CN101020059B - Medicine composition containing docetaxel matter and its preparation process - Google Patents

Abstract

Translated fromChinese

本发明涉及一种适合于静脉注射的含多烯紫杉醇类物质的药物组合物及其制备方法，该药物组合物以多烯紫杉醇或它的盐或水合物为活性成分，采用环糊精和氨基酸联合作为增溶剂。利用本发明药物组合物制成的注射剂具有稳定性良好，毒副作用低等优点。The invention relates to a pharmaceutical composition containing docetaxel suitable for intravenous injection and a preparation method thereof. The pharmaceutical composition uses docetaxel or its salt or hydrate as an active ingredient, and adopts cyclodextrin and amino acid Combined as a solubilizer. The injection prepared by using the pharmaceutical composition of the invention has the advantages of good stability, low toxic and side effects, and the like.

Description

A kind of pharmaceutical composition that contains docetaxel matter and preparation method thereof

Technical field

The present invention relates to a kind of pharmaceutical composition that contains docetaxel matter, refer to that more specifically a kind of is the medicinal composition for injections of active component with Docetaxel or its salt or hydrate.The invention still further relates to the preparation method of said composition.

Background technology

Docetaxel matter is the chemical compound that a known class has remarkable active anticancer, belong to mitotic inhibitor, by promoting the microtubule dimer to be assembled into microtubule, simultaneously by preventing that the multimerization process from making microtubule stable, the retardance cell is in G2 and M phase, thus the mitosis of anticancer and propagation.This class medicine is all effective for advanced breast cancer, nonsmall-cell lung cancer, ovarian cancer, carcinoma of prostate, cancer of pancreas, hepatocarcinoma, incidence cancer, gastric cancer etc.

Because Docetaxel is insoluble in water, the dissolubility of Docetaxel in water only is 0.5 μ g/ml under the room temperature, therefore at present commercially available Docetaxel injection mainly is that Tween 80 or polyoxyethylene hydrogenated Oleum Ricini with high concentration are the formulated concentrated solution of solubilizing agent, include ethanol or add the ethanol of (in the packing capsule subsidiary again 1) 13%, the injection back is to the human body toxic side effect.Tween 80 or polyoxyethylene hydrogenated Oleum Ricini are the surfactants of using always, but Tween 80 or polyoxyethylene hydrogenated Oleum Ricini have very strong toxicity and high anaphylaxis, tween 80 has hemolytic, be used for intravenous injection, can cause severe anaphylactic reaction, comprise shock, dyspnea, hypotension, angioedema, anaphylactoid reaction symptoms such as rubella, therefore before using the Docetaxel injection, must take Claritin or injection Claritin earlier and help to alleviate the toxic and side effects such as serious allergy that the use owing to the Docetaxel injection produces, usually bring misery, directly influenced the use of this medicine to patient.In addition, though tween 80 or polyoxyethylene hydrogenated Oleum Ricini can increase the dissolubility of Docetaxel, at the Docetaxel poor stability of solution state, effect duration is short; Subsidiary 1 13% ethanol has also increased production cost in the packing capsule.Therefore, change the dosage form of Docetaxel, increase the water solublity of Docetaxel, eliminate the focus that anaphylaxis that is caused by solvents such as tween 80s and the toxic and side effects that reduces Docetaxel self become present research.

Many researcheres are studied the untoward reaction that reduces docetaxel matter and cause in clinical practice and the toxic and side effects of himself, and for example Chinese patent application 02155476.5 discloses and adopted hyaluronic acid, glucose or mannitol, ethanol, propylene glycol, non-ionic surface active agent to prepare the injection of docetaxel matter; It is the compositions that solvent prepares docetaxel matter that United States Patent (USP) 97/196,934 discloses with dimethyl acetylamide, Polyethylene Glycol, is used for parenterai administration; Australian Patent 35544/93 discloses the dissolubility that the branched cyclodextrin that uses unsaturated cyclodextrin and enzyme modification can appropriateness increases docetaxel matter; Chinese patent ZL98811010.5 adopts acetylation cyclodextrin and HP-and other water solublity adjuvant to prepare the injection preparation of docetaxel matter; Chinese patent application 03112082.2 is adopting cyclodextrin, water solublity adjuvant and organic acid and acylate to prepare the injection preparation of docetaxel matter as adjuvant, the stability of resulting injection makes moderate progress, but the content of docetaxel matter descends still near 10% in 6 hours, and add organic acid and acylate owing to contain, the pH value of preparation low excessively (pH value is 3～5) easily produces tangible blood vessel irritation when intravenous injection is used.Utilize the injection of the docetaxel matter that prior art obtains mostly to exist stable dissatisfactory defective.

