CN102784105A - RLT polypeptide-mediated docetaxel tumor targeting submicroemulsion and preparation method thereof - Google Patents
RLT polypeptide-mediated docetaxel tumor targeting submicroemulsion and preparation method thereof Download PDFInfo
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Abstract
The invention relates to a polypeptide (RLT polypeptide)-mediated docetaxel tumor targeting submicroemulsion combined with a low density lipoprotein binding domain in an apolipoprotein B, and a preparation method thereof, which belong to the medical technology field. The submicroemulsion is the tumor targeting submicroemulsion prepared from oil for injection, emulsifying agents, auxiliary emulsifying agents, antioxygen, isoosmotic adjusting agents, bi-stearoyl phosphatidyl ethanolamine-polyethylene glycol 2000-maleimide, RLT polypeptide and docetaxel. The weight volume concentration range of the docetaxel is 0.5-20mg/mL. The tumor targeting submicroemulsion has uniform grain size, and the average grain size is 182nm. The docetaxel submicroemulsion is modified by tumor targeting cells, so that the tumor targeting efficiency is obviously improved, the distribution of medicine in the tumor cells is increased, and other normal medicine dose is reduced. Therefore, due to the RLT polypeptide-mediated docetaxel tumor targeting submicroemulsion, the therapeutic effect of the medicine can be improved, and the toxic and side effects are reduced.
Description
Technical field
The invention belongs to medical technical field, be specifically related to the anticarcinogen Docetaxel is prepared into the Docetaxel cancer target submicronized emulsion that the RLT polypeptide mediates with proper pharmaceutical excipients, the invention still further relates to method of making preparation.
Background technology
Docetaxel (Docetaxel) belongs to the taxanes antineoplastic agent; Main interference cell mitosis and the required microtubule network of interkinesis play antitumous effect, and its untoward reaction mainly shows anaphylaxis, thrombocytopenia, the property a crossed tachycardia, hypotension, neurotoxicity reaction, gastrointestinal reaction and dermal toxicity reaction etc.The Docetaxel water solublity is little, and commercially available taxotere adopts tween 80 solubilising, ethanol hydrotropy, can cause hemolytic reaction, poor stability.How to reduce its side effect, improve clinical efficacy and become problem demanding prompt solution.
Publication 200510084055.6 and 200610012102.0 is prepared into submicronized emulsion with Docetaxel, has improved the content and the stability of Docetaxel, has reduced oil content and has increased its targeting property to tumor tissues.The superiority that Docetaxel tradition submicronized emulsion is compared normal injection is confirmed.But the disclosed Docetaxel submicronized emulsion of two patents belongs to passive targeted preparation; Vein is annotated the back by macrophage phagocytic; Be enriched in liver, kidney and position, spleen, because tumor and other tissue distribution macrophage system seldom are unfavorable for the tumor tissues ingestion of drugs like this; Targeting efficient is low, and medicine is still bigger to the toxicity of other normal organs.Therefore the cancer target sexual needs further improve.
Low density lipoprotein receptor (LDLR) is a kind of cell membrane glycoprotein, and major function is the low density lipoprotein, LDL (LDL) that combines to carry cholesteryl ester.Discover cancerous cell overexpression LDLR such as carcinoma of prostate, malignant brain tumor.Therefore, LDLR can be used as the molecular target of cancer therapy drug target administration.Apolipoprotein B100 (apoB) is a kind of glycoprotein, and 9 amino acid residues wherein (3359 – 3367) combine with the functional domain of LDLR.Based on this, designed and contain the RLT polypeptide that combines with the LDLR functional domain and adopt solid-phase synthesis to synthesize the RLT polypeptide.With the peptide modified submicronized emulsion emulsion droplet of synthetic RLT surface, make its tumor cell at molecular level identification high expressed LDLR.The Docetaxel cancer target submicronized emulsion of the RLT polypeptide mediation of the present invention's preparation is an active target tumor preparation, and it can significantly improve the tumor-targeting of Docetaxel.
