CN1961864A

CN1961864A - Anticancer composition

Info

Publication number: CN1961864A
Application number: CNA2006102012723A
Authority: CN
Inventors: 孙娟; 张婕; 邹会凤
Original assignee: Jinan Shuaihua Pharmaceutical Technology Co Ltd
Current assignee: Jinan Shuaihua Pharmaceutical Technology Co Ltd
Priority date: 2006-12-12
Filing date: 2006-12-12
Publication date: 2007-05-16

Abstract

Disclosed is an anti-cancer composition slow release injection which comprises slow release microspheres and dissolvent, the slow release microspheres comprise slow release auxiliary materials, Epothilone derivatives, glyoxaline piperazidine, Topo enzyme inhibitor combination selected from SN-38, CPT-11, HCPT, Topotecan, Irinotecan, Etoposide, Teniposide, Amrubicin, Valtaxin, XK469, AD312, ICRF-187 or ICRF-193, the dissolvent being specific dissolvent containing suspension adjuvant, the slow release auxiliary materials are selected from polylactic acid and its copolymer, polyethylene / polylactic acid, glycollic acid copolymer, aliphatic acid and sebacylic acid copolymer, the viscosity of the suspension adjuvant is 80-3000cp (at 20-30 deg C), and is selected from carboxymethylcellulose, The slow release microspheres can also be prepared into slow release implanting agent, for injection or placement in or around tumor with a release period of about 40 days. The slow release injection and slow release implanting agent can be used independently for effectively suppressing tumor accretion, or used in combination with non-operative methods such as chemotherapy and/or radiotheraphy with the function of improving their treatment effects.

Description

Anticancer composition

(I) technical field

The invention relates to an anticancer composition, and belongs to the technical field of medicines. Specifically, the invention provides a composition loaded with epothilone, derivatives thereof and anticancer drugs, which is mainly a sustained-release injection and a sustained-release implant. The anticancer sustained release agent is applied in or around tumor, which is beneficial to the drug to obtain and maintain the effective drug concentration in solid tumor and can increase the sensitivity of tumor cells to the drug.

(II) background of the invention

The treatment of cancer mainly comprises methods such as surgery, radiotherapy, chemotherapy and the like. Wherein the surgical treatment can not eliminate scattered tumor cells, so that the tumor cells are frequently relapsed or caused to spread and metastasize due to surgical stimulation; radiotherapy and traditional chemotherapy have no selectivity, are difficult to form effective drug concentration or therapeutic dose locally on tumors, have poor effect and high toxicity, and are limited by systemic toxicity reaction when the drug or radiation dose is simply increased. See, e.g., "Intratumoral Placement of cisplatin plus systemic Carmustine in rat brain tumors" J.Takema.69, pp 76-82, 1998 (Kong Q et al, J Surg Oncol.1998 Oct; 69 (2): 76-82).

Low dose anti-cancer drug therapy can not only increase drug tolerance of cancer cells, but also promote invasive growth thereof, see Beam et al, "anti-cancer drug pulsed screening increases drug tolerance and in vitro infiltration capacity of human lung cancer cells with concomitant changes in gene expression" [ International journal of cancer (Liang Y, et al, Int Jcancer. 2004; 111 (4): 484-93) ].

The solid tumor is composed of tumor cells and tumor stroma, wherein blood vessels in the tumor stroma not only provide a scaffold and essential nutrients for the growth of the tumor cells, but also influence the penetration and diffusion of chemotherapeutic drugs around the tumor and in the tumor tissue, see Niti et al, "influence of the condition of extracellular stroma on drug operation in the solid tumor" [ Cancer research ] No. 60, No. 2497 and No. 503, 2000 (Netti PA, Cancer Res.2000, 60 (9): 2497 and No. 503).

The components of fibrin and collagen in blood vessels and connective tissues in tumor stroma and hyperproliferative tumor cells cause high interstitial pressure (interstitial pressure), high interstitial viscosity (interstitial viscosity), high tissue tension coefficient (tissue tension module) and low interstitial fluid conductivity (hydralic con) of solid tumors. These factors greatly limit the effective diffusion of drugs into solid tumors and within tumors, and thus constitute a major obstacle to tumor chemotherapy.

The defects can be better overcome by local injection or placement of the anti-tumor drug, the local drug concentration of the tumor can be obviously improved, and the systemic toxic reaction can be obviously reduced. A number of in vitro and in vivo experiments have shown therapeutic efficacy against solid tumors, see Kongqing et al, "cisplatin placement in tumors plus systemic carmustine for treatment of rat brain tumors", J.Surg Oncol.1998 Oct (Kong Q et al, J.Surg.Oncol.1998 Oct; 69 (2): 76-82) and Kongqing et al, "cisplatin placement in tumors for cure of rat primary brain tumors", J.Surg.Oncol.1997 Oct, 64: 268-273 (1997) (Kong Q et al, J.Surg.Oncol.1997 Oct). See also Chinese patent (ZL 00111093.4; ZL 96115937.5; application Nos. 001111264, 001111272) and U.S. patent Nos. 6,376,525B 1; 5,651,986; 5,626,862).

However, single-drug chemotherapy often results in increased resistance of tumor cells to anticancer drugs, with consequent therapeutic failure.

Disclosure of the invention

Aiming at the defects of the prior art, the invention provides a novel pharmaceutical composition containing epothilone and an anticancer drug. More particularly, the sustained-release preparation is a sustained-release preparation for resisting solid tumors, mainly a sustained-release implant and a sustained-release injection. The local application can effectively inhibit or destroy the growth of the tumor and can also increase the sensitivity of the tumor cells to anticancer drugs; the epothilone and/or anticancer drug is prepared into sustained release agent (mainly sustained release injection and sustained release implant), which not only can greatly improve the drug concentration of the local tumor, reduce the drug concentration of the drug in the circulatory system and reduce the toxicity of the drug to the normal tissue, but also can greatly facilitate the drug injection, reduce the complications of the operation and reduce the cost of the patient. The anticancer medicine can inhibit tumor growth and raise the sensitivity of tumor cell to anticancer medicine.

The anti-solid tumor sustained release agent comprises an anti-cancer active ingredient and a pharmaceutical adjuvant, wherein the anti-cancer active ingredient is selected from the combination of epothilone derivatives, topoisomerase inhibitors and/or tetrazine drugs, and preferably the combination of the epothilone derivatives and the topoisomerase inhibitors and/or tetrazine drugs.

After a great deal of research, the invention discovers that the combined application of the epothilone derivative and the topoisomerase inhibitor and/or the tetrazine medicine can mutually enhance the anti-tumor effect, and the selected slow-release auxiliary materials are used as carriers to prepare slow-release implant and slow-release injection to achieve unexpected effects when the slow-release implant and the slow-release injection are locally applied. The above unexpected findings constitute the subject of the present invention.

The compound pharmaceutical composition of the present invention can be prepared into any preparation form, such as, but not limited to, capsules, sustained release preparations, granules, pills, tablets, powders, injections, ointments, patches, implants, sustained release injections, etc. Among them, sustained release preparations are preferable, and sustained release implants and sustained release injections are most preferable.

Aiming at the defects of the prior art, the invention provides a novel sustained-release injection containing epothilone and anticancer drugs.

The slow released epothilone injection consists of slow released microsphere and solvent. Specifically, the anticancer sustained-release injection consists of the following components:

(A) a sustained release microsphere comprising:

0.5-60% of anticancer active ingredient

Sustained release auxiliary materials 40-99%

0.0 to 30 percent of suspending agent

The above are weight percentages

And

(B) the solvent is common solvent or special solvent containing suspending agent.

Wherein,

the anticancer active ingredient is the combination of epothilone and anticancer drugs, and the anticancer drugs are selected from topoisomerase inhibitors and/or tetrazines drugs.

The medicine prepared from the epothilone, the topoisomerase inhibitor and/or the tetrazine medicine is mainly administrated by conventional ways such as oral administration or intravenous injection, and the administration mode of the invention is local slow release administration, so that the systemic toxicity effect of the medicine is obviously reduced while the treatment effect of the medicine is obviously enhanced. In the case of epothilones with anticancer activity, not all sustained-release excipients can achieve a sustained-release effect of effective release. The medicinal auxiliary materials are more than hundreds of medicinal auxiliary materials with slow release function, in particular the medicinal auxiliary materials which can slowly release the selected epothilone in human bodies or animal bodies within a certain time can be obtained through a large number of creative experiments, and the selection of the combination of the specific slow release auxiliary materials and the slow-release medicines can be determined through a large number of creative labor. The related data, particularly the data of the release characteristics in animals, can be obtained through a large number of creative experiments in vivo and in vitro, can not be determined through limited experiments, and is non-obvious.

The above unexpected findings constitute the subject of the present invention.

Epothilones are, but are not limited to, epothilones (a-F) and epothilone derivatives, selected from one or a combination of the following: epothilone (Epothilone) or Epothilone derivatives selected from Epothilone A, Epothilone B (Patupilone, EP0-906), Epothilone C (desoxyepothilone, desoxyepothilone D), Epothilone D (Epothilone D, 12, 13-desoxyepothilone B, dEpoB, KOS-862 or NSC-703147), Epothilone E, Epothilone F, and the like, as well as derivatives thereof.

Among them, derivatives of epothilone C such as, but not limited to, 4-desmethyl-9-one-epothilone C, 12, 13-dihydro-13-oxoepothilone C (12, 13-dihydro-13-oxoepothilone C);

derivatives of epothilone B such as, but not limited to, epothilone B with amino substitutions at positions 21 and 26, respectively or simultaneously, dehydroepothilone B with hydrogen at positions 9, 10, 11, 26, 27 halogen substituted epothilone B, epothilone B with hydroxyl substitutions at positions 9, 10, 11, 14, 21, 26, respectively, 21-dihydroxy epothilone B, 21-hydroxy-10, 11 dehydroepothilone B, 4-demethyl-9-one-epothilone B, 4-demethyl-9, 10-didehydro epothilone B, 4-demethyl-10, 11-didehydro epothilone B, 6-demethyl-10, 11-didehydro epothilone B, 21-amino epothilone B, 21-hydroxy epothilone B, 21-dehydroepothilone B, 26-hydroxyepothilone B, 26-fluoroepothilone B, 26-aminoepothilone B, 12, 13-cyclopropylepothilone B, 12, 13-cyclobutyl epothilone B, ixabepilone (BMS-247550), Azaepothilone B (Azaepothilone B, the oxygen in the lactone ring is replaced by nitrogen), 26-Trifluoro- (E) -9, 10-dehydro-12, 13-desoxyepothilone B (26-trifluo- (E) -9, 10-dehydro-12, 13-desoxyepothilone B [ Fludulenone (Flu) ]; derivatives of epothilone D such as, but not limited to, epothilone D with amino substitution at positions 21 and 26, dehydroepothilone D with amino substitution at positions 9 and 10, dehydroepothilone D with hydrogen at positions 10, 11, 26, 27, halogen substitution at positions 9, 10, 11, 14, 21, 26, respectively, epothilone D with hydroxy substitution at positions 21, 26, 21-hydroxy-10, 11, dehydroepothilone D, 4-demethyl-9-one-epothilone D, 4-demethyl-9, 10-didehydro epothilone D, 4-demethyl-10, 11-didehydro epothilone D, 6-demethyl-10, 11-didehydro epothilone D, 21-hydroxy epothilone D, 21-aminoepothilone D, and, 26-hydroxyepothilone D, 26-aminoepothilone D, 26-fluoroepothilone D, 6-ethyl, 16-fluoro, 17-pyridinylesoprazole (or isoepothilone), isoepothilone D, 9, 10-dehydroepothilone D, 10, 11-dehydroepothilone D, furaetheromycin D (furano-epothilone D), (E) -9, 10-dehydro-12, 13-desoxyepothilone D), BMS-310705, 6-ethyl, 16-fluoro, 17-pyridinyloxyepothilone (ZK-EPO), 11, 12-dehydro-12, 13-dehydro-13-epothilone D11, 12, 13-dehydro-13-desoxyepothilone D (12, 13-dihydro-13-oxoepothilone D), 9-oxoepothilone D (9-oxoepothilone D), 8-epi-9-oxoepothilone D (8-epi-9-oxoepothilone D),

The epothilone and epothilone derivatives are preferably one or a combination of epothilone, epothilone A, epothilone B, epothilone C, epothilone D, isoepothilone D, epothilone E, epothilone F, BMS-247550, azaepothilone B, and furan epothilone D, BMS-310705. Epothilone B, epothilone D, isoepothilone D, BMS-247550, azaepothilone B, furan epothilone D, and BMS-310705 are preferred.

