CN1927178A - Slow release injection loading both tetrazole violet and cytotoxic drug - Google Patents

Abstract

Disclosed is an anticancer slow release injection carrying both tetrazole Ionone and cytotoxic drugs, which comprises slow release micro-balloons and dissolvent, wherein the slow release microballoons comprise anti-cancer active constituents and slow release auxiliary materials, the dissolvent being specific dissolvent containing suspension adjuvant. The anticancer active constituents include tetrazole lonone and cytotoxic drugs selected from adriamycin, pidorubicin, Melphalan, 4-hydroperoxycyclophosphamide, actinomycin D, Vinorelbine, Tamoxifen, Tallimustine, Atrimustine, Semustine, ranimustine, the slow release auxiliary materials are selected from polylactic acid and its copolymer, polyethylene glycol mono methyl, polyethylene glycol / polylactic acid copolymer, EVAc, aliphatic acid and sebacylic acid copolymer, the viscosity of the suspension adjuvant is 100-3000cp (at 25-30 deg C), and is selected from sodium carboxymethylcellulose, the slow release microspheres can also be prepared into slow release implanting agent, for injection or placement in or around tumor with the period of local drug release can be about 40-50 days, as a result, the agent can be employed individually or used along with non-operative treatment methods such as chemotherapy pharmaceuticals and radiation.

Description

The slow releasing injection of a kind of loading both tetrazole violet and cell toxicity medicament

(1) technical field

The present invention relates to slow releasing injection of a kind of loading both tetrazole violet and cell toxicity medicament and preparation method thereof, belong to technical field of pharmaceuticals.Particularly, the invention provides a kind of anticancer medicine slow-release preparation containing that contains tetrazole violet and cell toxicity medicament, be mainly slow releasing injection and sustained-release implant.

(2) background technology

As one of cancer conventional treatments, chemotherapeutics has been widely used in the treatment of multiple malignant tumor, and action effect is comparatively obvious.Yet its tangible general toxicity has greatly limited the application of this medicine.

Because entity tumor excessive expansion hypertrophy, the viscosity of matter was high than its normal surrounding tissue all between matter pressure, tissue elasticity pressure, fluid pressure reached therebetween, therefore, conventional chemotherapy, be difficult to tumor by local and form effective drug level, referring to " placing cisplatin adding system carmustine treatment rat brain tumor in the tumor " " surgery tumor magazine " 69 phase 76-82 pages or leaves (1998) (Kong Q et al., J Surg Oncol.1998 Oct such as Kong Qingzhongs; 69 (2): 76-82), improve the restriction that dosage is subjected to general reaction again merely.Pharmaceutical topical application may solve the problem of drug level to a certain extent, yet operation techniques such as medicine implantation are complicated, traumatic big, the various complication such as, infection hemorrhage, immunity reduction, also can cause or quicken the diffusion and the transfer of tumor except that easily causing.In addition, the preparation of perioperatively itself and expensive expense usually influence its effective enforcement.In addition, the cancer drug therapy of low dosage not only can increase the Drug tolerance of cancerous cell, but also can promote its infiltrative growth " (referring to beam etc. " increased the Drug tolerance of human lung carcinoma cell and external wetting capacity after the cancer therapy drug pulse screening and with the change of gene expression " " international journal of cancer " 111 phase 484-93 page or leaf (2004) (Liang Y; et al., Int J Cancer.2004; 111 (4): 484-93)).

Moreover, blood vessel in the mesenchyma stroma of tumors, connective tissue, stromatin, fibrin and collagen protein etc. not only provide support and requisite nutrient substance for the growth of tumor cell, also influenced chemotherapeutics around tumor and the infiltration in the tumor tissues and diffusion (carry and to wait " situation of extracellular matrix to entity tumor in the medicine influence of turning round " " cancer research " 60 phase 2497-503 page or leaf (2000) (Netti PA referring to the Buddhist nun, Cancer Res.2000,60 (9): 2497-503)).Therefore, be convenient to keep high drug level and increase tumor cell the preparation and the method for the sensitivity of medicine just become an important subject at tumor by local.

(3) summary of the invention

Tetrazole violet and cell toxicity medicament have been widely used in multiple entity tumor of treatment or hematological system tumor etc. both at home and abroad as new or commonly used cancer therapy drug.Yet in application process, its tangible general toxicity has greatly limited the application of this medicine.

Be effectively to improve tumor by local drug level, reduce the drug level of medicine in blood circulation, people have studied the slow-released system that contains cancer therapy drug, comprise that sustained-release micro-spheres (capsule) (sees: Chinese patent (patent No. ZL00809160.9; Application number 91109723.6), Ciftci K etc. " with the polylactic acid microsphere treatment entity tumor that contains fluorouracil and the research of drug release " " drug development technology " (Pharm Dev Technol.) 2 (2): 151-60,1997), sustained-release implant (sees: China Patent No. ZL96115937.5; ZL97107076.8) etc.Yet, solid sustained-release implant (China Patent No. ZL96115937.5; ZL97107076.8) and existing as be used for the treatment of the cerebral tumor (ZL00809160.9) sustained-release micro-spheres or United States Patent (USP) (US5,651,986) and all have problem such as be not easy more than operation, weak curative effect, the complication.In addition, the sensitivity that many entity tumors are drawn together tetrazole violet to anticancer medicated bag is relatively poor, and is easy to generate drug resistance in therapeutic process.The present invention finds that cell toxicity medicament of mentioning among the present invention and tetrazole violet share and can make its antitumaous effect strengthen (the following cell toxicity medicament that the tetrazole violet antitumaous effect will be increased mutually is referred to as the tetrazole violet synergist) mutually.In addition, with the assembly packaging of tetrazole violet or tetrazole violet and its synergist in specific slow-release auxiliary material and be equipped with special solvent and make drug level that anti-cancer medicine sustained-release injection not only can greatly improve tumor by local, reduce the drug level of medicine in blood circulation, reduce the toxicity of medicine to normal structure, can also greatly make things convenient for the medicine injection, reduce operation technique complication, reduce patient's expense.The cell toxicity medicament decapacitation suppresses can also increase the sensitivity of tumor cell to cancer therapy drug outside the tumor growth.

Conventional route administrations such as main oral administration of tetrazole violet or intravenous injection, administering mode of the present invention is the local sustained release administration, obviously reduces the toxic action of its whole body in the therapeutic effect that significantly strengthens medicine.The cancer therapy drug of having reported of using through the slow release approach is increasing day by day, finds to have only several chemical compounds to enter clinical research yet existing countless chemical compounds are entered the screening back, has only 2-4 chemical compound to get the Green Light and comes into the market.Therefore, in having the medicine of active anticancer, be not all slow release effects that all can in slow-release auxiliary material, reach effective release yet.

Pharmaceutic adjuvant have hundreds of more than, pharmaceutic adjuvant with slow releasing function, particularly can slowly release ratio be non-apparent in the regular hour in human body or animal body with selected tetrazole violet among the present invention, but specific slow-release auxiliary material need could be determined through a large amount of creative works with the selection of slow releasing pharmaceutical combination.The data of release characteristics need could obtain through a large amount of creationary experiments in inside and outside in the related data, particularly animal body, are not just can determine to have unobviousness through limited experiment.

The above unexpected main contents of the present invention of finding to constitute.

The present invention is directed to the deficiencies in the prior art, a kind of new slow releasing injection that contains tetrazole violet and/or cell toxicity medicament is provided.

Tetrazole violet slow releasing injection of the present invention is made up of sustained-release micro-spheres and solvent.Particularly, this slow-releasing anticarcinogen injection is grouped into by following one-tenth:

(A) sustained-release micro-spheres comprises:

Anticancer effective component 0.5-60%

Slow-release auxiliary material 40-99%

Suspending agent 0.0-30%

More than be weight percentage

With

(B) solvent is for common solvent or contain the special solvent of suspending agent.

