CN104510715A - Excipient-containing lyophilized paclitaxel powder preparation and preparation method thereof - Google Patents

Abstract

The invention belongs to the technical field of medicinal preparations, and relates to an excipient-containing lyophilized paclitaxel powder preparation and a preparation method thereof. The preparation adopts paclitaxel as a medicinal effective component, adopts mPEG-PLA-aspartic acid as a matrix, and contains an excipient; and the excipient comprises one or more of lactose, mannitol, dextran, glycine and glucose. The preparation method comprises the following steps: 1, mixing mPEG-PLA-aspartic acid with paclitaxel; 2, fully dissolving the above obtained mixture in an organic solvent; 3, removing the organic solvent; 4, adding water, and hydrating to form a micelle solution; and 5, adding the excipient accounting for 5% or less of the total weight of materials added in step 1 to the medicine-loaded micelle solution, fully dissolving the excipient, lyophilizing the above obtained mixed micelle solution through a routine lyophilizing technology to prepare the excipient-containing lyophilized paclitaxel powder preparation. The excipient-containing lyophilized paclitaxel powder preparation has the advantages of simple preparation method, simple formula, high safety, good dissolution, rapid re-dissolving in water, and realization of small average particle size and easy performance of the ERP effect after the re-dissolving.

Description

A kind of paclitaxel freeze drying powder preparation containing excipient and preparation method thereof

Technical field

The invention belongs to pharmaceutical preparations technology field, be specifically related to paclitaxel freeze drying powder preparation and preparation method.

Background technology

Taxane molecule formula is: C ₄₇h ₅₁nO ₁₄, molecular weight 853.9, fusing point 213 ~ 216 DEG C, has height lipotropy, water insoluble, is the natural anti-cancer drugs being separated a kind of efficient, low toxicity, the wide spectrum that obtain at first from Chinese yew genus plants.It is the natural secondary metabolite of a kind of complexity in Chinese yew genus plants, molecular structure See Figure.

Paclitaxel is the specific medicine had compared with high anti-cancer activity of a kind of microtubule, and sales volume ranks the umber one of anti-cancer agent throughout the year, and with 20% speed increase.

1977, Horwit Z find paclitaxel anticancer mechanism be can with tubulin binding, promote that tubulin polymerization is assembled into microtubule dimer, thus the normal physiological depolymerization of microtubule in T suppression cell, make cell mitogen stop at G2 phase and M phase, prevent the Fast-propagation of cancerous cell.Further research finds that paclitaxel can activating macrophage, causes minimizing and the release thereof of Tumor Necrosis Factor Receptors, kills and wounds or inhibition tumor cell; Paclitaxel can cell death inducing, causes apoptosis; Paclitaxel can also the migration of inhibition tumor cell.Clinical research shows, paclitaxel has wide spectrum and efficient active anticancer, and give prominence to for treatment ovarian cancer, breast carcinoma, uterus carcinoma, gastric cancer etc. curative effect, recent research finds that it also has certain curative effect to rheumatoid arthritis, senile dementia etc.

Because paclitaxel is insoluble in water, usually paclitaxel is dissolved in polyoxyethylene castor oil (cremophor EL) and dehydrated alcohol mixed solvent to increase its water solublity in current clinical practice.But ethanol has certain cytotoxicity, and during polyoxyethylene castor oil degradation in vivo, discharge histamine, cause anaphylaxis in various degree, neurocyte endoparticle also can be caused to discharge and disentwining angle velocity and increase the weight of the peripheral nervous toxicity of paclitaxel.

For the toxicity reducing formulation for paclitaxel improves its curative effect, develop the novel form of paclitaxel in recent years clinically successively.Wherein at home and abroad do not go on the market containing polyoxyethylene castor oil injection paclitaxel albumin nano suspension (ABI-007).To have without antiallergic pretreatment, curative effect better, the feature such as toxicity is lower; The Paclitaxel liposome now carrying out the research and development of clinical research China just has at home started the prodrug (DHA-PTX) of the paclitaxel in clinical practice and polymeric dosage form genexol-PM and xyotax also just before clinical and in clinical I-III phase research, shows good prospect.