The ratio of epothilone and epothilone derivatives in the composition can be, depending on the particular case, 0.1% to 60%, preferably 1% to 40%, most preferably 5% to 20%.

The topoisomerase inhibitor is selected from one or a combination of the following: camptothecin (CPT), derivatives of camptothecin, Lurtotecan (Lurtocan), topotecan (10-hydroxy-9-methylenethonyl- (S) -camptothecin, topotecan), irinotecan (irinotecan, IRT), gemecan (Gimatecan), 9-nitrocamptothecin (9-nitrocamptothecin, 9NC), 7-ethyl-10-camptothecin (7-ethyl-10-hydroxy-camptothecin, SN-38), 7-ethyl-10- [4- (1-piperidino) -1-piperidino ] carbonylcamptothecin (7-ethyl-10- [4- (1-piperidino) -1-piperidino ] carbonylcamptothecin, CPT-11), 10-Hydroxycamptothecin (10-Hydroxycamptothecin, HCPT), homocamptothecin (HCthecine, HCC), camptothecin (HCT-1-Hydroxycamptothecin, HCT-1-camptothecin), camptothecin (HCT-1, HCT-1-camptothecin), and pharmaceutically acceptable salts thereof, Methylenedioxycamptothecin (10, 11-methylenedioxy, MD-CPT), (RS) -methylenedioxycamptothecin (10, 11-MD-20(RS) -CPT), (S) methylenedioxycamptothecin glycinate (10, 11-MD-20(S) -cpT-glycinate ester (Gly). HCl), 9-amino- (S) -methylenedioxycamptothecin glycinate (9-amino-10, 11-MD-20(S) -CPT-Gly), N- [2 (dimethylamino) ethyl ] piperidine-4-carboxamide (N- [2- (dimethylamino) ethyl ] acridine-4-carboxamide, DACA) and its derivatives substituted at position 5 or 7, podophyllotoxin, Etoposide (Etoposide, epipodophyllotoxins, Etoposide, Etoposide, VP-16), Teniposide (Teniposide, etoposide, VM-26), podophyllinic acid, podophyllotoxin, trihydroxyisoflavone (Genistein), 14-hydroxydaunorubicin, Amrubicin (Amrubicin), doxorubicin (4 ' - (acetylamino) methansulfo-m-aniside (amsacrine, m-AMSA)), 4-demethoxydaunorubicin (4-demethoxydaunorubicin), ditobicin, 7-O-methylnocar-4 ' -epirubicin (7-O-methylnocalol-4 ' -epirubicin), Esorubicin (Esorubicin), carubicin, idarubicin (idarubicin, IDA), roxubicin, epirubicin (Leuriubicin), medubicin, nerubicin (Nemoulutin), pivalotoxin (N-14, N-trifloxystroburin, N-14-norubicin, N-14-pivalolide, N-14-norubicin, N-E, AD 32, valrubicin), 2- [4- (7-chloro-2-quinoxalinyloxyphenoxy ] -propionic acid ((2- [4- (7-chloro-2-quinoxalinyloxyphenyloxy ] -propionic acid, XK469), Zorubicin (Zorubicin), N- (2-chloroethane) -N-nitrosoureidodaunorubicin (N- (2-Chloroethyl) -N-nitrosoureidodaunorubicin, AD312), pyrazolo [1, 5-a ] indole derivatives, such as, but not limited to, GS-2, -3, -4, GS-5, dioxopiperazine derivatives, such as, but not limited to, (+) -1, 2-bis (3, 5-dioxopiperazinyl) propane ((+) -1, 2-bis (3, 5-dioxopiperazinyl-1-propyl) pane, ICRF-187), m-2, 3-bis (3, 5-dioxopiperazin-1-yl) butane (meso-2, 3-bis (3, 5-dioxopiperazine-1-yl) butane, ICRF-193), bis-dioxopiperazine (bisdioxopiperazine); suramin (Suramin), Deoxyguanosine (Deoxyguanosine), lithocholic acid (LCA) or sodium azide (sodium azide).

Among the above topoisomerase inhibitors, camptothecin, hydroxycamptothecin, 9-nitrocamptothecin, 7-ethyl-10-hydroxy-camptothecin (SN-38), 7-ethyl-10- [4- (1-piperidino) -1-piperidino ] carbonylcamptothecin (CPT-11), 10-hydroxy-camptothecin (HCPT), Homocamptothecins (Homocamptothecins), methylenedioxycamptothecin (MD-CPT), podophyllotoxin, trihydroxyisoflavone, lurtotecan, topotecan, irinotecan, etoposide, teniposide, 14-hydroxydaunorubicin, amrubicin, doxorubicin, ditorelbine, 7-O-methylcar-4' -epirubicin, esorubicin, carrubicin, doxorubicin, and the like, Idarubicin, epirubicin, medroxobin, nemorubicin, doxorubicin, roxobicin, valrubicin, zorubicin, 2- [4- (7-chloro-2-quinoxalinyloxyphenoxy ] -propionic acid (XK469), zorubicin, N- (2-chloroethane) -N-nitrosoureidodaunorubicin (AD 312), (+) -1, 2-bis (3, 5-dioxopiperazinyl) propane (ICRF-187), m-2, 3-bis (3, 5-dioxopiperazin-1-yl) butane (ICRF-193) are preferred.

The proportion of the topoisomerase inhibitor in the composition is determined by the particular circumstances, and in general, it may be from 0.01% to 50%, preferably from 1% to 40%, and most preferably from 5% to 30%. All are weight percent.

The tetrazine compound is selected from one or a combination of the following: imidazotetrazine, imidazopiperazine, imidazopyridine, Procarbazine (PCB), Mitozolomide (MTZ), Temozolomide (Temozolomide ornSC 362856)), 4-carboxytemozolomide, 3-N-methyltemozolomide [3-N-methyl ] Temozolomide). Of these, imidazotetrazine, imidazopiperazine, imidazopyridine, procarbazine, mitozolamide, 4-carboxy temozolomide, 3-N-methyl temozolomide and temozolomide are preferable.

The above compounds also include their salts such as, but not limited to, sulfate, phosphate, hydrochloride, lactobionate, acetate, aspartate, nitrate, citrate, purine or pyrimidine salts, succinate, maleate and the like.

The proportion of the tetrazine compound in the sustained release agent is determined by specific conditions, generally speaking, can be from 0.1% to 50%, preferably from 1% to 40%, and most preferably from 5% to 30%. All are weight percent.

The viscosity range IV (dl/g) of the sustained-release auxiliary material is 0.1-0.8, and the sustained-release auxiliary material is selected from racemic polylactic acid (D, L-PLA), racemic polylactic acid/glycollic acid copolymer (D, L-PLGA), monomethyl polyethylene glycol/polylactic acid (MPEG-PLA), monomethyl polyethylene glycol/polylactic acid copolymer (MPEG-PLGA), polyethylene glycol/polylactic acid (PLA-PEG-PLA), polyethylene glycol/polylactic acid copolymer (PLGA-PEG-PLGA), carboxyl-terminated polylactic acid (PLA-COOH), carboxyl-terminated polylactic acid/glycollic acid copolymer (PLGA-COOH), polifeprosan, difatty fatty acid and sebacic acid copolymer (PFAD-SA), poly (erucic acid dimer-sebacic acid) [ P (EAD-SA) ], poly (fumaric acid-sebacic acid) [ P (FA-SA) ], poly (FA-sebacic acid) ], and the like, Polylactic acid (PLA), polyglycolic acid and glycolic acid copolymer (PLGA), Polydioxanone (PDO), polytrimethylene carbonate (PTMC), xylitol, oligosaccharide, chondroitin, chitin, chitosan, hyaluronic acid, collagen, gelatin, poloxamer, protein glue or their combination; the suspending agent is selected from one or more of sodium carboxymethylcellulose, (iodine) glycerol, dimethicone, propylene glycol, carbomer, mannitol, sorbitol, surfactant, Tween 20, Tween 40 and Tween 80.

When the anticancer active ingredient in the drug sustained-release microspheres is only epothilone or anticancer drug, the anticancer sustained-release injection is mainly used for increasing the effect of the epothilone or the anticancer drug applied by other ways or for the synergy of radiotherapy or other therapies. When the anticancer drug in the drug sustained-release microspheres is only epothilone or an anticancer drug, the application and the synergy mode of the anticancer sustained-release injection are as follows:

(1) the slow release injection containing epothilone is injected locally, while the anticancer drug is applied by other ways;

(2) the slow release injection containing the anti-cancer drugs is locally injected, and the epothilone is applied in other ways;

(3) locally injecting a slow release injection containing epothilone and a slow release injection containing anticancer drugs; or

(4) Local injection is slow released injection containing epothilone and anticancer medicine.

The slow released anticancer injection for local application may be also used in raising the effect of radiotherapy and other treatment. Other routes refer, but are not limited to, arterial, venous, intraperitoneal, subcutaneous, intraluminal administration.

The weight percentage of the anticancer active ingredient epothilone and/or anticancer drug in the drug sustained release microsphere is 0.1-60%, preferably 1-40%, and most preferably 2-30%. The weight ratio of epothilone to anti-cancer agent is 1-19: 1 to 1: 1-19, preferably 1-9: 1 to 1: 1-9, most preferably 1-5: 1 to 1: 1-5.