Summary of the invention
The objective of the invention is to overcome the shortcoming of existing injectable emulsion and lipomul; A kind of new Docetaxel submicronized emulsion agent with active target tumor is provided; Promptly provide in a kind of apolipoprotein B and to combine the bonded polypeptide in territory (RLT polypeptide) with low density lipoprotein, LDL; The said preparation curative effect is superior to existing aqueous injection property, has the characteristic of excellent drug stability and active target tumor simultaneously.
The objective of the invention is to realize through following technical scheme:
The Docetaxel cancer target submicronized emulsion of RLT polypeptide mediation of the present invention contains Docetaxel 0.5～20mg, injection vegetable oil 50～200 mg, emulsifying agent 8～24 mg, co-emulsifier 1～20 mg, antioxidant vitamin E 0.2mg, 18-amine. 0.6mg, DSPE-Macrogol 2000-maleimide 0.1～1.0mg, RLT polypeptide 0.08～0.8 mg and isoosmotic adjusting agent glycerol 22.5 mg in every milliliter of submicron emulsion.
Mean diameter＜the 200nm of submicron emulsion decentralized photo of the present invention.
Used injection vegetable oil is selected from the mixture a kind of or two or more material in the middle of them in soybean oil, Oleum Camelliae, Oleum Sesami, safflower oil, the olive oil among the present invention.
Used emulsifying agent is selected from one or more in Ovum Gallus domesticus Flavus lecithin, soybean phospholipid, hydrogenated soya phosphatide, the synthetic phospholipid.
Used co-emulsifier is selected from the mixture of poloxamer, one or both materials of polyglycol distearate.
Described RLT polypeptide is cysteine-glutamic-lysine-leucine-LYS-GLU-alanine-phenylalanine-arginine-leucine-threonine-arginine-lysine-arginine-glycine-leucine-lysine-leucine-alanine; HS-glutamic-lysine-leucine-LYS-GLU-alanine-phenylalanine-arginine-leucine-threonine-arginine-lysine-arginine-glycine-leucine-lysine-leucine-alanine; Glutamic-lysine-leucine-LYS-GLU-alanine-phenylalanine-arginine-leucine-threonine-arginine-lysine-arginine-glycine-leucine-lysine-leucine-alanine-SH, a kind of in glutamic-lysine-leucine-LYS-GLU-alanine-phenylalanine-arginine-leucine-threonine-arginine-lysine-arginine-glycine-leucine-lysine-leucine-alanine-cysteine.
Used antioxidant and isoosmotic adjusting agent are respectively vitamin E and glycerol.
The method for preparing of the Docetaxel cancer target submicronized emulsion of above-mentioned RLT polypeptide mediation provided by the invention comprises the following steps:
(1) preparation oil phase: under 55～75 ℃ of conditions, emulsifying agent, Docetaxel, antioxidant vitamin E, 18-amine., DSPE-Macrogol 2000-maleimide and injection vegetable oil joined in the dehydrated alcohol dissolve; Under 30～45 ℃, ethanol is removed in decompression, is heated to 55～75 ℃, stirs, and forms oil phase;
(2) preparation water: under 55～75 ℃ of conditions, co-emulsifier, glycerol etc. are added in the water for injection, stir, form water;
(3) preparation colostrum: remain in temperature under the prerequisite of 55～75 ℃ of constant temperature and constantly stirring, oil phase slowly is injected into water, stir fast; Regulate pH value 7.2～7.4; Standardize solution, homogenizing is 2～15 minutes under even matter speed 6000～12000rpm, makes colostrum;
(4) preparation of breast eventually: the colostrum that makes is crossed the microjet appearance, under the pressure of 3000～18000 PSI, even matter 6～20 times; Make the Docetaxel submicron emulsion;
(5) preparation of the Docetaxel cancer target submicronized emulsion of RLT polypeptide mediation: the RLT polypeptide is joined in the Docetaxel submicronized emulsion 37 ℃ of stirrings hatched 15～60 minutes, the filter membrane of crossing 0.22 μ m can make the Docetaxel cancer target submicron emulsion of RLT polypeptide mediation.
