CN104306332B

CN104306332B - Camptothecin phospholipid compound, and medicinal composition and application thereof

Info

Publication number: CN104306332B
Application number: CN201410495174.XA
Authority: CN
Inventors: 李新松; 方硕
Original assignee: Southeast University
Current assignee: Southeast University
Priority date: 2014-09-24
Filing date: 2014-09-24
Publication date: 2017-02-15
Anticipated expiration: 2034-09-24
Also published as: CN104306332A; WO2016045505A1

Abstract

The invention discloses a camptothecin phospholipid compound, and a preparation method, a medicinal composition and a use thereof. The medicinal composition is a camptothecin phospholipid compound or a combined medicinal composition composed of the camptothecin phospholipid compound and a pharmacodynamically acceptable carrier, and can be a liquid preparation, a solid preparation, a semisolid preparation, a capsule, a granule, a gel or an injection. The medicinal composition is camptothecin phospholipid compound or a liposome nanoparticle prepared by using the camptothecin phospholipid compound and an assistant, and the particle size of the nanoparticle is 10-1000nm. The camptothecin phospholipid compound and its liposome nanoparticle can be used as a liquid preparation, a solid preparation, a semisolid preparation, a sterilized preparation or an aseptic preparation, have low toxicity, and can be used for efficient treatment of various tumors.

Description

A kind of camptothecin phosphatide cpd, its pharmaceutical composition and application

Technical field

The present invention is a kind of camptothecin phosphatide cpd with antitumor action and its pharmaceutical composition and purposes, relates to And pharmaceutical technology field.

Background technology

Camptothecine (Camptothecin, CPT) is a kind of alkaloid, has stronger cytotoxic activity, can be used for treating Several Kinds of Malignancy, such as gastric cancer, hepatocarcinoma, bladder cancer and leukemia etc..Scientist finds the unique anticancer of camptothecine compounds Mechanism：The triplet complex stable by forming camptothecine-topoisomerase I-DNA, after so that DNA double helical structure is ruptured not Reproducible.Camptothecine compounds are the specific inhibitors of Topo I the most classical.Topo I has become design new type anticancer One of the most popular target spot of medicine.Topo I inhibitor is analyzed network system by national cancer institute (NCI) drug Mechanism System is classified as one of six big series antineoplastic medicaments of primary study.

Camptothecine and the like can extract from plant, also can carry out chemosynthesis.This kind of camptothecine and its similar Thing etc. also produces the serious side effects such as bone marrow depression, vomiting and diarrhoea while playing its anti-tumor activity.Camptothecine and its The greatest drawback that analog is used for oncotherapy is poorly water-soluble it is difficult to obtain the solution of high concentration.In order to improve camptothecine Defect, many research institutions and enterprise have carried out modification to the chemical constitution of camptothecine, have synthesized hundreds of camptothecin derivatives. So far there are 2 kinds of camptothecin derivatives to be approved for clinical cancer therapy, be according to for vertical health irinotecan with open up respectively Flutter vertical health topotecan, but its water solublity is still relatively low.Additionally, 9-AC, 9-NC, G7147211, DX-8951f, Bay38- 3441, BN80915 and CKD-602 etc. are respectively at clinical experimental stage.More camptothecin analogues are not applied.Portion Point reason is because that water solublity is too poor, and camptothecine and the like has the defects such as side reaction is big for oncotherapy.

Four hexatomic rings and a five-membered ring is contained, wherein E ring is a α-hydroxyl in the molecule of camptothecine and the like Base lactone, and have a chiral centre, it is 20S configuration.E cyclocarbonyl activity is high, captures it is easy to open in entering of nucleopilic reagent Ring.Because camptothecine has the balance between the lactone form of closing and the carboxylate form of open loop in vivo, and open loop shape Formula depends on pH value and animal species.Particularly human serum albumin (HSA) is preferentially combined with the open loop form of camptothecine, is formed Stable complex, makes balance to open loop form displacement so that the lactone content in vivo with anti-tumor activity is too low, leads to Anti-tumor activity in human body is very low.It is said that the serious toxicity (mice maximum tolerated dose MTD 12mg/kg) of camptothecine is main Be because when medicine enter human body in after camptothecine open loop form combine closely with HSA, make camptothecine balance in vivo from Close lactone form to shift to open loop form, camptothecine open loop (half-life is less than half an hour) quickly.

Because camptothecine and 10-hydroxycamptothecine are not modified natural products, toxicity is larger, and anti-tumor activity is low, Limit them in clinical application；Furthermore, these medicines of patient's life-time service are also easy to produce drug resistance, in the urgent need to improving Fructus seu radix camptothecae acuminatae (Fructus Camptothecae Acuminatae) Alkali or the water solublity of its analog, find the camptothecin antineoplastic agents of high-efficiency low-toxicity.

It is therefore desirable to by structural modification is carried out to camptothecin analogues, improving drug solubility, target to reach Tropism, prevents lactone open loop, improves curative effect, drops hypotoxic purpose, also the study hotspot of always drug world.

Liposome is a kind of new medicinal preparation with target administration function, has extremely in cancer therapy drug research field It is widely applied.Phospholipid is the main chemical compositions constituting liposome, and by having, the connected substituent group of phosphoric acid is hydrophilic because constitute The hydrophobic tail composition of head and long hydrocarbyl chain composition.When being scattered in aqueous phase, the hydrophobic tail of molecule is tended to flock together, and keeps away Boiled water phase, and hydrophilic head is exposed to aqueous phase, formed have bilayer structure vesicle, form liposome.Due to Contain unsaturated bond natural phospholipid hydrophobic tail more, very big impact is caused to the stability of liposome, therefore phospholipid is dredged Water afterbody carries out structural modification and is increasingly becoming study hotspot to improve its stability.During as carrier, hydrophobic drug generally exists In double lipid layers of liposome, hydrophilic medicament is in the aqueous phase in lipid body.Due to the mobility of liposome membrane, lead to medicine It is easy to leak out, make the medicine of parcel be difficult to play good drug effect.

The present invention is led to by the use of camptothecine and the like the chemical moieties of 2 molecules as hydrophobic tail and phospholipid hydrophilic head Cross covalent bond be connected, prepare camptothecin phosphatide cpd, dissolubility is significantly better than camptothecin analogues, can individually or Person and phospholipid are mixed and made into elaioplast nanometer particle；The present invention camptothecin phosphatide cpd is had can be formed liquid preparation, The characteristic of solid preparation, semi-solid preparation, sterilization preparation and sterile preparation；Camptothecin phosphatide cpd is prepared into by the present invention Nano-particle, has the characteristic of liposome, has and can form liquid preparation, solid preparation, semi-solid preparation, sterilization preparation and nothing The characteristic of bacteria preparation, for oncotherapy.This camptothecin phosphatide cpd is degraded by esterase in vivo, discharges medicine, no It is only a kind of prodrug, be also the brand-new drug release carrier of camptothecine；The camptothecin phosphatide cpd of the present invention, tool There are relatively low toxic and side effects, liposome has target function.

Content of the invention

Technical problem：It is an object of the invention to provide a kind of improve camptothecine or the deliquescent camptothecin of its analog Phosphatide cpd, is simultaneously based on the pharmaceutical composition of this camptothecin phosphatide cpd, and this camptothecin phosphatide cpd Application in preparing antitumor drug.

Technical scheme：In order to Compound nomenclature and positioning, the position defining each atom of camptothecine is as follows：

The female ring of all camptothecines according to the present invention and the like or its phosphatide cpd is all according to the position in formula (A) Put and be defined, and 20 carbon atoms are S configuration.