The anticancer active ingredients in the anticancer drug composition are one or a plurality of epothilones and/or one or a plurality of anticancer drugs, and the anticancer active ingredients are preferably combined as follows:

(1) 1-40% of epothilone B, epothilone D, isoepothilone D, BMS-247550, azaepothilone B, furaetheromycin D or BMS-310705 in combination with 1-40% of camptothecin, hydroxycamptothecin, 9-nitrocamptothecin, SN-38, CPT-11, HCPT, homocamptothecin, MD-CPT, podophyllotoxin, trihydroxyisoflavone, lurtotecan, topotecan, irinotecan, etoposide, teniposide, 14-hydroxydaunorubicin, amrubicin, doxorubicin, ditobicin, 7-O-methylnoca-4' -epirubicin, esorubicin, carubicin, idarubicin, epirubicin, medroxobin, nemorubicin, doxorubicin, roxobicin, valrubicin, zorubicin, XK469, zorubicin, doxorubicin, roxburgiximab, valrubicin, doxycycline, a combination of AD312, ICRF-187 or ICRF-193; or

(2) 1-40% epothilone B, epothilone D, isoepothilone D, BMS-247550, azaepothilone B, furazaepothilone D or BMS-310705 in combination with 1-40% imidazotetrazine, imidazopiperazine, imidazopyridine, procarbazine, mitozolamide, temozolomide, 4-carboxy temozolomide or 3-N-methyl temozolomide.

The slow release auxiliary material of the invention can be hydrolyzed or degraded by enzyme, acid and alkali or tissue fluid, and comprises one or the combination of the following components:

(1) biocompatible polymers, including biodegradable or non-biodegradable polymers and mixtures or copolymers thereof;

(2) a water-soluble low-molecular compound; or/and

(3) suitable additives and excipients for realizing injection, sustained release and other drug dosage forms.

The slow release auxiliary materials can be various water-soluble or non-water-soluble high molecular polymers, and are selected from one or a combination of racemic polylactic acid, a racemic polylactic acid/glycollic acid copolymer, monomethyl polyethylene glycol/polylactic acid, a polyethylene glycol/polylactic acid copolymer, terminal carboxyl polylactic acid, a terminal carboxyl polylactic acid/glycollic acid copolymer, polifeprosan, a copolymer of di-fatty acid and sebacic acid, poly (erucic aciddipolymer-sebacic acid), poly (fumaric acid-sebacic acid), an ethylene vinyl acetate copolymer, polylactic acid, a copolymer of polyglycolic acid and glycolic acid, xylitol, oligosaccharide, chondroitin, chitin, chitosan, hyaluronic acid, collagen, gelatin and albumin glue.

The most preferable sustained-release auxiliary materials in the sustained-release microspheres and the weight percentage thereof are as follows:

(1) 55-90% PLA;

(2) 50-90% PLGA;

(3) 50-85% of the polyphenyl is endogenic;

(4) 55-90% of a copolymer of di-fatty acid and sebacic acid;

(5) mixing 30-60% polifeprosan with PLA or PLGA;

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

(7) 40-95% of racemic polylactic acid, racemic polylactic acid/glycollic acid copolymer, monomethyl polyethylene glycol/polylactic acid copolymer, polyethylene glycol/polylactic acid copolymer, carboxyl-terminated polylactic acid or carboxyl-terminated polylactic acid/glycollic acid copolymer.

Among the various polymers, preferred are polylactic acid, sebacic acid, and a mixture or copolymer of polylactic acid and sebacic acid, and the mixture or copolymer can be selected from, but not limited to, PLA, PLGA, a mixture of glycolic acid and hydroxycarboxylic acid, and a mixture or copolymer of sebacic acid and an aromatic polyanhydride or an aliphatic polyanhydride. The blending ratio of glycolic acid and hydroxycarboxylic acid is 10/90-90/10 (by weight), preferably 25/75-75/25 (by weight). The method of blending is arbitrary. The contents of glycolic acid and hydroxycarboxylic acid in copolymerization are 10-90 wt% and 90-10 wt%, respectively. Representative of aromatic polyanhydrides are polifeprosan [ poly (1, 3-di (P-carboxyphenoxy) propane-sebacic acid) (P (CPP-SA)), di-fatty acid-sebacic acid copolymer (PFAD-SA) ], poly (erucic acid dimer-sebacic acid) [ P (EAD-SA) ], and poly (fumaric acid-sebacic acid) [ P (FA-SA) ], and the like. The content of p-carboxyphenoxy propane (p-CPP) and sebacic acid in copolymerization is 10-60 wt% and 20-90 wt%, respectively, and the blending weight ratio is 10-40: 50-90, preferably 15-30: 65-85.

The molecular weight peak of polylactic acid may be, but is not limited to, 5000-100,000, but is preferably 20,000-60,000, and most preferably 5,000-30,000; the molecular weight of polyglycolic acid may be, but is not limited to, 5000-; the polyhydroxy acids can be selected singly or in multiple ways. When selected individually, polylactic acid (PLA) or a copolymer of hydroxycarboxylic acid and glycolic acid (PLGA) is preferred, and the molecular weight of the copolymer can be, but is not limited to, 5000-l00,000, but is preferably 20,000-60,000, and is most preferably 30,000-50,000; when more than one choice is selected, the polymer or the composite polymer or copolymer of different polymers is preferred, and the composite polymer or copolymer of polylactic acid or sebacic acid with different molecular weight is most preferred, such as, but not limited to, polylactic acid with molecular weight of 1000 to 30000 mixed with polylactic acid with molecular weight of 20000 to 50000, polylactic acid with molecular weight of 10000 to 30000 mixed with PLGA with molecular weight of 30000 to 80000, polylactic acid with molecular weight of 20000 to 30000 mixed with sebacic acid, PLGA with molecular weight of 30000 to 80000 mixed with sebacic acid. The polylactic acid used is preferably L-polylactic acid (L-PLA). The viscosity range IV (dl/g) of the L-polylactic acid (L-PLA) is 0.2-0.8, the glass transition temperature range is 55-65 ℃, and the melting point is 175-185 ℃.

In addition to the above sustained-release excipients, other substances can be selected and used as described in detail in U.S. Pat. Nos. 4757128, 4857311, 4888176 and 4789724 and "pharmaceutical excipients" in general (p. 123, published by Sichuan scientific and technical Press 1993, compiled by Roming and Gaoyun). In addition, Chinese patent (application No. 96115937.5; 91109723.6; 9710703.3; 01803562.0) and U.S. patent No. 5,651,986) also list some pharmaceutical excipients, including fillers, solubilizers, absorption promoters, film-forming agents, gelling agents, pore-forming agents, excipients or retarders.

In order to adjust the drug release rate or change other characteristics of the present invention, the monomer component or molecular weight of the polymer can be changed, and the composition and ratio of the pharmaceutical excipients can be added or adjusted, and water-soluble low molecular compounds such as, but not limited to, various sugars or salts can be added. Wherein the sugar can be, but is not limited to, xylitol, oligosaccharide, (chondroitin sulfate), chitin, etc., and the salt can be, but is not limited to, potassium salt, sodium salt, etc.; other pharmaceutical adjuvants such as, but not limited to, filler, solubilizer, absorption enhancer, film-forming agent, gelling agent, pore-forming agent, excipient or retarder can also be added

In the slow release injection, the drug slow release system can be prepared into microspheres, submicron spheres, micro emulsion, nanospheres, granules or spherical pellets, and then the injection is prepared after the drug slow release system is mixed with an injection solvent. The suspension type sustained-release injection is preferably selected from various sustained-release injections, the suspension type sustained-release injection is a preparation obtained by suspending a drug sustained-release system containing an anti-cancer component in injection, the used sustained-release auxiliary material is one or the combination of the sustained-release auxiliary materials, and the used solvent is a common solvent or a special solvent containing a suspending agent. Common solvents are, but not limited to, distilled water, water for injection, physiological saline, absolute ethanol or buffers formulated with various salts. The suspending agent is intended to effectively suspend the microspheres containing the drug, thereby facilitating injection. For convenient injection, the suspending agent has viscosity of 100-3000 cp (at 20-30 deg C), preferably 1000-3000 cp (at 20-30 deg C), and most preferably 1500-3000 cp (at 20-30 deg C). The suspending agent is selected from one or more of sodium carboxymethylcellulose, (iodine) glycerol, dimethicone, propylene glycol, carbomer, mannitol, sorbitol, surfactant, Tween 20, Tween 40 and Tween 80.

The content of the suspending agent in the common solvent depends on the characteristics of the suspending agent, and can be 0.1-30% according to the specific situation. Preferably, the suspending agent consists of:

A) 0.5-5% of sodium carboxymethylcellulose and 0.1-0.5% of Tween 80; or

B) 5-20% of mannitol and 0.1-0.5% of Tween 80; or

C)0.5 to 5 percent of sodium carboxymethylcellulose, 5 to 20 percent of sorbitol and 0.1 to 0.5 percent of Tween 80.

The preparation of the solvent depends on the kind of the solvent, and common solvents are commercially available or self-made, such as distilled water, water for injection, physiological saline, absolute ethanol or buffers prepared from various salts, but the preparation must strictly follow the relevant standards. The special solvent should consider the type and composition of suspending agent, the composition and properties of the drug suspended in the solvent, the sustained release microsphere (or microcapsule) and the required amount thereof, and the preparation method of the injection, for example, sodium carboxymethylcellulose (1.5%) + mannitol and/or sorbitol (15%) and/or tween 80 (0.1%) are dissolved in physiological saline to obtain the corresponding solvent with viscosity of 10-650 cp (at 20-30 deg.C).

The invention discovers that the key factor influencing the suspension and/or injection of the medicament and/or the sustained-release microspheres is the viscosity of the solvent, and the higher the viscosity is, the better the suspension effect is and the stronger the injectability is. This unexpected finding constitutes one of the main exponential features of the present invention. The viscosity of the solvent depends on the viscosity of the suspending agent, and the viscosity of the suspending agent is 100cp-3000cp (at 20-30 ℃), preferably 1000cp-3000cp (at 20-30 ℃), and most preferably 1500cp-3000cp (at 20-30 ℃). The viscosity of the solvent prepared according to the condition is 10cp-650cp (at 20-30 ℃), preferably 20cp-650cp (at 20-30 ℃), and most preferably 60cp-650cp (at 20-30 ℃).

The preparation of injection has several methods, one is that the slow release particles (A) whose suspending agent is '0' are directly mixed in special solvent to obtain correspondent slow release particle injection; the other is that the slow release particles (A) of which the suspending agent is not 0 are mixed in a special solvent or a common solvent to obtain the corresponding slow release particle injection; and the other one is that the slow release particles (A) are mixed in common dissolvent, then suspending agent is added and mixed evenly, and the corresponding slow release particle injection is obtained. Besides, the sustained-release particles (A) can be mixed in special solvent to prepare corresponding suspension, then the water in the suspension is removed by methods such as vacuum drying, and then the suspension is suspended by special solvent or common solvent to obtain the corresponding sustained-release particle injection. The above methods are merely illustrative and not restrictive of the invention. It is noted that the concentration of the suspended drug or the sustained release microspheres (or microcapsules) in the injection may be, but is not limited to, 10-400mg/ml, but is preferably 30-300mg/ml, and most preferably 50-200mg/ml, depending on the particular need. The viscosity of the injection is 50-1000 cp (at 20-30 deg C), preferably 100-1000 cp (at 20-30 deg C), and most preferably 200-650 cp (at 20-30 deg C). This viscosity is suitable for 18-22 gauge needles and specially made needles with larger (to 3 mm) inside diameters.