Wherein,

Anticancer effective component be tetrazole violet and be selected from melphalan, 4H-peroxide cyclophosphamide, vinorelbine, tamoxifen, methotrexate, amycin, epirubicin, actinomycin D, tallimustine, atrimustine, semustine and or the combination of the cell toxicity medicament of Ranimustine;

Slow-release auxiliary material range of viscosities IV (dl/g) is 0.1～0.8, be selected from poly-dl-lactide (D, L-PLA), poly-dl-lactide/ethanol copolymer (D, L-PLGA), monomethyl polyethylene glycol (MPEG-PLA), monomethyl polyethylene glycol copolymer (MPEG-PLGA), polyethylene glycol (PLA-PEG-PLA), polyethylene glycol copolymer (PLGA-PEG-PLGA), end carboxyl polylactic acid (PLA-COOH), end carboxyl polylactic acid/ethanol copolymer (PLGA-COOH), polifeprosan, bis-fatty acid and decanedioic acid copolymer [P (FAD-SA)], poly-(erucic acid dimer-decanedioic acid) [P (EAD-SA)], poly-(fumaric acid-decanedioic acid) [P (FA-SA)], ethylene vinyl acetate copolymer (EVAc), polylactic acid (PLA), the copolymer of polyglycolic acid and hydroxyacetic acid (PLGA), PPDO (PDO), PTMC (PTMC), xylitol, oligosaccharide, chrondroitin, chitin, hyaluronic acid, collagen protein, gelatin, one of albumin glue or its combination; Suspending agent is selected from one of sodium carboxymethyl cellulose, (iodine) glycerol, simethicone, propylene glycol, carbomer, mannitol, sorbitol, surfactant, soil temperature 20, soil temperature 40 and soil temperature 80 or its combination.

Tetrazole violet shared percentage by weight in slow releasing agent is decided because of concrete condition, can be 0.1%-50%, is good with 1%-40%, and 5%-30% is best.

The percentage by weight of above-mentioned cell toxicity medicament in slow releasing agent is good from 0.01%-99.99% with 1%-50%, is best with 5%-30%.

When the anticancer effective component in the medicament slow-release microsphere only is tetrazole violet or cell toxicity medicament, slow-releasing anticarcinogen injection is mainly used in the tetrazole violet of other approach application of increase or the action effect of cell toxicity medicament, or is used for the potentiation to radiotherapy or other therapies.When the anticancer effective component in the medicament slow-release microsphere only was tetrazole violet or its synergist (cell toxicity medicament), the application of slow-releasing anticarcinogen injection and potentiation mode were:

(1) contain the slow releasing injection local injection of tetrazole violet, and cell toxicity medicament is used through other approach;

(2) local injection contains the slow releasing injection of cell toxicity medicament, and other approach are used tetrazole violet;

(3) local injection contains the slow releasing injection and the slow releasing injection that contains cell toxicity medicament of tetrazole violet; Or

(4) local injection contains the slow releasing injection of tetrazole violet and cell toxicity medicament.

The slow-releasing anticarcinogen injection of topical application also is used for the potentiation to radiotherapy or other therapies.Other approach refer to, but, be not limited to tremulous pulse, vein, abdominal cavity, subcutaneous, intracavitary administration.

Anticancer effective component tetrazole violet and/or the cell toxicity medicament percentage by weight in medicament slow-release microsphere is 0.5%-60%, is good with 2%-40%, is best with 5%-30%.The weight ratio of tetrazole violet and cell toxicity medicament is 1-9: 1 to 1: 1-9, with 1-2: 1 serves as preferred.

Anticancer effective component in the slow-releasing anticarcinogen injection microsphere of the present invention is preferably as follows, and all is weight percentage:

The combination of the tetrazole violet of 2-40% and the melphalan of 2-40%, 4H-peroxide cyclophosphamide, vinorelbine, tamoxifen, methotrexate, amycin, epirubicin, actinomycin D, tallimustine, atrimustine, semustine or Ranimustine.

Slow-release auxiliary material can be various water solublity or water-insoluble macromolecule polymer, and slow-release auxiliary material is selected from poly-dl-lactide, poly-dl-lactide/ethanol copolymer, the monomethyl polyethylene glycol, monomethyl polyethylene glycol copolymer, polyethylene glycol, the polyethylene glycol copolymer, end carboxyl polylactic acid, end carboxyl polylactic acid/ethanol copolymer, polifeprosan, bis-fatty acid and decanedioic acid copolymer, poly-(erucic acid dimer-decanedioic acid), poly-(fumaric acid-decanedioic acid), ethylene vinyl acetate copolymer, polylactic acid, the copolymer of polyglycolic acid and hydroxyacetic acid, xylitol, oligosaccharide, chrondroitin, chitin, hyaluronic acid, collagen protein, one of gelatin and albumin glue or its combination.

Slow-release auxiliary material and percentage by weight thereof are most preferably as follows in the sustained-release micro-spheres of the present invention:

(1) PLA of 55-90%;

(2) PLGA of 50-90%;

(3) polifeprosan of 50-85%;

(4) bis-fatty acid of 55-90% and decanedioic acid copolymer;

(5) EVAc of 55-90%;

(6) xylitol of 40-95%, oligosaccharide, chrondroitin, chitin, hyaluronic acid, collagen protein, gelatin or white tempera; Or

(7) poly-dl-lactide of 40-95%, poly-dl-lactide/ethanol copolymer, monomethyl polyethylene glycol, monomethyl polyethylene glycol copolymer, polyethylene glycol, polyethylene glycol copolymer, end carboxyl polylactic acid or end carboxyl polylactic acid/ethanol copolymer.

In various high molecular polymers, with polylactic acid, decanedioic acid, the mixture or the copolymer that contain the macromolecule polymer of polylactic acid or certain herbaceous plants with big flowers diacid is first-selection, mixture and copolymer can be selected from, but be not limited to the mixture or the copolymer of the mixture of PLA, PLGA, glycolic and hydroxy carboxylic acid, certain herbaceous plants with big flowers diacid and fragrant polyanhydride or aliphatic polyanhydride.The blend ratio of glycolic and hydroxy carboxylic acid is 10/90-90/10 (weight), preferably 25/75-75/25 (weight).The method of blend is arbitrarily.Content when glycolic and hydroxy carboxylic acid copolymerization is respectively percentage by weight 10-90% and 90-10%.The representative of fragrance polyanhydride is polifeprosan [poly-(1,3-two (to the carboxyl phenoxy group) propane-decanedioic acid) (p (CPP-SA)), bis-fatty acid-decanedioic acid copolymer (PFAD-SA)], poly-(erucic acid dimer-decanedioic acid) [P (EAD-SA)] and poly-(fumaric acid-decanedioic acid) [P (FA-SA)] etc.Content during to carboxylic phenoxypropane (p-CPP) and decanedioic acid copolymerization is respectively percentage by weight 10-60% and 20-90%, and the blend weight ratio is 10-40: 50-90, preferably weight ratio 15-30: 65-85.

The molecular weight peak value of polylactic acid can be, but is not limited to, 5000-100, and 000, but with 20,000-60,000 is preferred, with 5,000-30,000 for most preferably; The molecular weight of polyglycolic acid can be, but is not limited to, 5000-100, and 000, but with 5,000-50,000 is preferred, with 10,000-30,000 for most preferably; Above polyhydroxy acid can singly select or multiselect.When singly selecting, serve as preferred with the copolymer (PLGA) of polylactic acid (PLA) or hydroxy carboxylic acid and glycolic, the molecular weight of copolymer can be, but is not limited to, 5000-100,000, but with 20,000-60,000 be preferably, with 30,000-50,000 for most preferably; When multiselect, compound polymer or the copolymer formed with macromolecule polymer or different macromolecule polymer serve as preferred, with the compound polymer that contains different molecular weight polylactic acid or decanedioic acid or copolymer for most preferably, as, but be not limited to, molecular weight is 1000 to 30000 polylactic acid with molecular weight is that 20000 to 50000 polylactic acid mixes, molecular weight is 10000 to 30000 polylactic acid with molecular weight is that 30000 to 80000 PLGA mixes, molecular weight is that 20000 to 30000 polylactic acid mixes with decanedioic acid, molecular weight is that 30000 to 80000 PLGA mixes with decanedioic acid.Used polylactic acid serves as preferred with Poly-L-lactic acid (L-PLA).Poly-L-lactic acid (L-PLA) range of viscosities IV (dl/g) is 0.2～0.8, and glass transition temperature range is 55～65 ℃, 175～185 ℃ of fusing points.

