in the open CN1302559 of Chinese patent; it is the curcumin molecular dispersoid by being prepared in the blendable organic solvent of water or in the mixture of water and the blendable organic solvent of water; then add protecting colloid aqueous solution to this solution, the hydrophobic of curcumin forms millimicro decentralized photo mutually.Final dosage form is aqueous dispersion or dry powder mainly, and particle diameter is less than 10 μm in the preferred case.
The method being extracted beta-elemene by secondary fractional distillation from citronella oil is disclosed in CN1200266A.
In the open CN1247756A of Chinese patent, disclose the Jiangyu injecta being used for the treatment of malignant tumor, wherein the weight ratio of Rhizoma Curcumae Longae and Radix Curcumae is 50-60: 50-40, and this injection is a kind of simple mixture.
The open CN1076613A of Chinese patent relates to elemene emulsion injection and preparation method thereof, and this injection is a kind of emulsion, and mean diameter is generally no more than 15 microns, is no more than 2 microns especially.
The open CN1244389A of Chinese patent relates to elemene injection and preparation method thereof, and this injection is a kind of simple mixtures, is not liposome.
The open CN1110134A of Chinese patent relates to liposomal preparation technique and preparation, the elemene liposome injecta wherein particularly describing elemene liposome injecta preparation method and obtained by the method, the envelop rate of elemene reaches more than 85%, but does not provide the data of the mean diameter of related lipase plastid.Can judge that this liposome has larger mean diameter from preparation method.
The open CN1221607A of Chinese patent relates to and utilizes CO
2critical or supercritical process prepare the technique of liposoluble medicinal liposome and prepared elemene liposome injecta, although the envelop rate of the liposome of elemene is very high, the mean diameter of liposome is equally too large.
Preparation above disclosed in these documents all also exists deficiency on bin stability and active anticancer.
In addition, in some patent documentations, also disclose various derivant and the elemene metal complex of elemene.Such as, in CN11531158A, a kind of elemene metal complex is disclosed and it is as the use of cancer therapy drug.CN1153167A relates to elemene derivatives containing nitrogen and the purposes as cancer therapy drug thereof.CN1153168A relates to elemene hydroxyls derivs and the purposes as cancer therapy drug thereof.
Known some isomer of elemene in prior art field, effect that Rhizoma Curcumae Longae extract, common turmeric extract possess Tumor suppression, certain references disclose the various dosage forms (or dosage form) of these natural drugs, the preparations such as such as liposome, Emulsion, fat milk.But the mean diameter due to these dosage forms is general comparatively large, be expelled to the various reactions that to cause after in human or animal body in human or animal body, such as zest is large, sometimes haemolysis occurs.
The present inventor is through years of researches work, find sesquiterpene compounds or the various injection types making mean diameter≤1 micron or≤100 nanometers containing the extract of sesquiterpene that extract from plant, comprising Lipidosome, nanometer liposome dosage form, pro-liposome dosage form, long circulating liposomes dosage form, long-circulating nanoliposome dosage form, nanometer microemulsion type, vein emulsion type, lipid nanometer microsphere dosage forms, micro-emulsion etc., the problems in the ejection preparation of prior art can be overcome, especially these dosage forms of the present invention greatly reduce zest in intravenous injection application, their stability is further enhanced.Novel form of the present invention significantly improves the drug effect of pharmaceutical preparation, but the mechanism of the anticancer of these dosage forms is not also fully aware of at present.The present inventor makes following supposition according to a large amount of result of the tests: have little mean diameter due to invention formulation and have special surface nature (such as hydrophile/lipophile balance, surface charge etc.), the medicine in these preparations (or active component) is made to have stronger affinity to cancerous cell, can fully be attached on cancerous cell, allow active component directly act on cancerous cell, reach the effect suppressing or kill cancerous cell.In addition, find, the preparation of nano particle size is rapid-action, prevents cancer cell metastasis aspect from having certain effect.
Meanwhile, although invention formulation has little mean diameter, the preparation of the various dosage forms obtained has high stability simultaneously, and they coagulation problems can not occur in long storage, makes it possible to the long period carry out storing and not reducing stability.
Common turmeric extract vein emulsion injection, described common turmeric extract vein emulsion injection prepares gained by following method: by the extract with total sesquiterpene alkene class content >=70wt% extracted from RADIX CURCUMAE plant, Refined Soybean phospholipid, cholesterol is with weight ratio 0.5-1.5: 2.5-4.0: 1.0-2.0, mix with water for injection, with the high pressure homogenizer emulsifying of routine, sampling utilizes microscope or the Coulter instrument inspection mean diameter of band scale, behind mean diameter≤1 micron, regulate pH, pH value is made to be 4.5 ~ 6.5, through 1.2 μ filtering with microporous membranes, bottling, through 100 DEG C ~ 120 DEG C sterilizings, obtain extract vein breast, the ultimate density that gas phase or liquid chromatography record sesquiterpene active component is 1-10mg/ml.
The invention provides the preparation method of common turmeric extract vein emulsion injection, it comprises the following steps:
The extract of total sesquiterpene alkene class content >=70wt% will be extracted from RADIX CURCUMAE plant, Refined Soybean phospholipid, cholesterol is with weight ratio 0.5-1.5: 2.5-4.0: 1.0-2.0, mix with water for injection, with the high pressure homogenizer emulsifying of routine, sampling utilizes microscope or the Coulter instrument inspection mean diameter of band scale, behind mean diameter≤1 micron, regulate pH, pH value is made to be 4.5 ~ 6.5, through 1.2 μ filtering with microporous membranes, bottling, through 100 DEG C ~ 120 DEG C sterilizings, obtain extract vein breast, the ultimate density that gas phase or liquid chromatography record sesquiterpene is 1-10mg/ml.
It is to be noted, various raw materials in the present invention for the preparation of the preparation of various dosage form can use the product be purchased as elemene product (extract), beta-elemene product (extract), Rhizoma Curcumae Longae extract or common turmeric extract, and 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. extract product.If need in this application oneself to prepare these extracts, the various volatile oil for the preparation of these extracts can use commercial products, also can use the volatile oil obtained 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 is (also referred to as Flos Caryophylli alkene, caryophyllene), gima ethylenic (large 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 containing more than 60wt% sesquiterpene extracted from plant all can be used for preparing novel form of the present invention, and sesquiterpene is the active component of all dosage forms.Extract containing more than 70wt% sesquiterpene is desirable, extract containing more than 75wt% sesquiterpene is preferred, extract containing more than 85wt% sesquiterpene is preferred, is particularly preferred containing more than 90wt% or the extract even containing more than 95wt% sesquiterpene.
