A kind of taxol compound and the pharmaceutical composition containing this taxol compound
Technical field
The present invention relates to a kind of taxol compound, the preparation method of this taxol compound, also relate to the pharmaceutical composition containing this taxol.
Background technology
At present, risen year by year both at home and abroad by the mortality ratio of the initiation of cancer, China accounts for 20% of death toll because of number of cancer deaths.Mammary cancer, lung cancer, prostate cancer and colorectal carcinoma etc. are the most common, and conventional cancer therapy drug has taxol, Docetaxel, capecitabine etc.
Taxol (paclitaxel) is a kind of diterpene-kind compound with highly effective antineoplastic activity of extraction and isolation from taxaceae Chinese yew genus plants Ramulus et folium taxi cuspidatae, it has unique mechanism of action, by inducing and promoting tubulin polymerization, assembling and stable microtubule effect, stoping the growth of tumour cell, is spectrum cancer therapy drug.Be be used for the treatment of one of best medicine of advanced ovarian cancer, mammary cancer curative effect clinically at present, to non-small cell carcinoma, prostate cancer, tumor of head and neck, rheumatoid arthritis etc., there is good curative effect.Its structure is complicated diterpene-kind compound, and its chemical name is 5 β, 20-epoxy-1,2 α, 4,7 β, 10 β, 13 α-hexahydroxy-Taxan-11-alkene-9-ketone-4,10-diacetate esters-2-benzoic ether-13-(2 ' R, 3 ' S)-N-benzoyl-3-phenylisoserine ester, molecular formula C
47h
51nO
14, its skeleton symbol is as follows:
Taxol water-soluble very poor, water-soluble hardly, the effective bioavailability in human body is low.The paclitaxel injection of current Clinical practice uses polyoxyethylenated castor oil and dehydrated alcohol to dissolve taxol with the ratio mixed solvent of 1:1, makes the liquid of 5ml containing 30mg taxol.But, discharge histamine during polyoxyethylenated castor oil degradation in vivo, cause serious anaphylaxis, some patients within several minutes after instillation, just there will be Drug as fash, be short of breath, the anaphylaxis such as bronchospasm, ypotension.Also there are some researches show, the divinyl hexyl phthalate in polyoxyethylenated castor oil solubilized polyethylene infusion device, cause serious toxic reaction, the safety affecting medicine is drunk.
The approach of existing solution taxol soluble difference mainly comprises change formulation and modifying for chemical structure, the method wherein changing formulation comprises prepares Paclitaxel liposome, yew alcohol micro-emulsion, taxol microballoon and effect of nano-paclitaxel etc., above-mentioned preparation improves the water-soluble of medicine to a certain extent, but also there is many defects, change formulation and can only alleviate untoward reaction, but fundamentally cannot improve the drug effect of taxol.Modifying for chemical structure can introduce the little group of solubility promoter, such as carboxylic acid, sulfonic acid etc. on 2 '-hydroxyl or 7-hydroxy position, also can in conjunction with water-soluble polymer carriers such as polyoxyethylene glycol, polyglutamic acid, poly aspartic acids on taxol.But there is the defects such as synthetic method, separation and purification requirement is high, yield is low in this chemically modified, cannot industrializing implementation.
Chinese patent application CN200810034331.1 discloses a kind of polyenic taxusol sesquihydrate crystal its preparation method, not containing organic solvent in this crystal, filtering solution, the purity of product is high, impurity is few, although not containing organic solvent in this Docetaxel, the water-soluble of this Docetaxel does not improve.Given this, special proposition the present invention.
Summary of the invention
The object of this invention is to provide a kind of taxol compound crystal, this compound be water-soluble making moderate progress, and the process ratio preparing this taxol be easier to operation.
Another object of the present invention provides a kind of preparation method of taxol compound crystal,
Of the present inventionly be to provide a kind of taxol drug composition an object, it comprises active ingredient taxol and pharmaceutically acceptable carrier, can be prepared into the formulation such as injection liquid, oral tablet.
A kind of taxol compound, uses Cu-K α
1as shown in Figure 1, described taxol contains 3.5 crystal water to the X-ray powder diffraction of radionetric survey.