Advantage of the present invention is: because the present invention is prepared into submicronized emulsion with emulsifying agent, oil for injection and Docetaxel, with the matrix material of co-emulsifier, stabilizing agent, isoosmotic adjusting agent modification submicron emulsion, increase the dissolubility of Docetaxel.Thereby prolong drug action time in vivo, has slow-releasing and controlled-releasing action.The peptide modified submicronized emulsion of RLT makes it have good active target tumor characteristic, has improved the therapeutic effect of medicine, reduces toxic and side effects; The preparation technology that the present invention adopts is simple, drug loading is high, quality controllable, be fit to suitability for industrialized production.
Description of drawings
The RLT polypeptide mass spectrum of Fig. 1 solid-phase synthesis preparation.
Docetaxel cancer target submicron emulsion and other two kinds of Emulsion Re in each tissue of the mediation of Fig. 2 RLT polypeptide.
Docetaxel cancer target submicron emulsion and other two kinds of Emulsion Re in each tissue of the mediation of Fig. 3 RLT polypeptide.
The Docetaxel cancer target submicron emulsion of Fig. 4 RLT polypeptide mediation and the cancer target efficient of other two kinds of Emulsions.
The relative targeting efficient of tumor of the Docetaxel cancer target submicron emulsion of Fig. 5 RLT polypeptide mediation.
The specific embodiment
The used RLT polypeptide of the present invention is situated between for the solid-phase synthesis preparation, at first uses chloromethyl resin to make carrier, reuse sodium hydride solution and 2-methylol-3; The 4-dihydropyran is room temperature and chloromethyl resin reaction in acetic acid dimethylamide, obtains 3,4-dihydropyran hydroxymethyl resin; Further synthetic then RLT peptide, at first with 3,4-dihydropyran hydroxymethyl resin adds the protection base in the synthetic RLT peptide process; Promptly under the effect of p-methyl benzenesulfonic acid pyridiniujm, fluorenylmethyloxycarbonyl-uncle's 0-butyl ether Soviet Union ammonia alcohol is linked 3; 4-dihydropyran hydroxymethyl resin gets on, and with 3,4-dihydropyran hydroxymethyl resin joins among the THF of new steaming and soaks into; According to the synthetic method of Fmoc/tBu solid-phase polypeptide aminoacid is connected one by one then; Use trichloroacetic acid at last: water: thioanisole: the mixed solution of dithioglycol downcuts peptide from resin, and the bullion that obtains is through the freezing RLT peptide that obtains of preparation HPLC purification.
Below in conjunction with embodiment the present invention is elaborated, but protection scope of the present invention is not limited to this.
1. the raw material and the proportioning raw materials of the Docetaxel cancer target submicronized emulsion of preparation RLT polypeptide mediation are as shown in table 1.
Described RLT polypeptide is glutamic-lysine-leucine-LYS-GLU-alanine-phenylalanine-arginine-leucine-threonine-arginine-lysine-arginine-glycine-leucine-lysine-leucine-alanine-cysteine.
2. the method for the Docetaxel cancer target submicronized emulsion of preparation RLT polypeptide mediation is following:
(1) preparation oil phase: under 55 ℃ of conditions, emulsifying agent, Docetaxel, antioxidant vitamin E, 18-amine., DSPE-Macrogol 2000-maleimide and injection vegetable oil joined in the dehydrated alcohol dissolve; Under 30 ℃, ethanol is removed in decompression, is heated to 55 ℃, stirs, and forms oil phase;
(2) preparation water: under 55 ℃ of conditions, co-emulsifier, glycerol etc. are added in the water for injection, stir, form water;
(3) preparation colostrum: remain in temperature under the prerequisite of 55 ℃ of constant temperature and constantly stirring, oil phase slowly is injected into water, stir fast, regulate pH value 7.2, standardize solution, homogenizing 15min under even matter speed 6000rpm makes colostrum;
(4) preparation of breast eventually: the colostrum that makes is crossed the microjet appearance, under the pressure of 3000 PSI, even matter 20 times; Make the Docetaxel submicronized emulsion;
(5) the RLT polypeptide is joined in the Docetaxel submicronized emulsion 37 ℃ of stirrings and hatched 15 minutes, the filter membrane of crossing 0.22 μ m can make the Docetaxel cancer target submicronized emulsion of RLT polypeptide mediation.