The extract containing elemene extracted from natural plants contains α-elemene usually, beta-elemene, δ-elemene, γ-elemene, bourbonene etc.In the present invention as specific embodiment, for raw material, elemene is refined out through material filling type vacuum rectifying apparatus or molecular distillation instrument with RADIX CURCUMAE Curcuma wenyujin Y.H.Chen et C.Ling volatile oil, citronella oil Cymbopoqon citratus (DC.) Stapt submember (the mixing grease that citronella oil is remaining after removing citral, citronellol).
In addition, each elemene isomer is as α-elemene, and beta-elemene, as long as δ-elemene or γ-elemene purity also can be used in making novel form of the present invention at more than 85wt%.
At present according to domestic elemene quality standard (see the national drug standards WS that the National Drug Administration of 2000 issue
1-(X-094)-2000Z), if isolated product (extract) is with gas chromatography, (carbowax-20M is immobile phase, white diatomite carrier 60 ~ 80 order, column temperature is 135 ~ 140 DEG C, number of theoretical plate calculates should be not less than 1000 by beta-elemene) detect total elemene enantiomeric purity and reach>=85% this product is called elemene, if detect beta-elemene purity reach>=95%, product is beta-elemene, in fact this is inaccurate, because use chromatographic apparatus such as 30 meters of long capillary gas chromatograph-mass spectrometer (GC-MS) (chromatographs: Bio-Rad FTS-65A that sensitivity is higher, HP5890II, chromatographic column: BP-5) detect this>=the beta-elemene sample of 95% purity, can find that it still contains the bourbonene of at least 6% (at the chromatographic chromatogram of muting sensitivity, the peak of beta-elemene becomes a peak with the peak overlapping of bourbonene), elemene sample is the mixture of multiple sesquiterpene, namely containing beta-elemene, δ-elemene, γ-elemene, bourbonene, Flos Caryophylli alkene, gima ethylenic etc.
Rhizoma Curcumae Longae Rhizoma Curcuma Longa L. extract is the product extracted 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 refined 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%, the sesquiterpene of more preferably >=80wt%, particularly preferably >=85wt%, even more preferably >=90wt% is as the curcumin of curcumene, zingiberene etc. and a small amount of (being equivalent to about 1/25 ~ 1/5 of whole sesquiterpene gross weight).
RADIX CURCUMAE Curcuma wenyujin Y.H.Chen et C.Ling extract comprises the product extracted in the volatile oil obtained from RADIX CURCUMAE plant.In this application as specific embodiment, the product that common turmeric extract refers to the component that the essential oil from Curcuma wenyujin goes out through the rectification of vacuum material filling type rectifier unit or refines out through molecular distillation instrument, detect through GC-MS, product contains >=70wt%, preferably >=75wt%, more preferably >=80wt%, the sesquiterpene of particularly preferably >=85wt%, even more preferably >=90wt% is as various elemene isomer, gima ethylenic, Flos Caryophylli alkene etc. and the curcumin of small amount (being equivalent to the about 1/50-1/10 of whole sesquiterpene gross weight).
The explanation of term:
Particle diameter described in the present invention or granularity refer to particle diameter or the granularity of decentralized photo.
In this application, the component that common turmeric extract refers to the component that the essential oil from Curcuma wenyujin goes out through the rectification of vacuum material filling type rectifier unit or refines out through molecular distillation instrument, detect through GC-MS, common turmeric extract contains the sesquiterpene of primary amount and the curcumin of small amount.
If add " nanometer " to modify before dosage form, then refer to mean diameter≤150 nanometer of this dosage form, in ideal conditions mean diameter≤100 nanometer.Injection and ejection preparation are used interchangeably.
Lipidosome, refer to the Lipid Bilayer Structure be made up of phospholipid and cholesterol etc. be dispersed or suspended in water-bearing media in the present invention, this Lipid Bilayer Structure is similar to membrane structure.
Nanometer liposome dosage form, refers to the liposome of mean diameter less than or equal to 150 nanometers in the present invention.
Pro-liposome dosage form, refers in the present invention and add excipient again on the formula basis of liposome, by the lyophilized powder prepared by vacuum lyophilization (vacuum freeze-drying) method.This lyophilized powder by adding water for injection, through jolting or vibration, made it to revert to liposome before clinical practice.
Long circulating liposomes dosage form, briefly, refers to the liposome increased again on the formula basis of liposome prepared by Polyethylene Glycol PHOSPHATIDYL ETHANOLAMINE in the present invention.It is longer that it is expelled to the time of playing effectiveness in vivo than common liposome after in body, or eliminate in vivo slowly.
Nanometer microemulsion type, refers to mean diameter≤150 nanometer, the Emulsion of in the ideal case≤100 nanometers.
Vein emulsion type, refers to the injection breast of mean diameter≤1 micron in the present invention.
Lipid nanometer microsphere dosage forms, refers to the fat milk of mean diameter≤150 nanometer in the present invention.
Micro-emulsion, requires its mean diameter≤1 micron in the present invention; Also mean diameter≤1 micron is specified in addition in China national pharmacopeia.
Nm refers to nanometer, and μ refers to micron.Purity refers to percent purity by weight in this application, except as otherwise noted.
Raw material used in this application is common turmeric extract, and its total sesquiterpene alkene class content >=70wt%, it is desirable to 75wt%, preferably >=80wt%, more preferably >=85wt%, particularly preferably >=90wt%, particularly preferably >=95wt%.This extract contains≤curcumin of 25wt%, and the curcumin ideally containing≤15wt%, the curcumin preferably containing≤10wt%, the curcumin more preferably containing≤5wt%.
Use 30 meters of long capillary gas chromatograph-mass spectrometer (GC-MS) Detection and Extraction things described here, the extract containing >=70wt% sesquiterpene just can be used in the present invention.The common turmeric extract used in the application can buy acquisition by commercial sources with extract (various isomer mixture) or pure isomer (requiring that purity is higher than 85wt%) Product Form, also can oneself preparation as required, for the preparation method of common turmeric extract see preparation example.
The common turmeric extract raw material used in the application, reagent have no particular limits, as long as this extract be from natural plants RADIX CURCUMAE extract and total sesquiterpene alkene class purity >=70wt% just can use in the present invention, more preferably, this purity >=75wt%, better >=80wt%, preferably >=85wt%, particularly preferably >=90wt%.