The camptothecin phosphatide cpd of the present invention, is the compound of general formula (1) or the compound of described formula (1) The pharmaceutically acceptable salt being formed with counter ion counterionsl gegenions：

In formula (1), R₁For spacerarm, be alkylene, described alkylene be defined as straight chain that carbon number is 1～20 or Alkylene alkyl/Asia the alkylene of chain or the aryl that carbon number is 6～10；L represents 2- amino -2- carboxy ethyl, 2- amino Ethyl, 2- trimethyl amido ethyl cation or 2,3- dihydroxypropyl；

R_xIt is camptothecine compounds (20S) 20 bit substituents, the camptothecin of formula (3) that structure is following formula (2) 7 methylene substituent of camptothecine compounds (20S) of compound (20S) 10 bit substituent or formula (4)：

In formula (2) and (3) and (4), R₇、R₉、R₁₀、R₁₁And R₁₂It is any one group following：Hydrogen, halogen, hydroxyl, C₁- C₆The alkyl/alkoxyl of straight or branched, cyano group, nitro, amino, C₁-C₆Straight or branched haloalkyl/alkyl halide Epoxide, C₁-C₆Three substituted silane bases of straight or branched alkyl, C₁-C₆Straight or branched alkylene, carboxaldehyde radicals, C₁-C₆'s Straight or branched alkyl ester group, C₁-C₆Straight or branched alkyl carbonyl, C₁-C₆The hydroxy alkyl of straight or branched, C₁-C₆ Straight or branched alkoxyl methylene, phenyoxymethylene, benzene methyl methylene, dimethylamino methyl, dimethyl amido Ethyl, there is piperazine structureSubstituent group, there is structureSubstituent group, have Connection piperazine carboxyl structureSubstituent group and-CH₂NR₁₃R₁₄, described-CH₂NR₁₃R₁₄In, R₁₃And R₁₄For C₁-C₆Straight chained alkyl, C₁-C₆Branched alkyl or R₁₁And R₁₀Form structureSubstituent group；

In formula (3) and (4), R represents C₁-C₁₈Straight chain/branched alkyl or C₆-C₁₀Aryl.

In formula (2) and (3) and (4), 20 carbon atoms are S configuration.

In the present invention, R_xPreferably 20 hydroxyl substituents of camptothecine (CPT), 20 hydroxyl substituents of 10-hydroxycamptothecine, 10 hydroxyl substituents of 10-hydroxycamptothecine, 20 hydroxyl substituents of irinotecan (CPT11 joins piperazine camptothecine), the vertical health of topology Vertical health (9- dimethylamino -10- the hydroxy-camptothecin of (9- dimethylamino -10- hydroxy-camptothecin seals) 10 hydroxyl substituents, topology Seal) 20 hydroxyl substituent, 20 hydroxyl substituents of SN38, SN38s 10 Hydroxyl substituent,

In the preferred version of camptothecin phosphatide cpd of the present invention, counter ion counterionsl gegenions be cation proton, sodium ion, potassium from Son, calcium ion, iron ion, magnesium ion, ammonium ion, zinc ion, anion chloride ion, sulfate ion, sulfate ion, nitric acid Radical ion, carboxylic acid ion, carbanion, bromide ion, phosphate anion, formate, acetate, citrate, lactate, Fumaric acid radical, tartrate anion, any one or any two combination of gluconic acid radical ion, preferably proton, sodium ion, chlorine from Son.

The method preparing above-mentioned camptothecin phosphatide cpd of the present invention, first by camptothecine or camptothecin analogues Hydroxyl and dicarboxylic anhydride or carboxylic acid reaction, prepare the intermediate containing carboxyl, then further by described intermediate and glycerol Phosphocholine carries out condensation reaction under condensation reagent acts on, that is, obtain camptothecin phosphatide cpd.

In the preferred version of preparation method of the present invention, using condensation reagent preferably 1,3 dicyclohexylcarbodiimides, two pyridines Carbonic ester, 1- (3- dimethylamine propyl) -3- ethyl carbimide hydrochlorate, 1,3- diisopropylcarbodiimide or N, N- carbonyl diurethane One of imidazoles and tertiary amine, the preferred DMAP of tertiary amine or 4- pyrollidinopyridine.

The pharmaceutical composition of the present invention, including above-mentioned camptothecin phosphatide cpd or described camptothecin phospholipid Compound and acceptable carrier on pharmacodynamicss.

The pharmaceutical composition of the present invention is liquid preparation, solid preparation, semi-solid preparation, capsule, granule, gel Agent, injection, slow releasing preparation or controlled release preparation.

In the preferred version of pharmaceutical composition of the present invention, pharmaceutical composition is the liposome nanometer of particle diameter 10-1000 nanometer Granule, also includes auxiliary agent in this pharmaceutical composition.

In the above-mentioned preferred version of pharmaceutical composition of the present invention, auxiliary agent is phospholipid.

Application in preparing antitumor drug for the camptothecin phosphatide cpd of the present invention, by camptothecin phospholipid chemical combination Thing or its pharmaceutically acceptable salt, are prepared into medicament with acceptable carrier on pharmacodynamicss.

The compounds of this invention can be presented in isomer, and generally described " the compounds of this invention " includes this change The isomer of compound.

The compounds of this invention there may be asymmetric center and has S configuration or R configuration, and the present invention includes all possible solid Isomer and the mixture of two or more isomers.

The invention still further relates to helping containing as the compounds of this invention of active ingredient or the compounds of this invention and conventional medicine The pharmaceutical composition of agent.Generally pharmaceutical composition of the present invention contains the compounds of this invention of 0.1-100 weight %.

The pharmaceutical composition of the compounds of this invention can be prepared according to method well known in the art.When for this purpose, if Need, can be by the compounds of this invention or the compounds of this invention and one or more solid or liquid pharmaceutical excipients and/or adjuvant In conjunction with making the suitable administration form that can use as people's medicine or dosage form.

The compounds of this invention or can be administered in a unit containing its pharmaceutical composition, route of administration can be intestinal Or non-bowel, as oral, muscle, subcutaneous, nasal cavity, oral mucosa, skin, peritoneum or rectum etc..

The compounds of this invention or the route of administration containing its pharmaceutical composition can be drug administration by injection, including intravenous injection, Intramuscular injection, subcutaneous injection, intradermal injection and acupoint injection therapy etc..Form of administration can be liquid dosage form, solid dosage formss.As liquid Dosage form can be true solution class, colloidal type, particulate formulations, emulsion dosage form, mixed suspension form.Other dosage forms for example tablet, capsule, drip Ball, aerosol, pill, powder, solution, suspensoid, Emulsion, granule, suppository, lyophilized injectable powder etc..

The compounds of this invention can be made ordinary preparation, can also be slow releasing preparation, controlled release preparation, targeting preparation and various Particulate delivery system.

In order to unit dosage forms for administration is made tablet, various carrier well known in the art can be widely used.With regard to carrier Example be, such as diluent and absorbent, such as starch, dextrin, calcium sulfate, Lactose, Mannitol, sucrose, sodium chloride, Fructus Vitis viniferae Sugar, carbamide, Calcium Carbonate, kaolin, Microcrystalline Cellulose, aluminium silicate etc.；Wetting agent and binding agent, such as water, glycerol, Polyethylene Glycol, Ethanol, propanol, starch slurry, dextrin, syrup, Mel, glucose solution, mucialga of arabic gummy, gelatine size, sodium carboxymethyl cellulose, Lac, methylcellulose, potassium phosphate, Polyvinylpyrrolidone etc.；Disintegrating agent, be for example dried starch, alginate, agar powder, Laminaran, sodium bicarbonate and citric acid, Calcium Carbonate, polyoxyethylene sorbitol fatty acid ester, dodecyl sodium sulfate, methyl Cellulose, ethyl cellulose etc.；Disintegration inhibitor, such as sucrose, glyceryl tristearate, cocoa butter, hydrogenated oil and fat etc.；Absorb and promote Enter agent, such as quaternary ammonium salt, sodium lauryl sulphate etc.；Lubricant, such as Pulvis Talci, silicon dioxide, corn starch, stearic acid Salt, boric acid, liquid paraffin, Polyethylene Glycol etc..Tablet can also be made further coated tablet, such as sugar coated tablet, thin film bag Garment piece, ECT, or double-layer tablet and multilayer tablet.