1. the raw material and the proportioning raw materials of the Docetaxel cancer target submicronized emulsion of preparation RLT polypeptide mediation are as shown in table 1.
Described RLT polypeptide is glutamic-lysine-leucine-LYS-GLU-alanine-phenylalanine-arginine-leucine-threonine-arginine-lysine-arginine-glycine-leucine-lysine-leucine-alanine-SH.
2. the method for the Docetaxel cancer target submicronized emulsion of preparation RLT polypeptide mediation is following:
(1) preparation oil phase: under 60 ℃ of conditions, emulsifying agent, Docetaxel, antioxidant vitamin E, 18-amine., DSPE-Macrogol 2000-maleimide and injection vegetable oil joined in the dehydrated alcohol dissolve; Under 35 ℃, ethanol is removed in decompression, is heated to 60 ℃, stirs, and forms oil phase;
(2) preparation water: under 60 ℃ of conditions, co-emulsifier, glycerol etc. are added in the water for injection, stir, form water;
(3) preparation colostrum: remain in temperature under the prerequisite of 60 ℃ of constant temperature and constantly stirring, oil phase slowly is injected into water, stir fast, regulate pH value 7.2, standardize solution, homogenizing 12min under even matter speed 8000rpm makes colostrum;
(4) preparation of breast eventually: the colostrum that makes is crossed the microjet appearance, under the pressure of 6000 PSI, even matter 16 times; Make the Docetaxel submicron emulsion;
(5) the RLT polypeptide is joined in the Docetaxel submicronized emulsion 37 ℃ of stirrings and hatched 30 minutes, the filter membrane of crossing 0.22 μ m can make the Docetaxel cancer target submicronized emulsion of RLT polypeptide mediation.
1. the raw material and the proportioning raw materials of the Docetaxel cancer target submicronized emulsion of preparation RLT polypeptide mediation are as shown in table 1.
Described RLT polypeptide is cysteine-glutamic-lysine-leucine-LYS-GLU-alanine-phenylalanine-arginine-leucine-threonine-arginine-lysine-arginine-glycine-leucine-lysine-leucine-alanine.
2. the method for the Docetaxel cancer target submicronized emulsion of preparation RLT polypeptide mediation is following:
(1) preparation oil phase: under 65 ℃ of conditions, emulsifying agent, Docetaxel, antioxidant vitamin E, 18-amine., DSPE-Macrogol 2000-maleimide and injection vegetable oil joined in the dehydrated alcohol dissolve; Under 35 ℃, ethanol is removed in decompression, is heated to 65 ℃, stirs, and forms oil phase;
(2) preparation water: under 65 ℃ of conditions, co-emulsifier, glycerol etc. are added in the water for injection, stir, form water;
(3) preparation colostrum: remain in temperature under the prerequisite of 65 ℃ of constant temperature and constantly stirring, oil phase slowly is injected into water, stir fast, regulate pH value 7.4, standardize solution, homogenizing 10min under even matter speed 10000rpm makes colostrum;
(4) preparation of breast eventually: the colostrum that makes is crossed the microjet appearance, under the pressure of 8000 PSI, even matter 12 times; Make the Docetaxel submicron emulsion;
(5) the RLT polypeptide is joined in the Docetaxel submicronized emulsion 37 ℃ of stirrings and hatched 30 minutes, the filter membrane of crossing 0.22 μ m can make the Docetaxel cancer target submicronized emulsion of RLT polypeptide mediation.
1. the raw material and the proportioning raw materials of the Docetaxel cancer target submicronized emulsion of preparation RLT polypeptide mediation are as shown in table 1.
Described RLT polypeptide is HS-glutamic-lysine-leucine-LYS-GLU-alanine-phenylalanine-arginine-leucine-threonine-arginine-lysine-arginine-glycine-leucine-lysine-leucine-alanine.