Summary of the invention

The purpose of this invention is to provide a kind of have good stability and toxic and side effects little, be suitable for the intravenous pharmaceutical composition of docetaxel matter that contains to overcome the existing in prior technology defective.

Another object of the present invention is to provide this preparation of drug combination method.

The characteristics of pharmaceutical composition of the present invention are with cyclodextrin and aminoacid as solubilizing agent.Use aminoacid and cyclodextrin to increase the dissolubility of Docetaxel in water by uniting, stop medicine to separate out gathering, increase its physical stability and chemical stability.

By weight, the pharmaceutical composition that contains docetaxel matter of the present invention comprises:

1 part of docetaxel matter

Cyclodextrin 40-200 part

0.2～10 part in aminoacid

The said docetaxel matter of the present invention is meant chemical compound Docetaxel or its salt or hydrate.Said cyclodextrin is HP-or acetyl-gamma-cyclodextrin, and said aminoacid is glycine, valine, aspartic acid, glutamic acid, arginine, ornithine, lysine, tyrosine or histidine.

The pharmaceutical composition that contains docetaxel matter of the present invention refers to the dried frozen aquatic products after aqueous liquid medicine composition or its lyophilization.Preparation of drug combination method of the present invention may further comprise the steps:

(1) successively aminoacid and cyclodextrin are dissolved in the water;

(2) add docetaxel matter or be dissolved in alcoholic acid docetaxel matter;

(3) ultrasonic or be stirred to the solution clear and obtain fluid composition, or

(4) fluid composition is carried out lyophilization and obtain dried frozen aquatic products.

In the said method, by weight, the consumption of water is 50-1000 a times of docetaxel matter, and consumption of ethanol is 1-100 a times of docetaxel matter; Can also use caffolding agent during the preparation dried frozen aquatic products, caffolding agent can be selected mannitol, lactose, glucose, sucrose, dextran or its mixture for use; The dried frozen aquatic products that said method obtains can be quickly dissolved in the aqueous solution of water for injection or sodium chloride-containing, glucose, mannitol, dextran etc. for loose block or powder, obtains the solution of clear.

Pharmaceutical composition of the present invention is suitable for making intravenous injection, and by the administration of intravenous drip mode, the administration process of its lyophilized formulations can be finished in 2 hours.Effective using dosage of pharmaceutical composition of the present invention is the 2mg/kg body weight.

Beneficial effect

1, pharmaceutical composition of the present invention is solubilizing agent with aminoacid and cyclodextrin, toxic and side effects is low, need not treat in advance before the administration by Claritin or other medicine, not only made things convenient for administration, also increased compliance of patients, reduced cost, pharmaceutical composition of the present invention is to the blood vessel nonirritant, and intravenous administration suits.

2, having good stability of pharmaceutical composition of the present invention, fluid composition was placed 28 hours down at 4 ℃, and the changes of contents of its Docetaxel is less than 2%; The pharmaceutical composition of dried frozen aquatic products form the aqueous solution room temperature under can stablize and deposit at least 24 hours, Docetaxel does not have to assemble separates out, its concentration change is less than 2%.

3, simple, the easily control of preparation of drug combination technology of the present invention is fit to industrialized great production.

The specific embodiment

The present invention is further elaborated below in conjunction with specific embodiment, but do not limit the present invention.

Among the following embodiment, adopt the HPLC method to measure Docetaxel content, chromatographic condition: mobile phase is acetonitrile one water (60: 40, v/v), flow velocity is 1.0ml/min, and sample size is 15 μ l, column temperature is 25 ℃, adopt UV-detector, the detection wavelength is 229nm, chromatographic column adopting octadecylsilane chemically bonded silica post.