The chemical structure skeleton symbol of described taxol is:
(1) paclitaxel solid is added in the mixing solutions of toluene, methylene dichloride and methyl alcohol, reflux, until paclitaxel solid all dissolves;
(2) control temperature is under the condition of 35-40 DEG C, adds the aqueous solution of ethanol in the solution that step (1) obtains, and wherein, the mass percent containing ethanol in the aqueous solution of ethanol is 10%-15%;
(3) after adding the aqueous solution of ethanol, be that 2-3 DEG C/10min is cooled to-5-0 DEG C with speed, at-5-0 DEG C of standing 12-18 hour, crystallize out, filter, filter cake washed with diethylether, 2-4 hour after vacuum-drying, obtains taxol.
It is preferred, and in step (1), toluene, volume between methylene dichloride and methyl alcohol are 2.0:(1.5-2.0): (2.0:-3.5); Preferred, described toluene, the volume between methylene dichloride and methyl alcohol are 2.0:2.0:(2.0-3.0).
Described taxol and the mass volume ratio of mixing solutions are (4-5) g:10ml.
A kind of paclitaxel composition, can be oral preparations or liquid preparation, described oral preparations be conventional tablet, capsule, granule etc., and described liquid preparation is oral liquid, freeze-dried powder, injection liquid etc.
Described pharmaceutically acceptable carrier, comprises thinner conventional in preparation, weighting agent, tackiness agent, disintegrating agent, lubricant, wetting agent or stablizer etc.
Described weighting agent comprises N.F,USP MANNITOL, lactose, ethylene glycol etc.; Described tackiness agent comprises starch or Microcrystalline Cellulose etc.; Described disintegrating agent comprises carboxymethyl cellulose, low-substituted hydroxypropyl cellulose etc.; Described lubricant comprises talcum powder or Magnesium Stearate; Described wetting agent comprises propylene glycol or ethanol; Described stablizer comprises EDTA-2Na, Sulfothiorine, sodium carbonate etc.
The specification of tablet has 25mg, 30mg, 40mg or 50mg.
Wherein, described paclitaxel injection comprises taxol, dehydrated alcohol, lactic acid and Cremophor EL.
Described dehydrated alcohol, between lactic acid and Cremophor EL volume be 1:(0.007-0.0085): (1.8-2.0); Described taxol and the mass volume ratio of dehydrated alcohol are 1g/ (80-85) ml.
The preparation method of described paclitaxel injection comprises:
(1) get dehydrated alcohol, lactic acid, the Cremophor EL of recipe quantity respectively, add the gac of 0.1%, stir, place 10 minutes, filtering decarbonization, for subsequent use;
(2) taxol of recipe quantity is dissolved in dehydrated alcohol, after stirring and dissolving, then adds lactic acid, then add Cremophor EL to full dose, filter with G3 sand core funnel;
(3) by gained filtrate after filtering by quality standard measure the content of pH value and taxol qualified after, filling, tamponade, roll aluminium lid;
(4) 100 DEG C of flowing steam sterilizations 30 minutes, lamp inspection, Quan Jian, packaging, warehouse-in.
Further describe to summary of the invention of the present invention below:
Taxol compound of the present invention, uses Cu-K α
1as shown in Figure 1, described taxol contains 3.5 crystal water to the X-ray powder diffraction of radionetric survey.
This paclitaxel crystal compound is prepared from by the following method:
(1) paclitaxel solid is added in the mixing solutions of toluene, methylene dichloride and methyl alcohol, reflux, until paclitaxel solid all dissolves;
(2) control temperature is under the condition of 35-40 DEG C, adds the aqueous solution of ethanol in the solution that step (1) obtains, and wherein, the mass percent containing ethanol in the aqueous solution of ethanol is 10%-15%;
(3) after adding the aqueous solution of ethanol, be that 2-3 DEG C/10min is cooled to-5-0 DEG C with speed, at-5-0 DEG C of standing 12-18 hour, crystallize out, filter, filter cake washed with diethylether, 2-4 hour after vacuum-drying, obtains taxol.