2. the method for the Docetaxel cancer target submicronized emulsion of preparation RLT polypeptide mediation is following:
(1) preparation oil phase: under 70 ℃ of conditions, emulsifying agent, Docetaxel, antioxidant vitamin E, 18-amine., DSPE-Macrogol 2000-maleimide and injection vegetable oil joined in the dehydrated alcohol dissolve; Under 35 ℃, ethanol is removed in decompression, is heated to 70 ℃, stirs, and forms oil phase;
(2) preparation water: under 70 ℃ of conditions, co-emulsifier, glycerol etc. are added in the water for injection, stir, form water;
(3) preparation colostrum: remain in temperature under the prerequisite of 70 ℃ of constant temperature and constantly stirring, oil phase slowly is injected into water, stir fast, regulate pH value 7.4, standardize solution, homogenizing 8min under even matter speed 700rpm makes colostrum;
(4) preparation of breast eventually: the colostrum that makes is crossed the microjet appearance, under the pressure of 12000 PSI, even matter 10 times; The Docetaxel submicron emulsion;
(5) the RLT polypeptide is joined in the Docetaxel submicronized emulsion 37 ℃ of stirrings and hatched 45 minutes, the filter membrane of crossing 0.22 μ m can make the Docetaxel cancer target submicronized emulsion of RLT polypeptide mediation.
1. the raw material and the proportioning raw materials of the Docetaxel cancer target submicronized emulsion of preparation RLT polypeptide mediation are as shown in table 1.
Described RLT polypeptide is cysteine-glutamic-lysine-leucine-LYS-GLU-alanine-phenylalanine-arginine-leucine-threonine-arginine-lysine-arginine-glycine-leucine-lysine-leucine-alanine.
2. the method for the Docetaxel cancer target submicronized emulsion of preparation RLT polypeptide mediation is following:
(1) preparation oil phase: under 75 ℃ of conditions, emulsifying agent, Docetaxel, antioxidant vitamin E, 18-amine., DSPE-Macrogol 2000-maleimide and injection vegetable oil joined in the dehydrated alcohol dissolve; Under 35 ℃, ethanol is removed in decompression, is heated to 75 ℃, stirs, and forms oil phase;
(2) preparation water: under 75 ℃ of conditions, co-emulsifier, glycerol etc. are added in the water for injection, stir, form water;
(3) preparation colostrum: remain in temperature under the prerequisite of 75 ℃ of constant temperature and constantly stirring, oil phase slowly is injected into water, stir fast, regulate pH value 7.4, standardize solution, homogenizing 2～5min under even matter speed 11000rpm makes colostrum;
(4) preparation of breast eventually: the colostrum that makes is crossed the microjet appearance, under the pressure of 16000 PSI, even matter 8 times; Make the Docetaxel submicron emulsion;
(5) the RLT polypeptide is joined in the Docetaxel submicronized emulsion 37 ℃ of stirrings and hatched 45 minutes, the filter membrane of crossing 0.22 μ m can make the Docetaxel cancer target submicronized emulsion of RLT polypeptide mediation.
1. the raw material and the proportioning raw materials of the Docetaxel cancer target submicronized emulsion of preparation RLT polypeptide mediation are as shown in table 1.
Described RLT polypeptide is cysteine-glutamic-lysine-leucine-LYS-GLU-alanine-phenylalanine-arginine-leucine-threonine-arginine-lysine-arginine-glycine-leucine-lysine-leucine-alanine.
2. the method for the Docetaxel cancer target submicronized emulsion of preparation RLT polypeptide mediation is following:
(1) preparation oil phase: under 75 ℃ of conditions, emulsifying agent, Docetaxel, antioxidant vitamin E, 18-amine., DSPE-Macrogol 2000-maleimide and injection vegetable oil joined in the dehydrated alcohol dissolve; Under 35 ℃, ethanol is removed in decompression, is heated to 75 ℃, stirs, and forms oil phase;
(2) preparation water: under 75 ℃ of conditions, co-emulsifier, glycerol etc. are added in the water for injection, stir, form water;
(3) preparation colostrum: remain in temperature under the prerequisite of 75 ℃ of constant temperature and constantly stirring, oil phase slowly is injected into water, stir fast, regulate pH value 7.4, standardize solution, homogenizing 2min under even matter speed 12000rpm makes colostrum;
(4) preparation of breast eventually: the colostrum that makes is crossed the microjet appearance, under the pressure of 18000 PSI, even matter 6 times; Make the Docetaxel submicron emulsion;
(5) the RLT polypeptide is joined in the Docetaxel submicronized emulsion 37 ℃ of stirrings and hatched 60 minutes, the filter membrane of crossing 0.22 μ m can make the Docetaxel cancer target submicronized emulsion of RLT polypeptide mediation.