Among the following embodiment, remove and specify, the method that the lyophilization fluid composition obtains dried frozen aquatic products is: behind the packing fluid composition, do not seal the metal dish of putting into freeze drying box, and temperature is reduced to-30 ℃, the metal dish that preparation will be housed is rapidly put into freeze drying box, keeps 7 hours, making the vacuum in the case then is 0.2Pa, again shelf is heated up, and be controlled at below the eutectic point-3 ℃, kept 12 hours, be warming up to 4 ℃ at last, and kept 8 hours, formulation temperature is overlapped with shelf temperature, put into aseptic filtering gas, the water content of control preparation is sealed rapidly and can be obtained dried frozen aquatic products 3%.

Embodiment 1

Precision takes by weighing the 40mg Docetaxel and is dissolved in 1ml ethanol.In addition precision takes by weighing the 30mg glycine and is dissolved in the 5ml water, adds the alcoholic solution that 3g HP-dissolving back adds Docetaxel, ultrasonic 20min, stir clear solution, be fluid composition.

Embodiment 2

Precision takes by weighing the 40mg Docetaxel and is dissolved in 0.5ml ethanol.Precision takes by weighing the 20mg aspartic acid and is dissolved in the 10ml water, adds the alcoholic solution that 4g HP-dissolving back slowly adds Docetaxel, ultrasonic 30min, placed again 1 hour, obtain clear solution, after every bottle of quantitative 2ml packing, lyophilization obtains dried frozen aquatic products.

Embodiment 3

Precision takes by weighing the 6g HP-and is dissolved in the 20ml 0.2% arginine aqueous solution, places on the magnetic force heating stirrer.Precision takes by weighing Docetaxel 80mg in addition, behind the 1ml dissolve with ethanol, slowly be added drop-wise in the mixed solution of HP-and smart glycine, continue to stir 5h under the room temperature, add 7.5% mannitol and 5% lactose (w/v, aqueous solution), after every bottle of quantitative 5ml packing, lyophilization obtains dried frozen aquatic products.

Embodiment 4

Take by weighing Docetaxel 40mg, add the 0.5ml dissolve with ethanol; Other precision takes by weighing the 2g HP-and is dissolved in the histidine aqueous solution of 10ml 0.2%, put 20 ℃ of constant temperature water-soluble in, stir and inject the Docetaxel alcoholic solution down, continue to stir 2h,-30 ℃ of following pre-freezes, put dry 24h in the vacuum freeze dryer, get the pulverous dried frozen aquatic products of white solid.

Embodiment 5

Taking by weighing the 20g HP-is dissolved in the glutamic acid aqueous solution of 50ml 0.2%, other precision takes by weighing the 200mg Docetaxel and is dissolved in the 5ml ethanol, slowly be injected in the mixed solution of above-mentioned cyclodextrin and glutamic acid, supersound process 30min, placed again 1 hour, obtain fluid composition, after every bottle of quantitative 1ml packing, lyophilization obtains dried frozen aquatic products.

Embodiment 6

Precision takes by weighing 400mg sulphuric acid Docetaxel and is dissolved in 10ml ethanol.In addition precision takes by weighing the 300mg valine and is dissolved in the 5ml water, add 30g acetyl-gamma-cyclodextrin dissolving back and add the alcoholic solution of sulphuric acid Docetaxel, ultrasonic 20min, stir clear solution, be fluid composition.

Embodiment 7

The three water Docetaxels that precision takes by weighing 200mg are dissolved in 5ml ethanol.In addition precision takes by weighing 300mg lysine and is dissolved in the 5ml water, add 30g acetyl-gamma-cyclodextrin dissolving back and add the alcoholic solution of three water Docetaxels, ultrasonic 20min, stir clear solution, be fluid composition.

Embodiment 8

The Docetaxel that precision takes by weighing 200mg is dissolved in 5ml ethanol.In addition precision takes by weighing 300mg tyrosine and is dissolved in the 5ml water, add 30g acetyl-gamma-cyclodextrin dissolving back and add the alcoholic solution of Docetaxel, ultrasonic 20min, stir clear solution, be fluid composition.

