CN105326782A - Controlled-release preparation for treating cancer and preparation method thereof - Google Patents

Abstract

The invention relates to a controlled-release preparation for treating cancer and a preparation method thereof. The controlled-release preparation comprises a compound (I) or a salt thereof and polylactide-co-glycolide copolymer or a salt thereof, the compound (I) is shown as Ac-D-Tyr-Hyp-Asn-Thr-Phe-AzaGly-Leu-Arg (Me)-Trp-NH2, and weight-average molecular weight of the polylactide-co-glycolide copolymer is 10000-30000. The controlled-release preparation slowly and stably releases the compound (I) or the salt thereof for a long time and brings efficacy of the compound (I) or the salt thereof into play for a long time. In addition, the controlled-release preparation is an excellent preparation as clinical drug due to the fact that convenience is brought to a patient by lowering administration frequency.

Description

A kind of slow releasing preparation being used for the treatment of cancer and preparation method thereof

Technical field

The present invention relates to medicament slow release preparation technical field, particularly relate to a kind of slow releasing preparation being used for the treatment of cancer and preparation method thereof.

Background technology

Patent documentation WO2007/72997 discloses the stable tumor migration chalone derivant with excellent tumor migration chalone sample activity (metastin-likeactivity).At present, the tumour medicines such as tumor migration chalone derivant need could obtain good therapeutic effect with higher dosage usually, the using of tumour medicine of high dose brings serious side effect, because tumour medicine also can kill the cell of part health while killing tumor cell.It is must frequent drug administration that current tumour medicine also exists a serious problem, mainly because the problem that consumes rapidly of the too fast medicine brought of release of active constituents of medicine.Frequent drug administration not only increases the generation of side effect, and all brings very large burden to the physiology of patient and economy.

Therefore, develop a kind of can slow releasing tumor migration chalone derivant and the slow releasing preparation had as the advantageous property of clinical medicine has important value for current using of tumor migration chalone derivant for a long time.Patent documentation WO02/85399 discloses about the slow releasing preparation containing tumor migration chalone or derivatives thereof.But its sustained release performance is still lower.

Patent documentation WO2011/162413 discloses a kind of slow releasing preparation, and it comprises lactic acid polymer or its salt that tumor migration chalone derivant and weight average molecular weight are about 10000 to about 30000.Described slow releasing preparation can improve the slow release effect of tumor migration chalone derivant or its salt, also can the long drug effect playing tumor migration chalone derivant or its salt temporally, improves the convenience of patient to a certain extent.But its slow release effect can't meet the demand of this area completely.

Summary of the invention

Reduce side effect in order to obtain by requiring to obtain drug effect with not high dosage, improve the convenience of patient, overcoming pain and the long-time benefit producing drug effect by reducing administration frequency, it is desirable to develop a kind of can slow releasing tumor migration chalone derivant and there is the slow releasing preparation of the advantageous property as clinical medicine for a long time.Especially, iting is desirable to develop one can long-time stable and the preparation of slow release compounds (I) or its salt.

In order to solve foregoing problems, the present inventor furthers investigate, found that to be the Poly(D,L-lactide-co-glycolide (poly (lactic-co-glycolicacid) of about 10000 to about 30000 containing tumor migration chalone derivant or its salt and weight average molecular weight according to the application, or the slow releasing preparation of its salt PLGA), it is in drug effect, safety, stability, dosage, dosage form and use the advantageous property had needed for clinical medicine, finally completes the present invention.

Particularly, the present invention relates to the manufacture method of following slow releasing preparation and this slow releasing preparation.

A kind of slow releasing preparation, it comprises the compound shown in following formula:

Ac-D-Tyr-Hyp-Asn-Thr-Phe-AzaGly-Leu-Arg (Me)-Trp-NH2 (I) (in this manual, sometimes referred to as compound (I)) or its salt and weight average molecular weight are Poly(D,L-lactide-co-glycolide or its salt of about 10000 to about 30000.

Preferably, foregoing slow releasing preparation, wherein, the weight average molecular weight of this Poly(D,L-lactide-co-glycolide or its salt is about 13000 to about 18000.

Preferably, foregoing slow releasing preparation, it is the preparation of 3 to 6 months slow release.

Preferably, foregoing slow releasing preparation, it is treatment or the preventive of cancer.

Preferably, foregoing slow releasing preparation, it is parenteral administration.

A method for treatment or prophylaxis of cancer, comprises and gives the aforesaid slow releasing preparation of effective dose to mammal.

Manufacture a method for aforesaid slow releasing preparation, the method comprises makes following emulsion carry out seasoning (in-waterdryingmethod) in water:

(1) W/O/W emulsion, it is obtained by emulsifying W/O emulsion, and described W/O emulsion comprises interior aqueous phase containing compound (I) or its salt and the oil phase containing Poly(D,L-lactide-co-glycolide or its salt, or

(2) O/W emulsion, it is obtained by emulsifying oil phase, described oil phase inclusion compound (I) or its salt and Poly(D,L-lactide-co-glycolide or its salt.

Preferably, method as the aforementioned, the method comprises makes O/W emulsion carry out seasoning in water, and described O/W emulsion is obtained by emulsifying oil phase, and described oil phase contains compound (I) or its salt and Poly(D,L-lactide-co-glycolide or its salt.

In addition, the present invention also relates to a kind of slow releasing preparation and the Therapeutic Method according to following aspect.

A kind of inclusion compound (I) or its salt and weight average molecular weight are the Poly(D,L-lactide-co-glycolide of about 10000 to about 30000 or the slow releasing preparation of its salt, and this slow releasing preparation uses in the following manner: make compound (I) or its salt with the dosage of about 0.01 to about 4mg/kg body weight and with every 3 weeks or more for a long time (preferably 1 month) intervals once give to patient.

A kind of inclusion compound (I) or its salt and weight average molecular weight are the Poly(D,L-lactide-co-glycolide of about 10000 to about 30000 or the slow releasing preparation of its salt, and this slow releasing preparation uses in the following manner: make compound (I) or its salt with the dosage of about 0.03 to about 12mg/kg body weight and with every 2 months or more for a long time the intervals (being preferably 3 months) once give to patient.

A kind of inclusion compound (I) or its salt and weight average molecular weight are the Poly(D,L-lactide-co-glycolide of about 10000 to about 30000 or the slow releasing preparation of its salt, and this slow releasing preparation uses in the following manner: make compound (I) or its salt with the dosage of about 0.06 to about 24mg/kg body weight and with every 4 months or more for a long time the intervals (being preferably 6 months) once give to patient.

Preferably, foregoing slow releasing preparation, it is by using the method manufacture of (1) W/O/W emulsion or (2) O/W emulsion.

Preferably, foregoing slow releasing preparation, its for be used for the treatment of or prophylaxis of cancer (such as, pulmonary carcinoma, gastric cancer, hepatocarcinoma, cancer of pancreas, colorectal cancer, rectal cancer, colon cancer, carcinoma of prostate, ovarian cancer, cervical cancer, breast carcinoma, renal cancer, bladder cancer, cerebroma), pancreatic disease (such as, acute or chronic pancreatitis, cancer of pancreas), choriocarcinoma, hydatidiform mole, invasive mole, miscarriage, asyntaxia, abnormal carbohydrate metabolism, lipidosis and abnormal labor preparation.

Preferably, foregoing slow releasing preparation, it is microcapsule formulation; And

A kind for the treatment of or prophylaxis of cancer are (such as, pulmonary carcinoma, gastric cancer, hepatocarcinoma, cancer of pancreas, colorectal cancer, rectal cancer, colon cancer, carcinoma of prostate, ovarian cancer, cervical cancer, breast carcinoma, renal cancer, bladder cancer, cerebroma), pancreatic disease (such as, acute or chronic pancreatitis), choriocarcinoma, hydatidiform mole, invasive mole, miscarriage, asyntaxia, abnormal carbohydrate metabolism, lipidosis and abnormal labor method, the method comprises: give the aforesaid slow releasing preparation of effective dose to mammal.

Beneficial effect of the present invention: slow releasing preparation of the present invention long-time (such as, 3 weeks or more of a specified duration) slowly and stably discharge compound (1) or its salt, and also plays the drug effect of compound (1) or its salt for a long time.In addition, slow releasing preparation of the present invention improves the convenience of patient by reducing administration frequency, be the elegant formulations as clinical medicine.

Detailed description of the invention

Below in conjunction with embodiment, embodiment of the present invention are described in detail.It will be understood to those of skill in the art that following examples are only the preferred embodiments of the present invention, so that understand the present invention better, thus should not be considered as limiting scope of the present invention.For a person skilled in the art, the present invention can have various modifications and variations, within the spirit and principles in the present invention all, and any amendment done, equivalent replacement or improvement etc., all should be included within protection scope of the present invention.Experimental technique in following embodiment, if no special instructions, is conventional method; Experiment material used, if no special instructions, is and is purchased available from routine biochemistry chemical reagent work.

The present invention will more specifically illustrate below.