In order to administration unit is made pill, various carrier well known in the art can be widely used.Example with regard to carrier Son is, such as diluent and absorbent, such as glucose, Lactose, starch, cocoa butter, hydrogenated vegetable oil, Polyvinylpyrrolidone, Gelucire, Kaolin, Pulvis Talci etc.；Binding agent, such as arabic gum, Tragacanth, gelatin, ethanol, Mel, liquid sugar, rice paste or Batter etc.；Disintegrating agent, such as agar powder, starch, alginate, dodecyl sodium sulfate, methylcellulose, ethyl cellulose are dried Deng.

In order to administration unit is made capsule, effective ingredient the compounds of this invention is mixed with above-mentioned various carriers, and Thus obtained mixture is placed in hard gelatine capsule or soft capsule.Also effective ingredient the compounds of this invention can be made micro- Wafer, is suspended in aqueous medium and forms suspensoid, also can load in hard capsule or make injection application.

The compounds of this invention is made injection preparation, such as solution, suspensoid solution, Emulsion, lyophilized injectable powder, this Kind of preparation can be aqueous or non-aqueous, can contain acceptable carrier on a kind of and/or multiple pharmacodynamicss, diluent, binding agent, Lubricant, preservative, surfactant or dispersant.As diluent is selected from water, ethanol, Polyethylene Glycol, 1,3-PD, second The isooctadecanol of epoxide, polyoxygenated isooctadecanol, Polyoxyethylene Sorbitol Fatty Acid Esters etc..In addition, it is isotonic in order to prepare Injection, can add appropriate sodium chloride, glucose or glycerol in injection preparation, further, it is also possible to add routine Cosolvent, buffer agent, pH adjusting agent etc..These adjuvants are commonly used in the art.

If desired, coloring agent, preservative, spice, correctivess, sweeting agent or other material can also be added in pharmaceutical preparation Material.

Elaioplast nanometer particle, grain diameter 10-1000 nanometer are made by the compounds of this invention.

Elaioplast nanometer particle, particle diameter 10-1000 nanometer are made by the compounds of this invention and auxiliary agent, the auxiliary agent of use is two Dimyristoylphosphatidycholine DMPC, Distearoyl Phosphatidylcholine DSPC, dipalmitoyl phosphatidyl choline DPPC, L- α-phospholipid Phatidylcholine-β-arachidonic acyl-γ-stearoyl, two caprylyl glyceryl -3- phosphocholine DDPC, 1,2- dioleoyl phosphatidyl Choline DOPC, 2- oleoyl -1- palmitoyl glycerol -3- phosphocholine POPC, 1- stearoyl -2- oleoyl glycerol-3-phosphocholine SOPC, 1- stearoyl -2- palmitoyl glycerol -3- phosphocholine SPPC, 1- myristoyl -2- stearoyl phosphatidyl choline MSPC, 1- palmityl -2- myristoyl phosphatidylcholine PMPC, 1- palmityl -2- stearoyl phosphatidyl choline PSPC, 1- stearyl -2- myristoyl phosphatidylcholine SMPC, 1- myristoyl -2- oleolyl phosphatidyl choline MOPC, 1- palmityl -2- oleolyl phosphatidyl choline POPC, 1- oleoyl -2- palmitoyl glycerol -3- phosphocholine OPPC, 1- Oleoyl -2- monostearoyl glyceride -3- phosphocholine OSPC, 1,2- DLPC DLPC, 1,2- bis- arachidonic acyl Base phosphatidylcholine DAPC, 1- palmityl -2- myristoyl-glycerol-3-phosphocholine PMPC, 1- myristoyl -2- are hard Phosphatidyl glycerol -3- phosphocholine MSPC, 1- myristoyl -2- palmitoyl glycerol -3- phosphocholine MPPC, 1- palmityl -2 flower Raw tetraene acyl-glycerol-3-phosphocholine PAPC, 1- stearoyl -2- arachidonic acyl glycerol-3-phosphocholine SAPC, 1- oleoyl - 2- myristoyl glycerol-3-phosphocholine OMPC, 1- palmityl -2- monostearoyl glyceride -3- phosphocholine HSPC, two mustard acyls Base glycerol -3- phosphocholine DEPC, 1,2- bis- lauroyl PHOSPHATIDYL ETHANOLAMINE, two myristoyl PHOSPHATIDYL ETHANOLAMINE DMPE, DPPE DPPE, DSPE DSPE, DOPE DOPE, two mustard acylphosphatidyl ethanolamine DEPE, 1- palmityl -2- oleoyl base PHOSPHATIDYL ETHANOLAMINE DLPE, 1- palmityl Base -2- oleoyl base PHOSPHATIDYL ETHANOLAMINE POPE, DSPE-PEG DSPE-PEG2000 is (poly- Molecular weight glycol 400-3000), distearoylphosphatidyl poly glycol monomethyl ether (molecular weight polyethylene glycol 400-3000) or One or more of cholesterol.Auxiliary agent also contains a kind of targeting group Folic Acid, galactose, antibody, biotin or polypeptide.

The medicine compound liposome nano-particle of the present invention, is liquid preparation, solid preparation, semi-solid preparation, capsule Agent, granule, gel, injection, slow releasing preparation or controlled release preparation.

From the point of view of anti tumor activity in vitro screening, the good anti-tumor activity of the compounds of this invention performance.Test shows this The toxicity in vivo of invention compound, less than camptothecine, topotecan.Therefore animal can be used for as antitumor drug, preferably use In mammal, particularly people.

From the point of view of anti tumor activity in vitro screening, the good antitumor of the compounds of this invention elaioplast nanometer particle performance is lived Property.Test shows that the toxicity in vivo of the compounds of this invention is much smaller than camptothecine, topotecan.Therefore can use as antitumor drug In animal, it is preferred for mammal, particularly people.

The preparation method of the camptothecin phosphatide cpd elaioplast nanometer particle of the present invention, is to be liked by the compounds of this invention Tree bases phosphatide cpd or the compounds of this invention and the mixture of auxiliary agent, are done by film dispersion method, reverse phase evaporation, freezing Prepared by the methods such as dry method, ultrasonic dispersion, spray drying method, film squeezing and pressing method or high pressure homogenization method.

The present invention passes through covalent bond by the use of the camptothecine of 2 molecules or camptothecin analogues as hydrophobic tail and phospholipid hydrophilic head It is connected, prepare camptothecin phosphatide cpd, dissolubility is significantly better than camptothecine or camptothecin analogues；The present invention is by camptothecine Class phosphatide cpd is prepared into nano-particle, has the characteristic of liposome, has and can form liquid preparation, solid preparation, partly consolidate The characteristic of body preparation, sterilization preparation and sterile preparation, for oncotherapy.This camptothecin phosphatide cpd nano-particle is not It is only a kind of prodrug, be also the brand-new drug release carrier of camptothecine, there is target function；The camptothecin of the present invention Phosphatide cpd and its elaioplast nanometer particle are also a kind of prodrug of brand-new camptothecine, are also pharmaceutical carrier, Play drug effect through esterase hydrolyzed in vivo, there is low toxicity side effect.