Embodiment 9

The Docetaxel that precision takes by weighing 200mg is dissolved in 5ml ethanol.Other precision takes by weighing 300mg tyrosine and is dissolved in the 5ml water, add 30g acetyl-gamma-cyclodextrin dissolving back and add the alcoholic solution of Docetaxel, ultrasonic 20min, stir clear solution, obtain fluid composition, add 5% mannitol (w/v, aqueous solution) again, after every bottle of quantitative 1ml packing, lyophilization obtains dried frozen aquatic products.

Embodiment 10

The Docetaxel that precision takes by weighing 300mg is dissolved in 5ml ethanol.In addition precision takes by weighing the 200mg glycine and is dissolved in the 5ml water, add 30g acetyl-gamma-cyclodextrin dissolving back and add the alcoholic solution of Docetaxel, ultrasonic 20min, stir clear solution, be fluid composition.Add 5% mannitol (w/v, aqueous solution) again, after every bottle of quantitative 1ml packing, lyophilization obtains dried frozen aquatic products.

The mensuration of the saturation solubility of embodiment 11 fluid compositions

Taking by weighing the 3g HP-is dissolved in the glycine solution of 10ml 0.2%, other precision takes by weighing the 40mg Docetaxel and is dissolved in the 0.5ml ethanol, slowly be injected in the mixed solution of above-mentioned cyclodextrin and glycine, supersound process 30min, placed again 1 hour, obtain fluid composition, carry out the mensuration of saturation solubility: get above-mentioned fluid composition and each 1mL of Docetaxel raw material, put respectively under 30 ℃, the 40r/min 48h that vibrates, lyophilization adds water 1mL, vortex 30s, 3000r/min is centrifugal, behind 0.22 μ m membrane filtration, suitably carry out liquid phase analysis after the dilution with mobile phase, sample size is 20 μ L, mobile phase is acetonitrile: water (60: 40), flow velocity is 1mL/min, measures peak area, calculates the saturated concentration of Docetaxel in water.The results are shown in Table 1.

Table 1

As shown in Table 1, the present composition is compared with the Docetaxel raw material, and the dissolubility of Docetaxel in water increased by 8000 times, with the Docetaxel of the script slightly solubility Yi Rong that becomes.Experiment shows, among the present invention HP-and glycine unite the dissolubility that use can significantly improve Docetaxel, solved the difficult problem in the preparation Docetaxel injection.

The stability test of embodiment 12 fluid compositions

Precision takes by weighing the 400mg Docetaxel and is dissolved in 1ml ethanol.In addition precision takes by weighing the 200mg glycine and is dissolved in the 40ml water, adds the alcoholic solution that 20g HP-dissolving back adds Docetaxel, ultrasonic 30min, stir clear solution, be fluid composition of the present invention.

Above-mentioned prepared fluid composition adds in an amount of distilled water and dilutes, and is mixed with the settled solution that Docetaxel content is 1mg/ml.4 ℃, 25 ℃ and 35 ℃ of placements, respectively at 8,12,16,20,24 and 28h sampling, the HPLC method is measured Docetaxel content, the results are shown in Table 2 with this solution.

Table 2

The result shows that fluid composition of the present invention is placed 28h, changes of contents＜2% at 4 ℃; Deposit 24h at ambient temperature, changes of contents＜2%; Place 24h at 35 ℃, lowering of concentration is lower than 2%, is not higher than in ambient temperature under 35 ℃ the situation, and this fluid composition can keep the above stability of 24h, shows having good stability of fluid composition of the present invention.

The stability test of embodiment 13 dried frozen aquatic products state combination things

Precision takes by weighing the 300mg glycine and is dissolved in the 4ml water, add 30g HP-dissolving back and add the 400mg Docetaxel, 25 ℃ are stirred 5h, obtain clear solution, add 10% mannitol (w/v again, aqueous solution), after every bottle of quantitative 0.5ml packing, lyophilization promptly gets dried frozen aquatic products.From in appearance, dried frozen aquatic products is off-white color lyophilizing block or powder.

It is an amount of to get the above-mentioned dried frozen aquatic products that makes, and sealing is placed in 25 ℃ and to be carried out accelerated test June, in 0,1,2,3, measure indexs such as outward appearance, related substance, content June again, estimates the stability of dried frozen aquatic products.The results are shown in Table 3.

In the table, related substance is pressed the HPLC method and is measured, and content is for being equivalent to labelled amount.