By mixed solvent toluene, methylene dichloride and methyl alcohol, preferred toluene, volume between methylene dichloride and methyl alcohol are 2.0:(1.5-2.0): (2.0:-3.5); Preferred, described toluene, the volume between methylene dichloride and methyl alcohol are 2.0:2.0:(2.0-3.0).After paclitaxel solid dissolves by the mixing solutions of this ratio, and then use aqueous ethanolic solution recrystallization, obtain paclitaxel crystal compound.The water-soluble paclitaxel solid compared to existing technology of this paclitaxel crystal compound, its solubility property increases, and stability and other performances also keep good.
The paclitaxel crystal of gained can be prepared into different formulations.Can be oral preparations or liquid preparation, described oral preparations be conventional tablet, capsule, granule etc., and described liquid preparation is oral liquid, freeze-dried powder, injection liquid etc.
Relative proportions those skilled in the art between the auxiliary material that various formulation is used and each component can go adjustment according to the actual requirements.
Wherein, described paclitaxel injection comprises taxol, dehydrated alcohol, lactic acid and Cremophor EL.
Described dehydrated alcohol, between lactic acid and Cremophor EL volume be 1:(0.007-0.0085): (1.8-2.0); Described taxol and the mass volume ratio of dehydrated alcohol are 1g/ (80-85) ml.
Due in paclitaxel injection of the present invention, auxiliary material is dehydrated alcohol, lactic acid and ethoxylate castor oil, and the polyoxyethylene groups Viscotrol C consumption contained can lack, and keep high in vivo relatively for a long time and need concentration, its toxic side effect reduces greatly.
That is, the stable chemical nature of paclitaxel crystal provided by the invention, water-solublely improves a lot, and the bioavailability when treating tumour improves.Paclitaxel crystal of the present invention is used for, in paclitaxel composition, being prepared into Taxol injection liquid formulation, and its component is simple, and the consumption of auxiliary material reduces greatly, improves the security that it uses.
Accompanying drawing explanation
The XRD figure of Fig. 1 taxol compound
The TG figure of Fig. 2 taxol compound of the present invention
The pharmacokinetic curve of Fig. 3 paclitaxel injection of the present invention
Embodiment
Further describe preparation process and the effect of paclitaxel crystal compound of the present invention below by specific examples, but be not limited to following examples.
Embodiment 1:
The preparation of taxol compound:
Added by 40g paclitaxel solid in the mixing solutions 100ml of toluene, methylene dichloride and methyl alcohol, reflux, until paclitaxel solid all dissolves, wherein, toluene, volume ratio between methylene dichloride and methyl alcohol are 2.0:1.5:2.0; Control temperature is under the condition of 40 DEG C, add the aqueous solution of the ethanol of 250ml, wherein, the mass percent containing ethanol is in the aqueous solution of 10%, is then that 3 DEG C/10min is cooled to-5 DEG C with speed, 17 hours are left standstill at-5 DEG C, crystallize out, filters, filter cake washed with diethylether, after vacuum-drying 4 hours, obtain taxol.This taxol compound adopts Cu-K α
1the X-ray powder diffraction of radionetric survey as shown in Figure 1, and containing 3.5 crystal water (see such as Fig. 2, the content of this taxol compound crystal water is 6.89%).
Embodiment 2:
The preparation of taxol compound:
Added by 50g paclitaxel solid in the mixing solutions 100ml of toluene, methylene dichloride and methyl alcohol, reflux, until paclitaxel solid all dissolves, wherein, toluene, volume between methylene dichloride and methyl alcohol are 2.0:2.0:3.0; Control temperature, under the condition of 35 DEG C, adds the aqueous solution of the ethanol of 200ml, and wherein, the mass percent containing ethanol is in the aqueous solution of 15%; Then be that 2.5 DEG C/10min is cooled to 0 DEG C with speed, leave standstill 12 hours at 0 DEG C, crystallize out, filter, filter cake washed with diethylether, after vacuum-drying 2.5 hours, obtain taxol.This taxol compound adopts Cu-K α
1the X-ray powder diffraction of radionetric survey shows, conforms to the result shown in accompanying drawing 1, and containing 3.5 crystal water (matching with Fig. 2).