The invention provides the slow releasing preparation containing the tumor migration chalone derivant (in this description, sometimes referred to as compound (I)) shown in following formula or its salt:

Ac-D-Tyr-Hyp-Asn-Thr-Phe-AzaGly-Leu-Arg (Me)-Trp-NH2 (I) (serial number: 1).

Compound used in the present invention (I) can exist in a salt form.About the salt that compound (I) is formed, particularly preferably pharmacologically acceptable salt.The example of described salt comprises the salt formed with mineral acid (such as, hydrochloric acid, phosphoric acid, hydrobromic acid, sulphuric acid); The salt formed with organic acid (such as, acetic acid, formic acid, propanoic acid, fumaric acid, maleic acid, succinic acid, tartaric acid, citric acid, malic acid, oxalic acid, benzoic acid, methanesulfonic acid, benzenesulfonic acid); Salt (such as, the alkali metal salt such as sodium salt and potassium salt formed with inorganic base; Alkali salt such as calcium salt and magnesium salt; Aluminum salt; Ammonium salt); And the salt to be formed with organic base (such as, Trimethylamine, triethylamine, pyridine, picoline, ethanolamine, diethanolamine, triethanolamine, dicyclohexylamine, N, N-benzhydryl ethylenediamine).

The preferred embodiment that can be used in the salt that compound of the present invention (I) is formed comprises the salt formed with acetic acid.

Compound used in the present invention (I) or its salt manufacture, more specifically, by the method manufacture described in WO2007/72997 by known peptide symthesis method.

Slow releasing preparation of the present invention is except containing except compound (I) or its salt, be also Poly(D,L-lactide-co-glycolide (in this description, sometimes referred to as Poly(D,L-lactide-co-glycolide) or its salt of about 10000 to about 30000 containing weight average molecular weight.

In the present invention, Poly(D,L-lactide-co-glycolide refers to the polymer be made up of lactic acid and hydroxyacetic acid copolymerization.

The weight average molecular weight of Poly(D,L-lactide-co-glycolide used in the present invention or its salt is about 10000 to about 30000, be preferably about 10000 to about 25000, more preferably about 12000 to about 20000, and be more preferably about 13000 to about 18000 further.

Polydispersity (polydispersity) (weight-average molecular weight/number-average molecular weight) of Poly(D,L-lactide-co-glycolide or its salt is preferably about 1.2 to about 4.0, and more preferably about 1.5 to about 3.5.

Weight average molecular weight as used in this specification and polydispersity refer to measures by gel permeation chromatography (GPC) value obtained.The weight average molecular weight of each polymer and content are respectively polystyrene-equivalent weight average molecular weight (polystyrene-equivalentweightaveragemolecularweight) and the content by the calculated each polymer of this polystyrene-equivalent weight average molecular weight, wherein, this polystyrene-equivalent weight average molecular weight obtained as the gpc measurement of standard substance by using such as monodisperse polystyrene.The weight average molecular weight of each polymer and content such as can pass through high speed GPC equipment (HLC-8120GPC is manufactured by TosohCorporation) and measure.About post, SuperH4000 × 2 and SuperH2000 (being manufactured by TosohCorporation respectively) can be used.About mobile phase, oxolane can be used, and flowrate settable is 0.6mL/ minute.About detection method, differential refractive index can be used.

It should be noted that about Poly(D,L-lactide-co-glycolide and salt thereof, commercially available product can be used.

In the present invention, Poly(D,L-lactide-co-glycolide can exist in a salt form.The example of this salt comprises and inorganic base (such as, alkali metal such as sodium and potassium; Alkaline-earth metal such as calcium and magnesium) and organic base (such as, organic amine such as triethylamine; Basic amino acid is arginine such as) salt that formed; Or the salt to be formed with transition metal (such as, zinc, ferrum, copper); And complex salt (complexsalt).

Slow releasing preparation of the present invention is such as manufactured by following method: mixing cpd (I) or its salt and Poly(D,L-lactide-co-glycolide or its salt, and if desired, molded thus obtained mixture.Relative to Poly(D,L-lactide-co-glycolide or its salt, the use amount of compound (I) or its salt is such as about 0.01 to about 50% (w/w), is preferably about 0.1 to about 30% (w/w).

Now, the manufacture method of slow releasing preparation of the present invention will be more specifically described below.

(1) manufacture method of bar-shaped (rod-form) mechanograph etc.

(1-a)

Poly(D,L-lactide-co-glycolide or its salt are dissolved in organic solvent (being preferably dichloromethane etc.), and add the aqueous solution of compound (I) or its salt, then emulsifying.The emulsion of vacuum drying gained, has compound (I) or its salt and Poly(D,L-lactide-co-glycolide or its salt be dispersed in powder wherein to obtain.This powder warm, and cooling, to obtain lamellar, the mechanograph such as membranaceous, bar-shaped.Warm temperatures is such as about 50 to about 100 DEG C, and chilling temperature is such as about 0 to about 40 DEG C.The use amount of compound (I) or its salt changed according to persistent period etc. of the type of compound (I) or its salt, expection pharmacological action and effect; But, relative to Poly(D,L-lactide-co-glycolide or its salt, the use amount of compound (I) or its salt is such as about 0.01 to about 50% (w/w), is preferably about 0.1 to about 30% (w/w), is particularly preferably about 1 to about 20% (w/w).

(1-b)

Poly(D,L-lactide-co-glycolide or its salt are dissolved in organic solvent (being preferably dichloromethane etc.), and dispersed compound (I) or its salt.The dispersion liquid of vacuum drying gained, is dispersed in powder in Poly(D,L-lactide-co-glycolide or its salt to obtain compound (I) or its salt.This powder warm, and cooling, to obtain lamellar, the mechanograph such as membranaceous, bar-shaped.The use amount of warm temperatures, chilling temperature and compound (I) or its salt is identical with described in upper joint (1-a).

(2) manufacture method of microcapsule (also referred to as microsphere)

(2-a) seasoning in water

Microcapsule carries out seasoning in water by making (i) W (interior aqueous phase)/O (oil phase)/W (outer aqueous phase) emulsion or (ii) O (oil phase)/W (outer aqueous phase) emulsion and obtains, wherein, this W/O/W emulsion is obtained by emulsifying W (interior aqueous phase)/O (oil phase) emulsion, described W (interior aqueous phase)/O (oil phase) emulsion comprises interior aqueous phase containing compound (I) or its salt and the oil phase containing Poly(D,L-lactide-co-glycolide or its salt, and this O/W emulsion contains the oil phase of compound (I) or its salt and Poly(D,L-lactide-co-glycolide or its salt by emulsifying and obtains.

Above-mentioned emulsion (i), is namely comprised the W/O emulsion of interior aqueous phase containing compound (I) or its salt and the oil phase containing Poly(D,L-lactide-co-glycolide or its salt, is obtained by following method.

First, compound (I) or its salt are dissolved, disperse or suspend in water, with obtained interior aqueous phase.Compound (I) or the concentration of its salt in water are such as 0.001 to 90% (w/w), are preferably 0.01 to 80% (w/w).

The use amount of compound (I) or its salt changed according to persistent period etc. of the type of compound (I) or its salt, expection pharmacological action and effect; Relative to Poly(D,L-lactide-co-glycolide or its salt, the use amount of compound (I) or its salt is such as about 0.01 to about 50% (w/w), be preferably about 0.1 to about 30% (w/w), more preferably about 1 to about 20% (w/w).

In order to improve the encapsulating of compound in microcapsule (I) or its salt, if desired, medicine can be retained material (drugretainingsubstance) such as gelatin, agar, sodium alginate, polyvinyl alcohol or basic amino acid (such as, arginine, histidine, lysine) to be added in interior aqueous phase.Medicine retains the addition of material, by weight, is generally compound (I) or its salt about 0.01 to about 10 times.

Can be powder by the lyophilization for the time being of interior aqueous phase, then be dissolved in water to obtain suitable concentration, then place for subsequent use again.

In addition, Poly(D,L-lactide-co-glycolide or its salt are dissolved in organic solvent with obtained oil phase.

The example of organic solvent comprises halogenated hydrocarbon (such as, dichloromethane, chloroform, ethyl chloride, trichloroethane, carbon tetrachloride), fatty acid ester (such as, ethyl acetate, butyl acetate) and aromatic hydrocarbon (such as, benzene,toluene,xylene).Wherein, dichloromethane is preferably.

Concentration in organic solvent of Poly(D,L-lactide-co-glycolide or its salt changes according to Poly(D,L-lactide-co-glycolide or the type of its salt and the type of weight average molecular weight and organic solvent; Usually, the value represented by following formula:

[weight/(weight+Poly(D,L-lactide-co-glycolide of organic solvent or the weight of its salt) of Poly(D,L-lactide-co-glycolide or its salt] × (100%) is about 0.01 to about 90% (w/w), is preferably about 0.01 to about 70% (w/w).Oil phase is not preferably containing insoluble matter.

Homo-mixer etc. is utilized the aqueous solution of compound (I) or its salt, dispersion liquid or suspension (interior aqueous phase) to be added, disperse and be emulsifiable in the organic solvent solution (oil phase) of obtained Poly(D,L-lactide-co-glycolide or its salt, to manufacture W/O emulsion.