Table 3

The result shows that Docetaxel composition freeze-drying product were placed 6 months at 25 ℃, and its outward appearance, relevant medicine and content do not have significant change, show that Docetaxel composition freeze-drying product have good stability.

The stability test of embodiment 14 dried frozen aquatic products state combination things

Test sample: the dried frozen aquatic products state combination thing of pressing the preparation of embodiment 5 methods

Reference substance: press the liquid Docetaxel compositions of Chinese patent 98811010.5 methods preparation, add 5% mannitol (w/v, aqueous solution), obtain dried frozen aquatic products by every bottle of quantitative 1ml packing postlyophilization.The preparation method of its fluid composition is: take by weighing the 20g HP-and be dissolved in the 20ml water, add the 400mg Docetaxel, 25 ℃ are stirred 3h, obtain clear solution, obtain fluid composition.

Docetaxel injection (Docetaxel for Injection, 20mg/ props up, available from Shanghai City pharmaceuticals)

Get each 3 of test sample, reference substance and Docetaxel for Injection, be dissolved in respectively in the 5ml water, be diluted to 1mg/ml with the sodium-chloride water solution of 0.9%w/v respectively then; Other gets each 3 of test sample, reference substance and Docetaxel for Injection, be dissolved in respectively behind the 5ml water respectively that the D/W with 5%w/v is diluted to 1mg/ml, placing the clarity of checking test sample, reference substance and Docetaxel for Injection on the the 0th, 2,4,8,12,24 hour respectively.The results are shown in Table 4.

Table 4

As can be seen from Table 4, dried frozen aquatic products state combination thing of the present invention does not see after 24 hours that through the dilution of sodium-chloride water solution and D/W medicine separates out, and reference substance dilutes with 0.9% sodium-chloride water solution and began to have small amount of drug to separate out in back 2 hours, commercially available Docetaxel injection also began have small amount of drug separate out in 2 hours with sodium-chloride water solution dilution 4 hours with the D/W dilution.Show that dried frozen aquatic products state combination thing diluent of the present invention has good physical stability, is enough to satisfy the needs of vein treatment.

The stability test of embodiment 15 dried frozen aquatic products state combination things

Taking by weighing the 20g HP-is dissolved in the histidine aqueous solution of 50ml 0.3%, other takes by weighing the 200mg Docetaxel and is dissolved in the 1.5ml ethanol, slowly be injected in the mixed solution of above-mentioned cyclodextrin and histidine, supersound process 30min, placed again 1 hour, obtain the Docetaxel composition solution, can obtain dried frozen aquatic products by every bottle of quantitative 5ml (containing Docetaxel 20mg) packing postlyophilization.

Each 3 of the above-mentioned dried frozen aquatic productses that makes are dissolved in respectively in the sodium chloride or 5%w/v D/W of 20ml0.9%w/v, prepare the settled solution that Docetaxel concentration is about 1mg/ml respectively.The sodium-chloride water solution that contains the present composition and the D/W that prepare are contained in the sealed glass container, placed 6 hours in 35 ℃, in the concentration of sampling in the 0th, 2,4,8,12,24 hour by Docetaxel in the HPLC analytic sample, analysis result sees Table 5 respectively.

Table 5

Experimental result shows that the present composition was placed 24 hours down at 35 ℃, Docetaxel concentration almost constant (＜2%), and promptly the Docetaxel composition stable is good.

The stability test of embodiment 16 dried frozen aquatic products state combination things

Taking by weighing the 40g HP-is dissolved in the ornithine aqueous solution of 10ml 0.3%, other takes by weighing the 400mg Docetaxel and is dissolved in the 1.5ml ethanol, slowly be injected in the mixed solution of above-mentioned cyclodextrin and ornithine, supersound process 30min, placed again 1 hour, obtain the Docetaxel composition solution, can obtain dried frozen aquatic products by every bottle of quantitative 5ml (containing Docetaxel 20mg) packing postlyophilization.

Reference substance: press the liquid Docetaxel compositions of Chinese patent 98811010.5 methods preparation, add 5% mannitol (w/v, aqueous solution), after every bottle of quantitative 1ml packing, lyophilization obtains dried frozen aquatic products.The preparation method of its fluid composition is with embodiment 14.