Except as otherwise noted, otherwise other composition used in the present invention such as cholesterol, soybean phospholipid, lecithin, glycerol, linoleic acid, Polyethylene Glycol PHOSPHATIDYL ETHANOLAMINE, aliphatic amine, Semen sojae atricolor wet goods General Requirements meet national medicinal standard, solvent also should meet national medicinal standard, and these are mainly for the consideration of healthy aspect; Similarly, for some raw materials with " refine " modification, show that this raw material must meet national medicinal standard, such as Refined Soybean phospholipid be injection can product, refined soybean oil be injection can product.
Unless otherwise stated, gas chromatograph used in this application is: Bio-RadFTS-65A, HP5890II, chromatographic column is: BP-5.
Various dosage form of the present invention can be used for the treatment of object with intravenous injection, topical modes.
The preferred embodiments of the invention
In following embodiment, mainly use extract " elemene ", extract " beta-elemene ", Rhizoma Curcumae Longae extract, common turmeric extract as raw material, 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. can also be used.Obviously, wherein the product of the purity of a certain sesquiterpene isomer higher (such as >=85wt%) can both as raw material, and the various sesquiterpene utilizing chemical method to synthesize and derivant thereof can both be used in the present invention.
The invention provides the preparation method of liposome or nanometer liposome, it comprises: by 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, particularly preferably mix at 1: 3.2: 1.5, add solvent (as ether or acetone or chloroform), addition is enough to mixture is fully dissolved, insert desolvation film forming on Rotary Evaporators, add the water for injection (calculating this surplus according to the ultimate density that will prepare) of surplus, then insert in conventional ultrasonic cell disruptor and carry out ultrasonic Treatment, sampling utilizes the microscope of band scale, Coulter instrument, mean diameter checked by ultramicroscope, reaching the mean diameter such as≤1 micron (liposome) of regulation, or after≤100 nanometers (nanometer liposome), regulate PH, pH value is made to be 4.5 ~ 6.5, through 0.8 μ filtering with microporous membrane with then bottle, 100 DEG C of vapor stream sterilizings, the ultimate density (weight (mg)/total volumes of formulation (ml) of sesquiterpene active component) 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, namely, utilize SephadexG25 tomographic system, extract nano-liposome preparation is divided into two stream parts, i.e. extract nano-liposome stream part and free extract flow part, detects the extractive content contained in two stream parts through gas chromatograph.The extractive content ÷ of envelop rate computing formula=extract nano-liposome stream part (extractive content of extract nano-liposome stream part+free extractive content).Whether microscope or transmission electron microscope checking are liposome.
The invention provides the preparation method of pro-liposome: by extract and Refined 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, particularly preferably mix at 1: 3.2: 1.5, add solvent (as ether or acetone or chloroform), addition is enough to fully dissolve this mixture, insert desolvation film forming on Rotary Evaporators, add the water for injection (calculating this surplus according to the ultimate density that will prepare) of surplus, then insert ultrasonic cell disruptor and carry out supersound process, sampling utilizes the microscope of band scale or Coulter instrument to check mean diameter, when behind mean diameter≤1 micron, regulate PH, PH value is made to be 4.5 ~ 6.5, through 0.8 μ filtering with microporous membrane with then bottle, add the excipient (such as lactose or average molecular weight Mw are the low molecular dextran of about 4000-40000) of the 1-2wt% accounting for total amount of formulation, 100 DEG C of vapor stream sterilizings, lyophilization under aseptic condition, embedding.Add injection water before the use to reach final activity component concentration (weight (the mg)/volumes of formulation (ml) of sesquiterpene active component) for 1 ~ 10mg/ml, be preferably 2 ~ 8mg/ml, be more preferably 2 ~ 7.5mg/ml, be more typically 5 ~ 7.5mg/ml.
The detection of envelop rate: use sieve method detection method, that is: according to the final formulation concentrations (mg/ml) that will prepare, by even for the water jolting that a certain amount of extract proliposome preparation first adds surplus volume, now become liposome again, Sephadex G25 tomographic system is utilized to be divided into two stream parts, i.e. extract liposome stream part and free extract flow part, detects the extract amount contained in two stream parts through gas chromatograph.The extractive content ÷ of envelop rate computing formula=extract liposome stream part (extractive content of extract liposome stream part+free extract amount).Microscope or transmission electron microscope confirm whether be pro-liposome.
The invention provides the preparation method of long circulating extract nano-liposome, the method comprises: by extract and Refined 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, particularly preferably mix with weight ratio 1: 2.6-2.7: 0.7-0.9: 1.3-1.5, add solvent (as ether or acetone or chloroform), addition is enough to fully dissolve this mixture, insert desolvation film forming on Rotary Evaporators, add the water for injection (calculating this surplus according to the ultimate density that will prepare) of surplus, then insert conventional ultrasonic cell disruptor and carry out ultrasonic Treatment, sampling utilizes the microscope of band scale, mean diameter checked by Coulter instrument or ultramicroscope, after reaching the mean diameter (such as≤100 nanometer) of regulation, regulate PH, pH value is made to be 4.5 ~ 6.5, through 0.8 μ filtering with microporous membrane with then bottle, 100 DEG C of vapor stream sterilizings, the ultimate density (weight (mg)/total volumes of formulation (ml) of sesquiterpene active component) that (gas phase or liquid phase) chromatography records active component is 1 ~ 10mg/ml, be preferably 2 ~ 7.5mg/ml, be more preferably 2 ~ 8mg/ml, be 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 lipidosome injection above.
The invention provides the preparation method of electronegative and positively charged extract nano microemulsion:
Preparation method 1: the preparation of electronegative nanometer microemulsion
By extract and Refined 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, particularly preferably mix at 1: 3.2: 1.5, add solvent (as ether or acetone or chloroform), addition is enough to fully dissolve this mixture, insert desolvation film forming on Rotary Evaporators, add the water for injection (calculating this surplus according to the ultimate density that will prepare) of surplus, then insert conventional ultrasonic cell disruptor and carry out ultrasonic Treatment, sampling utilizes the microscope of band scale, mean diameter checked by Coulter instrument or ultramicroscope, after reaching the mean diameter (such as≤100 nanometer) of regulation, regulate PH, pH value is made to be 4.5 ~ 6.5, through 0.8 μ filtering with microporous membrane with then bottle, 100 DEG C of vapor stream sterilizings, the ultimate density (weight (mg)/total volumes of formulation (ml) of sesquiterpene active component) of 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 nanometer microemulsion.