Embodiment 7～16
1. the raw material and the proportioning raw materials of the Docetaxel cancer target submicronized emulsion of preparation RLT polypeptide mediation are as shown in table 1.
Described RLT polypeptide is cysteine-glutamic-lysine-leucine-LYS-GLU-alanine-phenylalanine-arginine-leucine-threonine-arginine-lysine-arginine-glycine-leucine-lysine-leucine-alanine.
2. the method for the Docetaxel cancer target submicronized emulsion of preparation RLT polypeptide mediation adopts the method for preparing in the foregoing description 1～6.
Among each embodiment of table 1 the Docetaxel cancer target submicronized emulsion of preparation RLT polypeptide mediation and proportioning raw materials
Used antioxidant can also adopt the antioxidant of other commercially available type in the foregoing description, and isoosmotic adjusting agent can adopt the isoosmotic adjusting agent of other commercially available type.
The Docetaxel cancer target submicronized emulsion that the RLT polypeptide for preparing with embodiment 3 mediates is an example, and result of study is following:
The RLT polypeptide that adopts mass spectrograph to detect the solid-phase synthesis preparation is seen Fig. 1.
Adopt Nano-ZS90 type laser nano particle size analyzer to measure emulsion droplet particle diameter and zeta current potential.The emulsion droplet of the Docetaxel cancer target submicronized emulsion of RLT polypeptide mediation as a result is evenly distributed, and mean diameter is 182.2 ± 10nm, and average zeta current potential is 39.62 ± 2.41mV.
Medicine assay:The accurate Docetaxel cancer target submicronized emulsion of drawing the mediation of 20 μ LRLT polypeptide adds the methanol ultrasound destruction and is diluted to scale in the 5mL volumetric flask, measures with HPLC, and the peak area substitution standard curve that obtains calculates its medicament contg.
Entrapment efficiency determination:The accurate Docetaxel cancer target submicronized emulsion of drawing the mediation of 0.2mLRLT polypeptide places the ultrafiltration pipe; 10000r/min, centrifugal 20min makes water all centrifugal to the ultrafiltration tube outside as far as possible; High performance liquid chromatogram is measured the content of ultrafiltrate Chinese medicine, utilizes the computes envelop rate.
The result records that Docetaxel content is 1.410 ± 0.14mg/ml in the Docetaxel cancer target submicronized emulsion of RLT polypeptide mediation, envelop rate>98%.
The tumor-targeting evaluation of the Docetaxel cancer target submicronized emulsion of polypeptide mediation:27 lotus S180 tumor mices are divided into three groups at random; Every group 9; Three groups of mices are the Docetaxel cancer target submicronized emulsions of tail vein injection Docetaxel tradition submicronized emulsion, Docetaxel cation submicronized emulsion and the mediation of RLT polypeptide respectively, and 3 mices of each time point are taken out mice when 0.5h, 1h, three time points of 3h after the administration from each group; Anatomical isolation obtains the heart, liver, spleen, lung, kidney, brain and tumor; With each tissue back homogenate of normal saline flushing, employing HPLC mensuration is individual organizes Chinese medicine content, and calculates its AUC and C
MaxTo AUC and C
MaxHandle, adopt the tumor-targeting of peak concentration ratio, relative uptake ratio, targeting efficient and relative targeting efficiency rating preparation:
Peak concentration ratio: Ce=(C
Max)
C/ (C
Max)
B or ARelative uptake ratio: Re=(AUC
i)
C/ (AUC
i)
B or A
Targeting efficient: Te=(AUC
i)
Target/ (AUC
i)
Non-targetRelative targeting efficient: R
Te=(Te)
C/ (Te)
B or A
AUC is the area under the drug-time curve of i tissue in the following formula; Subscript A, B and C represent the Docetaxel cancer target submicronized emulsion of Docetaxel tradition submicronized emulsion, Docetaxel cation submicronized emulsion and the mediation of RLT polypeptide respectively.