Respectively above-mentioned dried frozen aquatic products that makes and reference substance are added in an amount of normal saline or the 5%w/v D/W, preparation Docetaxel concentration is about the settled solution of 1mg/ml, the normal saline that contains the present composition or reference substance and the D/W that prepare are at room temperature deposited, measured the concentration of Docetaxel in each solution with HPLC every 12 hours, analysis result sees Table 6.

Table 6

Experimental result shows, at room temperature placed 24 hours, contain the normal saline of compositions of Chinese patent 98811010.5 methods preparations and the Docetaxel concentration in the 5% glucose diluent and all descended 10%, Docetaxel concentration does not almost change in the sample diluting liquid of the present composition and contain.Illustrate that the stability of the present composition is better than the compositions of prior art report.

The inhibition test of embodiment 17 dried frozen aquatic products state combination things

Trial drug: press the dried frozen aquatic products state combination thing of embodiment 13 methods preparation, being diluted to Docetaxel concentration with an amount of normal saline during test is 8mg/ml.

Positive reference substance: Docetaxel injection (Docetaxel for Injection, 20mg/ props up, available from Shanghai City pharmaceuticals)

Negative control product: sterile saline

Preparation mouse mastopathy cell EMT-6 suspension makes every milliliter of suspension contain 5 * 107 cancerous cell.Get BALB/C mice, 6 every group, every Corium Mus is inoculation 0.2ml down, carries out administration by dosage shown in the table 7 after 24 hours behind the inoculated tumour, once a day, and continuous 5 days.Wherein the subcutaneous or abdominal cavity of test group gives trial drug, subcutaneous or the abdominal cavity of positive controls gives the Docetaxel injection, subcutaneous or the abdominal cavity of negative control group gives isopyknic sterile saline, each treated animal is execution in the 12nd day behind inoculated tumour, get the tumor back of weighing and calculate Docetaxel compositions and the Docetaxel injection suppression ratio to tumor, it is heavy that average tumor is organized in the average tumor weight-treatment of suppression ratio %=[(matched group)/the average tumor weight of matched group] * 100%.The result is as shown in table 7:

Table 7

Result of the test shows, the subcutaneous tumour inhibiting rate that gives the present composition is 44.7～53.0%, tumour inhibiting rate behind the intraperitoneal administration is 41.8～44.3%, the tumour inhibiting rate that gives the present composition of same dose and Docetaxel injection under the identical administering mode is suitable, shows that the present composition has the good antitumor effect.

The inhibition test of embodiment 18 dried frozen aquatic products state combination things

Trial drug: press the dried frozen aquatic products state combination thing of embodiment 13 methods preparation, being diluted to Docetaxel concentration with an amount of normal saline during test is 8mg/ml;

Positive reference substance: the taxotere concentrated injection, being diluted to Docetaxel concentration with normal saline during test is 8mg/ml;

Negative control product: normal saline

Lewis lung cancer is inoculated in 30 BDF1 mices, press dosage shown in the table 8 behind the inoculated tumour respectively once at the 1st, 3,5,7 each intraperitoneal injection of day interval, wherein test group is injected Docetaxel composition freeze-drying product, positive controls injection taxotere injection, the isopyknic normal saline of negative control group lumbar injection, the execution in the 12nd day behind inoculated tumour of each treated animal is dissected, and gets and calculates the suppression ratio of trial drug to tumor after tumor is weighed.The result is as shown in table 8:

Table 8

Body is interior but the tumor result shows Docetaxel pharmaceutical composition of the present invention to be compared with the taxotere injection, has suitable antitumor activity.

The irritation test of embodiment 19 dried frozen aquatic products state combination things

Trial drug: press the dried frozen aquatic products state combination thing of embodiment 13 methods preparation, being diluted to Docetaxel concentration with an amount of normal saline during test is 1mg/ml;

Reference substance 1: the liquid Docetaxel compositions by the preparation of Chinese patent application CN1535678A method, add 5% mannitol (w/v, aqueous solution), obtain dried frozen aquatic products by every bottle of quantitative 1ml packing postlyophilization.The preparation method of its fluid composition is: take by weighing the 20g HP-and be dissolved in the 20ml water, add the 400mg Docetaxel, 25 ℃ are stirred 3h, obtain clear solution, add Chinese holly edge acid adjust pH to 3.5 again, obtain fluid composition behind the mixing.