The method of preparation of the sustained release injection is arbitrary and can be prepared by several methods: such as, but not limited to, mixing, melting, dissolving, spray drying to prepare microspheres, dissolving in combination with freezing (drying) and pulverizing to form fine powders, liposome-encapsulating, and emulsifying. Among them, a dissolving method (i.e., solvent evaporation method), a drying method, a spray drying method and an emulsification method are preferable. The microspheres can be used for preparing the various sustained-release injections, and the method is arbitrary. The microspheres used may have a particle size in the range of 5-400um, preferably 10-300um, most preferably 20-200 um.

The microspheres can also be used for preparing other sustained-release injections, such as gel injections and block copolymer micelle injections. The block copolymer micelle is formed by a hydrophobic-hydrophilic block copolymer in an aqueous solution and has a spherical core-shell structure, wherein the hydrophobic block forms a core, and the hydrophilic block forms a shell. The drug-loaded micelle is injected into the body to achieve the purpose of controlling the release of the drug or targeting therapy. The drug carrier is any one of the above or the combination thereof. Of these, polyethylene glycol (PEG) having a molecular weight of 1000-15000 is preferable as the hydrophilic block of the micelle copolymer, and biodegradable polymers such as PLA, polylactide, polycaprolactone and copolymers thereof (molecular weight 1500-25000) are preferable as the hydrophobic block of the micelle copolymer. The block copolymer micelles may have a particle size in the range of 10 to 300um, preferably 20 to 200 um. The gel injection is prepared by dissolving biodegradable polymer (such as PLA, PLGA or DL-LA and epsilon-caprolactone copolymer) in certain amphiphilic solvent, adding the medicine, mixing (or suspending) with the solvent to form gel with good fluidity, and can be injected around tumor or in tumor. Once injected, the amphiphilic solvent diffuses into the body fluid quickly, and the water in the body fluid permeates into the gel, so that the polymer is solidified and the drug is released slowly.

The sustained-release microspheres can also be used for preparing sustained-release implants, the used pharmaceutic adjuvant can be any one or more of the above pharmaceutic adjuvants, but the water-soluble high polymer is selected as the main choice, and the mixture or copolymer of polylactic acid, sebacic acid, and high polymer containing polylactic acid or sebacic acid is selected as the first choice among various high polymers, and the mixture and copolymer can be selected from, but are not limited to, PLA, PLGA, mixture of PLA and PLGA, mixture or copolymer of sebacic acid and aromatic polyanhydride or aliphatic polyanhydride, fatty acid dimer-sebacic acid [ P (EAD-SA) ], poly (fumaric acid-sebacic acid) [ P (FA-SA) ]. The blending ratio of polylactic acid (PLA) to polyglycolic acid is 10/90 to 90/10 (by weight), preferably 25/75 to 75/25 (by weight). The method of blending is arbitrary. The contents of glycolic acid and lactic acid in copolymerization are respectively 10-90% and 90-10% by weight. The aromatic polyanhydride is represented by p-carboxyphenylpropane (p-CPP), the content of the p-carboxyphenylpropane (p-CPP) and sebacic acid in copolymerization is respectively 10-60% and 20-90% by weight, and the blending weight ratio is 10-40: 50-90, preferably 15-30: 65-85.

Still another form of the anticancer drug sustained-release preparation of the present invention is that the anticancer drug sustained-release preparation is a sustained-release implant. The effective components of the anticancer implant can be uniformly packaged in the whole pharmaceutic adjuvant, and also can be packaged in the center of a carrier support or on the surface of the carrier support; the active principle can be released by direct diffusion and/or by degradation via polymers.

The slow release implant is characterized in that the slow release auxiliary material contains any one or more of the other auxiliary materials besides the high molecular polymer. The added pharmaceutic adjuvants are collectively called as additives. The additives can be classified into fillers, pore-forming agents, excipients, dispersants, isotonic agents, preservatives, retarding agents, solubilizers, absorption enhancers, film-forming agents, gelling agents, etc. according to their functions.

The main components of the sustained-release implant can be prepared into various dosage forms. Such as, but not limited to, capsules, sustained release formulations, implants, sustained release implants, and the like; in various shapes such as, but not limited to, granules, pills, tablets, powders, spheres, chunks, needles, rods, columns, and films. Among various dosage forms, slow release implants in vivo are preferred. It can be in the form of rod of 0.1-5mm (thick) × 1-10mm (long), or in the form of sheet.

The optimal dosage form of the sustained-release implant is biocompatible, degradable and absorbable sustained-release implant, and can be prepared into various shapes and various dosage forms according to different clinical requirements. The packaging method and procedure for its main ingredients are described in detail in US patent (US5651986) and include several methods for preparing sustained release formulations: such as, but not limited to, (i) mixing a carrier support powder with a drug and then compressing into an implant, a so-called mixing process; (ii) melting the carrier support, mixing with the drug to be packaged, and then cooling the solid, the so-called melt process; (iii) dissolving the carrier support in a solvent, dissolving or dispersing the drug to be packaged in a polymer solution, and then evaporating the solvent and drying, the so-called dissolution method; (iv) spray drying; and (v) freeze-drying method.

The anticancer active ingredients of the sustained-release implant are preferably as follows, and the weight percentages are as follows:

(1) 1-40% of epothilone B, epothilone D, isoepothilone D, BMS-247550, azaepothilone B, furaetheromycin D or BMS-310705 in combination with 1-40% of camptothecin, hydroxycamptothecin, 9-nitrocamptothecin, SN-38, CPT-11, HCPT, homocamptothecin, MD-CPT, podophyllotoxin, trihydroxyisoflavone, lurtotecan, topotecan, irinotecan, etoposide, teniposide, 14-hydroxydaunorubicin, amrubicin, doxorubicin, ditobicin, 7-O-methylnoca-4' -epirubicin, esorubicin, carubicin, idarubicin, epirubicin, medroxobin, nemorubicin, doxorubicin, roxobicin, valrubicin, zorubicin, XK469, zorubicin, doxorubicin, roxburgiximab, valrubicin, doxycycline, a combination of AD312, ICRF-l87, or ICRF-l 93; or

The slow release microspheres can also be used for preparing slow release implanting agent, the slow release adjuvant can be any one or more of the above medicinal adjuvants, and water soluble high molecular polymer is selected from various high molecular polymers.

The sustained-release auxiliary materials in the sustained-release implant and the weight percentage thereof are most preferably as follows:

(1) 55-90% PLA;

(2) 50-90% PLGA;

(3) 50-85% of polifeprosan;

(4) 55-90% of a copolymer of di-fatty acid and sebacic acid;

(5) 55-90% EVAc;

The route of administration depends on a variety of factors, and in order to achieve effective concentrations at the site of the primary or metastatic tumor, the drug may be administered by a variety of routes, such as subcutaneous, intraluminal (e.g., intraperitoneal, thoracic, and intravertebral), intratumoral, peritumoral injection or placement, selective arterial injection, intralymph node, and intramedulary injection. Selective arterial injection, intracavitary, intratumoral, peritumoral injection or placement is preferred.

The application and the synergy mode of the sustained-release implant are the same as those of an anticancer sustained-release injection, namely the combination of locally-placed epothilone and anticancer drugs administered by other routes, the combination of locally-placed anticancer drugs and epothilone administered by other routes, and the combination of locally-placed anticancer drugs and locally-placed epothilone. Wherein the locally applied anticancer drug and epothilone can be produced, packaged, sold, and used separately or in combination. The package refers to the loading process of the drug for the auxiliary materials and the internal and external package of the drug-containing sustained release agent for transportation and/or storage.

The invention can be used for preparing pharmaceutical preparations for treating various tumors of human and animals, mainly sustained-release injections or sustained-release implants, wherein the tumors comprise primary or metastatic cancers or sarcomas or carcinosarcomas originated from brain, central nervous system, kidney, liver, gall bladder, head and neck, oral cavity, thyroid, skin, mucous membrane, gland, blood vessel, bone tissue, lymph node, lung, esophagus, stomach, mammary gland, pancreas, eye, nasopharynx, uterus, ovary, endometrium, cervix, prostate, bladder, colon and rectum.

The sustained-release injection or the sustained-release implant prepared by the invention can also be added with other medicinal components, such as, but not limited to, antibiotics, analgesic drugs, anticoagulant drugs, hemostatic drugs and the like.

The techniques of the anti-solid tumor compositions of the present invention are further described by the following assays and examples:

the in vivo tumor inhibition effect of the epothilone B and the anticancer drug is tested.

Using white rat as test object, 2X 10⁵Each colon cancer cell was injected subcutaneously into the quaternary costal region, and was divided into the following 10 groups 14 days after the tumor growth (see Table 1). The first group is control and groups 2 to 10 are treatment groups, wherein group 2 is epothilone B and groups 3 to 6 are camptothecin, hydroxycamptothecin, lurtotecan, topotecan, respectively. Groups 7 to 10 are combinations of epothilone B with camptothecin, hydroxycamptothecin, lurtotecan, topotecan, respectively. Except for epothilone B, which is placed intratumorally, camptothecin, hydroxycamptothecin, lurtotecan, topotecan are all administered intraperitoneally. 15mg/kg, except for 2.5mg/kg of epothilone B. Tumor volume was measured on day 30 after treatment and the treatment effect was compared (see table 1).

TABLE 1

Test set (n)	Is treated by	Tumor volume (cm)³)	P value
Test set (n)	Is treated by	Tumor volume (cm)³)	P value	1(6)	Control	56±14
2(6)	Epothilone B	38±10	＜0.05	1(6)	Control	56±14
2(6)	Epothilone B	38±10	＜0.05	3(6)	Camptothecin	42±8	＜0.05
4(6)	Hydroxycamptothecin	40±10	＜0.05	3(6)	Camptothecin	42±8	＜0.05
4(6)	Hydroxycamptothecin	40±10	＜0.05	5(6)	Lurtotecan	44±10	＜0.01
6(6)	Topotecan	36±8	＜0.01	5(6)	Lurtotecan	44±10	＜0.01
6(6)	Topotecan	36±8	＜0.01	7(6)	Epothilone B + camptothecin	26±4	＜0.001
8(6)	Epothilone B + hydroxycamptothecin	28±6	＜0.001	7(6)	Epothilone B + camptothecin	26±4	＜0.001
8(6)	Epothilone B + hydroxycamptothecin	28±6	＜0.001	9(6)	Epothilone B + lurtotecan	16±4	＜0.001
10(6)	Epothilone B + topotecan	14±4	＜0.001	9(6)	Epothilone B + lurtotecan	16±4	＜0.001

Note: camptothecin, hydroxycamptothecin, lurtotecan, topotecan are all topoisomerase inhibitors. The results show that epothilone B (group 2) and the topoisomerase inhibitor (groups 3 to 6) have a certain anti-tumor effect (P < 0.05) when applied alone, especially when administered topically, compared to the control group. The combination (groups 7 to 10) had a significant synergistic effect (P < 0.001).