Fig. 2 is Docetaxel cancer target submicron emulsion and other two kinds of Emulsion Re in each tissue of the mediation of RLT polypeptide, and Fig. 3 is Docetaxel cancer target submicron emulsion and other two kinds of Emulsion Re in each tissue of RLT polypeptide mediation.Can find out by Fig. 2 and Fig. 3: in tumor tissues; Two kinds of preparations of Docetaxel cancer target submicronized emulsion and other of RLT polypeptide mediation are compared; Re and Ce explain that all greater than 1 its distribution in tumor tissues is higher than other two kinds of submicronized emulsions, and tumor-targeting is relatively good.
Fig. 4 is the Docetaxel cancer target submicron emulsion of RLT polypeptide mediation and the cancer target efficient of other two kinds of Emulsions; Can find out by Fig. 4: if with the heart, liver, spleen, lung and kidney as non-target tissue; The tumor-targeting of three kinds of preparations not obvious (Te < 1), the targeting efficient size of each tissue is followed successively by: the Docetaxel cancer target submicronized emulsion of RLT polypeptide mediation>traditional submicronized emulsion.
The relative targeting efficient of tumor of the Docetaxel cancer target submicron emulsion of Fig. 5 RLT polypeptide mediation; Can find out by Fig. 5: if with the heart, liver, spleen, lung, kidney and brain as non-target tissue, the Docetaxel cancer target submicronized emulsion and the relative targeting efficient of other two kinds of submicronized emulsions: the R of RLT polypeptide mediation
Te>1, explain that the targeting property of the Docetaxel cancer target submicronized emulsion that the RLT polypeptide mediates is better than other two kinds of preparations, especially compare it and have better tumor-targeting with traditional submicronized emulsion.
Claims (9)
1. the Docetaxel cancer target submicronized emulsion of a RLT polypeptide mediation is characterized in that: contain Docetaxel 0.5～20mg, injection vegetable oil 0.05～0.2 g, emulsifying agent 8～24 mg, co-emulsifier 0～20 mg, antioxidant 0.2mg, 18-amine. 0.6mg, DSPE-Macrogol 2000-maleimide 0.1～1.0mg, RLT polypeptide 0.08～0.8 mg, isoosmotic adjusting agent 22.5 mg in every milliliter of submicronized emulsion.
2. the Docetaxel cancer target submicronized emulsion of RLT polypeptide mediation according to claim 1 is characterized in that: the mean diameter＜200nm of submicronized emulsion decentralized photo.
3. the Docetaxel cancer target submicronized emulsion of RLT polypeptide mediation according to claim 1 and 2 is characterized in that: described injection vegetable oil is selected from the mixture a kind of or two or more material in the middle of them in soybean oil, Oleum Camelliae, Oleum Sesami, safflower oil, the olive oil.
4. the Docetaxel cancer target submicronized emulsion of RLT polypeptide mediation according to claim 3, it is characterized in that: described emulsifying agent is selected from one or more in Ovum Gallus domesticus Flavus lecithin, soybean phospholipid, hydrogenated soya phosphatide, the synthetic phospholipid.
5. according to the Docetaxel cancer target submicronized emulsion of claim 1 or 2 or 4 described RLT polypeptide mediations, it is characterized in that: described co-emulsifier is selected from the mixture of one or both materials of poloxamer, polyglycol distearate.
6. the Docetaxel cancer target submicronized emulsion of RLT polypeptide mediation according to claim 1; It is characterized in that: described RLT polypeptide is cysteine-glutamic-lysine-leucine-LYS-GLU-alanine-phenylalanine-arginine-leucine-threonine-arginine-lysine-arginine-glycine-leucine-lysine-leucine-alanine; HS-glutamic-lysine-leucine-LYS-GLU-alanine-phenylalanine-arginine-leucine-threonine-arginine-lysine-arginine-glycine-leucine-lysine-leucine-alanine; Glutamic-lysine-leucine-LYS-GLU-alanine-phenylalanine-arginine-leucine-threonine-arginine-lysine-arginine-glycine-leucine-lysine-leucine-alanine-SH, a kind of in glutamic-lysine-leucine-LYS-GLU-alanine-phenylalanine-arginine-leucine-threonine-arginine-lysine-arginine-glycine-leucine-lysine-leucine-alanine-cysteine.