Reference substance 2: Docetaxel injection (Docetaxel for Injection, 20mg/ props up, available from Shanghai City pharmaceuticals)

Get 15 of rabbit, be divided into three test group, inject (2mg/kg) trial drug, reference substance 1 and reference substance 2 respectively, after the sacrificed by exsanguination, observe the response situation of injection site at auricular vein or the quadriceps femoris of each test group rabbit.The rabbit erythrocyte suspension of preparation 2% adds trial drug, observes it to erythrocytic influence.The results are shown in Table 9.

Table 9

Annotate: the blood vessel irritation reaction is not found in "-" expression in the table; "+" expression has the blood vessel irritation reaction.

Result of the test shows that irritative response in various degree all appears in the rabbit after reference substance 1 and reference substance 2 injections, as redness, hyperemia, inflammatory infiltration etc.And to behind rabbit vein or the intramuscular injection pharmaceutical composition of the present invention, perusal there is no irritant reaction such as redness, hyperemia, through pathological examination, symptoms such as fibrinoid necrosis, inflammatory infiltration do not appear, and the present composition is no haemolysis coacervation in observing time, and the present composition intravenous administration that suits is described.

The toxicity test of embodiment 20 dried frozen aquatic products state combination things

Trial drug: press the Docetaxel pharmaceutical composition dried frozen aquatic products of embodiment 13 methods preparation, being diluted to Docetaxel concentration with an amount of normal saline during test is 8mg/ml;

Reference substance: the taxotere concentrated injection, being diluted to Docetaxel concentration with normal saline during test is 8mg/ml;

Get 60 of the kunming mices of body weight 18～22g sex unanimity, be divided into 6 groups at random, 10 every group, respectively get 3 groups and be included into test group and matched group respectively, an intravenous injection present composition of test group dried frozen aquatic products, an intravenous injection taxotere of matched group injection.Dosage is arranged by geometric progression between group, ratio observe at 1: 0.8 and record administration 10 days in the reaction and the mortality rate of each treated animal, calculate LD, its credible being limited to of 95% with simplifying probit method:

Taxotere injection LD50=107mg ± 32mg/kg,

Docetaxel composition freeze-dried powder LD50=219mg ± 30mg/kg of the present invention.

The experimental result of LD50 shows that present composition dried frozen aquatic products is compared with the taxotere injection, and toxicity obviously reduces.

The sensitivity test of embodiment 21 dried frozen aquatic products state combination things

Trial drug and reference substance are with embodiment 20.

Medication: get 12 of Cavia porcelluss, be divided into 2 groups at random, 6 every group, male and female half and half.For test group and matched group, with the dosage of 10mg/kg body weight, intramuscular injection trial drug and reference substance 0.2～0.3mL inject every other day respectively, continuous 3 times, observe the untoward reaction of Cavia porcellus behavior and appearance.The injection back is the 7th day first, respectively gets 3 Cavia porcelluss in test group and the matched group, and lumbar injection 2～3mL trial drug and reference substance are attacked respectively, and observation injection back Cavia porcellus has or not and allergic symptom occurs.Remaining second half Cavia porcellus is injected the trial drug or the reference substance of back 14 days lumbar injection same doses in first, and carries out allergic symptom and observe.

Judge index: the allergic symptom difference according to two groups of Cavia porcellus reactions is divided into 1～4 grade.0 grade: no significant reaction; One-level: have and slightly grab nose, perpendicular hair; Secondary: have and significantly grab nose, perpendicular hair, tremble or sneeze; Three grades: repeatedly continuously sneeze, with dyspnea or spasm, tic; Level Four: spasm, tic, gatism, shock, death.