And secondly, the in vivo tumor inhibition effect of the epothilone D and the anticancer drugs is tested.

The in vivo tumor-inhibiting effect of epothilone D and anti-cancer drugs on pancreatic cancer was determined according to the test one, and the results show that, epothilone D significantly enhances the tumor-suppressing effect (P < 0.05) of camptothecin, hydroxycamptothecin, 9-nitrocamptothecin, SN-38, CPT-11, HCPT, homocamptothecin, MD-CPT, podophyllotoxin, trihydroxyisoflavone, lurtotecan, topotecan, irinotecan, etoposide, teniposide, 14-hydroxydaunorubicin, amrubicin, doxorubicin, ditobicin, 7-O-methylnocardia-4' -epirubicin, esorubicin, casrubicin, idarubicin, epirubicin, medubicin, nemorubicin, doxorubicin, roxobicin, valrubicin, zorubicin, XK469, zorubicin, AD312, ICRF-187 or ICRF-193. The two have a certain tumor inhibiting effect (P is less than 0.05) on pancreatic cancer when being applied independently, particularly when being applied locally, and have obvious synergistic effect (P is less than 0.001) when being applied together.

Further research shows that the two drugs have certain tumor inhibiting effect (P is less than 0.05) on prostatic cancer, lung cancer, breast cancer, liver cancer and colorectal cancer when being applied independently, particularly when being applied locally, and the combined application has obvious synergistic effect (P is less than 0.001).

And thirdly, the tumor inhibition effect of the locally applied anticancer drug and the epothilone D is tested.

Using white rat as test object, 2X 10⁵Individual breast cancer cells were injected subcutaneously into the quaternary costal region and were divided into the following 10 groups 14 days after tumor growth (see table 2). The first group was control and groups 2 to 10 were treatment groups, where group 2 was epothilone D and groups 3 to 6 were control. Groups 7 to 10 are combinations of epothilone D with irinotecan, etoposide, teniposide, amrubicin, respectively. All drugs were placed intratumorally, 15mg/kg except 5mg/kg lysozyme. Tumor volume was measured on day 30 after treatment and the treatment effect was compared (see table 2).

TABLE 2

Test set (n)	Is treated by	Tumor volume (cm)³)	P value
Test set (n)	Is treated by	Tumor volume (cm)³)	P value	1(6)	Control	58±12
2(6)	Epothilone D	32±6	＜0.05	1(6)	Control	58±12
2(6)	Epothilone D	32±6	＜0.05	3(6)	Irinotecan	38±10	＜0.01
4(6)	Etoposide	42±8	＜0.01	3(6)	Irinotecan	38±10	＜0.01
4(6)	Etoposide	42±8	＜0.01	5(6)	Teniposide	38±8	＜0.01
6(6)	Aminorubicin	36±8	＜0.01	5(6)	Teniposide	38±8	＜0.01
6(6)	Aminorubicin	36±8	＜0.01	7(6)	Epothilone D + irinotecan	26±4.0	＜0.001
8(6)	Epothilone D + etoposide	22±4.2	＜0.001	7(6)	Epothilone D + irinotecan	26±4.0	＜0.001
8(6)	Epothilone D + etoposide	22±4.2	＜0.001	9(6)	Epothilone D + teniposide	18±3.4	＜0.001
10(6)	Epothilone D + amrubicin	14±3.8	＜0.001	9(6)	Epothilone D + teniposide	18±3.4	＜0.001

Note: irinotecan, etoposide, teniposide and amrubicin are all topoisomerase inhibitors. The results show that compared with the control group, the epothilone D (group 2) and the topoisomerase inhibitor (groups 3 to 6) have a certain tumor inhibition effect (P is less than 0.05) when used alone. However, the combination (groups 7 to 10) had a significant synergistic effect (P < 0.001).

Fourth, the in vivo tumor suppression effect of epothilone (BMS-247550) and topoisomerase inhibitors was tested.

The in vivo tumor suppression effect of BMS-247550 and topoisomerase inhibitor was examined according to the method of test three. The result shows that BMS-247550 and the anticancer drugs selected from SN-38, CPT-11, HCPT, MD-CPT, trihydroxyisoflavone, 14-hydroxydaunorubicin, 7-O-methylnocar-4' -epirubicin, isosbiracin, valrubicin, XK469, zorubicin, AD312, ICRF-187 or ICRF-193 have obvious tumor inhibition effect (P is less than 0.05) when being singly applied. The combined application has obvious synergistic effect (P is less than 0.001).

Fifth, the in vivo tumor suppression effect of isoepothilone D and anticancer drugs was tested.

Using white rat as test object, 2X 10⁵Each ovarian cancer cell was injected subcutaneously into the quaternary costal region and divided into the following 10 groups 14 days after tumor growth (see Table 3). Group 1 is a control and groups 2 to 10 are treatment groups, wherein group 2 is isoepothilone D; groups 3 to 6 are etoposide, roxobicin, doxorubicin, zorubicin, respectively. Groups 7 to 10 are combinations of isoepothilone D with different anti-cancer drugs, respectively. Except for the intratumoral placement of isoepothilone D, all were administered intraperitoneally. 5mg/kg except for 10mg/kg of isoepothilone D. Tumor volume was measured on day 30 after treatment and the treatment effect was compared (see table 3).

TABLE 3

Test set (n)	Is treated by	Tumor volume (cm)³)	P value
Test set (n)	Is treated by	Tumor volume (cm)³)	P value	1(6)	Control	56±10
2(6)	Isoepothilone D	42±8	＜0.05	1(6)	Control	56±10
2(6)	Isoepothilone D	42±8	＜0.05	3(6)	Etoposide	44±8	＜0.01
4(6)	Rodobicin	42±8	＜0.01	3(6)	Etoposide	44±8	＜0.01
4(6)	Rodobicin	42±8	＜0.01	5(6)	Epirubicin	42±9	＜0.01
6(6)	Zorubicin	42±6	＜0.01	5(6)	Epirubicin	42±9	＜0.01
6(6)	Zorubicin	42±6	＜0.01	7(6)	Isoepothilone D + etoposide	28±4	＜0.001
8(6)	Isoepothilone D + Rodobisin	20±3.4	＜0.001	7(6)	Isoepothilone D + etoposide	28±4	＜0.001
8(6)	Isoepothilone D + Rodobisin	20±3.4	＜0.001	9(6)	Isoepothilone D + epirubicin	18±3.0	＜0.001
10(6)	Isoepothilone D + zorubicin	14±2.6	＜0.001	9(6)	Isoepothilone D + epirubicin	18±3.0	＜0.001

Note: etoposide, roxobicin, doxorubicin, zorubicin are all topoisomerase inhibitors.

The in vivo tumor inhibition effect of the hexaazaepothilone B and the anticancer drug is tested.

Using white rat as test object, 2X 10⁵One lung cancer cell was injected subcutaneously into the quaternary costal region, and the tumor was divided into the following 10 groups 14 days after growth (see table 4). Group 1 is a control and groups 2 to 10 are treatment groups, wherein group 2 is azaepothilone B; groups 3 to 6 are each topoisomerase inhibitors. Groups 7 to 10 are combinations of azaepothilone B with different anti-cancer drugs, respectively. DeazazepineThe pristinamycin B is placed in the tumor and is externally applied by the abdominal cavity with topotecan, irinotecan, mitozolomide and temozolomide. 16mg/kg except 4mg/kg of azaepothilone B. Tumor volume was measured on day 30 after treatment and the effect was compared (see table 4).

TABLE 4

Test set (n)	Is treated by	Tumor volume (cm)³)	P value
Test set (n)	Is treated by	Tumor volume (cm)³)	P value	1(6)	Control	62±14
2(6)	Azaetheriomycin B	46±12	＜0.05	1(6)	Control	62±14
2(6)	Azaetheriomycin B	46±12	＜0.05	3(6)	Topotecan	42±6.3	＜0.05
4(6)	Irinotecan	46±6.0	＜0.05	3(6)	Topotecan	42±6.3	＜0.05
4(6)	Irinotecan	46±6.0	＜0.05	5(6)	Mitozolomide	42±6.4	＜0.05
6(6)	Temozolomide	40±4.8	＜0.05	5(6)	Mitozolomide	42±6.4	＜0.05
6(6)	Temozolomide	40±4.8	＜0.05	7(6)	Epothilone B + topotecan	28±6.8	＜0.001
8(6)	Epothilone B + irinotecan	26±4.6	＜0.001	7(6)	Epothilone B + topotecan	28±6.8	＜0.001
8(6)	Epothilone B + irinotecan	26±4.6	＜0.001	9(6)	Epothilone B + mitozolomide	18±2.8	＜0.001
10(6)	Epothilone B + temozolomide	14±2.6	＜0.001	9(6)	Epothilone B + mitozolomide	18±2.8	＜0.001

Note: topotecan, irinotecan, mitozolomide and temozolomide are all anticancer drugs.

Experiment on the in vivo tumor inhibiting effect of the furan epothilone D and the anticancer drugs.

Using white rat as test object, 2X 10⁵Subcutaneous injection of gastric cancer tumor cellsIt was irradiated to the quaternary rib, and 14 days after the tumor growth, it was divided into the following 10 groups (see Table 5). The first group was control and groups 2 to 10 were treatment groups, wherein group 2 was furan epothilone D and groups 3 to 6 were procarbazine, mitozolomide, temozolomide, 4-carboxy temozolomide, respectively. Groups 7 to 10 are combinations of furan epothilone D with procarbazine, mitozolomide, temozolomide, 4-carboxy temozolomide, respectively. Except for the intratumoral placement of furan epothilone D, procarbazine, mitozolomide, temozolomide, 4-carboxy temozolomide were all administered intraperitoneally. 2.5mg/kg, except for 15mg/kg of furan epothilone D. Tumor volume was measured on day 30 after treatment and the treatment effect was compared (see table 5).

TABLE 5

Test set (n)	Is treated by	Tumor volume (cm)³)	P value
Test set (n)	Is treated by	Tumor volume (cm)³)	P value	1(6)	Control	56±12
2(6)	Furan epothilone D	42±8	＜0.05	1(6)	Control	56±12
2(6)	Furan epothilone D	42±8	＜0.05	3(6)	Methyl benzyl hydrazine	42±8.6	＜0.05
4(6)	Mitozolomide	50±10	＜0.05	3(6)	Methyl benzyl hydrazine	42±8.6	＜0.05
4(6)	Mitozolomide	50±10	＜0.05	5(6)	Temozolomide	44±10	＜0.01
6(6)	4-carboxy temozolomide	38±6	＜0.01	5(6)	Temozolomide	44±10	＜0.01
6(6)	4-carboxy temozolomide	38±6	＜0.01	7(6)	Furan epothilone D + Methylbenzylhydrazine	28±4.8	＜0.001
8(6)	Furan epothilone D + mitozolomide	24±4.6	＜0.001	7(6)	Furan epothilone D + Methylbenzylhydrazine	28±4.8	＜0.001
8(6)	Furan epothilone D + mitozolomide	24±4.6	＜0.001	9(6)	Furan epothilone D + temozolomide	22±4.2	＜0.001
10(6)	Furan epothilone D + 4-carboxy temozolomide	16±3.4	＜0.001	9(6)	Furan epothilone D + temozolomide	22±4.2	＜0.001

Note: methyl benzylhydrazine, mitozolomide, temozolomide and 4-carboxyl temozolomide are anticancer drugs. The results show that furan epothilone D (group 2) and the anticancer drugs (groups 3 to 6) have a certain anti-tumor effect (P < 0.05) when applied alone, especially when administered topically, compared to the control group. The combination (groups 7 to 10) had a significant synergistic effect (P < 0.001).