7. according to the Docetaxel cancer target submicronized emulsion of claim 1 or 2 or 6 described RLT polypeptide mediations, it is characterized in that: described antioxidant is a vitamin E.
8. the Docetaxel cancer target submicronized emulsion of RLT polypeptide mediation according to claim 7, it is characterized in that: described isoosmotic adjusting agent is a glycerol.
9. the method for preparing of the Docetaxel cancer target submicronized emulsion of the described RLT polypeptide of claim 1 mediation, it is characterized in that: method for preparing may further comprise the steps:
(1) preparation oil phase: under 55～75 ℃ of conditions, emulsifying agent, Docetaxel, antioxidant vitamin E, 18-amine., DSPE-Macrogol 2000-maleimide and injection vegetable oil joined in the dehydrated alcohol dissolve; Under 30～45 ℃, ethanol is removed in decompression, is heated to 55～75 ℃, stirs, and forms oil phase;
(2) preparation water: under 55～75 ℃ of conditions, co-emulsifier, glycerol etc. are added in the water for injection, stir, form water;
(3) preparation colostrum: remain in temperature under the prerequisite of 55～75 ℃ of constant temperature and constantly stirring, oil phase slowly is injected into water, stir fast; Regulate pH value 7.2～7.4; Standardize solution, homogenizing is 2～15 minutes under even matter speed 6000～12000rpm, makes colostrum;
(4) preparation of breast eventually: the colostrum that makes is crossed the microjet appearance, under the pressure of 3000～18000 PSI, even matter 6～20 times; Make the Docetaxel submicronized emulsion;
(5) preparation of the Docetaxel cancer target submicronized emulsion of RLT polypeptide mediation: the RLT polypeptide is joined in the Docetaxel submicron emulsion 37 ℃ of stirrings hatched 15 ~ 60 minutes, the filter membrane of crossing 0.22 μ m can make the Docetaxel cancer target submicronized emulsion of RLT polypeptide mediation.
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| CN1709236A (en) * | 2005-07-18 | 2005-12-21 | 山东鲁抗辰欣药业有限公司 | Fat emulsion containing docetaxel and its preparing method |
| CN1857222A (en) * | 2006-06-05 | 2006-11-08 | 中国医药研究开发中心有限公司 | Submicron docetaxel emulsion for intravenous injection and its preparing process |
| CN100998559A (en) * | 2006-12-27 | 2007-07-18 | 沈阳药科大学 | Submicroemulsion injection containing polyene paclitaxel and its preparing method |
| CN101401941A (en) * | 2008-11-21 | 2009-04-08 | 首都医科大学 | Preparation and uses of tumor-targeted carrier material RGD-fatty alcohol series of compounds |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1709236A (en) * | 2005-07-18 | 2005-12-21 | 山东鲁抗辰欣药业有限公司 | Fat emulsion containing docetaxel and its preparing method |
| CN1857222A (en) * | 2006-06-05 | 2006-11-08 | 中国医药研究开发中心有限公司 | Submicron docetaxel emulsion for intravenous injection and its preparing process |
| CN100998559A (en) * | 2006-12-27 | 2007-07-18 | 沈阳药科大学 | Submicroemulsion injection containing polyene paclitaxel and its preparing method |
| CN101401941A (en) * | 2008-11-21 | 2009-04-08 | 首都医科大学 | Preparation and uses of tumor-targeted carrier material RGD-fatty alcohol series of compounds |
Non-Patent Citations (1)
| Title |
|---|
| M.NIKANJAM，ET AL: "Synthetic nano-LDL with paclitaxel oleate as a targeted drug delivery vehicle for glioblastoma multiforme", 《JOURNAL OF CONTROLLED RELEASE》, vol. 124, 26 September 2007 (2007-09-26), pages 163 - 171, XP022347965, DOI: doi:10.1016/j.jconrel.2007.09.007 * |
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