Preparation method 2: the preparation of positively charged nanometer microemulsion
By 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, particularly preferably mix at 1: 2.8: 0.4: 1.5, add solvent (as ether or acetone or chloroform), addition is enough to fully dissolve this mixture, insert desolvation film forming on Rotary Evaporators, then the water for injection (calculating this surplus according to the densitometer that will prepare) of surplus is added, then insert conventional ultrasonic cell disruptor and carry out supersound process, sampling utilizes the microscope of band scale, mean diameter checked by Coulter instrument or ultramicroscope, after reaching the mean diameter (such as≤100 nanometer) of regulation, regulate PH, pH value is made to be 7.5 ~ 9.0, through 0.8 μ filtering with microporous membrane with then bottle, 100 DEG C of vapor stream sterilizings, 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 nanometer microemulsion.
The present invention also provides the preparation method of extract lipid nanospheres, and the method comprises: get extract and add together with soybean phospholipid in soybean oil, heats up and is controlled to oil phase (remaining oil phase), joined in glycerinated aqueous solution by this oil phase.Its concrete part by weight is: extract accounts for 0.08 ~ 0.8wt% of final preparaton total amount, soybean phospholipid 0.8 ~ 2.5wt%, soybean oil 8-12wt%, glycerol 1.8-2.8wt%, and 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%, soybean oil 10wt%, and glycerol 2.25 ~ 2.5wt%, water for injection adds to 5000ml.Then high-speed stirred, PH is regulated to be 4.5 ~ 6.5, again through common high pressure homogenizer (such as M110-E/H type (Japanese MIZUHO produces)) process, then foregoing ultrasonic Treatment is carried out, sampling utilizes the microscope of band scale, Coulter instrument or ultramicroscope to check mean diameter, the mean diameter of its lipid ball reach setting such as≤100nm after, then through 0.8 μ filtering with microporous membrane with then bottle, 100 DEG C ~ 120 DEG C sterilizings.The ultimate density that (gas phase or liquid phase) chromatography records sesquiterpene active component is 1 ~ 10mg/ml, and be preferably 2 ~ 7.5mg/ml, be more typically 2 ~ 5mg/ml, better is 2.0 ~ 3.0mg/ml.Appropriate cholesterol, linoleic acid etc. can be added as required in this final preparation.
The invention provides the preparation method of extract intravenous injection breast, the method comprises: by extract, Refined 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, particularly preferably 1: 3.2: 1.5, mix with the water for injection (calculating this surplus according to the ultimate density that will prepare) of surplus, with high pressure homogenizer (such as M110-E/H type (Japanese MIZUHO the produces)) emulsifying of routine, sampling utilizes the microscope of band scale or Coulter instrument to check mean diameter, after reaching the mean diameter such as≤1 micron of regulation, regulate PH, pH value is made to be 4.5 ~ 6.5, through 1.2 μ filtering with microporous membranes with then bottle, through 100 DEG C ~ 120 DEG C sterilizings, obtain extract vein breast, the ultimate density that (gas phase or liquid phase) chromatography records 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, the method comprises: get extract and soybean phospholipid adds in soybean oil, heats up and is controlled to oil phase (remaining oil phase), joined in glycerinated aqueous solution by this oil phase.Its concrete weight ratio is: extract accounts for 0.08 ~ 0.8wt% of final preparaton total amount, soybean phospholipid 0.8 ~ 2.5wt%, soybean oil 8-12wt%, glycerol 1.8-2.8wt%, and 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%, soybean oil 10wt%, and glycerol 2.25 ~ 2.5wt%, water for injection adds to 5000ml.Then high-speed stirred, regulate PH to be 4.5 ~ 6.5, then through common high pressure homogenizer (such as M110-E/H type (Japanese MIZUHO produces)) process, make its particle diameter be less than 1 μ, again through 1.2 μ filtering with microporous membranes with then bottle, 100 DEG C ~ 120 DEG C sterilizings.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.Appropriate cholesterol, linoleic acid etc. can be added as required in this final preparation.
As required, preparation of the present invention can filter with the microporous filter membrane of different size such as 1.2 μ, 1.0 μ, 0.9 μ, 0.8 μ, 0.7 μ, 0.6 μ, 0.5 μ etc., can intercept the decentralized photo of mean diameter≤0.8 μ ,≤0.7 μ ,≤0.6 μ ,≤0.5 μ ,≤0.45 μ ,≤0.4 μ ,≤0.3 μ etc.
Preparation example below and embodiment are used for illustrating 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: the preparation of common turmeric extract
Preparation method A: take the essential oil from Curcuma wenyujin as raw material, refine through Guangzhou Han Wei mechanical & electrical corporation MD-S80 type molecular distillation instrument, its process conditions: vapo(u)rizing temperature 35 ~ 45 DEG C, distillation pressure 70 ~ 90Pa, knifing speed 300 ~ 320 revs/min, cryosurface temperature 20 ~ 25 DEG C, system temperature 20 ~ 25 DEG C, material flow 15 ~ 30 mls/hour, only collects and heats up in a steamer excess, will heat up in a steamer excess and proceed molecular distillation.Vapo(u)rizing temperature is 45 ~ 60 DEG C, distillation pressure 30 ~ 70Pa, knifing speed 300 ~ 320 revs/min, cryosurface temperature 20 ~ 25 DEG C, system insulation 20 ~ 25 DEG C, material flow 15 ~ 30 mls/hour, collect distillation respectively and heat up in a steamer excess, excess will be heated up in a steamer and carry out molecular distillation.Vapo(u)rizing temperature is 30 ~ 70 DEG C, distillation pressure 10 ~ 50Pa, knifing speed 300 ~ 320 revs/min, cryosurface temperature 20 ~ 25 DEG C, system insulation 20 ~ 25 DEG C, material flow 15 ~ 30 mls/hour, collect distillation, bleed off and heat up in a steamer excess, by the secondary distillation mixing obtained, continue distillation, until (utilize 30 meters of long capillary gas chromatograph-mass spectrometer (GC-MS) gas chromatograpies to detect till gas chromatograph detection reaches total sesquiterpene alkene content >=75wt%, chromatograph: Bio-Rad FTS-65A, HP5890II, chromatographic column: BP-5).This extract detects through above-mentioned gas chromatograph, also containing be equivalent to whole sesquiterpene gross weight about 1/25 curcumin.Formulate for detection finger printing with gas chromatography.This product is called common turmeric extract A.