Eighth, antitumor effects of topical application of anticancer drugs and epothilone (BMS-310705).

Using white rat as test object, 2X 10⁵Each hepatoma cell was injected subcutaneously into the quaternary costal region, and the tumors were divided into the following 10 groups 14 days after growth (see Table 6). The first group was control, and groups 2 to 10 were treatment groups, wherein group 2 was BMS-310705, and groups 3 to 6 were mitozolomide, temozolomide, 4-carboxy temozolomide, and 3-N-methyl temozolomide, respectively. Groups 7 to 10 are BMS-310705 in combination with mitozolomide, temozolomide, 4-carboxy temozolomide, 3-N-methyl temozolomide, respectively. All the drugs were placed intratumorally, except BMS-310705, which was 2.5mg/kg, anticancer drugs were 17.5 mg/kg. Tumor volume was measured on day 30 after treatment and the treatment effect was compared (see table 6).

TABLE 6

Test set (n)	Is treated by	Tumor volume (cm)³)	P value
Test set (n)	Is treated by	Tumor volume (cm)³)	P value	1(6)	Control	58±12
2(6)	BMS-310705	38±8	＜0.05	1(6)	Control	58±12
2(6)	BMS-310705	38±8	＜0.05	3(6)	Mitozolomide	36±8	＜0.01
4(6)	Temozolomide	42±8.8	＜0.01	3(6)	Mitozolomide	36±8	＜0.01
4(6)	Temozolomide	42±8.8	＜0.01	5(6)	4-carboxy temozolomide	38±6	＜0.01
6(6)	3-N-methyl temozolomide	36±8	＜0.01	5(6)	4-carboxy temozolomide	38±6	＜0.01
6(6)	3-N-methyl temozolomide	36±8	＜0.01	7(6)	BMS-310705+ mitozolomide	28±5.6	＜0.001
8(6)	BMS-310705+ temozolomide	18±3.8	＜0.001	7(6)	BMS-310705+ mitozolomide	28±5.6	＜0.001
8(6)	BMS-310705+ temozolomide	18±3.8	＜0.001	9(6)	BMS-310705+ 4-carboxy temozolomide	14±4	＜0.001
10(6)	BMS-310705+ 3-N-methyl temozolomide	24±6	＜0.001	9(6)	BMS-310705+ 4-carboxy temozolomide	14±4	＜0.001

Note: mitozolomide, temozolomide, 4-carboxy temozolomide and 3-N-methyl temozolomide are all anticancer drugs. The results show that BMS-310705 (group 2) and the anticancer drugs (groups 3 to 6) have a certain tumor-inhibiting effect (P < 0.05) when used alone, compared with the control group. However, the combination (groups 7 to 10) had a significant synergistic effect (P < 0.001).

Similar synergy is seen with other epothilones in combination with other anticancer drugs, such as: a combination of mitozolomide or temozolomide with epothilone B or epothilone D. The tested tumor cells include brain tumor (CNS-1, C6, 9L), gastric gland epithelial cancer (SA), bone tumor (BC), breast cancer (BA), Papillary Adenocarcinoma of Thyroid (PAT), liver cancer, pancreatic cancer, renal cancer, esophageal cancer, etc.

Experiment nine, comparison of in vivo release of epothilone D sustained-release implant made of polylactic acid of different molecular weights

Rats were used as test subjects, and divided into groups (3/group) and subcutaneously administered equivalent amounts of epothilone D sustained release implants loaded with polylactic acid (PLA) of different Molecular Weights (MW). Then, the remaining amount of the drug in the implant was measured on days 1, 3, 7, 14, 21, 28 and 35, respectively, to obtain the in vivo release rate (%). The results show that the release with molecular weight 20000 is: 1 day (8%), 3 (28%), 7 (56%), 14 (82%), 21 (90%), 28(94) and 35 (98%). Comparing in vivo release of epothilone D sustained-release implants made with different molecular weights, it was found that the release slowed with increasing molecular weight, and compared to the systemic group, the tumor suppression rate increased with increasing molecular weight of polylactic acid, as exemplified by day 7, in the order of 68% (MW: 5000), 66% (MW: 15000), 54% (MW: 25000), 50% (MW: 40000) and 48 (MW: 60000).

Experiment ten, anti-tumor effect of local application of anti-cancer drugs and epothilone D.

Using white rat as test object, 2X 10⁵Individual esophageal cancer cells were injected subcutaneously into the quaternary costal region and divided into the following 10 groups 14 days after tumor growth (see table 7). The first group is control and groups 2 to 10 are treatment groups, where group 2 is epothilone D and groups 3 to 6 are camptothecin, hydroxycamptothecin, topotecan, irinotecan, respectively. Groups 7 to 10 are combinations of epothilone D with camptothecin, hydroxycamptothecin, topotecan, irinotecan, respectively. All the drugs were placed intratumorally, except for epothilone D at 2.5mg/kg, the anticancer drugs were 17.5 mg/kg. Tumor volume was measured on day 30 after treatment and the treatment effect was compared (see table 7).

TABLE 7

Test set (n)	Is treated by	Tumor volume (cm)³)	P value
Test set (n)	Is treated by	Tumor volume (cm)³)	P value	1(6)	Control	56±12
2(6)	Epothilone D	40±10	＜0.05	1(6)	Control	56±12
2(6)	Epothilone D	40±10	＜0.05	3(6)	SN-38	32±8	＜0.01
4(6)	CPT-11	36±6	＜0.01	3(6)	SN-38	32±8	＜0.01
4(6)	CPT-11	36±6	＜0.01	5(6)	HCPT	34±8	＜0.01
6(6)	MD-CPT	30±6	＜0.01	5(6)	HCPT	34±8	＜0.01
6(6)	MD-CPT	30±6	＜0.01	7(6)	Epothilone D + SN-38	24±4.8	＜0.001
8(6)	Epothilone D + CPT-11	22±4.2	＜0.001	7(6)	Epothilone D + SN-38	24±4.8	＜0.001
8(6)	Epothilone D + CPT-11	22±4.2	＜0.001	9(6)	Epothilone D + HCPT	26±4	＜0.001
10(6)	Epothilone D + MD-CPT	18±4	＜0.001	9(6)	Epothilone D + HCPT	26±4	＜0.001

Note: SN-38, CPT-11, HCPT, MD-CPT are all anticancer drugs. The results show that compared with the control group, the epothilone D (group 2) and the anticancer drugs (groups 3 to 6) have a certain tumor inhibition effect (P is less than 0.05) when being used alone. However, the combination (groups 7 to 10) had a significant synergistic effect (P < 0.001).

Eleven tests on the tumor-inhibiting effect of locally applied anticancer drugs and epothilone B.

Using white rat as test object, 2X 10⁵Individual esophageal cancer cells were injected subcutaneously into the quaternary costal region and divided into the following 10 groups 14 days after tumor growth (see table 8). The first group was control and groups 2 to 10 were treatment groups, where group 2 was epothilone B and groups 3 to 6 were control. Groups 7 to 10 are combinations of epothilone B with mitozolomide, temozolomide, camptothecin, topotecan, respectively. All the drugs were placed intratumorally, except for epothilone B at 5mg/kg, the anticancer drugs were 15 mg/kg. Tumor volume was measured on day 30 after treatment and the treatment effect was compared (see table 8).

TABLE 8

Test set (n)	Is treated by	Tumor volume (cm)³)	P value
Test set (n)	Is treated by	Tumor volume (cm)³)	P value	1(6)	Control	54±10
2(6)	Epothilone B	46±10	＜0.05	1(6)	Control	54±10
2(6)	Epothilone B	46±10	＜0.05	3(6)	Valrubicin	34±10	＜0.01
4(6)	XK469	44±6	＜0.01	3(6)	Valrubicin	34±10	＜0.01
4(6)	XK469	44±6	＜0.01	5(6)	AD312	38±8	＜0.01
6(6)	ICRF-187	32±8	＜0.01	5(6)	AD312	38±8	＜0.01
6(6)	ICRF-187	32±8	＜0.01	7(6)	Epothilone B + valrubicin	24±4.6	＜0.001
8(6)	Epothilone B + XK469	18±6.2	＜0.001	7(6)	Epothilone B + valrubicin	24±4.6	＜0.001
8(6)	Epothilone B + XK469	18±6.2	＜0.001	9(6)	Epothilone B + AD312	26±4.8	＜0.001
10(6)	Epothilone B + ICRF-187	18±4.6	＜0.001	9(6)	Epothilone B + AD312	26±4.8	＜0.001

Note: valrubicin, XK469, AD312 and ICRF-187 are all anticancer drugs. The results show that compared with the control group, the epothilone B (group 2) and the anticancer drugs (groups 3 to 6) have certain tumor inhibition effects (P is less than 0.05) when being singly applied. However, the combination (groups 7 to 10) had a significant synergistic effect (P < 0.001).

The same synergistic effect can also be seen in epothilone B, epothilone D, isoepothilone D, BMS-247550, azaepothilone B, furan epothilone D or the combination of BMS-310705 and ICRF-193, and the tested tumors are lung cancer, brain tumor, gastric cancer, breast cancer and rectal cancer.

Particularly, the sustained-release preparation, particularly the sustained-release injection, has simple and convenient operation and good repeatability. Not only has good curative effect, but also has little toxic and side effect.

Different drug packages have different drug release characteristics from different biodegradable polymers. Further research finds that the slow-release auxiliary materials most suitable for the slow release of the medicament are one of or a combination of racemic polylactic acid, racemic polylactic acid/glycolic acid copolymer, monomethyl polyethylene glycol/polylactic acid copolymer, polyethylene glycol/polylactic acid copolymer, terminal carboxyl polylactic acid/glycolic acid copolymer, polifeprosan, di-fatty acid and sebacic acid copolymer, poly (erucic aciddipolymer-sebacic acid), poly (fumaric acid-sebacic acid), polylactic acid, polyglycolic acid and glycolic acid copolymer, xylitol, oligosaccharide, chondroitin, chitosan, hyaluronic acid, collagen, gelatin, poloxamer and albumin glue; the most suitable suspending agent is one or more of methylcellulose, hydroxymethyl cellulose, sodium carboxymethylcellulose, (iodine) glycerol, dimethicone, propylene glycol, carbomer, mannitol, sorbitol, surfactant, Tween 20, Tween 40, Tween 80, or their combination.