When manufacturing W/O emulsion in room temperature (about 19 to 25 DEG C), the W/O emulsion of gained can be passed through along with the time and change the state (such as, gel state) being unfavorable for second emulsifying (being illustrated in hereafter) into.In in the case, be sometimes difficult to manufacture microcapsule with high yield (productive rate used herein refers to the ratio of the weight of compound (I) or its salt used in the weight of compound (I) contained in microcapsule or its salt and W/O emulsion).

In order to prevent this transformation, preferably by the temperature adjusting of manufactured W/O emulsion 31 DEG C or higher (are preferably 31 to 33 DEG C).

On the other hand, above-mentioned emulsion (ii), namely contains the oil phase of compound (I) or its salt and Poly(D,L-lactide-co-glycolide or its salt, is obtained by following method.

First, the organic solvent solution of Poly(D,L-lactide-co-glycolide or its salt is manufactured.About organic solvent, use the identical organic solvent for the manufacture of above-mentioned W/O emulsion.

Concentration in organic solvent solution of Poly(D,L-lactide-co-glycolide or its salt changes according to Poly(D,L-lactide-co-glycolide or the type of its salt and the type of weight average molecular weight and organic solvent; Usually, the value represented by following formula:

[weight/(weight+Poly(D,L-lactide-co-glycolide of organic solvent or the weight of its salt) of Poly(D,L-lactide-co-glycolide or its salt] × (100%) is about 0.01 to about 70% (w/w), is preferably about 1 to about 60% (w/w).

Then, compound (I) or its salt be dissolved in or be suspended in the organic solvent solution of Poly(D,L-lactide-co-glycolide or its salt, to prepare oil phase.This oil phase also manufactures by following method: be dissolved in compound (I) or its salt to prepare solution in alcohol, and then is dissolved in by this solution or is suspended in the organic solvent solution of Poly(D,L-lactide-co-glycolide or its salt.Example for the alcohol of dissolved compound (I) or its salt comprises methanol.

The use amount of compound (I) or its salt can be selected, to make the ratio of compound (I) or its salt and Poly(D,L-lactide-co-glycolide or its salt similar in appearance to the ratio used when manufacturing above-mentioned (i) W/O emulsion.

Then, above-mentioned (i) W/O emulsion or (ii) oil phase are added in outer aqueous phase, by dispersion also emulsifyings (second emulsifying) such as homo-mixers, with obtained emulsion (hereinafter, the emulsion obtained by W/O emulsion is sometimes referred to as W/O/W emulsion, and the emulsion obtained by (ii) oil phase is sometimes referred to as O/W emulsion).

The use amount of outer aqueous phase, is generally about 1 to about 10000 times of volume relative to W/O emulsion or oil phase, is preferably about 10 to 5000 times of volumes and is particularly preferably about 50 to about 1000 times of volumes.

Usually emulsifying agent is added in outer aqueous phase.It about emulsifying agent, such emulsifying agent can be used, as long as can form stable W/O/W emulsion or O/W emulsion usually.The example of emulsifying agent comprises anion surfactant, non-ionic surface active agent, castor oil derivatives, PVP, polyvinyl alcohol, carboxymethyl cellulose, lecithin, gelatin and hyaluronic acid.Wherein, polyvinyl alcohol is preferably.The concentration of emulsifying agent in outer aqueous phase is generally about 0.001 to about 20% (w/w), is preferably about 0.01 to about 10% (w/w), is particularly preferably about 0.05 to about 5% (w/w).

Obtained W/O/W emulsion or O/W emulsion (hereinafter, it separately sometimes can referred to as emulsion) is made to carry out seasoning in water, to remove the organic solvent be contained in emulsion.Mode according to this, can obtain microcapsule.

In addition, except using the method for aforementioned W/O/W emulsion or O/W emulsion, one is also had to make S (solid phase)/O (oil phase) emulsion carry out the manufacture method of seasoning in water, wherein, this S (solid phase)/O (oil phase) emulsion comprises the solid phase containing compound (I) or its salt and Poly(D,L-lactide-co-glycolide or its salt.

First, Poly(D,L-lactide-co-glycolide or its salt are dissolved in organic solvent.Decentralized compound (I) or its salt in gained organic solvent solution.Now, the use amount of compound (I) or its salt and Poly(D,L-lactide-co-glycolide or its salt can be selected, become identical with during manufacture above-mentioned (i) W/O emulsion to make compound (I) or its salt with the ratio of Poly(D,L-lactide-co-glycolide or its salt.In addition, in order to be dispersed in organic solvent by compound (I) or its salt, such as ultrasonic irradiation, turbine type agitator or homogenizer is used.

Then, prepared S/O emulsion is added in outer aqueous phase further, and by using such as ultrasonic irradiation, turbine type agitator or homogenizer to carry out disperseing and emulsifying, with obtained emulsion (hereinafter, being sometimes called S (solid phase)/O (oil phase)/W (aqueous phase) emulsion).Afterwards, oil phase solvent is evaporated, with obtained microcapsule.Now, the volume of aqueous phase is typically chosen in about 1 times to about 10000 times of the volume of oil phase, more preferably about 10 times to about 5000 times, is particularly preferably about 50 times to about 1000 times.

Above-mentioned emulsifying agent can be added in outer aqueous phase.The addition of outer aqueous phase and the type that is added into the emulsifying agent in outer aqueous phase with use during concentration and manufacture above-mentioned W/O/W emulsion identical.

The S/O/W emulsion of gained is made to carry out seasoning in water, to remove organic solvent.Mode according to this, can obtain microcapsule.

The microcapsule obtained by using W/O/W emulsion, O/W emulsion, S/O/W emulsion by centrifugal, sieving or filtering is separated, and then, if desired, re-uses distilled water wash, to remove the emulsifying agent etc. being attached to microcapsule surface.Afterwards, microcapsule is scattered in distilled water etc., lyophilization, and if desired, more warm to remove water in microcapsule and organic solvent further.Warm can in decompression under carry out.About the condition of warm step, heat drying carries out in following temperature: this temperature is not less than the glass transition temperature of Poly(D,L-lactide-co-glycolide used herein or its salt, and microcapsule granule can't adhere to mutually in this temperature.

Preferably, heat drying carries out in following temperature range: from the glass transition temperature of Poly(D,L-lactide-co-glycolide or its salt to the temperature higher than this glass transition temperature about 30 DEG C.Glass transition temperature used herein refers to the mid point being measured the temperature obtained by use differential scanning calorimetry (DSC) with the heating rate of 10 to 20 DEG C/min.

(2-b) phase separation method

When manufacturing microcapsule by the method, flocculating agent (coacervationagent) is progressively added in the W/O emulsion described in above-mentioned (2-a) " in water seasoning " under stirring.Mode according to this, microcapsule is through precipitating and solidifying.The amount of flocculating agent is chosen as about 0.01 to about 1000 times of oil phase volume, is preferably about 0.05 to about 500 times, is particularly preferably about 0.1 to about 200 times.

There is no particular restriction for flocculating agent, as long as it is can with immiscible organic solvent and the polymer compound that biodegradable polymer of the present invention can not be dissolved, mineral oils compound or plant oil compound etc.The instantiation of spendable flocculating agent comprises silicone oil, Oleum sesami, soybean oil, Semen Maydis oil, Oleum Gossypii semen, Oleum Cocois, Semen Lini oil, mineral oil, normal hexane and normal heptane.These materials can use in the form of mixtures of two or more.

After the microcapsule of gained is separated, use the repeated washing microcapsule such as heptane, so that the flocculating agent except biological active substances and the constituent that formed by biodegradable polymer of the present invention is removed, then drying under reduced pressure.Selectively, wash with the same way described in above-mentioned (2-a) " in water seasoning ", then lyophilization warm drying again.

(2-c) spray drying method

When manufacturing microcapsule by the method, the W/O emulsion described in above-mentioned (2-a) " in water seasoning " is sprayed in the dewatering station of spray dryer by nozzle, the organic solvent in microlayer model is volatilized in very short time, and obtained microcapsule.The example of nozzle comprises two-fluid spray nozzle type, pressure nozzle type and rotating disk type.Afterwards, if desired, wash with the same way described in above-mentioned (2-a) " in water seasoning ", then can lyophilization again, and by warm further drying.

About other dosage form except above-mentioned microcapsule, microparticle can be mentioned, it is obtained by following method: by such as rotary evaporator, the W/O emulsion described in above-mentioned (2-a) " in water seasoning " is dried to solid, by regulation and control vacuum, organic solvent and water are evaporated simultaneously, and then pulverized by jet mill etc.

In addition, by the microparticle through pulverizing with the same way washing described in above-mentioned (2-a) " in water seasoning ", then can lyophilization again, and by warm further drying.