It is pharmaceutically acceptable that the camptothecin phosphatide cpd of formula (1) of the present invention and these compounds are formed The salt containing counter ion counterionsl gegenions, there is good water solublity, there is antitumor action, the drug release half-life can be extended, and have There are relatively low toxic and side effects.

Beneficial effect：The present invention compared with prior art, has advantages below：

The camptothecin phosphatide cpd of the present invention has following general structure (1)：

Wherein, R₁For spacerarm, it is alkylene, described alkylene is defined as the straight or branched that carbon number is 1～20 Alkylene alkyl/Asia alkylene or aryl that carbon number is 6～10；L represents 2- amino -2- carboxy ethyl, 2- amino second Base, 2- trimethyl amido ethyl cation, molecular weight are N- Polyethylene Glycol-amino-ethyl or 2, the 3- dihydroxy third of 200-2000 Base, R_xIt is the substituent group of camptothecine or camptothecin analogues, be specifically shown in technical scheme.

Contain hydrophilic head part in formula (1) structure, make phosphatide cpd water solublity be significantly better than camptothecine, irinotecan (CPT11 joins piperazine camptothecine), topology vertical health (9- dimethylamino -10- hydroxy-camptothecin seals), SN38, There is hypotoxicity and excellent anti-tumor activity；

R in formula (1)_xIt is the substituent group of camptothecine or camptothecin analogues, is connected with phospholipid hydrophilic head by ester bond, giving birth to By this ester bond of esterase hydrolyzed in object, a molecule camptothecin phosphatide cpd discharges camptothecine or the camptothecine of two molecules Analog, so that the antitumor drug effect of camptothecine or camptothecin analogues phosphatide cpd reaches camptothecine or camptothecin seemingly 2 times of thing；R_xSubstituent group has hydrophobic interaction, mutually collaborative, directly constitutes stable liposome, is of liposome structure Point, the shortcoming that when overcoming general liposome dewatering medicament, medicine easily leaks out, improves the efficiency of medicine parcel simultaneously.

R in formula (1)₁For spacerarm, this spacerarm and adjacent two ester bonds have the enhancing hydrophobic work of hydrophobic head With, the proper alignment to camptothecin phosphatide cpd and to be self-assembled into elaioplast nanometer particle highly beneficial, formed stable Liposome particles, thus extending release and the antitumor action time of camptothecine or the like medicine, make camptothecin phosphorus simultaneously Compound is easy to add intracellular, performance drug effect by liposomal form.

Two molecule camptothecines or camptothecin analogues bioactive molecule is contained in the camptothecin phosphatide cpd structure of the present invention, Antitumor drug effect is camptothecine, irinotecan (CPT11 joins piperazine camptothecine), the vertical health of topology (like by 9- dimethylamino -10- hydroxyl Tree seal), 2 times about of SN38；

The camptothecin phosphatide cpd of the present invention can adopt membrane process etc. to be easily self-assembled into liposome and receive Rice grain, particle diameter 10-1000 nanometer,

The camptothecin phosphatide cpd of the present invention can form compound system with phospholipid, is very easy to using membrane process etc. Be self-assembled into elaioplast nanometer particle, particle diameter 10-1000 nanometer,

The liposome structure of the camptothecin phosphatide cpd elaioplast nanometer particle of the present invention is had with membrane structure Similarity is it is easy to be phagocytized by cells, and discharges active drug molecule camptothecine or camptothecin analogues, plays antitumor action；

The camptothecin phosphatide cpd elaioplast nanometer particle of the present invention has passive target effect, and this is that liposome is released The common trait of medicine system；And the vertical health of camptothecine, irinotecan (CPT11, join piperazine camptothecine), topology (9- dimethylamino- 10- hydroxy-camptothecin seals), the camptothecin analogues such as SN38 all do not possess this characteristic；

The camptothecin phosphatide cpd of the present invention and its elaioplast nanometer particle are a kind of brand-new camptothecines Release vehicle, be also a kind of prodrug；

The compound of the compound of the present invention or the present invention and the pharmaceutical composition of Conventional pharmaceutical carriers, containing 0.1-100 The compounds of this invention of weight %, has hypotoxicity and excellent anti-tumor activity.

The camptothecin phosphatide cpd of the present invention and its elaioplast nanometer particle, can be used as liquid preparation, solid preparation, Semi-solid preparation, sterilization preparation and sterile preparation, can form under the aqueous phase systems such as water, phosphate buffer, citrate buffer solution Liposome；

The camptothecin phosphatide cpd liposome of the present invention combines the auxiliary agent containing targeting group, has active targeting and makees With；

The camptothecin phosphatide cpd of formula (1) involved in the present invention and its elaioplast nanometer particle preparation process is simple；

The camptothecin phosphatide cpd of formula (1) involved in the present invention and its elaioplast nanometer particle are also a kind of brand-new The prodrug of camptothecine, plays drug effect through esterase hydrolyzed in vivo, can extend the drug release half-life, and have relatively low Toxic and side effects, antitumor action is significantly better than camptothecine, irinotecan (CPT11 joins piperazine camptothecine), vertical health (the 9- diformazan of topology Base amino -10- hydroxy-camptothecin seals), the camptothecin analogues such as SN38.

Brief description

Fig. 1 is camptothecine -20- phosphatide cpd synthetic route chart.

Fig. 2 is the synthetic route chart of camptothecine -20- acetass -10- phosphatide cpd.

Fig. 3 is irinotecan -20- phosphatide cpd synthetic route chart.

Fig. 4 is camptothecine -7- methylene phosphatide cpd synthetic route chart.

Fig. 5 is the synthetic route chart of 9-nitrocamptothecin -20- phosphatide cpd.

Fig. 6 is the synthetic route chart of hexacyclic camptothecins (Lurtotecan) -20- phosphatide cpd.

Fig. 7 is the synthetic route chart of difluoro camptothecine (BN80915) -20- phosphatide cpd.

Fig. 8 is the synthetic route chart of CKD-602-20- phosphatide cpd.

Fig. 9 is the synthetic route chart of 9-nitrocamptothecin -20- phosphatidyl ethanol amines.

Figure 10 is the synthetic route of 9-nitrocamptothecin -20- PHOSPHATIDYL ETHANOLAMINE-N- polyethylene glycol compound.

Figure 11 is the synthetic route chart of camptothecine -20- Phosphatidylserine compound.

Figure 12 is the synthetic route chart of camptothecine -20- phosphatidyl glycerol compound.

Figure 13 is the synthetic route chart of 9-nitrocamptothecin -20- phthalic acid ester-phosphatidyl choline compounds.

Figure 14 is the synthetic route chart of 9-aminocamptothecin -20- phosphatide cpd.

Figure 15 is the synthetic route chart of camptothecin analogues DX-8951f-20- phosphatide cpd.

Figure 16 is the particle diameter distribution schematic diagram of camptothecine -20- phosphatide cpd liposome.

Figure 17 is the form (transmission electron microscope picture) of camptothecine -20- phosphatide cpd elaioplast nanometer particle.

Figure 18 is the particle diameter distribution schematic diagram of topological Li Kang -10- phosphatide cpd liposome.

Specific embodiment

With reference to Figure of description and embodiment to the present invention technical scheme be described in further details.

A kind of camptothecin phosphatide cpd involved in the present invention, is the compound of general formula (1) or described formula (1) the pharmaceutically acceptable salt that compound and counter ion counterionsl gegenions are formed：

R₇、R₉、R₁₀、R₁₁And R₁₂Can be identical group or mutually different group, can also be wherein one Part is identical group, and another part is different groups.