Embodiment 3:
The preparation of taxol compound:
Added by 45g paclitaxel solid in the mixing solutions 100ml of toluene, methylene dichloride and methyl alcohol, reflux, until paclitaxel solid all dissolves, wherein, toluene, volume between methylene dichloride and methyl alcohol are 2.0:1.5:2.0; Control temperature, under the condition of 40 DEG C, adds the aqueous solution of the ethanol of 250ml, and wherein, the mass percent containing ethanol is in the aqueous solution of 13%; Then be that 2.0 DEG C/10min is cooled to-3 DEG C with speed, leave standstill 15 hours at-3 DEG C, crystallize out, filter, filter cake washed with diethylether, after vacuum-drying 3.0 hours, obtain taxol.This taxol compound adopts Cu-K α
1the X-ray powder diffraction of radionetric survey shows, conforms to the result shown in accompanying drawing 1, and containing 3.5 crystal water (matching with accompanying drawing 2).
Embodiment 4:
The preparation of taxol compound:
Added by 40g paclitaxel solid in the mixing solutions 100ml of toluene, methylene dichloride and methyl alcohol, reflux, until paclitaxel solid all dissolves, wherein, toluene, volume between methylene dichloride and methyl alcohol are 2.0:2.0:3.5; Control temperature, under the condition of 35 DEG C, adds the aqueous solution of the ethanol of 300ml, and wherein, the mass percent containing ethanol is in the aqueous solution of 15%; Then be that 3 DEG C/10min is cooled to-5 DEG C with speed, leave standstill 12 hours at-5 DEG C, crystallize out, filter, filter cake washed with diethylether, after vacuum-drying 2 hours, obtain taxol.This taxol compound adopts Cu-K α
1the X-ray powder diffraction of radionetric survey shows, conforms to the result shown in accompanying drawing 1, and containing 3.5 crystal water (matching with Fig. 2).
Embodiment 5:
The formula of paclitaxel injection:
Taxol 30g
Dehydrated alcohol 2500ml
Lactic acid 20ml
Cremophor EL 4000ml.
Make 1000.
The preparation method of described paclitaxel injection comprises:
(1) get dehydrated alcohol, lactic acid, the Cremophor EL of recipe quantity respectively, add the gac of 0.1%, stir, place 10 minutes, filtering decarbonization, for subsequent use;
(2) taxol of recipe quantity is dissolved in dehydrated alcohol, after stirring and dissolving, then adds lactic acid, then add Cremophor EL to full dose, filter with G3 sand core funnel;
(3) by gained filtrate after filtering by quality standard measure the content of pH value and taxol qualified after, filling, tamponade, roll aluminium lid;
(4) 100 DEG C of flowing steam sterilizations 30 minutes, lamp inspection, Quan Jian, packaging, warehouse-in.
Embodiment 6:
The formula of paclitaxel injection:
Taxol 30g
Dehydrated alcohol 2550ml
Lactic acid 19ml
Cremophor EL 4000ml.
Be prepared into 1000.
The preparation method of described paclitaxel injection comprises:
(1) get dehydrated alcohol, lactic acid, the Cremophor EL of recipe quantity respectively, add the gac of 0.1%, stir, place 10 minutes, filtering decarbonization, for subsequent use;
(2) taxol of recipe quantity is dissolved in dehydrated alcohol, after stirring and dissolving, then adds lactic acid, then add Cremophor EL to full dose, filter with G3 sand core funnel;
(3) by gained filtrate after filtering by quality standard measure the content of pH value and taxol qualified after, filling, tamponade, roll aluminium lid;
(4) 100 DEG C of flowing steam sterilizations 30 minutes, lamp inspection, Quan Jian, packaging, warehouse-in.
Embodiment 7:
The formula of paclitaxel injection:
Taxol 30g
Dehydrated alcohol 2400ml
Lactic acid 20ml
Cremophor EL 3320ml
The preparation method of described paclitaxel injection comprises:
(1) get dehydrated alcohol, lactic acid, the Cremophor EL of recipe quantity respectively, add the gac of 0.1%, stir, place 10 minutes, filtering decarbonization, for subsequent use;
(2) taxol of recipe quantity is dissolved in dehydrated alcohol, after stirring and dissolving, then adds lactic acid, then add Cremophor EL to full dose, filter with G3 sand core funnel;
(3) by gained filtrate after filtering by quality standard measure the content of pH value and taxol qualified after, filling, tamponade, roll aluminium lid;
(4) 100 DEG C of flowing steam sterilizations 30 minutes, lamp inspection, Quan Jian, packaging, warehouse-in.