When the microcapsule above manufactured by chapters and sections (2-a), (2-b) or (2-c) being scattered in distilled water etc., in order to prevent particle aggregation (aggregation), aggregation-preventing agent (aggregationpreventingagent) can be added.The example of aggregation-preventing agent comprises water soluble polysaccharide such as mannitol, lactose, glucose, starch (such as, corn starch) and hyaluronic acid or its alkali metal salt; Protein is glycine, fibrin and collagen such as; And inorganic salt such as sodium chloride and dibastic sodium phosphate.Wherein, preferred mannitol.Relative to microcapsule (100 weight portion), the use amount of aggregation-preventing agent is preferably about 2 to about 100 weight portions, more preferably about 10 to about 25 weight portions.

In addition, can according to the same way described in example of above-mentioned (1-a), microcapsule is warm after cool again, to obtain plate-like, membranaceous and bar-shaped etc. mechanograph.

There is no particular restriction for compound (I) or the content of its salt in microcapsule; But the content of 3 months slow releasing preparation is such as 4% or more to 10% or less, be preferably 6% or more to 10% or less.In addition, the content of 6 months slow releasing preparation is such as 4% or more to 18% or less, is preferably 6% or more to 16% or less, is more preferred from 7% or more to 10% or less.

In view of the slow-release time of slow releasing preparation of the present invention, in aforementioned various manufacture method, be preferably following manufacture method: the method comprises to be made to contain the oil phase of compound (I) or its salt and Poly(D,L-lactide-co-glycolide or its salt by emulsifying and the O/W emulsion that obtains carries out seasoning in water.

In aforementioned various manufacture method, when Poly(D,L-lactide-co-glycolide or its salt are dissolved in organic solvent, zinc oxide can be added in organic solvent.

Relative to Poly(D,L-lactide-co-glycolide (100 weight portion), the use amount of zinc oxide is such as about 0.01 to about 100 weight portions, is preferably about 0.1 to about 20 weight portion.

In addition, the granularity of zinc oxide is generally about 0.001 to about 10 μm, is preferably about 0.005 to about 1 μm.

Similarly, by the slow releasing preparation using zinc oxide to obtain, there is excellent characteristic, such as " high encapsulation rate of medicine ", " ability of long-time slow releasing pharmaceutical " etc.

When manufacturing slow releasing preparation of the present invention, can compound (I) or its salt be dissolved in the aqueous solution of volatile salts such as ammonium acetate, place for subsequent use after lyophilization again.

Mode is by the operability using the lyophilization product of ammonium acetate process and the compound (I) that obtains or its salt to have small grain size and excellence according to this, is therefore conducive to manufacturing slow releasing preparation.

Slow releasing preparation of the present invention obtained thus, if necessary, suitably can contain medical additive (such as, stabilizing agent, antiseptic, soothing agent (soothingagent)).The Exemplary dosage forms of slow releasing preparation of the present invention comprises parenteral administration (such as, injection, implant, suppository) and oral administration agent (such as, solid preparation such as capsule, tablet, granule and powder agent; Liquid preparation is syrup, emulsion and suspensoid such as).The example of stabilizing agent comprises human serum albumin and Polyethylene Glycol.The example of antiseptic comprises benzyl alcohol and phenol.The example of soothing agent comprises benzalkonium chloride (benzalkoniumchloride) and ethocaine (procainehydrochloride).In slow releasing preparation of the present invention, the content of compound (I) or its salt suitably can be chosen as following ranges usually: relative to overall sustained release preparation, is about 0.01 to about 33% (w/w).

The advantage of slow releasing preparation of the present invention is that the blood drug concentration of compound (I) or its salt is stable between slow-release period.

Slow releasing preparation of the present invention is preferably parenteral administration, and more preferably injection.For example, when slow releasing preparation is microencapsulated form, by microcapsule and dispersant (such as, surfactant such as Tween80 and HCO-60; And polysaccharide such as carboxymethyl cellulose, sodium alginate and hyaluronic acid), antiseptic (such as, methyl parahydroxybenzoate and propyl p-hydroxybenzoate) and isotonic agent is (such as, sodium chloride, mannitol, Sorbitol and glucose) etc. combinationally use, to prepare aqueous suspension.Mode according to this, can obtain slow releasing injection.In addition, slow releasing injection also obtains by following method: microcapsule be scattered in vegetable oil (such as Oleum sesami and Semen Maydis oil) or be added with in the vegetable oil of phospholipid (such as lecithin), or be scattered in medium chain triglyceride (such as, Miglyol812) in, to obtain oil-based suspension.

When slow releasing preparation is in such as microencapsulated form, if be used as the granularity of the microcapsule of suspension injection to meet polydispersity by entry needle and scope, then the granularity of this microcapsule can be gratifying.About the particle mean size of microcapsule, for example, the scope of about 0.1 to about 300 μm can be addressed.The particle mean size of microcapsule preferably falls within following ranges: about 1 to about 150 μm, is particularly preferably about 2 to about 100 μm.

Aforementioned microcapsule carries out sterilization treatment by following method: under aseptic condition, carry out all manufacturing steps; Sterilize with gamma-rays; And interpolation biocide.There is no particular restriction for the method.

Because the toxicity of slow releasing preparation of the present invention is low, therefore can safely oral or parenteral to mammal (such as, the mankind, monkey, baboon, chimpanzee, pig, cattle, sheep, horse, Canis familiaris L., cat, mice, rat).

Slow releasing preparation of the present invention can be used for all diseases for the treatment of or prevention is relevant with the physiologically active of tumor migration chalone.Especially, slow releasing preparation of the present invention can effectively be used for the treatment of or prophylaxis of cancer (such as, pulmonary carcinoma, gastric cancer, hepatocarcinoma, cancer of pancreas, colorectal cancer, rectal cancer, colon cancer, carcinoma of prostate, ovarian cancer, cervical cancer, breast carcinoma, renal cancer, bladder cancer, cerebroma), pancreatic disease (such as, acute or chronic pancreatitis), choriocarcinoma, hydatidiform mole, invasive mole, miscarriage, asyntaxia, abnormal carbohydrate metabolism, lipidosis and abnormal labor.

Slow releasing preparation particularly suitable of the present invention is therapeutic agent or the preventive of cancer (being preferably carcinoma of prostate).

The dosage of slow releasing preparation of the present invention suitably can be selected according to the type of active component (that is, compound (I) or its salt) and content, dosage form, release duration, administration experimenter, route of administration, administration object, targeted condition and symptom etc.; But as long as active component can in the persistent period in vivo maintaining medicine effective concentration and reach expection, then this dosage can be gratifying.For example, in cancer patient is grown up in treatment, when slow releasing preparation of the present invention gives through such as injection and reaches the slow release of about 1 month, compound (I) or its salt in the use amount given at every turn are such as in following ranges: about 0.01 to about 4mg/kg body weight, and are preferably about 0.03 to 0.6mg/kg body weight.In addition, when slow releasing preparation of the present invention gives through injection and reaches the slow release of about 3 months, compound (I) or its salt in the use amount given at every turn are such as in following ranges: about 0.03 to about 12mg/kg body weight, and are preferably about 0.09 to about 1.8mg/kg body weight.In addition, when slow releasing preparation of the present invention gives through injection and reaches the slow release of about 6 months, compound (I) or its salt in the use amount given at every turn are such as in following ranges: about 0.06 to about 24mg/kg body weight, and are preferably about 0.18 to about 3.6mg/kg body weight.Administration frequency be such as every month 1 time, every 3 months once, every 6 months once, and can according to the content of compound (I) or its salt, dosage form, release duration, targeted condition and administration experimenter etc. and suitably select.About slow releasing preparation of the present invention, used be preferably 1 to 8 month slow releasing preparation (namely, the preparation of slow releasing compound (I) or its salt in during 1 to 8 month), be more preferably 1 to 6 month slow releasing preparation, more preferably 3 to 6 months slow releasing preparation, are more preferably 6 months slow releasing preparation further.

In addition, slow releasing preparation of the present invention can with the other medicines for various disease (various disease described in compound (I) or its salt pair have pharmaceutically effectively act on) (hereinafter, referred to as coupling medicine) combinationally use, especially following medicine, such as: be used for the treatment of the chemotherapeutant of cancer, hormone therapy agent and immunotherapeutic agent.In described situation, there is no particular restriction for the administration time of slow releasing preparation of the present invention and coupling medicine.Slow releasing preparation of the present invention and coupling medicine can simultaneously or certain interval of time give to administration experimenter.The dosage of coupling medicine suitably can be selected based on clinical dosage.In addition, the blending ratio of slow releasing preparation of the present invention and coupling medicine suitably can be selected according to administration experimenter, route of administration, targeted condition, symptom and combination etc.

The example of chemotherapeutant comprises alkylating agent (such as, cyclophosphamide, ifosfamide (ifosfamide), nimustine (nimustine), Ranimustine (ranimustine), carboquone (carboquone)), antimetabolite (such as, methotrexate (methotrexate), 5-fluorouracil, ftorafur (tegafur), carmofur (carmofur), UFT, doxifluridine (doxifluridine), cytosine arabinoside (cytarabine), enocitabine (enocitabine), mercaptopurine, tioinosine, thioguanine), anticancer antibiotic substance (such as, mitomycin (mitomycin), amycin (adriamycin), daunorubicin (daunorubicin), epirubicin (epirubicin), pirarubicin (pirarubicin), idarubicin (idarubicin), bleomycin (bleomycin), peplomycin (peplomycin), D actinomycin D (actinomycin)), anticarcinogen (such as, the vincristine (vincristine) of plant derivation, vinblastine (vinblastine), vindesine (vindesine), etoposide (etoposide), camptothecine (camptothecin), irinotecan (irinotecan)), cisplatin (cisplatin), carboplatin (carboplatin), nedaplatin (nedaplatin), paclitaxel (paclitaxel), docetaxel (docetaxel) and estramustine (estramustine).