In formula (2) and (3) and (4), 20 carbon atoms are S configuration.

R_xPreferably 20 hydroxyl substituents of camptothecine (CPT), 20 hydroxyl substituents of 10-hydroxycamptothecine, the happiness of 10- hydroxyl Tree 10 hydroxyl substituents of alkali, 20 hydroxyl substituents of irinotecan (CPT11 joins piperazine camptothecine), vertical health (the 9- dimethyl of topology Amino -10- hydroxy-camptothecin seals) 10 hydroxyl substituent, vertical 20 hydroxyls of health (9- dimethylamino -10- hydroxy-camptothecin seals) of topology Base substituent group, 20 hydroxyl substituents of SN38,10 hydroxyls of SN38 replace Base, 20 hydroxyl substituents of 9-aminocamptothecin, 20 hydroxyl substituents of 9-nitrocamptothecin, difluoro camptothecine (BN80915) 20 hydroxyl substituents, -20 hydroxyl substituents of hexacyclic camptothecins (Lurtotecan), camptothecin analogues DX-8951f-20 Position hydroxyl substituent, camptothecin analogues CKD-602-20 position hydroxyl substituent.

Counter ion counterionsl gegenions be cation proton, sodium ion, potassium ion, calcium ion, iron ion, magnesium ion, ammonium ion, zinc from Son, anion chloride ion, sulfate ion, sulfate ion, nitrate ion, carboxylic acid ion, carbanion, bromine from Son, phosphate anion, formate, acetate, citrate, lactate, fumaric acid radical, tartrate anion, gluconic acid radical ion Any one or any two combination, preferably proton, sodium ion, chloride ion.

The method of the described camptothecin phosphatide cpd of preparation, the method is first by camptothecine or camptothecin analogues Hydroxyl and anhydride or carboxylic acid reaction, prepare the intermediate containing carboxyl.Then, by described intermediate and choline glycerophosphatide Carry out condensation reaction, that is, obtain camptothecin phosphatidyl choline compounds；Or, by described intermediate and monohydroxy protected glycerol Condensation, through deprotection, and with phosphorus oxychloride reaction after, react with glycerol acetonide condensation of acetone, through take off acetone, that is, obtain camptothecin Phosphatidyl glycerol；Or, described intermediate and monohydroxy protected glycerol are condensed, through deprotection, and with phosphorus oxychloride reaction after, with Tertbutyloxycarbonyl Boc protection ethylaminoethanol hydroxyl condensation reaction, through Deprotection, that is, obtain camptothecin phosphatidyl ethanol Amine；Or, described intermediate and monohydroxy protected glycerol are condensed, through deprotection, and with phosphorus oxychloride reaction after, with amino and carboxylic The hydroxyl condensation reaction of the serine of base all protections, through Deprotection, that is, obtains camptothecin Phosphatidylserine.

Other method is will be single with the glyceryl dicarboxylic acids two of monohydroxy protection for the hydroxyl of camptothecine or camptothecin analogues Two carboxyls condensation of ester, dehydroxylation protection group, activate with phosphorus oxychloride reaction further, then, or with amido protecting Ethylaminoethanol hydroxyl condensation reaction, through Deprotection, that is, obtain camptothecin PHOSPHATIDYL ETHANOLAMINE；Or with glycerol acetonide acetone Condensation reaction, through de- acetone, that is, obtains camptothecin phosphatidyl glycerol；Or react with dimethylaminoethanol, and methylate, Obtain camptothecin phosphatidyl choline compounds；Or the hydroxyl reaction with amino and the serine of carboxyl all protections, through de- Protection group, that is, obtain camptothecin Phosphatidylserine.

Using condensation reagent preferably 1,3 dicyclohexylcarbodiimides, two pyridine carbonic esters, 1- (3- dimethylamine propyl) -3- Ethyl carbimide hydrochlorate, 1,3- diisopropylcarbodiimide or N, one of N- carbonyl dimidazoles and tertiary amine, three-level The preferred DMAP of amine or 4- pyrollidinopyridine.

The pharmaceutical composition of the present invention, is the elaioplast nanometer particle of particle diameter 10-1000 nanometer, is tied by having formula (1) The camptothecin phosphatide cpd self assembly of structure is constituted.

Particle diameter 10-1000 is constituted by the camptothecin phosphatide cpd and phospholipid co-assemble with formula (1) structure The elaioplast nanometer particle of nanometer.

Application in preparing antitumor drug for the camptothecin phosphatide cpd of the present invention it is characterized in that, the method will Described camptothecin phospholipid or its pharmaceutically acceptable salt, are prepared into medicament with acceptable carrier on pharmacodynamicss.Can be by The compounds of this invention or the compounds of this invention are combined with one or more solid or liquid pharmaceutical excipients and/or adjuvant, make The suitable administration form that can use as people's medicine or dosage form.Generally pharmaceutical composition of the present invention contains 0.1-100 weight The compounds of this invention of amount %.

The compounds of this invention or can be intestinal or non-bowel containing its pharmaceutical composition route of administration, such as oral, muscle, Subcutaneous, nasal cavity, oral mucosa, skin, peritoneum or rectum etc..Can be drug administration by injection, including intravenous injection, intramuscular injection, subcutaneous Injection, intradermal injection and acupoint injection therapy etc..Form of administration can be liquid dosage form, solid dosage formss.If liquid dosage form can be true Solution class, colloidal type, particulate formulations, emulsion dosage form, mixed suspension form.Other dosage forms such as tablet, capsule, drop pill, aerosol, ball Agent, powder, solution, suspensoid, Emulsion, granule, suppository, lyophilized injectable powder etc..

The carrier that the pharmaceutical composition of the present invention adopts is, such as diluent and absorbent, such as starch, dextrin, calcium sulfate, Lactose, Mannitol, sucrose, sodium chloride, glucose, carbamide, Calcium Carbonate, kaolin, Microcrystalline Cellulose, aluminium silicate etc.；Wetting agent With binding agent, such as water, glycerol, Polyethylene Glycol, ethanol, propanol, starch slurry, dextrin, syrup, Mel, glucose solution, Arab Rubber cement, gelatine size, sodium carboxymethyl cellulose, lac, methylcellulose, potassium phosphate, Polyvinylpyrrolidone etc.；Disintegrating agent, example As starch, alginate, agar powder, laminaran, sodium bicarbonate and citric acid, Calcium Carbonate, polyoxyethylene sorbitol are dried Fatty acid ester, dodecyl sodium sulfate, methylcellulose, ethyl cellulose etc.；Disintegration inhibitor, such as sucrose, three stearic acid Glyceride, cocoa butter, hydrogenated oil and fat etc.；Absorption enhancer, such as quaternary ammonium salt, sodium lauryl sulphate etc.；Lubricant, such as Talcum Powder, silicon dioxide, corn starch, stearate, boric acid, liquid paraffin, Polyethylene Glycol etc..Tablet can also be made further Coated tablet, such as sugar coated tablet, thin membrane coated tablet, ECT, or double-layer tablet and multilayer tablet.

The present invention passes through covalent bond by the use of the camptothecine of 2 molecules or camptothecin analogues as hydrophobic tail and phospholipid hydrophilic head It is connected, prepare camptothecin phosphatide cpd, dissolubility is significantly better than camptothecine or camptothecin analogues；The present invention is by camptothecine Class phosphatide cpd is prepared into nano-particle, has the characteristic of liposome, has and can form liquid preparation, solid preparation, partly consolidate The characteristic of body preparation, sterilization preparation and sterile preparation, for oncotherapy.This camptothecin phosphatide cpd nano-particle is not It is only a kind of prodrug, be also the brand-new drug release carrier of camptothecine, there is target function；The camptothecin of the present invention Phosphatide cpd and its elaioplast nanometer particle are also a kind of prodrug of brand-new camptothecine, are also pharmaceutical carrier, Play drug effect through esterase hydrolyzed in vivo, the drug release half-life can be extended, and there are relatively low toxic and side effects.