Preparation method B. by the essential oil from Curcuma wenyujin, through the rectification of material filling type vacuum rectifying apparatus,
Material filling type vacuum rectifying apparatus by filling the still of the essential oil from Curcuma wenyujin, stuffing rectification column, return channel, vacuum pump form; manufactured and designed by new packing technology development center of Shanghai Chemical Research Inst; filler is Dixon filler (Q net ring filler); vacuum is 1 ~ 3mmHg, collects the fraction of 82 DEG C/more than 3mmHg.Be sesquiterpenoid (utilize 30 meters long capillary gas chromatograph-mass spectrometer (GC-MS) gas chromatograpies detect) through chromatographic, total sesquiterpene alkene content >=75wt%.Extract detects through above-mentioned gas chromatograph, also containing be equivalent to whole sesquiterpene gross weight about 1/25 curcumin.Formulate for detection finger printing with gas chromatography.This product is called common turmeric extract B.
In addition, learn from else's experience CO
2supercritical extraction crosses the RADIX CURCUMAE medicinal residues of volatile oil, add methanol reflux 3 hours, obtain brown solution, methanol is removed with Rotary Evaporators, obtain viscous brown thing, this viscous brown paste silica gel is prepared chromatograph, with chloroform eluting, obtains the curcumin product of purity>=90wt%.Above-mentioned product A and product B are mixed with 50: 50 weight ratios, formed mixture is carried out mixing in the ratio of 20: 1 (weight) with this curcumin product and obtains another kind of mixture, being called extract C.
Summary of the invention
The object of this invention is to provide common turmeric extract vein emulsion injection.
Another object of the present invention is to provide the preparation method of common turmeric extract vein emulsion injection.
Another object of the present invention is to provide the purposes of common turmeric extract vein emulsion injection for the preparation of the drug injection of Therapeutic cancer or cerebral infarction, and these injection types are used for the purposes of tumor remission pain, and for suppressing and/or treat the purposes of tumor.
The invention provides the novel form of these medicines.Its vein emulsion type is prepared into respectively using common turmeric extract as raw material.The mean diameter of these dosage forms is generally less than or equals 1 micron, is even less than or equal to 100 nanometers.
Accompanying drawing is sketched
Fig. 1 is the transmission electron microscope photo of the 1A preparation of embodiment 1.
Detailed description of the present invention
Embodiment 1: the preparation of the nano liposome injection of common turmeric extract
Program 1A: by the product A of preparation example and Refined Soybean phospholipid, cholesterol is with weight ratio 1:3.2: 1.5 mixing, add ether, addition is enough to fully dissolve this mixture, insert on Rotary Evaporators and remove ether film forming, add the water for injection (calculating this surplus according to the ultimate density that will prepare and 5mg/ml) of surplus, then ultrasonic cell disruptor (model JCS-204) supersound process that Jining of Shandong Province ultrasonic electronic instrument plant produces is inserted, sampling utilizes the microscope of band scale, mean diameter checked by Coulter instrument or ultramicroscope, when after the nanometer of mean diameter≤100, PH is regulated with 1M biphosphate sodium water solution, pH value is made to be 4.5 ~ 6.5, through 0.8 μ microporous filter membrane, (new Asia, Shanghai purification device factory produces, specification 0.8 μ) filter and then bottle, 100 DEG C of vapor stream sterilizings, the injection product obtained is called for short 1A.The ultimate density that sampling gas chromatography (utilizing 30 meters of long capillary gas chromatograph-mass spectrometer (GC-MS) gas chromatograpies) records active component (weight (the mg)/volumes of formulation (ml) of total sesquiterpene alkene active component) is 5 ± 0.10mg/ml.
Envelop rate is detected with sieve method, namely, utilize Sephadex G25 tomographic system extract nano-liposome preparation to be divided into two stream parts, i.e. extract nano-liposome stream part and free extract flow part, detect the extractive content contained in two stream parts through gas chromatograph.Envelop rate is more than 85%.The extractive content ÷ of envelop rate computing formula=extract nano-liposome stream part [extractive content of extract nano-liposome stream part+free extract amount].Microscope, transmission electron microscope detect, and confirmation is nanometer liposome, provides the transmission electron microscope photo (scale 50nm) of this injection in fig. 1.
Program 1B: repeat above-mentioned 1A program, just replaces the product A of preparation example by the product B of preparation example.Microscope, transmission electron microscope detect, and confirm that preparation is nanometer liposome.In preparation, the concentration of total sesquiterpene alkene class is about 5 ± 0.10mg/ml.Envelop rate is higher than 85%.This injection is called for short 1B.
Embodiment 2: the preparation of the pro-liposome of common turmeric extract
2A: according to the method identical with the 1A program of embodiment 1 and proportioning raw materials, repeat the 1A program of embodiment 1, just require after ultrasonic Treatment decentralized photo particle diameter≤1 micron and after filtration and before bottling, add the lactose of the 1-2% accounting for total weight of formulation, then 100 DEG C of vapor stream sterilizings are utilized, lyophilization under aseptic condition, embedding (subpackage).Detect with Coulter instrument and confirm that preparation is pro-liposome.This injection product is called for short 2A.Envelop rate is higher than 85%.Said preparation added injection water to reach final active component (total sesquiterpene alkene class) concentration for 5 ± 0.10mg/ml before injection application.
2B: repeat above-mentioned 2A, just replaces the product A of preparation example with the B product of preparation example, replace lactose with the low molecular dextran that average molecular weight Mw is about 40000.This injection is called for short 2B.Detect with Coulter instrument and confirm that preparation is pro-liposome.Envelop rate is higher than 85%.Said preparation added injection water to reach final activity component concentration (weight (the mg)/volumes of formulation (ml) of total sesquiterpene alkene class) for 5 ± 0.10mg/ml before injection application.
Embodiment 3: the preparation of the long-circulating nanoliposome of common turmeric extract
3A: according to the method identical with the 1A program of embodiment 1, repeat the 1A program of embodiment 1, just raw material and their weight proportion are: common turmeric extract A: soybean phospholipid: Polyethylene Glycol (2000) PHOSPHATIDYL ETHANOLAMINE: cholesterol=1: 2.7: 0.8: 1.4.This injection is called for short 3A.Pick test mean diameter≤100nm.In preparation, the ultimate density of total sesquiterpene alkene class is 5 ± 0.10mg/ml.Envelop rate is higher than 85%.
3B: repeat above-mentioned 3A, just replaces the product A of preparation example by the product B of preparation example.The injection obtained is called for short 3B.Mean diameter≤100nm.In preparation, the ultimate density of total sesquiterpene alkene class is 5 ± 0.10mg/ml.Envelop rate is higher than 85%.