Except that above-mentioned slow-release auxiliary material, also can select for use other materials to see the United States Patent (USP) (patent No. 4757128; 4857311; 4888176; 4789724) and in " pharmaceutic adjuvant complete works " (the 123rd page, Sichuan science tech publishing house published in 1993, Luo Mingsheng and Gao Tianhui chief editor) have a detailed description.In addition, Chinese patent (application number 96115937.5; 91109723.6; 9710703.3; 01803562.0) and U.S.'s patent of invention (patent No. 5,651,986) also enumerated some pharmaceutic adjuvant, comprise filler, solubilizing agent, absorption enhancer, film former, gellant, system (or causing) hole agent, excipient or blocker etc.

For regulating drug releasing rate or changing other characteristic of the present invention, can change the composition and the proportioning of monomer component or molecular weight, interpolation or the adjusting pharmaceutic adjuvant of polymer, add the water-soluble low-molecular chemical compound, as, but be not limited to various sugar or salt etc.Wherein sugar can be, but is not limited to, xylitol, oligosaccharide, (sulphuric acid) chrondroitin and chitin etc., and wherein salt can be, but is not limited to, potassium salt and sodium salt etc.; Also can add other pharmaceutic adjuvant, as but be not limited to filler, solubilizing agent, absorption enhancer, film former, gellant, system (or causing) hole agent, excipient or blocker etc.

In the slow releasing injection, drug sustained release system can be made into microsphere, sub-micro ball, microemulsion, nanosphere, granule or spherical piller, makes the injection use then with after the injection solvent mixes.In various slow releasing injection, serve as preferred with the suspension type slow releasing injection, the suspension type slow releasing injection is the preparation that the drug sustained release system that will contain anticancer component is suspended in gained in the injection, used slow-release auxiliary material is a kind of or its combination in the above-mentioned slow-release auxiliary material, and used solvent is common solvent or the special solvent that contains suspending agent.Common solvent is, but is not limited to the buffer that distilled water, water for injection, physiology are prepared towards liquid, dehydrated alcohol or various salt.The purpose of suspending agent is the pastille microsphere that effectively suspends, thereby is beneficial to the usefulness of injection.Be convenient injection, the viscosity of suspending agent is 100cp-3000cp (20 ℃-30 ℃ time), preferred 1000cp-3000cp (20 ℃-30 ℃ time), most preferably 1500cp-3000cp (20 ℃-30 ℃ time).Suspending agent is selected from one of sodium carboxymethyl cellulose, (iodine) glycerol, simethicone, propylene glycol, carbomer, mannitol, sorbitol, surfactant, soil temperature 20, soil temperature 40 and soil temperature 80 or its combination.

The content of suspending agent in common solvent is decided because of its characteristic, can be 0.1-30% and decides because of concrete condition.Consisting of of preferred suspending agent:

A) 0.5-5% sodium carboxymethyl cellulose+0.1-0.5% soil temperature 80; Or

B) 5-20% mannitol+0.1-0.5% soil temperature 80; Or

C) 0.5-5% sodium carboxymethyl cellulose+5-20% sorbitol+0.1-0.5% soil temperature 80.

The kind of solvent is then depended in the preparation of solvent, and common solvent has commercially available, also can make by oneself, and as distilled water, water for injection, physiology buffer towards liquid, dehydrated alcohol or the preparation of various salt, but must be in strict accordance with related standards.Special solvent need be considered the kind of suspending agent and the medicine that composition, solvent suspended, composition, character and the requirement thereof of sustained-release micro-spheres (or microcapsule) and the preparation method of injection, as sodium carboxymethyl cellulose (1.5%)+mannitol and/or sorbitol (15%) and/or Tween 80 (0.1%) are dissolved in the normal saline mutually deserved solvent, viscosity is at 10cp-650cp (20 ℃-30 ℃ time).

The present invention finds to influence medicine and/or sustained-release micro-spheres suspends and/or the key factor of injection is the viscosity of solvent, and viscosity is big more, and suspension effect is good more, and syringeability is strong more.This unexpected one of main index characteristic of the present invention of finding to have constituted.The viscosity of solvent depends on the viscosity of suspending agent, and the viscosity of suspending agent is 100cp-3000cp (20 ℃-30 ℃ time), preferred 1000cp-3000cp (20 ℃-30 ℃ time), most preferably 1500cp-3000cp (20 ℃-30 ℃ time).According to the viscosity of the prepared solvent of this condition is 10cp-650cp (20 ℃-30 ℃ time), preferred 20cp-650cp (20 ℃-30 ℃ time), most preferably 60cp-650cp (20 ℃-30 ℃ time).

The preparation of injection has several different methods, and a kind of is that the sustained-release microparticle (A) of suspending agent for " 0 " directly mixed in special solvent, obtains corresponding sustained-release microparticle injection; Another kind is that suspending agent is not mixed in special solvent or common solvent for the sustained-release microparticle (A) of " 0 ", obtains corresponding sustained-release microparticle injection; Another is that sustained-release microparticle (A) is mixed in common solvent, adds the suspending agent mixing then, obtains corresponding sustained-release microparticle injection.Except, also can earlier sustained-release microparticle (A) be mixed and in special solvent, make corresponding suspension, with the moisture in ways such as the vacuum drying removal suspension, special solvent of reuse or common solvent suspendible obtain corresponding sustained-release microparticle injection afterwards then.Above method just is illustrative rather than definitive thereof the present invention.It should be noted that suspended drug or sustained-release micro-spheres (or microcapsule) concentration in injection decide because of specifically needing, can be, but be not limited to, 10-400mg/ml, but be preferably with 30-300mg/ml, with 50-200mg/ml most preferably.The viscosity of injection is 50cp-1000cp (20 ℃-30 ℃ time), preferred 100cp-1000cp (20 ℃-30 ℃ time), most preferably 200cp-650cp (20 ℃-30 ℃ time).This viscosity is applicable to 18-22 injection needle and special bigger (to 3 millimeters) injection needle of internal diameter.

The preparation method of slow releasing injection is arbitrarily, available some kinds of methods preparation: as, but be not limited to, mixing method, fusion method, dissolution method, spray drying method for preparation microsphere, dissolution method are made micropowder, liposome bag medicine method and emulsion process etc. in conjunction with freezing (drying) comminuting method.Serve as preferred wherein with dissolution method (being the solvent volatility process), seasoning, spray drying method and emulsion process.Microsphere then can be used for preparing above-mentioned various slow releasing injection, and its method is arbitrarily.The particle size range of used microsphere can be between 5-400um, serving as preferred between the 10-300um, with between the 20-200um for most preferably.

Microsphere also can be used for preparing other slow releasing injection, as gel injection, block copolymer micelle injection.Wherein, block copolymer micelle is formed in aqueous solution by hydrophobic-hydrophilic block copolymers, has spherical inner core-shell mechanism, and hydrophobic block forms kernel, and hydrophilic block forms shell.The carrier micelle injection enters the purpose that reaches control drug release or targeted therapy in the body.Used pharmaceutical carrier is above-mentioned any one or its combination.Wherein preferred molecular weight is the hydrophilic block of the Polyethylene Glycol (PEG) of 1000-15000 as the micelle copolymer, and preferred biological degradation polyalcohol (as PLA, polylactide, polycaprolactone and copolymer thereof (molecular weight 1500-25000)) is as the hydrophobic block of micelle copolymer.The particle size range of block copolymer micelle can be between 10-300um, between the 20-200um serving as preferred.Gel injection system is dissolved in some amphipathic solvent with biological degradation polyalcohol (as PLA, PLGA or DL-LA and epsilon-caprolactone copolymer), adds medicine miscible with it (or suspendible) back again and forms flowability gel preferably, can be through tumor week or intratumor injection.In case inject, amphipathic solvent diffuses to body fluid very soon, the moisture in the body fluid then infiltrates gel, makes polymer cure, slowly discharges medicine.