The example of hormone therapy agent comprises adrenocortical hormone (such as, prednisolone (prednisolone), prednisone (prednisone), dexamethasone (dexamethasone), cortisone acetate (cortisoneacetate)), estrogen (such as, estradiol (estradiol), ethinylestradiol (ethinylestradiol), fostestrol (fosfestrol), chlorotrianisene (chlorotrianisene)), estrogen antagonist (such as, epitiostanol (epitiostanol), mepitiostane (mepitiostane), tamoxifen (tamoxifen), clomifene (clomiphene)), progesterone (such as, hydroxyprogesterone caproate (hydroxyprogesteronecaproate), dydrogesterone (dydrogesterone), medroxyprogesterone (medroxyprogesterone), norethindrone (norethisterone), norethindrone (norethindrone)) and LHRH derivant (such as, TAP-144 (leuprorelinacetate)).

The example of immunotherapeutic agent comprises microorganism or bacterial component (such as, muramyldipeptide (muramyldipeptide) derivant, molten chain bacterium (picibanil)), there is the polysaccharide of immune-enhancing activity (such as, lentinan (lentinan), sizofiran (sizofiran), krestin (krestin)), the cytokine obtained by gene engineering method (such as, interferon, interleukin II (IL-2), interleukin 12 (IL-12), tumor necrosis factor (TNF)) and colony stimulating factor is (such as, granulocyte colony-stimulating factor, erythropoietin (erythropoietin)).

In addition, through confirming that there is the medicine improving cachexia effect in animal model or clinical practice, more specifically, cyclooxygenase-2 inhibitors (such as, indomethacin (indomethacin)) [CancerResearch, Vol.49, pages5935to5939,1989], derivatives of progesterone (such as, megestrol acetate (megesterolacetate)) [JournalofClinicalOncology, Vol.12, pages213to225,1994], glucocorticoid (such as, dexamethasone (dexamethasone)), metoclopramide (metoclopramide) class medicine, tetrahydrocannabinol (tetrahydrocannabinol) class medicine (identical with document mentioned above), lipid metabolism improving agent (such as, eicosapentaenoic acid) [BritishJournalofCancer, Vol.68, pages314to318,1993], growth hormone, IGF-1 or anti-antibody (that is, the TNF-α bringing out cachexia factor, LIF, IL-6, oncostatin M (oncostatinM)) can combinationally use with slow releasing preparation of the present invention.

In addition, be used for the treatment of or prevent the general medicine of Placenta Hominis and pancreatic disease also can be used as coupling medicine.The example of described medicine comprises general used clinically antiinflammatory, antipyretic/analgesics, antibacterial, antiviral agent and hormone preparation.

In this description, when alkali or aminoacid etc. represent with abbreviation, it represents according to the Conventional abbreviations used usual in IUPACIUB biochemical nomenclature commission or this area.The example is as follows.When there is amino acid whose optical isomer understandably, unless otherwise specified, otherwise be shown as L-type aminoacid.

Ac: acetyl group

AzaGly: azepine glycine

Hyp: trans-4-hydroxyproline

Leu: leucine

Thr: threonine

Arg (Me): N ω-methylarginine

Phe: phenylalanine

Tyr: tyrosine

Trp: tryptophan

Asn: agedoite

Embodiment

The present invention will more specifically be illustrated by embodiment and testing example below; But the present invention is not limited to described embodiment.

Except active component, by such as Japanese Pharmacopoeia the 15th edition (JapanesePharmacopeia, 15 ^threvision) applicable material listed in, illustrated in Japanese ingredient specification (JapaneseStandardsforPharmaceuticalIngredients) material or medicated premix specification 2003 uses as the composition (that is, additive) of prescription described in the following example.

Embodiment 1

Poly(D,L-lactide-co-glycolide (weight average molecular weight Mw:10000) (7.0417g) is dissolved in dichloromethane (13.186g).Take this solution (15.56g) and with the solution blending of acetate (0.7406g) being dissolved with compound (I) in methanol (2.819g), obtain oil phase.Then, oil phase is poured into 0.1% (w/w) polyvinyl alcohol (EG-40 being adjusted to about 18 DEG C in advance, by NipponSyntheticChemicalIndustryCo., Ltd. manufacture) in aqueous solution (1 liter), and use turbine type homo-mixer (being manufactured by Tokushukika) emulsifying, preparation O/W emulsion (whirling speed: about 7000rpm).Stir O/W emulsion about 3 hours (in water drying steps), and use 75 μm of standard screen clothes to sieve, (HIMACCR5DL, by Hitachi then to use centrifuge, Ltd. manufacture) collect microsphere by centrifugal (speed of rotation: about 2500rpm, 5 minutes).Microsphere is scattered in distilled water again, and centrifugal further, eccysis dissociant etc.By collected microsphere redispersion in a small amount of distilled water, and add mannitol (0.855g).By this mixture with freezer dryer (DF-01H, ULVAC) lyophilization, obtain microcapsule powder.The content of compound (I) in gained microcapsule powder is 8.0%.

Embodiment 2

Poly(D,L-lactide-co-glycolide (weight average molecular weight Mw:11000) (7.0530g) is dissolved in dichloromethane (13.269g).Take this solution (15.82g) and with the solution blending of acetate (0.7398g) being dissolved with compound (I) in methanol (2.832g), obtain oil phase.Then, oil phase is poured into 0.1% (w/w) polyvinyl alcohol (EG-40 being adjusted to about 18 DEG C in advance, by NipponSyntheticChemicalIndustryCo., Ltd. manufacture) in aqueous solution (1 liter), and use turbine type homo-mixer (being manufactured by Tokushukika) emulsifying, preparation O/W emulsion (whirling speed: about 7000rpm).Stir O/W emulsion about 3 hours (in water drying steps), and use 75 μm of standard screen clothes to sieve, (HIMACCR5DL, by Hitachi then to use centrifuge, Ltd. manufacture) collect microsphere by centrifugal (speed of rotation: about 2500rpm, 5 minutes).Microsphere is scattered in distilled water again, and centrifugal further, eccysis dissociant etc.By collected microsphere redispersion in a small amount of distilled water, and add mannitol (0.866g).By this mixture with freezer dryer (DF-01H, ULVAC) lyophilization, obtain microcapsule powder.The content of compound (I) in gained microcapsule powder is 8.5%.

Embodiment 3

Poly(D,L-lactide-co-glycolide (weight average molecular weight Mw:12000) (7.0405g) is dissolved in dichloromethane (13.184g).Take this solution (15.54g) and with the solution blending of acetate (0.7397g) being dissolved with compound (I) in methanol (2.82g), obtain oil phase.Then, oil phase is poured into 0.1% (w/w) polyvinyl alcohol (EG-40 being adjusted to about 18 DEG C in advance, by NipponSyntheticChemicalIndustryCo., Ltd. manufacture) in aqueous solution (1 liter), and use turbine type homo-mixer (being manufactured by Tokushukika) emulsifying, preparation O/W emulsion (whirling speed: about 7000rpm).Stir O/W emulsion about 3 hours (in water drying steps), and use 75 μm of standard screen clothes to sieve, (HIMACCR5DL, by Hitachi then to use centrifuge, Ltd. manufacture) collect microsphere by centrifugal (speed of rotation: about 2500rpm, 5 minutes).Microsphere is scattered in distilled water again, and centrifugal further, eccysis dissociant etc.By collected microsphere redispersion in a small amount of distilled water, and add mannitol (0.840g).By this mixture with freezer dryer (DF-01H, ULVAC) lyophilization, obtain microcapsule powder.The content of compound (I) in gained microcapsule powder is 8.5%.

Embodiment 4

Poly(D,L-lactide-co-glycolide (weight average molecular weight Mw:12500) (13.3028g) is dissolved in dichloromethane (24.15g).Take this solution (15.60g) and with the solution blending of acetate (0.7417g) being dissolved with compound (I) in methanol (2.83g), obtain oil phase.Then, oil phase is poured into 0.1% (w/w) polyvinyl alcohol (EG-40 being adjusted to about 18 DEG C in advance, by NipponSyntheticChemicalIndustryCo., Ltd. manufacture) in aqueous solution (1 liter), and use turbine type homo-mixer (being manufactured by Tokushukika) emulsifying, preparation O/W emulsion (whirling speed: about 7000rpm).Stir O/W emulsion about 3 hours (in water drying steps), and use 75 μm of standard screen clothes to sieve, (HIMACCR5DL, by Hitachi then to use centrifuge, Ltd. manufacture) collect microsphere by centrifugal (speed of rotation: about 2500rpm, 5 minutes).Microsphere is scattered in distilled water again, and centrifugal further, eccysis dissociant etc.By collected microsphere redispersion in a small amount of distilled water, and add mannitol (0.852g).By this mixture with freezer dryer (DF-01H, ULVAC) lyophilization, obtain microcapsule powder.The content of compound (I) in gained microcapsule powder is 8.6%.