Embodiment 4: the preparation of the nanometer microemulsion of electronegative and positively charged common turmeric extract
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 sesquiterpene alkene class is 5 ± 0.10mg/ml.Confirm it is nanometer microemulsion with 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: extract A: lecithin: n-octadecane base amine: cholesterol=1: 2.8: 0.4: 1.5, does not measure envelop rate.The ultimate density that sampling records total sesquiterpene alkene class is 5 ± 0.10mg/ml.Confirm it is nanometer microemulsion with 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 product A of preparation example by the product B of preparation example.The ultimate density that sampling records total sesquiterpene alkene class is 5 ± 0.10mg/ml.Confirm it is nanometer microemulsion with Coulter instrument.The injection obtained is called for short 4B-1.
4B-2: the preparation of positively charged nanometer microemulsion
Repeat above-mentioned 4A-2, just replace the product A of preparation example by the product B of preparation example.The ultimate density that sampling records total sesquiterpene alkene class is 5 ± 0.10mg/ml.Confirm it is nanometer microemulsion with Coulter instrument.The injection obtained is called for short 4B-2.
Embodiment 5: the preparation of the lipid nanospheres preparation of common turmeric extract
5A: the extract A of getting preparation example adds in soybean oil together with soybean phospholipid, heats up and is controlled to oil phase (namely remaining oil phase), joined in glycerinated aqueous solution by this oil phase.Its concrete weight ratio is: extract A accounts for 0.2% of total weight of formulation, soybean phospholipid 1.7%, soybean oil 10%, glycerol 2.5%, surplus is water for injection, then high-speed stirred, PH is regulated to be 4.5 ~ 6.5 with 1M biphosphate sodium water solution, again through M110-E/H type (Japanese MIZUHO produces) high pressure homogenizer process, then ultrasonic Treatment is carried out according to same procedure in embodiment 3, make the mean diameter≤100nm of its lipid ball, then through 0.8 μ filtering with microporous membrane with then bottle, 100 DEG C ~ 120 DEG C sterilizings.The ultimate density that sampling records total sesquiterpene alkene class is 2 ± 0.04mg/ml.The injection obtained is called for short 5A.Appropriate cholesterol, linoleic acid etc. can be comprised in the feed as required.
5B: repeat above-mentioned 5A program, just replaces the product A of preparation example by the product B of preparation example.Mean diameter≤the 100nm of lipid ball.The ultimate density that chromatography records total sesquiterpene alkene class is 2 ± 0.04mg/ml.The injection obtained is called for short 5B.
Embodiment 6: the preparation of the intravenous injection breast of common turmeric extract
6A: the common turmeric extract A of preparation example, Refined Soybean phospholipid, cholesterol are mixed with the water for injection (calculating this surplus according to required final concentration and 5mg/ml) of surplus with weight ratio 1: 3.2: 1.5, with M110-E/H type (Japanese MIZUHO produces) high pressure homogenizer emulsifying, till being less than 1 μ with Coulter instrument inspection particle diameter always.Then, regulate PH with 1M biphosphate sodium water solution, make pH value be 4.5 ~ 6.5, through 1.2 μ filtering with microporous membranes with then bottle, through 100 DEG C ~ 120 DEG C sterilizings, obtain common turmeric extract vein breast, the ultimate density that sampling records total sesquiterpene alkene class is 5 ± 0.10mg/ml.This injection is called for short 6A.
6B: repeat above 6A, just replaces the product A of preparation example by the product B of preparation example.1 micron is less than with Coulter instrument inspection particle diameter.In preparation, the ultimate density of total sesquiterpene alkene class is 5 ± 0.10mg/ml.The injection obtained is called for short 6B.
Embodiment 7: the preparation of common turmeric extract fat emulsion
7A: product A and the soybean phospholipid of getting preparation example add in soybean oil, heats up and is controlled to oil phase, joined in glycerinated aqueous solution by this oil phase.Its concrete weight ratio is: extract A accounts for 0.2% of total weight of formulation, soybean phospholipid 1.2%, soybean oil 10%, glycerol 2.5%, and surplus is water for injection.Then high-speed stirred, PH is regulated to be 4.5 ~ 6.5 with 1M biphosphate sodium water solution, again through M110-E/H type (Japanese MIZUHO produces) high pressure homogenizer process, until with till Coulter instrument inspection particle diameter≤1 μ, again through 1.2 μ filtering with microporous membranes with then bottle, 100 DEG C ~ 120 DEG C sterilizings.The ultimate density recording total sesquiterpene alkene class is 2 ± 0.04mg/ml.The injection obtained is called for short 7A.Appropriate cholesterol, linoleic acid etc. can be comprised in the feed as required.
7B: repeat above-mentioned 7A, just replaces the product A of preparation example by the product B of preparation example.The ultimate density recording total sesquiterpene alkene class is in the formulation 2 ± 0.04mg/ml.The injection obtained is called for short 7B.
Embodiment 8:CO
2supercritical methanol technology prepares liposome
8A: the chloroform of the 10wt% accounting for cholesterol weight is added in cholesterol and makes it to infiltrate, the extract A of soybean phospholipid and preparation example is added and wherein carries out mixing (extract A: soybean phospholipid: cholesterol=weight ratio 1.2: 3.2: 1.5), obtained mixture is put into CO
2in the extractor of supercritical extraction instrument (Guangzhou Han Wei mechanical & electrical corporation produces, model SFE100ml), pass into CO
2(pressure 50-80kg), system temperature 50 DEG C, carries out critical or supercritical dispersion, makes mixture Homogeneous phase mixing, and then slowly decompression discharges CO
2, allow CO
2chloroform is taken out of, take out extractor, the water for injection (calculating this surplus according to the ultimate density of injection) of surplus is added in extractor, stir with agitator, by formed dispersion ultrasonic Treatment, until till Dispersed Phase Size≤100 nanometer of liposome (being called nanometer liposome).Then liposome 1M biphosphate sodium water solution is regulated PH, make pH value be 4.5 ~ 6.5, filter through 0.8 μ microporous filter membrane (new Asia, Shanghai purification device factory produces, specification 0.8 μ) and then bottle, 100 DEG C of vapor stream sterilizings, the injection product obtained is called for short 8A.The ultimate density that sampling gas chromatography records total sesquiterpene alkene class is 5 ± 0.10mg/ml.Envelop rate is higher than 88%.
8B: repeat above-mentioned 8A, just replaces the product A of preparation example by the product B of preparation example, obtained liposome.The ultimate density recording total sesquiterpene alkene class is in the formulation 5 ± 0.10mg/ml.The injection obtained is called for short 8B.Envelop rate is higher than 88%.