It is below portion of reagent code name used in preparation process：

DMAP DMAP

CDI N, N'- carbonyl dimidazoles

DMSO dimethyl sulfoxide

GPC Phosphorylcholine glycerol

DBU 1,5- diazabicylo [5.4.0] 11-5- alkene

EDCI N, N'- thio-carbonyldiimidazole

HOBT I-hydroxybenzotriazole

Following the present invention is further illustrated by embodiment, but the invention is not restricted to following examples.

Embodiment 1：

The synthesis (synthetic route is shown in Fig. 1) of camptothecine -20- phosphatide cpd

Take camptothecine 1g, succinic anhydride 0.75g, DMAP 0.2g, triethylamine 0.2g, add dichloromethane or chloroform For reaction dissolvent, add back flow reaction, react 12h；Dilute hydrochloric acid washs three times, filters dilute hydrochloric acid layer, takes filter cake；Plus methanol is molten Solution, 4 DEG C of placement 12h, filter, take filter cake, be dried, obtain intermediate product camptothecine -20- succinate monoester.

Take intermediate product succinic acid camptothecine monoesters 0.5g, add CDI 0.5g, add dichloromethane, chloroform or DMSO, as reaction dissolvent, adds GPC 0.5g and DBU 0.5g room temperature reaction, and gained reactant liquor passes through column chromatography purification, obtains product Thing camptothecine -20- phosphatide cpd 0.32g.¹H NMR(500MHz,CD3OD:CDCl₃1:1):δ8.03-7.59(10H,m), 6.25(2H,s),4.71-4.55(7H,m),4.21(2H,t),3.86-3.55(8H,s),3.46(9H,s),2.67-2.41 (8H,t),1.98(4H,m),1.12(6H,s).[M+H]⁺M/z, 1119.34.

Embodiment 2：

The synthesis (synthetic route is shown in Fig. 2) of camptothecine -20- acetass -10- phosphatide cpd

10-hydroxycamptothecine 0.5g, acetic acid 1g, EDCI 1g, HOBT 0.5g, add dichloromethane or chloroform to be anti- Answer solvent, 40 DEG C of reaction 12h；Dilute hydrochloric acid washs three times, filters dilute hydrochloric acid layer, takes filter cake；Plus methanol dissolving, 4 DEG C of placement 12h, Filter, take filter cake, be dried, obtain 10-hydroxycamptothecine -20- acetass.

10-hydroxycamptothecine -20- acetass 0.3g, succinic anhydride 1g, DMAP 0.5g, triethylamine 0.5g, add dichloro Methane or chloroform are reaction dissolvent, add back flow reaction, react 12h；Dilute hydrochloric acid washs three times, filters dilute hydrochloric acid layer, takes Filter cake；Plus methanol dissolving, 4 DEG C of placement 12h, filter, take filter cake, be dried, obtain intermediate product camptothecine -20- acetass -10- fourth Monomethyl diester.

Take intermediate product camptothecine -20- acetass -10- succinate monoester 0.21g, add CDI 0.2g, add dichloromethane Alkane, chloroform or DMSO, as reaction dissolvent, add GPC 0.2g and DBU 0.2g room temperature reaction, and gained reactant liquor passes through Column chromatography purification, obtains product camptothecine -20- acetass -10- phosphatide cpd 0.13g.¹H NMR(500MHz,CD3OD:CDCl₃ 1:1):δ7.93-7.44(8H,m),6.23(2H,s),4.69-4.43(7H,m),3.61(2H,t),3.43(4H,s),3.22 (9H,s),3.09(4H,s),2.68-2.52(8H,m),2.14(6H,s),1.65(4H,m),1.05(6H,t).[M+H]⁺m/z, 1235.35.

Embodiment 3

The synthesis of 7- Ethyl-camptothecin -20- acetass -10- phosphatide cpd

SN38 0.5g, acetic acid 1g, EDCI 1g, HOBT 0.5g, add dichloromethane or trichlorine Methane is reaction dissolvent, 40 DEG C of reaction 12h；Dilute hydrochloric acid washs three times, filters dilute hydrochloric acid layer, takes filter cake；Plus methanol dissolving, 4 DEG C Place 12h, filter, take filter cake, be dried, obtain SN38-20- acetass.

SN38-20- acetass 0.35g, succinic anhydride 1g, DMAP 0.5g, triethylamine 0.5g, Add dichloromethane or chloroform to be reaction dissolvent, add back flow reaction, react 12h；Dilute hydrochloric acid washs three times, filters dilute salt Acid layer, takes filter cake；Plus methanol dissolving, 4 DEG C placement 12h, filter, take filter cake, be dried, obtain intermediate product 7- Ethyl-camptothecin- 20- acetass -10- succinate monoester.

Take intermediate product 7- ethyl-camptothecin -20- acetass -10- succinate monoester 0.23g, add CDI 0.5g, add Dichloromethane, chloroform or DMSO, as reaction dissolvent, add GPC 0.5g and DBU 0.5g room temperature reaction, and gained reacts Liquid passes through column chromatography purification, obtains product 7- Ethyl-camptothecin -20- acetass -10- phosphatide cpd 0.17g.By 7- ethyl-happiness Tree alkali -20- acetass -10- phosphatide cpd is dissolved in sodium-chloride water solution, lyophilizing, obtain containing counter ion counterionsl gegenions sodium ion and The SN38 phosphatide cpd salt of chloride ion.

Embodiment 4

The synthesis (synthetic route is shown in Fig. 3) of irinotecan (CPT11 joins piperazine camptothecine) -20- phosphatide cpd

Connection piperazine camptothecine 0.5g, succinic anhydride 0.5g, DMAP 0.5g, triethylamine 0.5g, add dichloromethane or three chloromethanes Alkane is reaction dissolvent, adds back flow reaction, reacts 12h；Dilute hydrochloric acid washs three times, filters dilute hydrochloric acid layer, takes filter cake；Plus methanol is molten Solution, 4 DEG C of placement 12h, filter, take filter cake, be dried, obtain intermediate product connection piperazine camptothecine -20- succinate monoester.

Take intermediate product connection piperazine camptothecine -20- succinate monoester 0.3g, add CDI 0.3g, add dichloromethane, trichlorine Methane or DMSO, as reaction dissolvent, add GPC 0.5g and DBU 0.5g room temperature reaction, and gained reactant liquor passes through column chromatography Purification, obtains product connection piperazine camptothecine -20- phosphatide cpd 0.16g.¹H NMR(500MHz,CD3OD:CDCl₃1:1):δ8.14- 7.39(6H,m),6.41(2H,s),4.73-4.51(7H,m),3.72(2H,t),3.49(4H,s),3.22(9H,s),3.04- 2.94(8H,s),2.68-2.52(12H,m),2.14(6H,s),1.75(4H,m),1.46-1.32(24H,m),1.15-1.08 (6H,tt).[M+H]⁺M/z, 1595.67.

Irinotecan (CPT11 joins piperazine camptothecine) -20- phosphatide cpd is dissolved in 0.1M calcium lactate aqueous solution, and lyophilizing obtains Solid to the irinotecan containing counter ion counterionsl gegenions calcium ion and lactate ion (CPT11 joins piperazine camptothecine) -20- phosphatide cpd Body powder.