Sustained-release micro-spheres also can be used for preparing sustained-release implant, used pharmaceutic adjuvant can be any or multiple material in the above-mentioned pharmaceutic adjuvant, but with the high molecular weight water soluble polymer is main separation, in various high molecular polymers, with polylactic acid, the certain herbaceous plants with big flowers diacid, the mixture or the copolymer that contain the macromolecule polymer of polylactic acid or certain herbaceous plants with big flowers diacid are first-selection, mixture and copolymer can be selected from, but be not limited to PLA, PLGA, the mixture of PLA and PLGA, the mixture or the copolymer of certain herbaceous plants with big flowers diacid and fragrant polyanhydride or aliphatic polyanhydride, bis-fatty acid and decanedioic acid copolymer (PFAD-SA), poly-(erucic acid dimer-decanedioic acid) [P (EAD-SA)], poly-(fumaric acid-decanedioic acid) [P (FA-SA)].Polylactic acid (PLA) is 10/90-90/10 (weight) with the blend ratio of polyglycolic acid, preferably 25/75-75/25 (weight).The method of blend is arbitrarily.Content when glycolic and lactic acid copolymerization is respectively percentage by weight 10-90% and 90-10%.The representative of fragrance polyanhydride is to carboxy phenyl propane (p-CPP), content during to carboxy phenyl propane (p-CPP) and the copolymerization of certain herbaceous plants with big flowers diacid is respectively percentage by weight 10-60% and 20-90%, the blend weight ratio is 10-40: 50-90, preferably weight ratio 15-30: 65-85.

Another form of anticancer medicine slow-release preparation containing of the present invention is that anticancer medicine slow-release preparation containing is a sustained-release implant.The effective ingredient of anticancer implant can be packaged in the whole pharmaceutic adjuvant equably, also can be packaged in carrier holder center or its surface; Can effective ingredient be discharged by direct diffusion and/or the mode of degrading through polymer.

The characteristics of sustained-release implant are that used slow-release auxiliary material removes the high molecular polymerization beyond the region of objective existence, also contain above-mentioned any one or multiple other adjuvant.The pharmaceutic adjuvant that adds is referred to as additive.Additive can be divided into filler, porogen, excipient, dispersant, isotonic agent, preservative agent, blocker, solubilizing agent, absorption enhancer, film former, gellant etc. according to its function.

The Main Ingredients and Appearance of sustained-release implant can be made into multiple dosage form.As, but be not limited to capsule, slow releasing agent, implant, slow releasing agent implant etc.; Be multiple shape, as, but be not limited to granule, pill, tablet, powder, sphere, bulk, needle-like, bar-shaped, column and membranaceous.In various dosage forms, serve as preferred slowly to discharge implant in the body.Can be the bar-shaped of 0.1-5mm (slightly) * 1-10mm (length), also can be other shapes such as lamellar.

The most preferred dosage form of sustained-release implant is that the slow releasing agent that biocompatibility, degradable absorb is implanted, and can make different shape and various dosage form because of the clinical needs of difference.The packing method of its Main Ingredients and Appearance and step in United States Patent (USP) (US5651986) have a detailed description, comprise the some kinds of methods that prepare slow releasing preparation: as, but be not limited to, (i) carrier holder powder and medicament mixed be pressed into implant then, promptly so-called mixing method; (ii) carrier holder fusing, mix solid cooled then, promptly so-called fusion method mutually with medicine to be packaged; (iii) the carrier holder is dissolved in the solvent, medicine dissolution to be packaged or be scattered in the polymer solution, evaporating solvent then, drying, promptly so-called dissolution method; (iv) spray drying method; And (v) freeze-drying etc.

The anticancer effective component of sustained-release implant is preferably as follows, and all is weight percentage:

The combination of the tetrazole violet of 2-40% and the melphalan of 2-40%, 4H-peroxide cyclophosphamide, vinorelbine, tamoxifen, methotrexate, amycin, epirubicin, actinomycin D, tallimustine, atrimustine, semustine or Ranimustine.

Sustained-release micro-spheres also can be used for preparing sustained-release implant, and used slow-release auxiliary material can be any or multiple material in the above-mentioned pharmaceutic adjuvant, is main separation with the high molecular weight water soluble polymer in various high molecular polymers.

Slow-release auxiliary material and percentage by weight thereof are most preferably as follows in the sustained-release implant of the present invention:

(1) PLA of 55-90%;

(2) PLGA of 50-90%;

(3) polifeprosan of 50-85%;

(4) bis-fatty acid of 55-90% and decanedioic acid copolymer;

(5) EVAc of 55-90%;

(6) xylitol of 40-95%, oligosaccharide, chrondroitin, chitin, hyaluronic acid, collagen protein, gelatin or albumin glue; Or

(7) poly-dl-lactide of 40-95%, poly-dl-lactide/ethanol copolymer, monomethyl polyethylene glycol, monomethyl polyethylene glycol copolymer, polyethylene glycol, polyethylene glycol copolymer, end carboxyl polylactic acid or end carboxyl polylactic acid/ethanol copolymer.

Route of administration depends on multiple factor, for obtain valid density in former or position, metastatic tumour place, medicine can give through number of ways, as in subcutaneous, intracavity (in abdominal cavity, thoracic cavity and canalis spinalis), the tumor, in all injections of tumor or placement, selective arterial injection, the lymph node and injection in the bone marrow.With in selective arterial injection, intracavity, the tumor, tumor week injection or be placed as preferred.When the cancer therapy drug in the medicament slow-release microsphere only is tetrazole violet or its synergist (cytotoxic drug), the application of anti-cancer sustained-released implantation agent and the same slow releasing injection of potentiation mode.

The present invention can be used to prepare the pharmaceutical preparation of the various tumors for the treatment of people and animal, be mainly slow releasing injection or sustained-release implant, the indication tumor comprises former or cancer or sarcoma or the carcinosarcoma that shifts that originates from brain, central nervous system, kidney, liver, gallbladder, incidence, oral cavity, thyroid, skin, mucosa, body of gland, blood vessel, osseous tissue, lymph node, lungs, esophagus, stomach, mammary gland, pancreas, eyes, nasopharynx part, uterus, ovary, endometrium, cervix uteri, prostate, bladder, colon, rectum.

Also can add other medicinal ingredient in slow releasing injection that the present invention is made or the sustained-release implant, as, but be not limited to antibiotics, antalgica, anticoagulant medicine, hemorrhage etc.

By following test and embodiment technology of the present invention is further described:

The local drug concentration that test 1, different modes are used behind the tetrazole violet compares

(Fisher344) is subjects with rat, with 2 * 10 ⁵Individual prostate tumor cells subcutaneous injection is in its hypochondrium, treats behind tumor growth to 1 cm diameter its grouping.Every group of dosage is the 5mg/kg tetrazole violet.Measure medicament contg (%) in the different time tumor, the result shows, the local drug concentration significant difference of tetrazole violet after different modes is used, topical can obviously improve and effectively keep the active drug concentration at position, tumor place, and is wherein best with the effect of placing sustained-release implant and intratumor injection slow releasing injection in the tumor.Yet, intratumor injection slow releasing injection operation most convenient, easy.This discovery constitutes key character of the present invention.Following relevant inhibition test has further confirmed this point.

The interior tumor-inhibiting action of body that test 2, different modes are used behind the tetrazole violet compares

(Fisher344) is subjects with rat, with 2 * 10 ⁵Individual breast tumor cell subcutaneous injection is in its hypochondrium, treats behind tumor growth to 0.5 cm diameter its grouping.Every group of dosage is the 2.5mg/kg tetrazole violet.The treatment back was measured gross tumor volume size, relatively therapeutic effect on the 20th day.The result shows, the tumor-inhibiting action significant difference of tetrazole violet after different modes is used, topical can obviously improve and effectively keep the active drug concentration at position, tumor place, and is wherein best with the effect of placing sustained-release implant and intratumor injection slow releasing injection in the tumor.Yet, intratumor injection slow releasing injection operation most convenient, easy.Good effect not only, toxic and side effects is also little.