In summary, the experimental results show the synergistic effect of the epothilones of the present invention on the listed topoisomerase inhibitors or tetrazine anticancer drugs. Therefore, the effective components of the anticancer compound are combined or packaged separately of the epothilone and any one (or more) topoisomerase inhibitor or tetrazine anticancer drug. The above effective components can be made into any dosage form or shape, but preferably slow release dosage form, mainly sustained release injection or sustained release implant.

(IV) detailed description of the preferred embodiments

Example 1.

80mg of polylactic acid (PLGA, 50: 50) with the molecular weight peak of 20000-40000 is put into a container, 100ml of dichloromethane is added, after being dissolved and mixed evenly, 10mg of epothilone B and 10mg of camptothecin are added, the mixture is shaken again and dried in vacuum to remove the organic solvent. Immediately forming the dried solid composition, subpackaging, and performing ray sterilization to obtain the anticancer sustained release preparation containing 10% epothilone B and 10% camptothecin. All are weight percent. The drug release time of the anti-solid tumor drug composition in vitro physiological saline is 15 to 25 days, and the drug release time under the skin of a mouse is 25 to 50 days.

Example 2.

As described in example 1, except that the molecular weight peak of polylactic acid is 40000-60000 (PLGA, 75: 25), the effective anticancer component and the weight percentage are one of the following: 10% of epothilone D, isoepothilone D, BMS-247550, azaepothilone B, furaetheromycin D or BMS-310705 in combination with 10% of camptothecin, hydroxycamptothecin, 9-nitrocamptothecin, SN-38, CPT-11, HCPT, homocamptothecin, MD-CPT, podophyllotoxin, trihydroxyisoflavone, lurtotecan, topotecan, irinotecan, etoposide, teniposide, 14-hydroxydaunorubicin, a combination of amrubicin, doxorubicin, etorubicin, mitorubicin, 7-O-methylnogad-4' -epirubicin, esorubicin, carrubicin, idarubicin, epirubicin, medroxobin, nemorubicin, doxorubicin, roxobicin, valrubicin, zorubicin, XK469, zorubicin, AD312, ICRF-187, or ICRF-193.

Example 3.

70mg of polifeprosan (p-carboxyphenylpropane (p-CPP): Sebacic Acid (SA) 20: 80) copolymer is put into a container, 100ml of dichloromethane is added, after dissolving and mixing uniformly, 10mg of epothilone D and 20mg of mitozolomide are added, after shaking uniformly again, the microspheres for injection containing 10% of epothilone D and 20% of mitozolomide are prepared by a spray drying method. Then suspending the microspheres in physiological saline containing 15 percent mannitol to prepare the corresponding suspension type sustained-release injection with the viscosity of 20-300 cp (at 20-30 ℃). The slow release injection has the release time in vitro physiological saline of 15-25 days and the release time under the skin of a mouse of about 30-40 days.

Example 4.

The steps of the method for processing the sustained-release injection are the same as the example 3, but the difference is that the polifeprosan is 20: 80, and the anticancer active ingredients and the weight percentage thereof are as follows: 10% epothilone B, epothilone D, isoepothilone D, BMS-247550, azaepothilone B, furaetheromycin D or BMS-310705 in combination with 20% imidazotetrazine, imidazopiperazine, imidazopyridine, procarbazine, mitozolomide, temozolomide, 4-carboxy temozolomide or 3-N-methyl temozolomide. The viscosity of the slow release injection is 200-600 cp (at 20-30 ℃).

Example 5.

70mg of polylactic acid (PLA) with the molecular weight peak of 20000-40000 is put into a container, 100ml of dichloromethane is added, after the mixture is dissolved and mixed evenly, 15mg of temozolomide and 15mg of isoepothilone D are added, the mixture is shaken up again and then is dried in vacuum to remove the organic solvent. The dried drug-containing solid composition is frozen and crushed into micro powder containing 15 percent of temozolomide and 15 percent of isoepothilone D, and then the micro powder is suspended in physiological saline containing 1.5 percent of sodium carboxymethylcellulose to prepare the corresponding suspension type sustained-release injection with the viscosity of 220cD-340cp (at the temperature of 20-30 ℃). The slow release injection has the release time of 10-15 days in-vitro physiological saline and the release time of about 20-30 days under the skin of a mouse.

Example 6

The steps of the method for processing the sustained-release injection are the same as the example 5, but the difference is polylactic acid with the molecular weight peak value of 40000-50000, and the anti-cancer active ingredients and the weight percentage thereof are as follows: a combination of 5% imidazotetrazine, imidazopiperazine, imidazopyridine, procarbazine, mitozolomide, temozolomide, 4-carboxy temozolomide or 3-N-methyl temozolomide and 25% BMS-247550, azaepothilone B, furaetheromycin D or BMS-310705. The viscosity of the prepared suspension type sustained-release injection is 120-240 cp (at 20-30 ℃).

The viscosity of the injection is 10-650 cp (at 20-30 deg C).

Example 7.

30mg of polifeprosan (20: 80) and 40mg of PLA (WM: 10000-20000) are placed in a container, 100ml of dichloromethane is added, after the mixture is dissolved and uniformly mixed, 20mg of procarbazine and 10mg of furan epothilone D are added, after the mixture is shaken again, the microspheres for injection containing 20% of procarbazine and 10% of furan epothilone D are prepared by a spray drying method. Then suspending the microspheres in physiological saline containing 1.5 percent of sodium carboxymethylcellulose and 0.5 percent of Tween 80 to prepare the corresponding suspension type sustained-release injection with the viscosity of 180cp-260cp (at the temperature of 25 ℃ -30 ℃). The slow release injection has the release time in vitro physiological saline of 14-25 days and the release time under the skin of a mouse of about 30-50 days.

Example 8.

The procedure of the process for preparing sustained release injection is the same as that of example 7, except that 30mg of polifeprosan (50: 50) and 40mg of PLA (WM: 20000-40000) contain the following anti-cancer effective ingredients: a combination of 5-23% epothilone B, epothilone D, isoepothilone D, BMS-247550, azaepothilone B, furaetheromycin D or BMS-310705 with 5-25% procarbazine, mitozolomide, temozolomide, 4-carboxy temozolomide or 3-N-methyl temozolomide; the viscosity of the slow release injection is 440-650 cp (at 25-30 ℃).

Example 9

50mg of polifeprosan (50: 50) and 20mg of PLGA (WM: 10000-20000, 50: 50) are placed in a container, 100ml of dichloromethane is added, after being dissolved and mixed evenly, 20mg of 4-carboxyl temozolomide and 10mg of BMS-310705 are added, after being shaken again, the microspheres for injection containing 20 percent of 4-carboxyl temozolomide and 10 percent of BMS-310705 are prepared by a spray drying method. Then suspending the microspheres in physiological saline containing 1.5 percent of sodium carboxymethylcellulose, 15 percent of sorbitol and 0.2 percent of Tween 80 to prepare the corresponding suspension type sustained-release injection with the viscosity of 100cp-160cp (at 25 ℃ -30 ℃). The slow release injection has the release time of 10-15 days in-vitro physiological saline and the release time of about 20-30 days under the skin of a mouse.

Example 10

The steps of the method for processing the sustained-release injection are the same as the example 9, but the difference is that the auxiliary materials are 40mg of polifeprosan (40: 60) and 30mg of PLGA (WM: 20000-40000, 75: 25), and the anticancer active ingredients are: a combination of 15% procarbazine, mitozolomide, temozolomide, 4-carboxy temozolomide or 3-N-methyl temozolomide with 15% epothilone B, epothilone D, isoepothilone D, BMS-247550, azaepothilone B, furaetheromycin D or BMS-310705; the viscosity is 560cp-640cp (at 25 ℃ -30 ℃).

Example 11

70mg of copolymer of di-fatty acid and sebacic acid (PFAD-SA) is put into a container, 100ml of dichloromethane is added, after the mixture is dissolved and mixed evenly, 10mg of 3-N-methyl temozolomide and 20mg of BMS-247550 are added, after the mixture is shaken up again, the microsphere for injection containing 10 percent of 3-N-methyl temozolomide and 20 percent of BMS-247550 is prepared by a spray drying method. Then the microspheres are prepared into the corresponding sustained-release implant by a tabletting method. The slow release implant has the release time in vitro physiological saline of 15-25 days and the release time under the skin of a mouse of about 30-40 days.

Example 12

The procedure of the method for processing the sustained release implant is the same as that of example 11, except that the sustained release excipients and the anticancer active ingredients are:

(1) 70% of racemic polylactic acid and 15% of epothilone B and 15% of temozolomide;

(2) 60% of racemic polylactic acid/glycolic acid copolymer and 15% of epothilone D and 25% of mitozolomide;

(3) 60% polyethylene glycol/polylactic acid and 15% isoepothilone D and 15% SN-38;

(4) 70% PFAD-SA and 10% azaepothilone B and 20% CPT-11;

(5) 70% P (EAD-SA) and 20% Furanepothilones D and 10% HCPT; or

(6) 70% P (FA-SA) and 5% BMS-247550 and 25% MD-CPT.

Example 13.

70mg of polifeprosan (50: 50) copolymer is put into a container, 100ml of dichloromethane is added, after being dissolved and mixed evenly, 10mg of epothilone D and 20mg of amrubicin are added, after shaking up again, the microspheres for injection containing 10% of epothilone D and 20% of amrubicin are prepared by a spray drying method. Then suspending the microspheres in physiological saline containing 15 percent mannitol to prepare the corresponding suspension type sustained-release injection with the viscosity of 200-300 cp (at 20-30 ℃). The slow release injection has the release time in vitro physiological saline of 15-25 days and the release time under the skin of a mouse of about 30-40 days.

Example 14

The procedure of the method for processing the sustained release implant is the same as that of example 13, except that the sustained release excipients and the anticancer active ingredients are:

(1) 40% of racemic polylactic acid (MW: 15K-25K), 30% of polifeprosan (50: 50), 15% of epothilone B and 15% of valrubicin, and the viscosity of the prepared sustained-release injection is 220cp-300 cp;

(2) 70% PLGA (MW: 15K-25K), 20% epothilone D and 10% mitozolomide, and the viscosity of the prepared sustained-release injection is 160cp-280 cp;

(3) 60% of polyethylene glycol/polylactic acid, 15% of isoepothilone B and 15% of XK469, and the viscosity of the prepared sustained-release injection is 220cp-300 cp;

(4) the viscosity of the prepared sustained-release injection is 260-380 cp by 70% of PFAD-SA, 15% of azaepothilone B and 15% of AD 312;

(5) 70% of P (EAD-SA), 15% of furan epothilone D and 15% of ICRF-187, and the viscosity of the prepared sustained-release injection is 320cp-420 cp; or

(6) 70% P (FA-SA), 25% BMS-310705 and 5% ICRF-193 to obtain the slow release injection with viscosity of 260-380 cp.

Example 15

70mg of polylactic acid (PLGA, 50: 50) with the molecular weight peak value of 15000-35000 is put into a container, 100ml of dichloromethane is added, after the materials are dissolved and mixed evenly, 25mg of mitozolomide and 5mg of epothilone B are added, after the materials are shaken up again, the microspheres for injection containing 25% of mitozolomide and 5% of epothilone B are prepared by a spray drying method. Then the microspheres are prepared into the corresponding sustained-release implant by a tabletting method. The slow release implant has the release time of 15-20 days in vitro physiological saline and the release time of about 35-50 days under the skin of a mouse.