8C: repeat above-mentioned 8A, just replaces the product A of preparation example by the extract C of preparation example, obtained liposome.The ultimate density recording total sesquiterpene alkene class is in the formulation 5 ± 0.10mg/ml.The injection obtained is called for short 8C.Envelop rate is higher than 88%.
Pharmacodynamics test
The evaluation methodology of the anti-tumor in vivo effect of pharmaceutical preparation has description (for example, see CN1153168A) in some patent documentation enumerated above.
In the present invention, the human tumor heteroplastic transplantation model QGY hepatocarcinoma of preparation to subcutaneous vaccination of various dosage form, the curative effect of MKN-45 gastric cancer, can pass through with the dosage of 80mg (sesquiterpene)/kg (body weight), 40mg/kg and 20mg/kg, every day tail intravenously administrable 1 time, the successive administration therapeutic scheme of 10 days confirms wherein do not have administration as a control group.The relevant regulations " the evaluating drug effect way of antitumor drug " of calculating National Drug Administration of (before referring to the applying date of the application) according at present of concrete operating process and tumor control rate (tumour inhibiting rate) is implemented.
Tumour inhibiting rate %=[tumor weight of (not having the tumor weight of the model of the tumor weight-administration of the model of administration)/do not have 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 | 30.3％ | 40 | 39.2％ | 80 | 45.3％ |
| 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 | 43.7％ |
| 3A | 20 | 29.7％ | 40 | 39.7％ | 80 | 44.5％ |
| 3B | 20 | 29.8％ | 40 | 38.7％ | 80 | 44.4％ |
| 4A-1 | 20 | 30.4％ | 40 | 38.5％ | 80 | 46.0％ |
| 4A-2 | 20 | 31.7％ | 40 | 37.9％ | 80 | 46.3％ |
| 4B-1 | 20 | 30.3％ | 40 | 37.4％ | 80 | 45.2％ |
| 4B-2 | 20 | 29.4％ | 40 | 38.7％ | 80 | 44.9％ |
| 5A | 20 | 29.2％ | 40 | 38.6％ | 80 | 44.2％ |
| 5B | 20 | 29.5％ | 40 | 38.7％ | 80 | 44.5％ |
| 6A | 20 | 29.8％ | 40 | 37.8％ | 80 | 43.3％ |
| 6B | 20 | 29.4％ | 40 | 37.9％ | 80 | 43.5％ |
| 7A | 20 | 29.3％ | 40 | 37.4％ | 80 | 44.0％ |
| 7B | 20 | 29.7％ | 40 | 37.3％ | 80 | 43.3％ |
| 8A | 20 | 29.4％ | 40 | 37.4％ | 80 | 44.0％ |
| 8B | 20 | 29.3％ | 40 | 37.3％ | 80 | 43.3％ |
| 8C | 20 | 29.4％ | 40 | 37.7％ | 80 | 44.3％ |
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 | 39.2％ | 80 | 45.3％ |
| 1B | 20 | 29.1％ | 40 | 39.1％ | 80 | 45.0％ |
| 2A | 20 | 29.2％ | 40 | 37.5％ | 80 | 44.0％ |
| 2B | 20 | 28.4％ | 40 | 38.5％ | 80 | 43.7％ |
| 3A | 20 | 29.1％ | 40 | 39.7％ | 80 | 44.5％ |
| 3B | 20 | 29.0％ | 40 | 38.7％ | 80 | 44.4％ |
| 4A-1 | 20 | 29.4％ | 40 | 38.5％ | 80 | 46.0％ |
| 4A-2 | 20 | 29.5％ | 40 | 37.9％ | 80 | 46.3％ |
| 4B-1 | 20 | 28.8％ | 40 | 37.4％ | 80 | 45.2％ |
| 4B-2 | 20 | 28.2％ | 40 | 38.7％ | 80 | 44.9％ |
Table 3: to the curative effect (tumour inhibiting rate) of intracranial original position Mice Inoculated cerebroma G422 model.
| Injection | Dosage (mg/kg) | Tumour inhibiting rate | Dosage (mg/kg) | Tumour inhibiting rate | Dosage (mg/kg) | Tumour inhibiting rate |
| 1A | 20 | 47.7％ | 40 | 59.5％ | 80 | 68.3％ |
| 1B | 20 | 46.8％ | 40 | 59.3％ | 80 | 68.5％ |
| 2A | 20 | 47.3％ | 40 | 57.7％ | 80 | 64.0％ |
| 2B | 20 | 48.3％ | 40 | 56.2％ | 80 | 63.7％ |
| 3A | 20 | 49.5％ | 40 | 59.7％ | 80 | 67.5％ |
| 3B | 20 | 48.4％ | 40 | 58.2％ | 80 | 66.4％ |
| 4A-1 | 20 | 49.2％ | 40 | 58.4％ | 80 | 67.0％ |
| 4A-2 | 20 | 49.4％ | 40 | 57.6％ | 80 | 67.3％ |
| 4B-1 | 20 | 48.7％ | 40 | 57.3％ | 80 | 66.2％ |
| 4B-2 | 20 | 49.2％ | 40 | 58.4％ | 80 | 67.9％ |
| 5A | 20 | 49.8％ | 40 | 58.6％ | 80 | 68.2％ |
| 5B | 20 | 47.8％ | 40 | 58.2％ | 80 | 65.5％ |
| 6A | 20 | 47.7％ | 40 | 56.8％ | 80 | 63.3％ |
| 6B | 20 | 46.9％ | 40 | 56.9％ | 80 | 63.5％ |
| 7A | 20 | 48.3％ | 40 | 56.4％ | 80 | 66.0％ |
| 7B | 20 | 46.8％ | 40 | 56.3％ | 80 | 63.3％ |
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 | 29.7％ | 80 | 35.3％ |
| 2A | 20 | 22.8％ | 40 | 27.8％ | 80 | 34.0％ |
| 2B | 20 | 22.8％ | 40 | 28.5％ | 80 | 33.2％ |
| 3A | 20 | 22.9％ | 40 | 29.8％ | 80 | 34.3％ |
| 3B | 20 | 23.0％ | 40 | 28.6％ | 80 | 34.8％ |
| 4A-1 | 20 | 24.3％ | 40 | 28.1％ | 80 | 36.3％ |
| 4A-2 | 20 | 24.4％ | 40 | 27.9％ | 80 | 36.3％ |
| 4B-1 | 20 | 23.7％ | 40 | 27.8％ | 80 | 35.3％ |
| 4B-2 | 20 | 23.3％ | 40 | 28.6％ | 80 | 34.4％ |
| 5A | 20 | 23.4％ | 40 | 28.6％ | 80 | 34.8％ |
| 5B | 20 | 24.1％ | 40 | 28.7％ | 80 | 34.5％ |
| 6A | 20 | 22.4％ | 40 | 27.9％ | 80 | 33.3％ |
| 6B | 20 | 22.7％ | 40 | 27.8％ | 80 | 33.4％ |
| 7A | 20 | 22.4％ | 40 | 27.5％ | 80 | 34.1％ |
| 7B | 20 | 22.3％ | 40 | 27.2％ | 80 | 33.7％ |
In addition, the injection of various dosage form improves 21% ~ 23% to the NK cells in mice of lotus Lewis lung cancer and matched group specific activity.