Tumor-inhibiting action in the body of test 3, tetrazole violet and cell toxicity medicament (slow releasing injection)

(Fisher344) is subjects with rat, with 2 * 10 ⁵Individual pancreatic tumor cell subcutaneous injection is in its hypochondrium, treats that tumor growth after 14 days is divided into it following 10 groups (seeing Table 1).First group is contrast, and the 2nd to 10 group is the treatment group, and medicine is through intratumor injection.The dosage of tetrazole violet is 5mg/kg, and the dosage of cell toxicity medicament is 15mg/kg.The treatment back was measured gross tumor volume size, relatively therapeutic effect (seeing Table 1) on the 20th day.

Table 1

Test group (n)	Suffered treatment	Gross tumor volume (cm 3)	The P value
				1(6)	Contrast	54±12
2(6)	Amycin	40±8.0	＜0.05
				3(6)	Tetrazole violet	42±8.6	＜0.01
4(6)	Melphalan	42±8.2	＜0.01
				5(6)	4H-peroxide cyclophosphamide	46±8.8	＜0.01
6(6)	Vinorelbine	38±6.0	＜0.01
				7(6)	Amycin+tetrazole violet	26±6.0	＜0.001
8(6)	Melphalan+tetrazole violet	30±5.6	＜0.001
				9(6)	4H-peroxide cyclophosphamide+tetrazole violet	22±5.2	＜0.001
10(6)	Vinorelbine+tetrazole violet	20±5.0	＜0.001

Above result shows, growth all has the obvious suppression effect to kinds of tumor cells when this concentration is used separately for tetrazole violet and its synergist (amycin, melphalan, 4H-peroxide cyclophosphamide, vinorelbine), can show significant potentiation when use in conjunction.This discovery constitutes the another key character of the present invention.

The tumor-inhibiting action of test 4, tetrazole violet and cell toxicity medicament (slow releasing injection)

Used tumor cell comprises CNS-1, C6,9L, gastric gland epithelial cancer (SA), bone tumor (BC), breast carcinoma (BA), pulmonary carcinoma (LH), papillary adenocarcinoma of thyroid (PAT), hepatocarcinoma etc.Tetrazole violet and cell toxicity medicament are added in 24 hours the various tumor cells of In vitro culture by 10ug/ml concentration, continue to cultivate counting cells sum after 48 hours.Its growth of tumour cell suppression ratio (%) is shown in Table 2.

Table 2

Oncocyte	Actinomycin D	Tamoxifen	Methotrexate	Tetrazole violet	Actinomycin D+tetrazole violet	Tamoxifen+tetrazole violet	Methotrexate+tetrazole violet
								CNS	42	42	38	22	74	70	80
C6	50	52	34	26	66	78	82
								SA	42	48	28	20	68	80	78
BC	52	46	26	24	66	84	80
								BA	54	46	20	22	66	70	78
LH	50	56	26	28	64	84	80
								PAT	50	52	28	18	78	72	76

Above result shows, growth all has the obvious suppression effect to kinds of tumor cells when this concentration is used separately for used cell toxicity medicament (actinomycin D, tamoxifen, methotrexate) and tetrazole violet, can show significant potentiation when use in conjunction.

The tumor-inhibiting action of test 5, tetrazole violet and cell toxicity medicament (slow releasing injection)

(Fisher344) is subjects with rat, with 2 * 10 ⁵Individual tumor cell of liver subcutaneous injection is in its hypochondrium, treats that tumor growth after 14 days is divided into it following 10 groups (seeing Table 3).First group is contrast, and the 2nd to 10 group is the treatment group, and sustained-release implant is placed in tumor.The dosage of tetrazole violet is 2.5mg/kg, and cell toxicity medicament dosage is 15mg/kg.The treatment back was measured gross tumor volume size, relatively therapeutic effect (seeing Table 3) on the 20th day.

Table 3

Test group (n)	Suffered treatment	Gross tumor volume (cm 3)	The P value
				1(6)	Contrast	50±12
2(6)	Tetrazole violet	42±8.0	＜0.05
				3(6)	Melphalan	32±6.0	＜0.01
4(6)	Tetrazole violet+melphalan	20±2.4	＜0.001
				5(6)	4H-peroxide cyclophosphamide	34±3.2	＜0.01
6(6)	Tetrazole violet+4H-peroxide cyclophosphamide	14±1.8	＜0.001
				7(6)	Vinorelbine	38±6.6	＜0.01
8(6)	Tetrazole violet+vinorelbine	22±2.4	＜0.001
				9(6)	Tamoxifen	42±8.4	＜0.01
10(6)	Tetrazole violet+tamoxifen	18±2.0	＜0.001

Above result shows, growth all has the obvious suppression effect to kinds of tumor cells when this concentration is used separately for used tetrazole violet and cell toxicity medicament (melphalan, 4H-peroxide cyclophosphamide, vinorelbine, tamoxifen), can show significant potentiation when use in conjunction.This discovery constitutes the another key character of the present invention.

The tumor-inhibiting action of test 6, tetrazole violet and cell toxicity medicament (slow releasing injection)

(Fisher344) is subjects with rat, with 2 * 10 ⁵Individual colon cancer cell subcutaneous injection is in its hypochondrium, treats that tumor growth after 14 days is divided into it negative control (blank), single therapy group (tetrazole violet or cell toxicity medicament) and therapeutic alliance group (tetrazole violet and cell toxicity medicament).Medicine is through intratumor injection.The dosage of tetrazole violet is 5mg/kg, and cell toxicity medicament dosage is 10mg/kg.The treatment back was measured the gross tumor volume size on the 20th day, made relatively therapeutic effect (seeing Table 4) of index with inhibition rate of tumor growth.

Table 4

Test group (n)	Suffered treatment	Tumor control rate (%)	The P value
				1(6)	Contrast	-
2(6)	Tetrazole violet	46	＜0.05
				3(6)	Semustine	44	＜0.01
4(6)	Ranimustine	40	＜0.01
				5(6)	Atrimustine	54	＜0.01
6(6)	Tallimustine	46	＜0.01
				7(6)	Tetrazole violet+semustine	82	＜0.001
8(6)	Tetrazole violet+Ranimustine	80	＜0.001
				9(6)	Tetrazole violet+atrimustine	76	＜0.001
10(6)	Tetrazole violet+tallimustine	84	＜0.001

Above result shows, growth all has the obvious suppression effect to kinds of tumor cells when this concentration is used separately for used tetrazole violet and cell toxicity medicament (semustine, Ranimustine, atrimustine, tallimustine), can show significant potentiation when use in conjunction.

The tumor-inhibiting action of test 7, tetrazole violet and cell toxicity medicament (slow releasing injection)

(Fisher344) is subjects with rat, with 2 * 10 ⁵Individual pulmonary carcinoma tumor cell subcutaneous injection is in its hypochondrium, treats that tumor growth after 14 days is divided into it negative control (blank), single therapy group, therapeutic alliance group.Medicine is through intratumor injection.The dosage of tetrazole violet is 7.5mg/kg, and cell toxicity medicament dosage is 2.5mg/kg.The treatment back was measured the gross tumor volume size on the 10th day, made relatively therapeutic effect (seeing Table 5) of index with inhibition rate of tumor growth.

Table 5

Test group (n)	Suffered treatment	Tumor control rate (%)	The P value
				1(6)	Contrast	-
2(6)	Tetrazole violet	48	＜0.05
				3(6)	Methotrexate	50	＜0.01
4(6)	Amycin	48	＜0.01
				5(6)	Epirubicin	48	＜0.01
6(6)	Actinomycin D	46	＜0.01
				7(6)	Tetrazole violet+methotrexate	80	＜0.001
8(6)	Tetrazole violet+amycin	74	＜0.001
				9(6)	Tetrazole violet+epirubicin	72	＜0.001
10(6)	Tetrazole violet+actinomycin D	82	＜0.001

Above result shows, growth all has the obvious suppression effect to kinds of tumor cells when this concentration is used separately for used tetrazole violet and cell toxicity medicament (methotrexate, amycin, epirubicin, actinomycin D), can show significant potentiation when use in conjunction.This discovery constitutes the another key character of the present invention.