The symptoms of allergic observed result: tangible anaphylaxis appears in the matched group Cavia porcellus, promptly occurs grabbing nose and gatism phenomenon after the 1st administration; Occur serious gatism, perpendicular hair and tic phenomenon after the 2nd administration, every Cavia porcellus all has proctoptosis phenomenon in various degree.The test group Cavia porcellus only occurs individually after the 3rd administration that gatism, lethargy are depressed, activity reduces phenomenon.Matched group is injected dead 3 of back for the first time, after the abdominal cavity was attacked in the 14th day 2 Cavia porcellus death is arranged, and has tangible anaphylaxis and signs of toxicity.Anaphylaxis does not appear in test group when intramuscular injection, even find no allergic symptom behind heavy dose of lumbar injection yet.The result shows that caused anaphylaxis degree of Docetaxel pharmaceutical composition of the present invention and mortality rate are starkly lower than the taxotere injection.

Claims (6)

Translated fromChinese

1.一种含多烯紫杉醇类物质的药物组合物，其特征在于，所述药物组合物以环糊精和氨基酸作为增溶剂，以重量计，包括：1. a pharmaceutical composition containing docetaxel substances, characterized in that, said pharmaceutical composition uses cyclodextrin and amino acid as solubilizer, by weight, comprising:多烯紫杉醇类物质    1份Docetaxel 1 part环糊精              40-200份Cyclodextrin 40-200 parts氨基酸              0.2～10份；Amino acid 0.2 to 10 parts;所述多烯紫杉醇类物质为多烯紫杉醇或它的盐或水合物；The docetaxel substance is docetaxel or its salt or hydrate;所述环糊精为羟丙基-β-环糊精或乙酰-γ-环糊精；The cyclodextrin is hydroxypropyl-β-cyclodextrin or acetyl-γ-cyclodextrin;所述氨基酸为甘氨酸、缬氨酸、天冬氨酸、谷氨酸、精氨酸、鸟氨酸、赖氨酸、酪氨酸或组氨酸。The amino acid is glycine, valine, aspartic acid, glutamic acid, arginine, ornithine, lysine, tyrosine or histidine.2.根据权利要求1所述的含多烯紫杉醇类物质的药物组合物，其特征在于，所述药物组合物为含水的液态药物组合物或其冷冻干燥后的冻干品。2 . The pharmaceutical composition containing docetaxel according to claim 1 , wherein the pharmaceutical composition is an aqueous liquid pharmaceutical composition or a freeze-dried product thereof. 3 .3.权利要求1所述含多烯紫杉醇类物质的药物组合物的制备方法，包括如下步骤：3. the preparation method of the pharmaceutical composition containing docetaxel class substance described in claim 1, comprises the steps:(1)依次将氨基酸和环糊精溶解于水中；(1) sequentially dissolving amino acid and cyclodextrin in water;(2)加入多烯紫杉醇类物质或溶于乙醇的多烯紫杉醇类物质；(2) adding docetaxel substances or docetaxel substances dissolved in ethanol;(3)超声或搅拌至溶液澄清透明得到液态组合物。(3) Ultrasound or stir until the solution is clear and transparent to obtain a liquid composition.4.权利要求1所述含多烯紫杉醇类物质的药物组合物的制备方法，包括如下步骤：4. the preparation method of the pharmaceutical composition containing docetaxel class substance described in claim 1, comprises the steps:(1)依次将氨基酸和环糊精溶解于水中；(1) sequentially dissolving amino acid and cyclodextrin in water;(2)加入多烯紫杉醇类物质或溶于乙醇的多烯紫杉醇类物质；(2) adding docetaxel substances or docetaxel substances dissolved in ethanol;(3)超声或搅拌至溶液澄清透明得到液态组合物，(3) Ultrasound or stir until the solution is clear and transparent to obtain a liquid composition,(4)将液态组合物进行冷冻干燥得到冻干品。(4) Freeze-drying the liquid composition to obtain a freeze-dried product.5.根据权利要求3或4所述含多烯紫杉醇类物质的药物组合物的制备方法，其特征在于，以重量计，水的用量为多烯紫杉醇类物质的50-1000倍，乙醇的用量为多烯紫杉醇类物质的1-100倍。5. according to the preparation method of the described pharmaceutical composition containing docetaxel substances of claim 3 or 4, it is characterized in that, by weight, the consumption of water is 50-1000 times of docetaxel substances, and the consumption of ethanol 1-100 times that of docetaxel.6.权利要求1所述含多烯紫杉醇类物质的药物组合物在制备静脉注射剂中的用途。6. The use of the pharmaceutical composition containing docetaxel according to claim 1 in the preparation of intravenous injections.