Embodiment 5

Poly(D,L-lactide-co-glycolide (weight average molecular weight Mw:11000) (13.3374g) is dissolved in dichloromethane (24.47g).Take this solution (15.54g) and blended with the aqueous solution of the acetate (0.7450g) being dissolved with compound (I) in distilled water (0.60g), then small-sized homogenizer (KINEMATICA) emulsifying is used, form W/O emulsion (speed of rotation: about 10000rpm, 30 seconds).Then, W/O emulsion is adjusted to 32 DEG C and pours 0.1% (w/w) polyvinyl alcohol (EG-40 being adjusted to about 18 DEG C in advance into, by NipponSyntheticChemicalIndustryCo., Ltd. manufacture) in aqueous solution (1 liter), then use turbine type homo-mixer (being manufactured by Tokushukika) to carry out second emulsifying, obtain W/O/W emulsion (whirling speed: about 7000rpm).Stir W/O/W emulsion about 3 hours (in water drying steps), and use 75 μm of standard screen clothes to sieve, (HIMACCR5DL, by Hitachi then to use centrifuge, Ltd. manufacture) collect microsphere by centrifugal (speed of rotation: about 2500rpm, 5 minutes).Microsphere is scattered in distilled water again, and centrifugal further, eccysis dissociant etc.By collected microsphere redispersion in a small amount of distilled water, and add mannitol (0.872g).By this mixture with freezer dryer (DF-01H, ULVAC) lyophilization, obtain microcapsule powder.The content of compound (I) in gained microcapsule powder is 7.3%.

Embodiment 6

Poly(D,L-lactide-co-glycolide (weight average molecular weight Mw:13000) (13.3334g) is dissolved in dichloromethane (24.30g).Take this solution (15.90g) and blended with the aqueous solution of the acetate (0.7554g) being dissolved with compound (I) in distilled water (0.60g), then small-sized homogenizer (KINEMATICA) emulsifying is used, form W/O emulsion (speed of rotation: about 10000rpm, 30 seconds).Then, W/O emulsion is adjusted to 32 DEG C and pours 0.1% (w/w) polyvinyl alcohol (EG-40 being adjusted to about 18 DEG C in advance into, by NipponSyntheticChemicalIndustryCo., Ltd. manufacture) in aqueous solution (1 liter), then use turbine type homo-mixer (being manufactured by Tokushukika) to carry out second emulsifying, obtain W/O/W emulsion (whirling speed: about 7000rpm).Stir W/O/W emulsion about 3 hours (in water drying steps), and use 75 μm of standard screen clothes to sieve, (HIMACCR5DL, by Hitachi then to use centrifuge, Ltd. manufacture) collect microsphere by centrifugal (speed of rotation: about 2500rpm, 5 minutes).Microsphere is scattered in distilled water again, and centrifugal further, eccysis dissociant etc.By collected microsphere redispersion in a small amount of distilled water, and add mannitol (0.864g).By this mixture with freezer dryer (DF-01H, ULVAC) lyophilization, obtain microcapsule powder.The content of compound (I) in gained microcapsule powder is 7.3%.

Embodiment 7

Poly(D,L-lactide-co-glycolide (weight average molecular weight Mw:13500) (13.2307g) is dissolved in dichloromethane (24.74g).Take this solution (15.57g) and blended with the aqueous solution of the acetate (0.7589g) being dissolved with compound (I) in distilled water (0.60g), then small-sized homogenizer (KINEMATICA) emulsifying is used, form W/O emulsion (speed of rotation: about 10000rpm, 30 seconds).Then, W/O emulsion is adjusted to 32 DEG C and pours 0.1% (w/w) polyvinyl alcohol (EG-40 being adjusted to about 18 DEG C in advance into, by NipponSyntheticChemicalIndustryCo., Ltd. manufacture) in aqueous solution (1 liter), then use turbine type homo-mixer (being manufactured by Tokushukika) to carry out second emulsifying, obtain W/O/W emulsion (whirling speed: about 7000rpm).Stir W/O/W emulsion about 3 hours (in water drying steps), and use 75 μm of standard screen clothes to sieve, (HIMACCR5DL, by Hitachi then to use centrifuge, Ltd. manufacture) collect microsphere by centrifugal (speed of rotation: about 2500rpm, 5 minutes).Microsphere is scattered in distilled water again, and centrifugal further, eccysis dissociant etc.By collected microsphere redispersion in a small amount of distilled water, and add mannitol (0.845g).By this mixture with freezer dryer (DF-01H, ULVAC) lyophilization, obtain microcapsule powder.The content of compound (I) in gained microcapsule powder is 6.9%.

Embodiment 8

Poly(D,L-lactide-co-glycolide (weight average molecular weight Mw:10000) (6.2666g) is dissolved in dichloromethane (10.975g).Take this solution (13.20g) and with the solution blending of acetate (1.4825g) in methanol (5.61g) being dissolved with compound (I), obtain oil phase.Then, oil phase is poured into 0.1% (w/w) polyvinyl alcohol (EG-40 being adjusted to about 18 DEG C in advance, by NipponSyntheticChemicalIndustryCo., Ltd. manufacture) in aqueous solution (1 liter), and use turbine type homo-mixer (being manufactured by Tokushukika) emulsifying, preparation O/W emulsion (whirling speed: about 7000rpm).Stir O/W emulsion about 3 hours (in water drying steps), and use 75 μm of standard screen clothes to sieve, (HIMACCR5DL, by Hitachi then to use centrifuge, Ltd. manufacture) collect microsphere by centrifugal (speed of rotation: about 2500rpm, 5 minutes).Microsphere is scattered in distilled water again, and centrifugal further, eccysis dissociant etc.By collected microsphere redispersion in a small amount of distilled water, and add mannitol (0.851g).By this mixture with freezer dryer (DF-01H, ULVAC) lyophilization, obtain microcapsule powder.The content of compound (I) in gained microcapsule powder is 16.8%.

Embodiment 9

Poly(D,L-lactide-co-glycolide (weight average molecular weight Mw:14000) (6.2621g) is dissolved in dichloromethane (10.948g).Take this solution (13.22g) and with the solution blending of acetate (1.4795g) being dissolved with compound (I) in methanol (5.60g), obtain oil phase.Then, oil phase is poured into 0.1% (w/w) polyvinyl alcohol (EG-40 being adjusted to about 18 DEG C in advance, by NipponSyntheticChemicalIndustryCo., Ltd. manufacture) in aqueous solution (1 liter), and use turbine type homo-mixer (being manufactured by Tokushukika) emulsifying, preparation O/W emulsion (whirling speed: about 7000rpm).Stir O/W emulsion about 3 hours (in water drying steps), and use 75 μm of standard screen clothes to sieve, (HIMACCR5DL, by Hitachi then to use centrifuge, Ltd. manufacture) collect microsphere by centrifugal (speed of rotation: about 2500rpm, 5 minutes).Microsphere is scattered in distilled water again, and centrifugal further, eccysis dissociant etc.By collected microsphere redispersion in a small amount of distilled water, and add mannitol (0.854g).By this mixture with freezer dryer (DF-01H, ULVAC) lyophilization, obtain microcapsule powder.The content of compound (I) in gained microcapsule powder is 16.3%.

Embodiment 10

Poly(D,L-lactide-co-glycolide (weight average molecular weight Mw:15000) (6.2617g) is dissolved in dichloromethane (10.971g).Take this solution (13.18g) and with the solution blending of acetate (1.4775g) being dissolved with compound (I) in methanol (5.63g), obtain oil phase.Then, oil phase is poured into 0.1% (w/w) polyvinyl alcohol (EG-40 being adjusted to about 18 DEG C in advance, by NipponSyntheticChemicalIndustryCo., Ltd. manufacture) in aqueous solution (1 liter), and use turbine type homo-mixer (being manufactured by Tokushukika) emulsifying, preparation O/W emulsion (whirling speed: about 7000rpm).Stir O/W emulsion about 3 hours (in water drying steps), and use 75 μm of standard screen clothes to sieve, (HIMACCR5DL, by Hitachi then to use centrifuge, Ltd. manufacture) collect microsphere by centrifugal (speed of rotation: about 2500rpm, 5 minutes).Microsphere is scattered in distilled water again, and centrifugal further, eccysis dissociant etc.By collected microsphere redispersion in a small amount of distilled water, and add mannitol (0.848g).By this mixture with freezer dryer (DF-01H, ULVAC) lyophilization, obtain microcapsule powder.The content of compound (I) in gained microcapsule powder is 15.8%.