The tumor-inhibiting action of test 8, tetrazole violet and cell toxicity medicament (slow releasing injection)

(Fisher344) is subjects with rat, with 2 * 10 ⁵Individual gastric cancer tumor cell subcutaneous injection treats that in its hypochondrium tumor growth divided photograph with it after 14 days.Sustained-release implant is placed in tumor.Dosage is 5mg/kg.The treatment back was measured the gross tumor volume size on the 20th day, made relatively therapeutic effect of index with inhibition rate of tumor growth.Above result shows, growth all has the obvious suppression effect to kinds of tumor cells when this concentration is used separately for used tetrazole violet and cell toxicity medicament (melphalan, 4H-peroxide cyclophosphamide, vinorelbine, tamoxifen, methotrexate, amycin, epirubicin, actinomycin D, tallimustine, atrimustine, semustine or Ranimustine), can show significant potentiation when use in conjunction.This discovery constitutes the another key character of the present invention.

Release ratio in the body of the tetrazole violet sustained-release implant that test 9, different molecular weight polylactic acid are made

With rat (Fisher344) is subjects, grouping (3/group) and the equivalent tetrazole violet sustained-release implant that carries in the subcutaneous polylactic acid (PLA) that contains different molecular weight (MW).Survey the surplus of medicine in implant respectively at 1,3,7,14,21,28 and 35 day then, and then draw rate of release (%) in its body.The result shows, molecular weight is 20000 is released to: 1 day (8%), 3 (28%), 7 (56%), 14 (82%), 21 (90), 28 (94) and 35 (98%).Discharge in the body of the tetrazole violet sustained-release implant that comparison different molecular weight polylactic acid is made and find, slack-off with the molecular weight increase, with the 7th day was example, compare with whole body administration group, tumor control rate increases with the polylactic acid molecule amount and improves, and is followed successively by 68% (MW:5000), 66% (MW:15000), 54% (MW:25000), 50% (MW:40000) and 48 (MW:60000).

It is the slow releasing agent of the compositions of adjuvant tetrazole violet, tetrazole violet, tetrazole violet, tetrazole violet, tetrazole violet or the tetrazole violet made and melphalan, 4H-peroxide cyclophosphamide, vinorelbine, tamoxifen, methotrexate, amycin, epirubicin, actinomycin D, tallimustine, atrimustine, semustine or Ranimustine that same result also sees with polylactic acid.

That pays special attention to is simple to operation, the good reproducibility of slow releasing agent of the present invention, particularly slow releasing injection.Good effect not only, toxic and side effects is little.

Different drug packages is to want characteristic different with different Biodegradable high moleculars.Discover that further the slow-release auxiliary material that is most appropriate to medicament slow release of the present invention is a poly-dl-lactide, poly-dl-lactide/ethanol copolymer, the monomethyl polyethylene glycol, monomethyl polyethylene glycol copolymer, polyethylene glycol, the polyethylene glycol copolymer, end carboxyl polylactic acid, end carboxyl polylactic acid/ethanol copolymer, polifeprosan, bis-fatty acid and decanedioic acid copolymer, poly-(erucic acid dimer-decanedioic acid), poly-(fumaric acid-decanedioic acid), ethylene vinyl acetate copolymer, polylactic acid, the copolymer of polyglycolic acid and hydroxyacetic acid, xylitol, oligosaccharide, chrondroitin, chitin, hyaluronic acid, collagen protein, gelatin, one of albumin glue or its combination; Optimum suspending agent is one of methylcellulose, hydroxy methocel, sodium carboxymethyl cellulose, (iodine) glycerol, simethicone, propylene glycol, carbomer, mannitol, sorbitol, surfactant, soil temperature 20, soil temperature 40, soil temperature 80 or its combination.

In a word, growth all had the obvious suppression effect to kinds of tumor cells when used tetrazole violet and various cell toxicity medicament were used separately, can show significant potentiation when use in conjunction.Therefore, effective ingredient of the present invention be tetrazole violet and with any one cell toxicity medicament.The medicine that contains above effective ingredient can be made into sustained-release micro-spheres, and then makes slow releasing injection and implant, serves as preferred with the suspensoid injectio that is combined to form with the special solvent that contains suspending agent wherein.

Slow releasing injection or sustained-release implant also can be further specified by following embodiment.Just the invention will be further described for the foregoing description and following examples, is not its content and use are imposed any restrictions.

(4) specific embodiment

Embodiment 1.

80mg polifeprosan (to carboxy phenyl propane (p-CPP): certain herbaceous plants with big flowers diacid (SA) is 20: 80) copolymer is put into container, add 100 milliliters of dichloromethane, behind the dissolving mixing, add 10mg actinomycin D and 10mg tetrazole violet, shake up the back contains 10% actinomycin D and 10% tetrazole violet with spray drying method for preparation injectable microsphere again.Then microsphere is suspended in the normal saline that contains 15% mannitol, makes corresponding suspension type slow releasing injection, viscosity is 220cp-460cp (20 ℃-30 ℃ time).The drug release time of this slow releasing injection in external normal saline is 10-15 days, is about 20-30 days at the subcutaneous drug release time of mice.

Embodiment 2.

The method step that is processed into slow releasing injection is identical with embodiment 1, but different is that contained anticancer effective component and percentage by weight thereof are:

The combination of the tetrazole violet of 2-40% and the melphalan of 2-40%, 4H-peroxide cyclophosphamide, vinorelbine, tamoxifen, methotrexate, amycin, epirubicin, actinomycin D, tallimustine, atrimustine, semustine or Ranimustine; Used slow-release auxiliary material is poly-dl-lactide, poly-dl-lactide/ethanol copolymer, monomethyl polyethylene glycol, monomethyl polyethylene glycol copolymer, polyethylene glycol, polyethylene glycol copolymer, end carboxyl polylactic acid or end carboxyl polylactic acid/ethanol copolymer.

Embodiment 3.

With 80mg molecular weight peak value is that 65000 polylactic acid (PLGA, 75: 25) is put into container, adds 100 milliliters of dichloromethane, behind the dissolving mixing, adds 5mg tetrazole violet and 15mg tallimustine, shakes up the dry organic solvent of removing of final vacuum again.Dried pastille solid composite freezing and pulverizing is made the micropowder that contains 5% tetrazole violet and 15% tallimustine, be suspended in then in the normal saline that contains 1.5% sodium carboxymethyl cellulose, make corresponding suspension type slow releasing injection, viscosity is 300cp-400cp (20 ℃-30 ℃ time).The drug release time of this slow releasing injection in external normal saline is 10-15 days, is about 20-30 days at the subcutaneous drug release time of mice.

Embodiment 4

The method step that is processed into slow releasing injection is identical with embodiment 3, but different is that contained anticancer effective component and percentage by weight thereof are: the combination of 5% tetrazole violet and 15% amycin, actinomycin D, melphalan, 4H-peroxide cyclophosphamide, vinorelbine, tamoxifen, methotrexate, amycin, epirubicin, actinomycin D, tallimustine, atrimustine, semustine or Ranimustine.

Embodiment 5.

(EVAc) puts into container with the 70mg ethylene vinyl acetate copolymer, after adding 100 milliliters of dichloromethane dissolving mixings, add 20mg vinorelbine and 10mg tetrazole violet, shake up the back contains 20% vinorelbine and 10% tetrazole violet with spray drying method for preparation injectable microsphere again.Then microsphere is suspended in the injection that contains the 5-15% sorbitol, makes corresponding suspension type slow releasing injection, viscosity is 100cp-200cp (20 ℃-30 ℃ time).The drug release time of this slow releasing injection in external normal saline is 10-15 days, is about 20-30 days at the subcutaneous drug release time of mice.

Embodiment 6.

The method step that is processed into slow releasing injection is identical with embodiment 5, but different is that contained anticancer effective component is: the combination of the tetrazole violet of 10-20% and the amycin of 10-20%, actinomycin D, melphalan, 4H-peroxide cyclophosphamide, vinorelbine, tamoxifen, methotrexate, amycin, epirubicin, actinomycin D, tallimustine, atrimustine, semustine or Ranimustine.

Embodiment 7.