The injection of various dosage form affects the mouse lymphocyte proliferation activity of lotus Lewis lung cancer, stimulate index to improve 1.4 ~ 1.55.
In a word, data as can be seen from above-mentioned table, various preparation is effective through QGY hepatocarcinoma, MKN-45 gastric cancer, mice cerebroma G422 model drug pharmacologic test.To Immune Function test, the NK cells in mice of lotus Lewis lung cancer with matched group than raising 21% ~ 23%.Lymphocyte proliferation activity influence, stimulation index improve 1.4 ~ 1.55.
So injection can be used in suppressing and/or treatment tumor.In addition, through overtesting, said preparation can also be used for anticancer transfer, for tumor remission pain, for prevention or treatment cerebral infarction.
Claims (6)
1. common turmeric extract vein emulsion injection, described common turmeric extract vein emulsion injection prepares gained by following method: by the extract with total sesquiterpene alkene class content >=70wt% extracted from RADIX CURCUMAE plant, Refined Soybean phospholipid, cholesterol is with weight ratio 0.5-1.5: 2.5-4.0: 1.0-2.0, mix with water for injection, with the high pressure homogenizer emulsifying of routine, sampling utilizes microscope or the Coulter instrument inspection mean diameter of band scale, behind mean diameter≤1 micron, regulate pH, pH value is made to be 4.5 ~ 6.5, through 1.2 μ filtering with microporous membranes, bottling, through 100 DEG C ~ 120 DEG C sterilizings, obtain extract vein breast, the ultimate density that gas phase or liquid chromatography record sesquiterpene active component is 1-10mg/ml,
The described extract component that to be the essential oil from Curcuma wenyujin go out through the rectification of vacuum material filling type rectifier unit or the component of refining out through molecular distillation instrument, this extract contain be equivalent to sesquiterpene gross weight 1/50 ~ 1/10 curcumin.
2. common turmeric extract vein emulsion injection according to claim 1, is characterized in that: the weight ratio of described extract and Refined Soybean phospholipid, cholesterol is 0.8-1.2: 2.8-3.6: 1.2-1.8.
3. common turmeric extract vein emulsion injection according to claim 1, is characterized in that: in described common turmeric extract vein emulsion injection, the ultimate density of sesquiterpene active component is 2 ~ 7.5mg/ml.
4. common turmeric extract vein emulsion injection according to claim 2, is characterized in that: in described common turmeric extract vein emulsion injection, the ultimate density of sesquiterpene active component is 2 ~ 7.5mg/ml.
5. the preparation method of common turmeric extract vein emulsion injection according to claim 1, it comprises the following steps:
The extract of total sesquiterpene alkene class content >=70wt% will be extracted from RADIX CURCUMAE plant, Refined Soybean phospholipid, cholesterol is with weight ratio 0.5-1.5: 2.5-4.0: 1.0-2.0, mix with water for injection, with the high pressure homogenizer emulsifying of routine, sampling utilizes microscope or the Coulter instrument inspection mean diameter of band scale, behind mean diameter≤1 micron, regulate pH, pH value is made to be 4.5 ~ 6.5, through 1.2 μ filtering with microporous membranes, bottling, through 100 DEG C ~ 120 DEG C sterilizings, obtain extract vein breast, the ultimate density that gas phase or liquid chromatography record sesquiterpene is 1-10mg/ml,
The described extract component that to be the essential oil from Curcuma wenyujin go out through the rectification of vacuum material filling type rectifier unit or the component of refining out through molecular distillation instrument, this extract contain be equivalent to sesquiterpene gross weight 1/50 ~ 1/10 curcumin.
6. the common turmeric extract vein emulsion injection in claim 1-4 described in arbitrary claim is for the preparation of suppressing or the purposes of medicine of Therapeutic cancer.
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| CNB021172188A CN100423713C (en) | 2002-04-17 | 2002-04-17 | Curcuma aromatica extract injection, and preparing method and use thereof |
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| CN2008101264177A Expired - Lifetime CN101306183B (en) | 2002-04-17 | 2002-04-17 | Common turmeric extract fat emulsion injection and its preparation method and use |
| CNB021172188A Expired - Lifetime CN100423713C (en) | 2002-04-17 | 2002-04-17 | Curcuma aromatica extract injection, and preparing method and use thereof |
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| CNB021172188A Expired - Lifetime CN100423713C (en) | 2002-04-17 | 2002-04-17 | Curcuma aromatica extract injection, and preparing method and use thereof |
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| CN100486567C (en) * | 2004-08-12 | 2009-05-13 | 山东绿叶天然药物研究开发有限公司 | Curcumin emulsion and its preparation process |
| CN101569606A (en) * | 2009-06-09 | 2009-11-04 | 沈阳药科大学 | Curcumol liposome and preparation method thereof |
| CN109444283A (en) * | 2018-12-12 | 2019-03-08 | 广西中医药大学 | The detection method of curcumin chemical compounds in a kind of osmanthus Radix Curcumae |
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| CN1076613A (en) * | 1993-02-15 | 1993-09-29 | 大连金港制药有限公司 | Elemene emulsion injection and preparation method thereof |
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| CN1084187C (en) * | 1997-04-14 | 2002-05-08 | 中国远大国际集团有限公司 | Method for preparing anti-cancer medicine using beta-elemene as main effective ingredient, and components of the medicine |
| CN1221607A (en) * | 1998-11-20 | 1999-07-07 | 大连市医药科学研究所 | Liposoluble medicinal liposome prodn. tech. and elemene liposome injecta |
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| CN101306183A (en) | 2008-11-19 |
| CN1451376A (en) | 2003-10-29 |
| CN101306184A (en) | 2008-11-19 |
| CN100423713C (en) | 2008-10-08 |
| CN101306183B (en) | 2011-06-29 |
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