But recent research finds, it is no matter the polymer micelle that the particle diameter that formed by heavy polymer is larger, or the polymer micelle that the particle diameter formed by low-molecular weight polymer is less, there is the shortcoming that blood stability is poor, micelle almost disintegrates and discharges the medicine carried at once after entering blood, the medicine distribution in vivo that final two kinds of micelles carry is almost as broad as long, does not show ERP effect.The research of the people such as Savic R finds, usually in 5 minutes, just there is the drug release more than 90% after paclitaxel micelle enters blood and spread (see Langmuir2006,22:3570-8. " Assessment of the integrity of poly (caprolactone)-b-poly (ethylene oxide) micelles under biological conditions:a fl uorogenic-based approach. ") toward tissue.

This result carries medicine with imagination by micelle particle, arrives tumor locus and is greatly differed from each other by the perfect condition of EPR effect release medicine, be i.e. non-maturation release (Premature release).Be directed at the non-ripe reason discharged on the one hand because the concentration of micelle after hemodilution causes micelle to disintegrate lower than its critical micelle concentration (CMC concentration), the protein meeting of a large amount of existence in blood on the other hand and micelle are had an effect and are caused it to reunite and discharge medicine.Therefore, must first consider its blood stability when designing intravenous injection cancer target nano-carrier, being only just expected to utilize EPR effect to improve the targeting of medicine under the prerequisite improving blood stability.

Lyophilized powder is a kind of effective ways of preservation medicine.To dry goods be needed to make the moisture contained by it freeze at low temperatures, dry under being then placed on the environment of vacuum, allow moisture directly be distilled by solid state and be steam and get rid of from goods to make goods active dry.The method effectively prevent the change of goods physics and chemistry and biological nature, the stability of available protecting thermal sensitivity medicament effective component; Freeze-dried products after the drying loose, the color of form does not change substantially, adds water or can rapid solution recover physicochemical property and the biological activity of original aqueous solution after hydrophilic organic solvent; Because drying is carried out under vacuum, for some oxidizable materials, there is good protective effect; Goods water content after lyophilizing is very low, and the stability of goods is improved, and contaminated chance reduces, and this not only facilitates transport and also extends the goods pot-life.

Because paclitaxel is water insoluble, be difficult to solve paclitaxel injection with the problem that water redissolves and after redissolution, particle diameter is bigger than normal, therefore the current achievement in research to paclitaxel injection maturation is less.The people such as Liu Tong disclose a kind of preparation method of paclitaxel injection in " solid nano-medicine and preparation method thereof " (CN100518831), but simultaneously also point out that above-mentioned paclitaxel injection adds water (or solvent for injection) when dissolving and separate out microgranule because the dissolubility in water reduces.The far refined preparation method waiting people to disclose a kind of paclitaxel injection in " lyophilized powder injection of taxol polymer bond drug and preparation method thereof " (number of patent application 200610016614) of scape, but its technique more complicated needs adds a large amount of stabilizing agents and cosolvent, above-mentioned paclitaxel injection redissolves rear particle diameter comparatively greatly simultaneously, is 100-150nm.

Summary of the invention

The object of the invention is to overcome above-mentioned the deficiencies in the prior art, providing one with mPEG-PLA-aspartic acid for carrier, take paclitaxel as freeze-dried powder preparation of active component and preparation method thereof.Its preparation technology is simple; The dissolubility that the preparation obtained redissolves, stability are better, and mean diameter is less, has a good application prospect.

The invention provides a kind of paclitaxel freeze drying powder preparation containing excipient, take paclitaxel as effective ingredient, with mPEG-PLA-aspartic acid for substrate, and containing excipient; Described excipient be selected from lactose, mannitol, dextran, glycine, glucose one or more.

MPEG-PLA-aspartic acid molecules structural formula is as follows:

Wherein, a=11-455, b=3-300.