70mg polifeprosan (to carboxy phenyl propane (p-CPP): certain herbaceous plants with big flowers diacid (SA) is 20: 80) copolymer is put into container, add 100 milliliters of dichloromethane, behind the dissolving mixing, add 20mg amycin and 10mg tetrazole violet, shake up the back contains 20% amycin and 10% tetrazole violet with spray drying method for preparation injectable microsphere again.Microsphere is suspended in the normal saline that contains 1.5% sodium carboxymethyl cellulose and 0.5% Tween 80 then, makes corresponding suspension type slow releasing injection, viscosity is 80cp-150cp (20 ℃-25 ℃ time).The drug release time of this slow releasing injection in external normal saline is 10-15 days, is about 20-30 days at the subcutaneous drug release time of mice.

Embodiment 8.

The method step that is processed into slow releasing injection is identical with embodiment 7, but different is that contained anticancer effective component is: the combination of 15% tetrazole violet and 5% amycin, actinomycin D, melphalan, 4H-peroxide cyclophosphamide, vinorelbine, tamoxifen, methotrexate, amycin, epirubicin, actinomycin D, tallimustine, atrimustine, semustine or Ranimustine.

Embodiment 9

80mg polifeprosan (to carboxy phenyl propane (p-CPP): certain herbaceous plants with big flowers diacid (SA) is 20: 80) copolymer is put into container, add 100 milliliters of dichloromethane, behind the dissolving mixing, add 10mg melphalan and 10mg tetrazole violet, shake up the back again and contain 10% melphalan and 10% tetrazole violet injectable microsphere with spray drying method for preparation.Then microsphere is suspended in the normal saline that contains 1.5% sodium carboxymethyl cellulose and 15% sorbitol and 0.2% Tween 80, makes corresponding suspension type slow releasing injection, viscosity is 560cp-640cp (20 ℃-30 ℃ time).The drug release time of this slow releasing injection in external normal saline is 10-15 days, is about 20-30 days at the subcutaneous drug release time of mice.

Embodiment 10

The method step that is processed into slow releasing injection is identical with embodiment 9, but different is that contained anticancer effective component is:

The combination of 15% tetrazole violet and 15% amycin, actinomycin D, melphalan, 4H-peroxide cyclophosphamide, vinorelbine, tamoxifen, methotrexate, amycin, epirubicin, actinomycin D, tallimustine, atrimustine, semustine or Ranimustine.

Embodiment 11

70mg polifeprosan (to carboxy phenyl propane (p-CPP): certain herbaceous plants with big flowers diacid (SA) is 20: 80) copolymer is put into container, add 100 milliliters of dichloromethane, behind the dissolving mixing, add 10mg tetrazole violet and 20mg epirubicin, shake up the back contains 10% tetrazole violet and 20% epirubicin with spray drying method for preparation injectable microsphere again.Then microsphere is made corresponding sustained-release implant through pressed disc method.The drug release time of this sustained-release implant in external normal saline is 10-15 days, is about 30-40 days at the subcutaneous drug release time of mice.

Embodiment 12

The method step that is processed into sustained-release implant is identical with embodiment 11, but different is that contained anticancer effective component is:

The combination of 10% tetrazole violet and 10% amycin, actinomycin D, melphalan, 4H-peroxide cyclophosphamide, vinorelbine, tamoxifen, methotrexate, amycin, epirubicin, actinomycin D, tallimustine, atrimustine, semustine or Ranimustine.

Embodiment 13

With 70mg molecular weight peak value 35000 polylactic acid (PLGA, 50: 50) put into container, add 100 milliliters of dichloromethane, behind the dissolving mixing, add 10mg Ranimustine and 20mg tetrazole violet, shake up the back contains 10% Ranimustine and 20% tetrazole violet with spray drying method for preparation injectable microsphere again.Then microsphere is made corresponding sustained-release implant through pressed disc method.The drug release time of this sustained-release implant in external normal saline is 10-15 days, is about 35-50 days at the subcutaneous drug release time of mice.

Embodiment 14

The method step that is processed into sustained-release implant is identical with embodiment 11,13, but different is that contained anticancer effective component is: the combination of 10% tetrazole violet and 15% amycin, actinomycin D, melphalan, 4H-peroxide cyclophosphamide, vinorelbine, tamoxifen, methotrexate, amycin, epirubicin, actinomycin D, tallimustine, atrimustine, semustine or Ranimustine.

Embodiment 15.

With 70mg molecular weight peak value is that 35000 polylactic acid (PLA) is put into container, adds 100 milliliters of dichloromethane, behind the dissolving mixing, adds 20mg4H-peroxide cyclophosphamide and 10mg tetrazole violet, shakes up the dry organic solvent of removing of final vacuum again.Dried pastille solid composite freezing and pulverizing is made the micropowder that contains 20%4H-peroxide cyclophosphamide and 10% tetrazole violet, be suspended in then in the normal saline that contains 1.5% sodium carboxymethyl cellulose, make corresponding suspension type slow releasing injection, viscosity is 220cp-260cp (25 ℃-30 ℃ time).The drug release time of this slow releasing injection in external normal saline is 10-15 days, is about 20-30 days at the subcutaneous drug release time of mice.

Embodiment 16

The method step that is processed into slow releasing injection is identical with embodiment 15, but different is that contained anticancer effective component and percentage by weight thereof are:

The combination of 10% tetrazole violet and 15% amycin, actinomycin D, melphalan, 4H-peroxide cyclophosphamide, vinorelbine, tamoxifen, methotrexate, amycin, epirubicin, actinomycin D, tallimustine, atrimustine, semustine or Ranimustine.

Embodiment 17.

With 70mg molecular weight peak value is that 30000 bis-fatty acid and certain herbaceous plants with big flowers diacid (SA) copolymer (bis-fatty acid: the certain herbaceous plants with big flowers diacid is 20: 80) are put into container, add 100 milliliters of dichloromethane, behind the dissolving mixing, add 15mg semustine and 15mg tetrazole violet, shake up the dry organic solvent of removing of final vacuum again.Dried pastille solid composite freezing and pulverizing is made the micropowder that contains 15% semustine and 15% tetrazole violet, be suspended in then in the normal saline that contains 1.5% sodium carboxymethyl cellulose, make corresponding suspension type slow releasing injection, viscosity is 380cp-460cp (25 ℃-30 ℃ time).The drug release time of this slow releasing injection in external normal saline is 10-15 days, is about 20-30 days at the subcutaneous drug release time of mice.

Embodiment 18.

The method step that is processed into slow releasing injection is identical with embodiment 17, but different is that contained anticancer effective component and percentage by weight thereof are:

The combination of 15% tetrazole violet and 15% melphalan, 4H-peroxide cyclophosphamide, vinorelbine, tamoxifen, methotrexate, amycin, epirubicin, actinomycin D, tallimustine, atrimustine, semustine, Ranimustine, amycin or actinomycin D.

Embodiment 19

The method step that is processed into slow releasing agent is identical with embodiment 1-18, but different is used slow-release auxiliary material is one of following or its combination:

A) polylactic acid (PLA), the molecular weight peak value is 10000-30000,300000-60000,60000-100000 or 100000-150000;

B) copolymer of polyglycolic acid and hydroxyacetic acid (PLGA), wherein, the ratio of polyglycolic acid and hydroxyacetic acid is 50-95: 50-50, the molecular weight peak value is 10000-30000,300000-60000,60000-100000 or 100000-150000;

C) ethylene vinyl acetate copolymer (EVAc);

D) polifeprosan, to carboxy phenyl propane (p-CPP): certain herbaceous plants with big flowers diacid (SA) is 10: 90,20: 80,30: 70,40: 60,50: 50 or 60: 40;

E) bis-fatty acid and decanedioic acid copolymer [P (FAD-SA)];

F) poly-(erucic acid dimer-decanedioic acid) [P (EAD-SA)];

G) poly-(fumaric acid-decanedioic acid) [P (FA-SA)];

H) xylitol, oligosaccharide, chrondroitin, chitin, hyaluronic acid, collagen protein, gelatin or white tempera;

I) poly-dl-lactide, poly-dl-lactide/ethanol copolymer, monomethyl polyethylene glycol, monomethyl polyethylene glycol copolymer, polyethylene glycol, polyethylene glycol copolymer, end carboxyl polylactic acid or end carboxyl polylactic acid/ethanol copolymer.