Embodiment 11

Poly(D,L-lactide-co-glycolide (weight average molecular weight Mw:14500) (13.3028g) is dissolved in dichloromethane (24.15g).Take this solution (13.46g) and with the solution blending of acetate (1.4821g) being dissolved with compound (I) in methanol (5.75g), obtain oil phase.Then, oil phase is poured into 0.1% (w/w) polyvinyl alcohol (EG-40 being adjusted to about 18 DEG C in advance, by NipponSyntheticChemicalIndustryCo., Ltd. manufacture) in aqueous solution (1 liter), and use turbine type homo-mixer (being manufactured by Tokushukika) emulsifying, preparation O/W emulsion (whirling speed: about 7000rpm).Stir O/W emulsion about 3 hours (in water drying steps), and use 75 μm of standard screen clothes to sieve, (HIMACCR5DL, by Hitachi then to use centrifuge, Ltd. manufacture) collect microsphere by centrifugal (speed of rotation: about 2500rpm, 5 minutes).Microsphere is scattered in distilled water again, and centrifugal further, eccysis dissociant etc.By collected microsphere redispersion in a small amount of distilled water, and add mannitol (0.857g).By this mixture with freezer dryer (DF-01H, ULVAC) lyophilization, obtain microcapsule powder.The content of compound (I) in gained microcapsule powder is 13.3%.

Embodiment 12

Poly(D,L-lactide-co-glycolide (weight average molecular weight Mw:12000) (13.3374g) is dissolved in dichloromethane (24.47g).Take this solution (13.42g) and blended with the aqueous solution of the acetate (1.4892g) being dissolved with compound (I) in distilled water (1.20g), then small-sized homogenizer (KINEMATICA) emulsifying is used, form W/O emulsion (speed of rotation: about 10000rpm, 30 seconds).Then, W/O emulsion is adjusted to 32 DEG C and pours 0.1% (w/w) polyvinyl alcohol (EG-40 being adjusted to about 18 DEG C in advance into, by NipponSyntheticChemicalIndustryCo., Ltd. manufacture) in aqueous solution (1 liter), then use turbine type homo-mixer (being manufactured by Tokushukika) to carry out second emulsifying, obtain W/O/W emulsion (whirling speed: about 7000rpm).Stir W/O/W emulsion about 3 hours (in water drying steps), and use 75 μm of standard screen clothes to sieve, (HIMACCR5DL, by Hitachi then to use centrifuge, Ltd. manufacture) collect microsphere by centrifugal (speed of rotation: about 2500rpm, 5 minutes).Microsphere is scattered in distilled water again, and centrifugal further, eccysis dissociant etc.By collected microsphere redispersion in a small amount of distilled water, and add mannitol (0.878g).By this mixture with freezer dryer (DF-01H, ULVAC) lyophilization, obtain microcapsule powder.The content of compound (I) in gained microcapsule powder is 13.5%.

Embodiment 13

Poly(D,L-lactide-co-glycolide (weight average molecular weight Mw:15000) (13.3334g) is dissolved in dichloromethane (24.30g).Take this solution (13.38g) and blended with the aqueous solution of the acetate (1.4866g) being dissolved with compound (I) in distilled water (1.21g), then small-sized homogenizer (KINEMATICA) emulsifying is used, form W/O emulsion (speed of rotation: about 10000rpm, 30 seconds).Then, W/O emulsion is adjusted to 32 DEG C and pours 0.1% (w/w) polyvinyl alcohol (EG-40 being adjusted to about 18 DEG C in advance into, by NipponSyntheticChemicalIndustryCo., Ltd. manufacture) in aqueous solution (1 liter), then use turbine type homo-mixer (being manufactured by Tokushukika) to carry out second emulsifying, obtain W/O/W emulsion (whirling speed: about 7000rpm).Stir W/O/W emulsion about 3 hours (in water drying steps), and use 75 μm of standard screen clothes to sieve, (HIMACCR5DL, by Hitachi then to use centrifuge, Ltd. manufacture) collect microsphere by centrifugal (speed of rotation: about 2500rpm, 5 minutes).Microsphere is scattered in distilled water again, and centrifugal further, eccysis dissociant etc.By collected microsphere redispersion in a small amount of distilled water, and add mannitol (0.885g).By this mixture with freezer dryer (DF-01H, ULVAC) lyophilization, obtain microcapsule powder.The content of compound (I) in gained microcapsule powder is 12.9%.

Embodiment 14

Poly(D,L-lactide-co-glycolide (weight average molecular weight Mw:13800) (13.2307g) is dissolved in dichloromethane (24.74g).Take this solution (13.43g) and blended with the aqueous solution of the acetate (1.4900g) being dissolved with compound (I) in distilled water (1.21g), then small-sized homogenizer (KINEMATICA) emulsifying is used, form W/O emulsion (speed of rotation: about 10000rpm, 30 seconds).Then, W/O emulsion is adjusted to 32 DEG C and pours 0.1% (w/w) polyvinyl alcohol (EG-40 being adjusted to about 18 DEG C in advance into, by NipponSyntheticChemicalIndustryCo., Ltd. manufacture) in aqueous solution (1 liter), then use turbine type homo-mixer (being manufactured by Tokushukika) to carry out second emulsifying, obtain W/O/W emulsion (whirling speed: about 7000rpm).Stir W/O/W emulsion about 3 hours (in water drying steps), and use 75 μm of standard screen clothes to sieve, (HIMACCR5DL, by Hitachi then to use centrifuge, Ltd. manufacture) collect microsphere by centrifugal (speed of rotation: about 2500rpm, 5 minutes).Microsphere is scattered in distilled water again, and centrifugal further, eccysis dissociant etc.By collected microsphere redispersion in a small amount of distilled water, and add mannitol (0.885g).By this mixture with freezer dryer (DF-01H, ULVAC) lyophilization, obtain microcapsule powder.The content of compound (I) in gained microcapsule powder is 13.1%.

Embodiment 15

Poly(D,L-lactide-co-glycolide (weight average molecular weight Mw:20000) (6.24g) is dissolved in dichloromethane (10.92g).Take this solution (13.28g) and with the solution blending of acetate (1.3619g) being dissolved with compound (I) in methanol (5.73g), obtain oil phase.Then, oil phase is poured into 0.1% (w/w) polyvinyl alcohol (EG-40 being adjusted to about 18 DEG C in advance, by NipponSyntheticChemicalIndustryCo., Ltd. manufacture) in aqueous solution (1 liter), and use turbine type homo-mixer (being manufactured by Tokushukika) emulsifying, preparation O/W emulsion (whirling speed: about 7000rpm).Stir O/W emulsion about 3 hours (in water drying steps), and use 75 μm of standard screen clothes to sieve, (HIMACCR5DL, by Hitachi then to use centrifuge, Ltd. manufacture) collect microsphere by centrifugal (speed of rotation: about 2500rpm, 5 minutes).Microsphere is scattered in distilled water again, and centrifugal further, eccysis dissociant etc.By collected microsphere redispersion in a small amount of distilled water, and add mannitol (0.740g).By this mixture with freezer dryer (DF-01H, ULVAC) lyophilization, obtain microcapsule powder.The content of compound (I) in gained microcapsule powder is 15.7%.

Embodiment 16

Poly(D,L-lactide-co-glycolide (weight average molecular weight Mw:22000) (6.26g) is dissolved in dichloromethane (10.98g).Take this solution (13.27g) and with the solution blending of acetate (1.3627g) being dissolved with compound (I) in methanol (5.61g), obtain oil phase.Then, oil phase is poured into 0.1% (w/w) polyvinyl alcohol (EG-40 being adjusted to about 18 DEG C in advance, by NipponSyntheticChemicalIndustryCo., Ltd. manufacture) in aqueous solution (1 liter), and use turbine type homo-mixer (being manufactured by Tokushukika) emulsifying, preparation O/W emulsion (whirling speed: about 7000rpm).Stir O/W emulsion about 3 hours (in water drying steps), and use 75 μm of standard screen clothes to sieve, (HIMACCR5DL, by Hitachi then to use centrifuge, Ltd. manufacture) collect microsphere by centrifugal (speed of rotation: about 2500rpm, 5 minutes).Microsphere is scattered in distilled water again, and centrifugal further, eccysis dissociant etc.By collected microsphere redispersion in a small amount of distilled water, and add mannitol (0.740g).By this mixture with freezer dryer (DF-01H, ULVAC) lyophilization, obtain microcapsule powder.The content of compound (I) in gained microcapsule powder is 15.9%.

Embodiment 17

Poly(D,L-lactide-co-glycolide (weight average molecular weight Mw:23000) (7.02g) is dissolved in dichloromethane (13.18g).Take this solution (15.64g) and with the solution blending of acetate (0.6799g) being dissolved with compound (I) in methanol (5.57g), obtain oil phase.Then, oil phase is poured into 0.1% (w/w) polyvinyl alcohol (EG-40 being adjusted to about 18 DEG C in advance, by NipponSyntheticChemicalIndustryCo., Ltd. manufacture) in aqueous solution (1 liter), and use turbine type homo-mixer (being manufactured by Tokushukika) emulsifying, preparation O/W emulsion (whirling speed: about 7000rpm).Stir O/W emulsion about 3 hours (in water drying steps), and use 75 μm of standard screen clothes to sieve, (HIMACCR5DL, by Hitachi then to use centrifuge, Ltd. manufacture) collect microsphere by centrifugal (speed of rotation: about 2500rpm, 5 minutes).Microsphere is scattered in distilled water again, and centrifugal further, eccysis dissociant etc.By collected microsphere redispersion in a small amount of distilled water, and add mannitol (0.740g).By this mixture with freezer dryer (DF-01H, ULVAC) lyophilization, obtain microcapsule powder.The content of compound (I) in gained microcapsule powder is 8.5%.