Preferred in the present invention, in described mPEG-PLA-aspartic acid, the molecular weight of methoxypolyethylene glycol block is 1000 ~ 3000; The molecular weight of polylactide block is 500 ~ 5000.

In the present invention, organic solvent content residual in described paclitaxel freeze drying powder preparation is less than 10ppm.

The present invention also provides a kind of preparation method of the paclitaxel freeze drying powder preparation containing excipient, comprises following step:

1. mPEG-PLA-aspartic acid is mixed with paclitaxel;

2. with an organic solvent dissolve said mixture fully to dissolve;

3. organic solvent is removed;

4. add water, aquation, makes it to form micellar solution;

5. add to polypeptide drug-loaded micelle solution and be no more than step and 1. feed intake the excipient of gross weight 5%, and make it abundant dissolving, then by the freeze-dry process lyophilizing routinely of above-mentioned mixed micellar solution, obtain paclitaxel freeze drying powder preparation; Wherein: described excipient be selected from lactose, mannitol, dextran, glycine, glucose one or more.

In above-mentioned preparation method, the 2. described organic solvent of step is ethanol or acetonitrile.

In the present invention, mPEG-PLA-aspartic acid is without obvious carcinogenecity, and without genotoxicity, without teratogenesis, mutagenicity, degradable is lactic acid, PEG, aspartic acid in vivo, all can directly excrete.Acute toxicity test is carried out to mice, the LD50>2.00g/kg of mice; In long term toxication, 1.00g/kg dosage, 1 times/day, successive administration 2 days weekly, drug withdrawal 5 days, successive administration, after 13 weeks, recovers to observe after 4 weeks, has no obvious toxic-side effects.

Cytotoxicity test shows, the cytotoxicity of mPEG-PLA-aspartic acid is lower than the block copolymer Mpeg-PLA of current generally recognized as safe (seeing the following form 1).

Table 1:MTT method hepatotoxicity test result

Compared with prior art, the present invention has following beneficial effect:

1, preparation technology is simple;

2, adopt the mPEG-PLA-aspartic acid of more low cytotoxicity to be carrier, formula is simple, and safety is higher;

3, dissolubility is better, redissolves rapidly in water;

4, the mean diameter after redissolving is less, more easily plays ERP effect.

Detailed description of the invention

Be described in detail the present invention below in conjunction with embodiment, the present embodiment is implemented under premised on technical solution of the present invention, give detailed embodiment and concrete operating process, but protection scope of the present invention is not limited to following embodiment.Material used in following embodiment, reagent etc., if no special instructions, all can obtain from commercial channels.

Embodiment (1-7)

A paclitaxel freeze drying powder preparation containing excipient, prepares according to following processing step.

1. raw material is taken according to different rate of charges (paclitaxel: mPEG-PLA-aspartic acid).Wherein, raw material paclitaxel (CAS33069-62-4) is produced by Rayleigh, Xi'an bio tech ltd, purity is greater than 95%, and raw material mPEG-PLA-aspartic acid, is prepared according to the technique described in patent No. PCT-CN-2013000453 voluntarily by inventor;

2. above-mentioned raw materials being dropped in container, at 15-45 DEG C of temperature, adding the organic solvent such as ethanol or acetonitrile to dissolving completely.Course of dissolution can adopt the means such as stirring or vibration.

3. by above-mentioned solution at 30-50 DEG C of rotary evaporation 2h, to organic solvent distilled-to-dryness.Vacuum drying > 12h removes residual organic solvent at 10-40 DEG C subsequently, obtains the polymer mixed film containing paclitaxel.

4. by above-mentioned hybrid films in 40-60 DEG C of water-bath to transparence, add the ultra-pure water of identical preheating temperature or add normal saline, phosphate buffer, shake well aquation, obtaining transparent polypeptide drug-loaded micelle solution.By described polypeptide drug-loaded micelle solution 0.45 μm of filtering with microporous membrane.

5. add to polypeptide drug-loaded micelle solution and be no more than step and 1. feed intake the excipient of gross weight 5%, and make it abundant dissolving.Excipient is lactose, mannitol, dextran, glycine, glucose wherein one or more.