Example 16

The steps of the method for processing the sustained-release implant are the same as the examples 13 to 15, but the difference is that the sustained-release implant comprises the following effective anticancer components in percentage by weight:

(1) 1-40% of epothilone B, epothilone D, isoepothilone D, BMS-247550, azaepothilone B, furaetheromycin D or BMS-310705 in combination with 1-40% of camptothecin, hydroxycamptothecin, 9-nitrocamptothecin, lurtotecan, topotecan, irinotecan, etoposide, teniposide, amrubicin, idarubicin, epirubicin, medroxobin, doxorubicin, roxobicin, valrubicin, zorubicin, XK469, AD312, ICRF-187 or ICRF-193; or

Example 17

The procedure of processing into sustained release preparation is the same as that of examples 1-16, except that the sustained release excipient is one or a combination of the following:

a) polylactic acid (PLA) with a molecular weight peak of 10000-;

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

c) polifeprosan, p-carboxyphenylpropane (p-CPP): sebacic Acid (SA) 10: 90, 20: 80, 30: 70, 40: 60, 50: 50 or 60: 40;

d) polifeprosan in combination with PLA or PLGA;

e) di-fatty acid and sebacic acid copolymer (PFAD-SA);

f) poly (erucic acid dimer-sebacic acid) [ P (EAD-SA) ];

g) poly (fumaric-sebacic acid) [ P (FA-SA) ];

h) xylitol, oligosaccharide, chondroitin, chitin, chitosan, hyaluronic acid, collagen, gelatin or albumin glue;

i) racemic polylactic acid, racemic polylactic acid/glycolic acid copolymer, monomethyl polyethylene glycol/polylactic acid copolymer, polyethylene glycol/polylactic acid copolymer, carboxyl-terminated polylactic acid or carboxyl-terminated polylactic acid/glycolic acid copolymer.

Example 18

The procedure for preparing a sustained release injection is the same as in examples 1 to 10, except that the suspending agent used is one or a combination of the following:

a) 0.5-3.0% carboxymethylcellulose (sodium);

b) 5-15% mannitol;

c) 5-15% sorbitol;

d) 0.1-1.5% of surface active substances;

e) 0.1-0.5% tween 20.

Example 19.

70mg of polylactic acid (PLGA, 75: 25) with the molecular weight peak of 20000-60000 is put into a container, 100ml of dichloromethane is added, after the mixture is dissolved and mixed evenly, 25mg of mitozolomide and 5mg of epothilone D are added, the mixture is shaken again and dried in vacuum to remove the organic solvent. Freeze-pulverizing the dried solid composition containing drug to obtain micropowder containing 25% mitozolomide and 5% epothilone D, and suspending in physiological saline containing 1.5% sodium carboxymethylcellulose to obtain suspension type sustained-release injection with viscosity of 220-340 cp (at 20-30 deg.C). The slow release injection has the release time in vitro physiological saline of 21-30 days and the release time under the skin of a mouse of about 20-30 days.

Example 20

The steps of the method for processing the sustained-release injection are the same as the example 19, but the difference is that the anticancer active ingredients and the weight percentage thereof are as follows:

(2) 1-40% epothilone B, epothilone D, isoepothilone D, BMS-247550, azaepothilone B, furazaepothilone D or BMS-310705 in combination with 1-40% imidazotetrazine, imidazopiperazine, imidazopyridine, procarbazine, mitozolamide, temozolomide, 4-carboxy temozolomide or 3-N-methyl temozolomide. The viscosity of the injection is 10-650 cp (at 20-30 deg C).

The above examples further describe the technical process of the present invention. Are intended to be illustrative and not limiting of the scope of the invention. Indeed, various modifications of the invention in addition to those shown and described herein will become apparent to those skilled in the art from the foregoing description and accompanying drawings. Such modifications are, of course, intended to be within the scope of the appended claims. It should be understood, therefore, that the foregoing description focuses on certain specific embodiments of the invention and that equivalent alterations and substitutions made thereto are within the spirit and scope of the appended claims.

Claims

1. An anti-solid tumor composition is characterized in that the anti-solid composition is a slow release injection and comprises the following components:

(A) a sustained release microsphere comprising:

0.5-60% of anticancer active ingredient

The sustained-release auxiliary material is 41 to 99.9 percent

0.0 to 30 percent of suspending agent

The above are weight percentages

And

Wherein,

the anticancer active ingredients are epothilone derivatives and anticancer drugs, and the anticancer drugs are selected from topoisomerase inhibitors and/or tetrazine drugs;

the slow release auxiliary material is selected from one or the combination of the following materials:

a) polylactic acid;

b) copolymers of polyglycolic acid and glycolic acid;

c) polifeprosan;

d) polifeprosan in combination with polylactic acid or a copolymer of polyglycolic acid and glycolic acid;

e) a di-fatty acid and sebacic acid copolymer;

f) poly (erucic acid dimer-sebacic acid) copolymer;

g) poly (fumaric acid-sebacic acid) copolymer;

The suspending agent is selected from sodium carboxymethylcellulose, iodoglycerol, dimethicone, propylene glycol, carbomer, mannitol, sorbitol, surfactant, Tween-20, Tween-40 and Tween-80 or their combination, and has viscosity of 100-3000 cp (at 20-30 deg.C).

2. The sustained-release anticancer injection according to claim 1, wherein the epothilone derivative is selected from the group consisting of epothilone, epothilone A, epothilone B, epothilone C, epothilone D, epothilone E, epothilone F, 4-demethyl-9-one-epothilone C, 12, 13-dihydro-13-oxoepothilone C, 21-aminoepothilone B, 26-aminoepothilone B, 21, 26 , and F-Truese ,9, 10-dehydroepothilone B, 10, 11-hydrogen epothilone B, 26, 27-halogenoepothilone B, 9, 10, 11, 14, 21, 26-hydroxy-substituted epothilone B, 21, 26-dihydroxy epothilone B, 21-hydroxy-10, 11-dehydroepothilone B, epothilone A, B, and C, B, and C, respectively, which are substituted with hydroxy groups at positions 9, 10, 11, 14, 21, 26, and 26, respectively, 4-demethyl-9-one-epothilone B, 4-demethyl-9, 10-didehydro-epothilone B, 4-demethyl-10, 11-didehydro-epothilone B, 6-demethyl-10, 11-didehydro-epothilone B, 21-aminoepothilone B, 21-hydroxyepothilone B, 26-fluoroepothilone B, 26-aminoepothilone B, 12, 13-cyclopropylepothilone B, 12, 13-cyclobutyl epothilone B, ixabepilone, azaepothilone B, 26-trifluoro- (E) -9, 10-dehydro-12, 13-desoxyepothilone B, epothilone D in which positions 21 and 26 are substituted by amino groups, epothilone D in positions 9 and 10, dehydroepothilone D in positions 9 and 10, respectively or simultaneously, 10, 11-dehydroepothilone D, 26, 27-halogen-substituted epothilone D, 9-epothilone D, 10-hydroxy-10, 11-dehydroepothilone D, 4-demethyl-9-one-epothilone D, 4-demethyl-9, 10-didehydro epothilone D, 4-demethyl-10, 11-didehydro epothilone D, 6-demethyl-10, 11-didehydro epothilone D, 21-hydroxyepothilone D, 21-aminopeptidase D, 26-hydroxyepothilone D, 26-aminoepothilone D, 26-fluoroepothilone D, isoepothilone, isoepothilone D, 9, 10-dehydroepothilone D, 10, 11-dehydroepothilone D, furaetheromycin D, (E) -9, 10-dehydro-12, 13-desoxyepothilone D, BMS-310705, 6-ethyl, 16-fluoro, 17-pyridinepothilone D, 11, 12-dehydro-12, 13-dehydro-13-desoxyepothilone D, 9-oxyepothilone D or 8-epi-9-oxyepothilone D.

3. The sustained-release anticancer injection according to claim 1, wherein the tetrazine drug is selected from one of imidazotetrazine, imidazopiperazine, imidazopyridine, procarbazine, mitozolomide, temozolomide, 4-carboxy temozolomide, 3-N-methyl temozolomide or a combination thereof.

4. The sustained-release anticancer injection according to claim 1, wherein the topoisomerase inhibitor is selected from one or a combination of camptothecin, hydroxycamptothecin, 9-nitrocamptothecin, SN-38, CPT-11, HCPT, homocamptothecin, MD-CPT, podophyllotoxin, trihydroxyisoflavone, lurtotecan, topotecan, irinotecan, etoposide, teniposide, 14-hydroxydaunorubicin, amrubicin, doxorubicin, ditobicin, 7-O-methylnocar-4' -epirubicin, esorubicin, carrubicin, idarubicin, epirubicin, medroxobin, nemorubicin, doxorubicin, roxobicin, valrubicin, zorubicin, XK469, zorubicin, AD312, ICRF-187, ICRF-193

5. The sustained-release anticancer injection according to claim 1, wherein the anticancer active ingredient is one of the following:

6. The sustained-release anticancer injection according to claim 1, wherein:

a) the molecular weight peak value of the polylactic acid is 10000-;

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

c) in polifeprosan, the ratio of p-carboxyphenylpropane to sebacic acid is 10: 90, 20: 80, 30: 70, 40: 60, 50: 50 or 60: 40.

7. The sustained-release anticancer injection according to claim 1, wherein the suspending agents are respectively one of the following:

a) 0.5-3.0% carboxymethylcellulose (sodium);

b) 5-15% mannitol;

c) 5-15% sorbitol;

d) 0.1-1.5% of surface active substances;

e) 0.1-0.5% tween 20;

f) (iodine) glycerol, dimethicone, propylene glycol or carbomer;

g) 0.5-5% of sodium carboxymethylcellulose and 0.1-0.5% of Tween 80;

h) 5-20% of mannitol and 0.1-0.5% of Tween 80;

i)0.5 to 5 percent of sodium carboxymethylcellulose, 5 to 20 percent of sorbitol and 0.1 to 0.5 percent of Tween 80.

8. The sustained-release anticancer injection according to claim 1, wherein the pharmaceutical preparation is a sustained-release implant made of sustained-release microspheres, and is administered by intratumoral or peritumoral injection or placement.

9. The sustained-release anticancer implant according to claim 7, characterized in that the anticancer active constituents and the weight percentages thereof are:

10. The sustained-release anticancer agent according to claims 1 and 8, characterized in that the sustained-release agent is used for preparing a drug for treating primary or secondary cancer, sarcoma or carcinosarcoma originated from brain, central nervous system, kidney, liver, gallbladder, head and neck, oral cavity, thyroid gland, skin, mucosa, gland, blood vessel, bone tissue, lymph node, lung, esophagus, stomach, breast, pancreas, eye, nasopharynx, uterus, ovary, endometrium, cervix, prostate, bladder, colon or rectum of human and animal, and is administered by intratumoral or peritumoral injection or placement.