Embodiment 18

Poly(D,L-lactide-co-glycolide (weight average molecular weight Mw:25000) (7.01g) is dissolved in dichloromethane (13.11g).Take this solution (15.50g) and with the solution blending of acetate (0.6870g) being dissolved with compound (I) in methanol (5.61g), obtain oil phase.Then, oil phase is poured into 0.1% (w/w) polyvinyl alcohol (EG-40 being adjusted to about 18 DEG C in advance, by NipponSyntheticChemicalIndustryCo., Ltd. manufacture) in aqueous solution (1 liter), and use turbine type homo-mixer (being manufactured by Tokushukika) emulsifying, preparation O/W emulsion (whirling speed: about 7000rpm).Stir O/W emulsion about 3 hours (in water drying steps), and use 75 μm of standard screen clothes to sieve, (HIMACCR5DL, by Hitachi then to use centrifuge, Ltd. manufacture) collect microsphere by centrifugal (speed of rotation: about 2500rpm, 5 minutes).Microsphere is scattered in distilled water again, and centrifugal further, eccysis dissociant etc.By collected microsphere redispersion in a small amount of distilled water, and add mannitol (0.768g).By this mixture with freezer dryer (DF-01H, ULVAC) lyophilization, obtain microcapsule powder.The content of compound (I) in gained microcapsule powder is 9.1%.

Comparative example 1

In this instance, to the dichloromethane solution of Poly(D,L-lactide-co-glycolide be dissolved with and in distilled water, be dissolved with compound (II) (Ac-D-Tyr-D-Trp-Asn-Thr-Phe-AzaGly-Leu-Arg (Me)-Trp-NH2) (serial number: the solution mixture of the aqueous solution of acetate 2) suspends with mini blender, this solution mixture gelation.Therefore, W/O emulsion cannot be obtained.

Testing example 1

By the microcapsule powder of embodiment 15 (in free cpds (I), for 9.6mg) be scattered in disperse medium (0.9mL) (being dissolved with the solution of carboxymethyl cellulose (5.50mg), polyoxyethylene sorbitan monoleate (0.90mg) and mannitol (45.0mg)), then use 22G entry needle through the subcutaneous back given to rat.In predetermined time interval after giving, from tail venous puncture blood sample, and measure the concentration of compound in blood plasma (I).As a result, the slow-release time of confirmation compound (I) is about 23 weeks.

Testing example 2

By the microcapsule powder of embodiment 16 (in free cpds (I), for 9.6mg) be scattered in disperse medium (0.9mL) (being dissolved with the solution of carboxymethyl cellulose (5.50mg), polyoxyethylene sorbitan monoleate (0.90mg) and mannitol (45.0mg)), then use 22G entry needle through the subcutaneous back given to rat.In predetermined time interval after giving, from tail venous puncture blood sample, and measure the concentration of compound in blood plasma (I).As a result, the slow-release time of confirmation compound (I) is about 26 weeks.

Testing example 3

By the microcapsule powder of embodiment 17 (in free cpds (I), for 9.6mg) be scattered in disperse medium (0.9mL) (being dissolved with the solution of carboxymethyl cellulose (5.50mg), polyoxyethylene sorbitan monoleate (0.90mg) and mannitol (45.0mg)), then use 22G entry needle through the subcutaneous back given to rat.In predetermined time interval after giving, from tail venous puncture blood sample, and measure the concentration of compound in blood plasma (I).As a result, the slow-release time of confirmation compound (I) is about 24 weeks.

Testing example 4

By the microcapsule powder of embodiment 5 (in free cpds (I), for 4.8mg) be scattered in disperse medium (0.9mL) (being dissolved with the solution of carboxymethyl cellulose (5.50mg), polyoxyethylene sorbitan monoleate (0.90mg) and mannitol (45.0mg)), then use 22G entry needle through the subcutaneous back given to rat.In predetermined time interval after giving, from tail venous puncture blood sample, and measure the concentration of compound in blood plasma (I).As a result, the slow-release time of confirmation compound (I) is about 18 weeks.

Testing example 5

By the microcapsule powder of embodiment 6 (in free cpds (I), for 4.8mg) be scattered in disperse medium (0.9mL) (being dissolved with the solution of carboxymethyl cellulose (5.50mg), polyoxyethylene sorbitan monoleate (0.90mg) and mannitol (45.0mg)), then use 22G entry needle through the subcutaneous back given to rat.In predetermined time interval after giving, from tail venous puncture blood sample, and measure the concentration of compound in blood plasma (I).As a result, the slow-release time of confirmation compound (I) is about 17 weeks.

Testing example 6

By the microcapsule powder of embodiment 7 (in free cpds (I), for 4.8mg) be scattered in disperse medium (0.9mL) (being dissolved with the solution of carboxymethyl cellulose (5.50mg), polyoxyethylene sorbitan monoleate (0.90mg) and mannitol (45.0mg)), then use 22G entry needle through the subcutaneous back given to rat.In predetermined time interval after giving, from tail venous puncture blood sample, and measure the concentration of compound in blood plasma (I).As a result, the slow-release time of confirmation compound (I) is about 21 weeks.

Slow releasing preparation of the present invention is long-time slowly and stably release tumor migration chalone derivant, and also plays the drug effect of tumor migration chalone derivant for a long time.In addition, slow releasing preparation of the present invention can reduce administration frequency, improves the convenience of patient thus, and can be used as clinical medicine.

Applicant states, the present invention illustrates detailed features of the present invention and method detailed by above-described embodiment, but the present invention is not limited to above-mentioned detailed features and method detailed, namely do not mean that the present invention must rely on above-mentioned detailed features and method detailed could be implemented.Person of ordinary skill in the field should understand, any improvement in the present invention, to equivalence replacement and the interpolation of auxiliary element, concrete way choice etc. that the present invention selects component, all drops within protection scope of the present invention and open scope.

Claims (10)

1. be used for the treatment of a slow releasing preparation for cancer, comprise the compound (I) shown in following formula or its salt:

Ac-D-Tyr-Hyp-Asn-Thr-Phe-AzaGly-Leu-Arg(Me)-Trp-NH ₂；

And weight average molecular weight is Poly(D,L-lactide-co-glycolide or its salt of 10000 to 30000.

2. slow releasing preparation according to claim 1, is characterized in that, the weight average molecular weight of described Poly(D,L-lactide-co-glycolide or its salt is 13000 to 18000.

3. slow releasing preparation according to claim 1 and 2, is characterized in that, the weight-average molecular weight/number-average molecular weight of described Poly(D,L-lactide-co-glycolide or its salt is 1.2 to 4.0, is preferably 1.5 to 3.5.

4. the slow releasing preparation according to any one of claim 1-3, is characterized in that, described slow releasing preparation is the preparation of 4 to 9 months slow release.

5. the slow releasing preparation according to any one of claim 1-4, is characterized in that, described slow releasing preparation is treatment or the preventive of cancer.

6. slow releasing preparation according to claim 5, is characterized in that, described cancer is pulmonary carcinoma, gastric cancer, hepatocarcinoma, cancer of pancreas, colorectal cancer, rectal cancer, colon cancer, carcinoma of prostate, ovarian cancer, cervical cancer, breast carcinoma, renal cancer, bladder cancer or cerebroma.

7. the slow releasing preparation according to any one of claim 1-6, it is characterized in that, relative to Poly(D,L-lactide-co-glycolide or its salt, the use amount of compound (I) or its salt is 0.01 to 50% (w/w), be preferably 0.1 to 30% (w/w), be particularly preferably 1 to 20% (w/w).

8. manufacture a method for the slow releasing preparation as described in any one of claim 1-7, comprise and make following emulsion carry out drying in water:

W/O/W emulsion, it is obtained by emulsifying W/O emulsion, and described W/O emulsion comprises interior aqueous phase containing the compound (I) shown in following formula or its salt and the oil phase containing Poly(D,L-lactide-co-glycolide or its salt;

Ac-D-Tyr-Hyp-Asn-Thr-Phe-AzaGly-Leu-Arg(Me)-Trp-NH ₂。

9. method according to claim 8, is characterized in that, in described interior aqueous phase, compound (I) or the concentration of its salt in water are 0.001 to 90% (w/w), is preferably 0.01 to 80% (w/w).

10. manufacture a method for the slow releasing preparation as described in any one of claim 1-7, comprise and make following emulsion carry out drying in water:

O/W emulsion, it is obtained by emulsifying oil phase, and described oil phase comprises the compound (I) shown in following formula or its salt and Poly(D,L-lactide-co-glycolide or its salt;

Ac-D-Tyr-Hyp-Asn-Thr-Phe-AzaGly-Leu-Arg(Me)-Trp-NH ₂。