Embodiment 5：

The synthesis of topological Li Kang -10- phosphatide cpd

9- dimethylamino -10-hydroxycamptothecine 0.5g, acetic acid 1g, EDCI 1g, HOBT 0.5g, add dichloromethane Or chloroform is reaction dissolvent, 40 DEG C of reaction 12h；Dilute hydrochloric acid washs three times, filters dilute hydrochloric acid layer, takes filter cake；Plus methanol is molten Solution, 4 DEG C of placement 12h, filter, take filter cake, be dried, obtain 9- dimethylamino -10-hydroxycamptothecine -20- acetass.

9- dimethylamino -10-hydroxycamptothecine -20- acetass 0.3g, phthalic anhydride 0.5g, DMAP 0.5g, Triethylamine 0.5g, adds dichloromethane or chloroform to be reaction dissolvent, adds back flow reaction, react 12h；Dilute hydrochloric acid washing three Time, filter dilute hydrochloric acid layer, take filter cake；Plus methanol dissolving, 4 DEG C of placement 12h, filter, take filter cake, be dried, obtain intermediate product 9- bis- Methylamino camptothecine -20- acetass -10- phthalic monoester.

Take intermediate product 9- dimethylamino camptothecine -20- acetass -10- phthalic monoester 0.16g, add CDI0.2g, adds dichloromethane, chloroform or DMSO as reaction dissolvent, adds GPC 0.2g and DBU 0.2g room temperature Reaction, gained reactant liquor passes through column chromatography purification, obtains product topology Li Kang -10- phosphatide cpd 0.11g.

Embodiment 6：

The synthesis (synthetic route is shown in Fig. 4) of camptothecine -7- methylene phosphatide cpd

Add 15mL methanol and 12.5mL water in the 100mL three neck round bottom flask being furnished with thermometer and stirring, in ice bath Cooling is lower to add 6.5mL concentrated sulphuric acid, is subsequently adding 0.5g (1.43mmol) camptothecine and 0.4g (1.43mmol) ferrous sulfate seven Hydrate, after mixture stirs 5 minutes, controls temperature in below 5C, is slowly added dropwise the peroxide of 0.24mL (0.21mmol) 30% Change hydrogen, solution colour gradually deepens, and after reactant is stirred at room temperature 10 hours, pours in 200mL water, aqueous phase stands overnight, mistake Filter, solid is washed, and is dried to obtain yellow solid 0.5g, and crude product DMF- re-crystallizing in ethyl acetate obtains the happiness of 0.3g 7- methylol Tree alkali, yield：274-276 DEG C of 55%, mp.1HNMR (CDCl3,300MHz)：δ 8.17 (t, 2H, Ar-H), 7.84 (t, 1H, Ar-H), 7.69 (t, 1H, Ar-H), 7.33 (s, 1H, Ar-H), 6.49 (s, 1H, 20OH), 5.79 (m, 1H, 7OH), 5.41d (d, 4H, H17,7-CH2), 5.26 (d, 2H, H5), 1.86 (m, 2H, 18-CH2), 0.87 (t, 3H, 19-CH3).

Take 7- methylol camptothecine 0.3g, succinic anhydride 2g, DMAP 0.1g, triethylamine 1g, add dichloromethane or trichlorine Methane is reaction dissolvent, adds back flow reaction, reacts 12h；Dilute hydrochloric acid washs three times, filters dilute hydrochloric acid layer, takes filter cake；Plus methanol Dissolving, 4 DEG C of placement 12h, filter, take filter cake, be dried, obtain intermediate product succinate monoester methylene -7- camptothecine 0.22g.

Take intermediate product succinate monoester methylene camptothecine 0.22g, add CDI 0.3g, add dichloromethane, three Chloromethanes or DMSO, as reaction dissolvent, add GPC 0.3g and DBU 0.2g room temperature reaction, and gained reactant liquor passes through post layer Analysis purification, obtains product camptothecine -7- methylene phosphatide cpd 0.16g.¹H NMR(500MHz,CD3OD:CDCl₃1:1):δ 8.13-7.61(8H,m),6.47(2H,s),5.26(4H,s),4.68-4.39(7H,m),3.79(2H,t),3.55(4H,s), 3.42(9H,s),3.12(4H,s),2.95-2.72(8H,tt),1.94(4H,m),0.98(6H,t).[M+H]⁺M/z, 1179.36.

Embodiment 7：

The synthesis (synthetic route is shown in Fig. 5) of 9-nitrocamptothecin -20- phosphatide cpd

Take adipic acid 1g, add CDI 1g, add dichloromethane, chloroform or DMSO to be reaction dissolvent, room temperature reaction 2h；Add 9-nitrocamptothecin 0.4g, DBU 0.4g；Room temperature reaction 2h；Add GPC 0.5g, DBU 0.5g, room temperature reaction 6h； Add 9-nitrocamptothecin 0.3g, DBU 0.3g, room temperature reaction 6h again.Gained reactant liquor passes through column chromatography purification, obtains product 9-nitrocamptothecin -20- phosphatide cpd 0.45g.¹H NMR(500MHz,CD3OD:CDCl₃1:1):δ8.22-6.95(10H, m),4.80(4H,s),4.63(4H,s),4.47-4.25(7H,m),3.81(2H,t),3.32(9H,s),2.54(8H,tt), 2.03(4H,m),1.56(8H,m),1.04(6H,t).[M+H]⁺M/z, 1165.11.

9-nitrocamptothecin -20- phosphatide cpd is dissolved in normal saline, lyophilizing obtain containing counter ion counterionsl gegenions sodium ion and 9-nitrocamptothecin -20- phosphatide cpd the pressed powder of chloride ion.

Embodiment 8：

The synthesis (synthetic route is shown in Fig. 6) of hexacyclic camptothecins (Lurtotecan) -20- phosphatide cpd

Take suberic acid 1g, add CDI 1.5g, add dichloromethane, chloroform or DMSO to be reaction dissolvent, room temperature is anti- Answer 2h；Add hexacyclic camptothecins (Lurtotecan) 0.5g, DBU 0.5g, room temperature reaction 2h；Add GPC 0.5g, DBU 0.5g, room temperature reaction 8h；Add hexacyclic camptothecins (Lurtotecan) 0.3g, DBU0.3g, room temperature reaction 4h again.Gained is anti- Answer liquid to pass through column chromatography purification, obtain product hexacyclic camptothecins (Lurtotecan) -20- phosphatide cpd 0.52g.¹H NMR (500MHz,CD3OD:CDCl₃1:1):δ7.56-6.98(6H,m),4.85(4H,s),4.66(2H,t),4.42(8H,tt), 4.31-4.16(5H,m),3.98(4H,s),3.65(6H,m),3.29(9H,s),2.56(16H,m),2.31(6H,s),2.01 (8H,m),1.82(4H,m),1.68(8H,m),1.33(8H,m),1.01(6H,t).[M+H]⁺M/z, 1571.35.

Hexacyclic camptothecins (Lurtotecan) -20- phosphatide cpd is dissolved in 0.1M sodium phosphate aqueous solution, and lyophilizing is contained There is hexacyclic camptothecins (the Lurtotecan) -20- phosphatide cpd pressed powder of counter ion counterionsl gegenions sulfate radical and sodium ion.