Experimental example 1:
This experimental example is the stability test of product of the present invention.
1. accelerated test
Three batches of products of Example 1, in temperature be 40 ± 2 DEG C, relative humidity be the condition of 75 ± 5% under place 6 months, respectively at 0,1,2,3,6 the end of month sampling once, measure by high spot reviews project.In table 1.
Table 1, accelerated test result (temperature 40 ± 2 DEG C, relative humidity 75% ± 5%)
Result shows, this product temperature be 40 ± 2 DEG C, relative humidity be 75 ± 5% conditions under place 6 months, related substance and other index have no significant change, and this product quality is basicly stable.
2. test of long duration
Example 5 product, in temperature be 25 DEG C, relative humidity be 60% ± 10% place, respectively at 0,3,6,9,12,18,24 the end of month sampling and measuring.Measure proterties, content and pH value according to relevant regulations under " quality standard " item, record result (the results are shown in following table 2), and compare with criticizing to record for 0 month.
Table 2, long-term test results (temperature 25 ± 2 DEG C, relative humidity 60% ± 5%)
Test-results
Shown by long-term test results: taxol compound of the present invention is 25 ± 2 DEG C in temperature, relative humidity is place under the condition of 60% ± 5% to stablize for 24 months, and indices is without considerable change.
Experimental example 2:
The water-soluble of taxol compound that this experimental example carries out testing embodiment of the present invention 1-4 is tested, compared with commercially available taxol.Specific experiment method is: in the low capacity bottle of constant temperature jacket, add appropriate distilled water, taxol is added to no longer dissolving at 25 DEG C, start magnetic stirrer, Keep agitation under constant temperature, in experimentation, system is in the state of two-phase coexistent all the time, after 70 minutes system liquid phase in paclitaxel concentration be solubleness at this temperature.Carry out sampling analysis after 2 hours, get the close mean value of adjacent two times result as measured value of experiment, before sampling, in order to make solid-liquid fully be separated, after stopping stirring, not molten Paclitaxel precipitation is to the bottom of low capacity bottle, a small amount of upper clear supernate is extracted with syringe, filter with the filter of 0.45 micron, sample thief from filtrate, measures the content of taxol by HPLC.Specifically in table 3.
Under table 3 room temperature, taxol compound of the present invention is water-soluble
Concentration (mg/ml) |
Embodiment 1 |
Embodiment 2 |
Embodiment 3 |
Embodiment 4 |
Commercially available |
Once |
0.0018 |
0.00182 |
0.00182 |
0.00178 |
0.00035 |
Secondary |
0.00178 |
0.00180 |
0.00179 |
0.00181 |
0.00034 |
Mean value |
0.00179 |
0.00181 |
0.00180 |
0.00179 |
0.000345 |
From upper table analysis, the water-soluble of taxol compound provided by the invention increases.
Experimental example 3:
This experimental example to the paclitaxel injection of the embodiment of the present invention 5 60 DEG C, high humidity (25 DEG C, humidity 90%), high light (4500lx ± 500lx) place 10 days, sampling in the 10th day detects sample respectively.
Table 4, high temperature, high light, high humidity test-results
Result shows: the paclitaxel injection prepared by the present invention is placed after ten days under illumination, high temperature, high humidity, and color is all unchanged, and content, related substance all change not quite.I.e. good stability.
Experimental example 4:
This experimental example is that the paclitaxel injection thinner of embodiment 5 dilutes later mixture, and this mixture joins in different infusion liquid, the time that insolubles occurs, refers to table 5:
After the dilution of table 5 paclitaxel composition, in infusion liquid, crystal separates out the comparison of time
As can be seen from the table, paclitaxel composition of the present invention, after thinner dilution, all just has crystal in the long time and separates out in conventional infusion liquid.