Embodiment 20

The method step that is processed into slow releasing injection is identical with embodiment 1-19, but different is used suspending agent is respectively one of following or its combination:

A) 0.5-3.0% carboxymethyl cellulose (sodium);

B) 5-15% mannitol;

C) 5-15% sorbitol;

D) 0.1-1.5% surfactant;

E) 0.1-0.5% polysorbas20.

Embodiment 21

The method step that is processed into slow releasing injection is identical with embodiment 1-20, but different is that contained anticancer effective component is: 10% tetrazole violet and 15% melphalan, 4H-peroxide cyclophosphamide, vinorelbine, tamoxifen, methotrexate, amycin, epirubicin, actinomycin D, tallimustine, atrimustine, semustine or Ranimustine.

Embodiment 22

The method step that is processed into slow releasing injection is identical with embodiment 1-20, but different is that contained anticancer effective component is:

(a) 10% tetrazole violet and 10% amycin, melphalan or epirubicin or combination;

(b) 15% tetrazole violet and 15% he do not seek the combination of sweet smell or 4H-peroxide cyclophosphamide;

(c) 5% tetrazole violet and 5% vinorelbine or the combination of methotrexate;

(d) combination of 20% tetrazole violet and 10% actinomycin D; Or

(e) combination of 10% tetrazole violet and 20% tallimustine, atrimustine, semustine or Ranimustine.

Above embodiment only is used for explanation, and is not limitation application of the present invention.

The present invention disclosed and the protection the content see claim.

Claims (10)

1. the slow releasing injection of loading both tetrazole violet and cell toxicity medicament is become to be grouped into by following:

(A) sustained-release micro-spheres comprises:

Anticancer effective component 0.5-60%

Slow-release auxiliary material 40-99%

Suspending agent 0.0-30%

More than be weight percentage

With

(B) solvent is for common solvent or contain the special solvent of suspending agent.

Wherein,

Anticancer effective component is tetrazole violet and the combination that is selected from the cell toxicity medicament of amycin, actinomycin D, melphalan, 4H-peroxide cyclophosphamide, vinorelbine, tamoxifen, methotrexate, amycin, epirubicin, actinomycin D, tallimustine, atrimustine, semustine or Ranimustine;

Slow-release auxiliary material is selected from one of following or its combination:

A) polylactic acid;

B) copolymer of polyglycolic acid and hydroxyacetic acid;

C) polifeprosan;

D) ethylene vinyl acetate copolymer;

E) bis-fatty acid and decanedioic acid copolymer;

F) poly-(erucic acid dimer-decanedioic acid) copolymer;

G) poly-(fumaric acid-decanedioic acid) copolymer;

H) poly-dl-lactide, poly-dl-lactide/ethanol copolymer, monomethyl polyethylene glycol, monomethyl polyethylene glycol copolymer, polyethylene glycol, polyethylene glycol copolymer, end carboxyl polylactic acid or end carboxyl polylactic acid/ethanol copolymer.

Suspending agent is selected from one of sodium carboxymethyl cellulose, iodine glycerol, simethicone, propylene glycol, carbomer, mannitol, sorbitol, surfactant, soil temperature 20, soil temperature 40 and soil temperature 80 or its combination,

The viscosity of suspending agent is 100cp-3000cp (20 ℃-30 ℃ time).

2. the slow-releasing anticarcinogen injection according to claim 1 is characterized in that tetrazole violet shared percentage by weight in slow releasing agent is 0.1%-50%.

3. the slow-releasing anticarcinogen injection according to claim 1, the weight ratio that it is characterized in that tetrazole violet and cell toxicity medicament is 1-9: 1 to 1: 1-9.

4. the slow-releasing anticarcinogen injection according to claim 1 is characterized in that the anticancer effective component of slow-releasing anticarcinogen injection and percentage by weight are: the combination of the tetrazole violet of 2-40% and the amycin of 2-40%, actinomycin D, melphalan, 4H-peroxide cyclophosphamide, vinorelbine, tamoxifen, methotrexate, amycin, epirubicin, actinomycin D, tallimustine, atrimustine, semustine or Ranimustine.

5. according to claim 1 and 6 described slow-releasing anticarcinogen injections, it is characterized in that in the slow-release auxiliary material,

A) polylactic acid molecule amount peak value is 10000-30000,300000-60000,60000-100000 or 100000-150000;

B) in the copolymer of polyglycolic acid and hydroxyacetic acid, the ratio of polyglycolic acid and hydroxyacetic acid is 50-95: 50-50, and the molecular weight peak value is 10000-30000,300000-60000,60000-100000 or 100000-150000;

C) in the polifeprosan, to carboxy phenyl propane: the certain herbaceous plants with big flowers diacid is 10: 90,20: 80,30: 70,40: 60,50: 50 or 60: 40.

6. according to claim 1 and 6 described slow-releasing anticarcinogen injections, it is characterized in that used suspending agent is respectively one of following or its combination:

A) 0.5-3.0% carboxymethyl cellulose (sodium);

B) 5-15% mannitol;

C) 5-15% sorbitol;

D) 0.1-1.5% surfactant;

E) 0.1-0.5% polysorbas20;

F) (iodine) glycerol, simethicone, propylene glycol or carbomer;

G) 0.5-5% sodium carboxymethyl cellulose+0.1-0.5% soil temperature 80;

H) 5-20% mannitol+0.1-0.5% soil temperature 80; Or

I) 0.5-5% sodium carboxymethyl cellulose+5-20% sorbitol+0.1-0.5% soil temperature 80.

7. the slow-releasing anticarcinogen injection according to claim 1, it is characterized in that sustained-release micro-spheres in the slow-releasing anticarcinogen injection also is used for the preparation treatment and originates from people and animal brain, the central nervous system, kidney, liver, gallbladder, incidence, the oral cavity, thyroid, skin, mucosa, body of gland, blood vessel, osseous tissue, lymph node, lungs, esophagus, stomach, mammary gland, pancreas, eyes, nasopharynx part, the uterus, ovary, endometrium, cervix uteri, prostate, bladder, former or the cancer of secondary of colon or rectum, the sustained-release implant of sarcoma or carcinosarcoma is in tumor or tumor week injection or place administration.

8. the anti-cancer sustained-released implantation agent according to claim 7 is characterized in that the constituent of sustained-release implant is:

Anticancer effective component is:

(a) 10% tetrazole violet and 10% amycin, melphalan or epirubicin or combination;

(b) 15% tetrazole violet and 15% tamoxifen or the combination of 4H-peroxide cyclophosphamide;

(c) 5% tetrazole violet and 5% vinorelbine or the combination of methotrexate;

(d) combination of 20% tetrazole violet and 10% actinomycin D; Or

(e) combination of 10% tetrazole violet and 20% tallimustine, atrimustine, semustine or Ranimustine.

9. the anti-cancer sustained-released implantation agent according to claim 7 is characterized in that slow-release auxiliary material is selected from one of following or its combination:

A) polylactic acid;

B) copolymer of polyglycolic acid and hydroxyacetic acid;

C) polifeprosan;

D) ethylene vinyl acetate copolymer;

E) bis-fatty acid and decanedioic acid copolymer;

F) poly-(erucic acid dimer-decanedioic acid) copolymer;

G) poly-(fumaric acid-decanedioic acid) copolymer;

H) poly-dl-lactide, poly-dl-lactide/ethanol copolymer, monomethyl polyethylene glycol, monomethyl polyethylene glycol copolymer, polyethylene glycol, polyethylene glycol copolymer, end carboxyl polylactic acid or end carboxyl polylactic acid/ethanol copolymer.

10. the anti-cancer sustained-released implantation agent according to claim 7 is characterized in that suspending agent is respectively one of following or its combination:

A) 0.0-3.0% carboxymethyl cellulose (sodium);

B) 0.0-15% mannitol;

C) 0.0-15% sorbitol;

D) 0.0-1.5% surfactant;

E) 0.0-0.5% polysorbas20.