6. prepare solid-state dry powder, step is as follows.

A. dividing plate puts into polypeptide drug-loaded micelle solution after being cooled to 0 DEG C in advance, is cooled to less than-45 DEG C, guarantees that polypeptide drug-loaded micelle solution temperature is down to-45 DEG C, maintains 2-4h.

B. above-mentioned steps terminates rear unlatching vacuum pump.Evacuation 1-1.5h.

C. baffle temperature setting rises to-25 DEG C by-45 DEG C, maintains more than 12h.Final vacuum is set as 0.013Bar.

D. baffle temperature setting rises to 0 DEG C by-25 DEG C, maintains more than 8h.Final vacuum is set as 0.013Bar.

E. baffle temperature setting rises to 5 DEG C by 0 DEG C, maintains more than 3h.Final vacuum is set as 0.013Bar.

F. baffle temperature setting rises to 20 DEG C by 5 DEG C, after products temperature reaches 20 DEG C, maintains more than 4h, then terminates lyophilizing, obtains loose block paclitaxel injection.

Embodiment 1-7 selects different rate of charges respectively, and selects the mPEG-PLA-aspartic acid of different block molecule amount respectively, obtains paclitaxel injection according to above-mentioned steps.Adopt HPLC method to measure the content of paclitaxel after redissolving, and calculate its drug loading, the results are shown in Table 1.

Table 1: the medicine carrying data of different embodiment

Carrier=mPEG-PLA-aspartic acid

The obtained paclitaxel injection of embodiment 1-7 and water is weighed according to the ratio of paclitaxel concentration 3mg/ml, add in water for injection, normal saline or glucose injection after at room temperature above-mentioned paclitaxel injection being pulverized, the all paclitaxel injections after 20 seconds that vibrate dissolve all completely, illustrate that it has good dissolubility.Use Dynamic Light Scattering Determination particle size distribution, record the particle size distribution of redissolution solution between 10-100nm, mean diameter is (be shown in table 2) between 21-39nm.Respectively at 15 DEG C, 25 DEG C, 30 DEG C, under normal indoor illumination condition, every 2h perusal solution, until find that solution occurs muddy or finds that there is Precipitation, then illustrate that solution finishes steady statue.The results are shown in Table 2, illustrate that the present invention has good stability.

Table 2: particle diameter and stability test result after redissolving

Claims (6)

1. the paclitaxel freeze drying powder preparation containing excipient, it is characterized in that, said preparation take paclitaxel as effective ingredient, with mPEG-PLA-aspartic acid for substrate, and containing excipient; Described excipient be selected from lactose, mannitol, dextran, glycine, glucose one or more.

2. paclitaxel freeze drying powder preparation according to claim 1, is characterized in that: in described mPEG-PLA-aspartic acid, and the molecular weight of methoxypolyethylene glycol block is 1000 ~ 3000.

3. paclitaxel freeze drying powder preparation according to claim 1, is characterized in that: in described mPEG-PLA-aspartic acid, and the molecular weight of polylactide block is 500 ~ 5000.

4. paclitaxel freeze drying powder preparation according to claim 1, is characterized in that: organic solvent content residual in described paclitaxel freeze drying powder preparation is less than 10ppm.

5. a preparation method for the paclitaxel freeze drying powder preparation containing excipient, is characterized in that comprising following step:

1. mPEG-PLA-aspartic acid is mixed with paclitaxel;

2. with an organic solvent dissolve said mixture fully to dissolve;

3. organic solvent is removed;

4. add water, aquation, makes it to form micellar solution;

5. add to polypeptide drug-loaded micelle solution and be no more than step and 1. feed intake the excipient of gross weight 5%, and make it abundant dissolving, then by the freeze-dry process lyophilizing routinely of above-mentioned mixed micellar solution, obtain paclitaxel freeze drying powder preparation; Wherein: described excipient be selected from lactose, mannitol, dextran, glycine, glucose one or more.

6. preparation method according to claim 5, is characterized in that: the 2. described organic solvent of step is ethanol or acetonitrile.