Table 6, table 7 are that the paclitaxel composition thinner of embodiment 5 dilutes later mixture respectively, and this mixture joins the investigation of 0.9% sodium chloride solution, 0.5% glucose solution Chinese traditional medicine stability.
Table 6 taxol is stability in 0.9% sodium chloride solution
Table 7 taxol is stability in 0.5% glucose solution
From 7, the experimental result of table 6 can draw, paclitaxel composition of the present invention can stable existence in 10 hours in infusion liquid, and relative content is stablized.
The present invention carries out above-mentioned experiment to the paclitaxel composition of other embodiments except embodiment 5, and its result matches with it.
Experimental example 5:
Anaphylaxis is tested
The paclitaxel injection of the preparation in embodiment 5,6,7 is got two batches and carries out parallel anaphylaxis test.After injection liquid group three intraperitoneal injections the 14th day and 21 days, intravenous injection original liquid is attacked, result shows, the cavy of paclitaxel injection group of the present invention after three abdominal injection sensitization the 14th day and the 21st day, intravenous injection original liquid is attacked, have cough or gently grab nose, tremble, the minor response symptom such as perpendicular hair, some does not even significantly react, paclitaxel composition of the present invention obviously can improve symptoms of allergic as can be seen here, and concrete outcome is see table 8.
Cavy anaphylaxis standards of grading: 0 is divided into without significant reaction; 1 is divided into only having and slightly grabs nose, trembles and perpendicular hair; 2 are divided into having and cough several times, grab nose, tremble or perpendicular hair; 3 are divided into and repeatedly or continuously cough, with expiratory dyspnea or spasm or tic; 4 are divided into spasm, twitch, gatism, shock death.Negative control group is one group without any reaction; Positive group shows as spasm, tic, gatism, shock death according to organizing for after drug administration.
The anaphylaxis test-results of table 8 paclitaxel injection
By above-mentioned, scratch test is carried out to the paclitaxel injection in the embodiment of the present invention, can draw from upper table, its supersensitivity improves significantly, contriver infers, because composition in this injection liquid is taxol hydrate crystal, greatly reduce the consumption of Cremophor EL, thus alleviate the anaphylaxis of cavy.Improve the security of medication.
Experimental example 6:
This experimental example carries out the pharmacokinetics of the paclitaxel injection (test article) testing the embodiment of the present invention 5, and commercially available paclitaxel injection is reference substance.
Concrete grammar is, selects 12 rabbit (male and female half and half), body weight and 2.0kg, and paclitaxel injection intravenously administrable dosage 1.2mg/kg, administration fasting in eve, freely drinks water.Upon administration 0,0.083,0.152,0.25,0.5,0.75,1,2,4,6,8,10,12h respectively at side ear edge vein exploitating blood 1-2ml, be placed in heparinization centrifuge tube, blood plasma is got in centrifugation.
Accurate absorption 50 μ l plasma sample is placed in 96 orifice plates, and add mark (Docetaxel) working solution in 5 μ l1 μ g/ml, after whirlpool mixing 5min, add t-butyl methyl ether, vortex oscillation again, after centrifugal, then after freezing 1h, take out 800 μ l organic solution layer to another 96 orifice plate, and this 96 orifice plate is placed in 30 C water bath's nitrogen gas stream dries up, after adding 50 μ l moving phases redissolution, vortex mixes, and after centrifugal, gets 20 μ l sample introductions.
Chromatographic condition during detection is, C18 chromatographic column (1.6mm × 150mm, 5 μm) moving phase acetonitrile/water volume is 65:35, and column temperature is room temperature, and flow velocity is 0.05ml/min.
As accompanying drawing 3,
represent the Plasma Concentration change curve in time of commercially available Taxol injection,
represent the Plasma Concentration curve over time of the paclitaxel injection of the embodiment of the present invention 5.Can draw through above-mentioned experiment, in rabbit body, paclitaxel injection of the present invention has different significantly from its pharmacokinetics behavior of common commercially available paclitaxel injection, paclitaxel injection of the present invention distributes faster in vivo, but eliminate slower, the effective prolong drug residence time in vivo, add curative effect, improve the bioavailability of taxol.