Embodiment 9：

The synthesis (synthetic route is shown in Fig. 7) of difluoro camptothecine (BN80915) -20- phosphatide cpd

Take adipic acid 1g, add CDI 1g, add dichloromethane, chloroform or DMSO to be reaction dissolvent, room temperature reaction 2h；Add difluoro camptothecine (BN80915) 0.3g, DBU 0.3g, room temperature reaction 2h；Add GPC 0.3g, DBU 0.3g, room temperature Reaction 4h；Add difluoro camptothecine (BN80915) 0.3g, DBU0.3g, room temperature reaction 4h again.Gained reactant liquor passes through post layer Analysis purification, obtains product difluoro camptothecine (BN80915) -20- phosphatide cpd 0.21g.¹H NMR(500MHz,CD3OD:CDCl₃ 1:1):δ7.76-6.94(8H,m),4.91(4H,s),4.65-4.26(7H,m),4.07(4H,m),3.75(2H,t),3.31 (9H,s),2.37(8H,t),2.07(4H,m),1.67(8H,m),1.02(6H,t).[M+H]⁺M/z, 1246.36.

Difluoro camptothecine (BN80915) -20- phosphatide cpd is dissolved in the aqueous acetic acid of 0.1M, and lyophilizing is contained Difluoro camptothecine (the BN80915) -20- phosphatide cpd pressed powder of acetate counter ion counterionsl gegenions.

Embodiment 10：

The synthesis (synthetic route is shown in Fig. 8) of CKD-602-20- phosphatide cpd

Take decanedioic acid 1g, add CDI 1g, add dichloromethane, chloroform or DMSO to be reaction dissolvent, room temperature reaction 2h；Add CKD-602 0.5g, DBU 0.5g, room temperature reaction 2h；Add GPC 0.5g, DBU0.5g, room temperature reaction 12h；Again Add CKD-602 0.3g, DBU 0.3g, room temperature reaction 4h.Gained reactant liquor passes through column chromatography purification, obtains product CKD-602- 20- phosphatide cpd 0.55g.¹H NMR(500MHz,CD3OD:CDCl₃1:1):δ8.10-7.03(10H,m),4.93(4H, s),4.71-4.43(7H,m),4.23(4H,s),3.75(2H,t),3.38(9H,s),3.04(2H,m),2.89(4H,t), 2.61(4H,t),2.25(8H,tt),1.99(4H,m),1.72(8H,m),1.34(16H,m),1.05(18H,m).[M+H]⁺m/ Z, 1458.71.

Embodiment 11：

The synthesis (synthetic route is shown in Fig. 9) of 9-nitrocamptothecin -20- phosphatidyl ethanol amines

Take 3- (4- methoxybenzyl epoxide) propane -1,2- glycol 1g, succinic anhydride 2.5g, DMAP 0.1g, triethylamine 1.5g, adds dichloromethane, chloroform or DMSO to be reaction dissolvent, adds back flow reaction, react 12h；Dilute hydrochloric acid washing three Time, filter dilute hydrochloric acid layer, take filter cake；Add CDI 0.5g, add dichloromethane, chloroform or DMSO to be reaction dissolvent, room Temperature reaction 2h；Add 9-nitrocamptothecin 0.5g, DBU 1g, room temperature reaction 8h, revolve solvent evaporated；Add spirit of vinegar, room temperature is anti- Answer 2h, remove solvent；Add phosphorus oxychloride 1g, triethylamine 0.8g, add dichloromethane, chloroform or THF to be reaction dissolvent, 10 DEG C of reaction 24h, revolve solvent evaporated；N- t-butoxycarbonyl-amino ethanol 1g, triethylamine 0.3g, add dichloromethane, three chloromethanes Alkane or DMSO are reaction dissolvent, 10 DEG C of reaction 24h, revolve solvent evaporated；Add isopropanol, aqueous acetic acid, reacting by heating 24h. Gained reactant liquor passes through column chromatography purification, obtains product 9-nitrocamptothecin -20- phosphatidyl ethanol amines 1.4g.

¹H NMR(500MHz,CD3OD:CDCl₃1:1):δ8.22-6.95(10H,m),4.80(4H,s),4.63(4H, s),4.47-4.25(7H,m),3.81(2H,t),3.32(9H,s),2.54(8H,tt),2.03(4H,m),1.56(2H,m).[M +H]⁺M/z, 1181.02.

Embodiment 12：

The synthesis (synthetic route is shown in Figure 10) of 9-nitrocamptothecin -20- PHOSPHATIDYL ETHANOLAMINE-N- polyethylene glycol compound

Take mPEG2000 2g, CDI 0.2g, dichloromethane, chloroform or DMSO are reaction dissolvent, room temperature reaction 2h； Add the 9-nitrocamptothecin -20- phosphatidyl ethanol amines 0.3g in embodiment 11, DBU 0.3g, reacting by heating 24h. Gained reactant liquor passes through column chromatography purification, obtains product 9-nitrocamptothecin -20- PHOSPHATIDYL ETHANOLAMINE-N- polyethylene glycol compound 0.15g.

Embodiment 13：

The synthesis (synthetic route is shown in Figure 11) of camptothecine -20- Phosphatidylserine compound

Take camptothecine 0.5g, adipic anhydride 2.5g, DMAP 0.1g, triethylamine 1.5g, add dichloromethane, chloroform Or DMSO is reaction dissolvent, heating reflux reaction, reacts 12h；Dilute hydrochloric acid washs three times, filters dilute hydrochloric acid layer, takes filter cake；Add CDI 2g, adds dichloromethane, chloroform or DMSO to be reaction dissolvent, room temperature reaction 2h；Add 3- (4- methoxyl group benzyloxy Base) propane -1,2- glycol 0.6g, DBU 1g, room temperature reaction 8h, revolve solvent evaporated；Add spirit of vinegar, room temperature reaction 2h, remove Solvent；Add compound A and B, tertbutanol peroxide is reaction dissolvent, reacting by heating；Palladium carbon catalytic hydrogenating reduction.Gained reacts Liquid passes through column chromatography purification, obtains product camptothecine -20- Phosphatidylserine compound 0.4g.

¹H NMR(500MHz,CD3OD:CDCl₃1:1):δ8.10-7.24(12H,m),4.93(4H,s),4.74(4H, s),4.56-4.14(8H,m),2.69(8H,tt),2.14(4H,m),1.78(8H,m),1.21(6H,t).

[M+H]⁺M/z, 1177.67.

Embodiment 14：

The synthesis (synthetic route is shown in Figure 12) of camptothecine -20- phosphatidyl glycerol compound

Take camptothecine -20- adipate monoester 0.3g, CDI 0.3g, add dichloromethane, chloroform or DMSO to be reaction Solvent, room temperature reaction 2h, addition 3- (4- methoxybenzyl epoxide) propane -1,2- glycol 0.3g, DBU 0.5g, room temperature reaction 8h, Rotation solvent evaporated；Add spirit of vinegar, room temperature reaction 2h, remove solvent；Add phosphorus oxychloride 0.3g, triethylamine 0.3g, add two Chloromethanes, chloroform or THF are reaction dissolvent, 10 DEG C of reaction 24h, revolve solvent evaporated；Add glycerol acetonide acetone 0.3g, three second Amine 0.3g, adds dichloromethane, chloroform or DMSO to be reaction dissolvent, and 10 DEG C of reaction 24h revolve solvent evaporated；Add isopropyl Alcohol, aqueous acetic acid, reacting by heating 24h.Gained reactant liquor passes through column chromatography purification, obtains product camptothecine -20- phosphatidyl glycerol Compound 0.22g.¹H NMR(500MHz,CD3OD:CDCl₃1:1):δ8.10-7.24(12H,m),4.93(4H,s),4.74 (4H,s),4.56-4.14(7H,m),3.71(2H,d),3.38(1H,m),2.69(8H,tt),2.14(4H,m),1.78(8H, m),1.21(6H,t).[M+H]⁺M/z, 1164.23.

Embodiment 15：

The synthesis (synthetic route is shown in Figure 13) of 9-nitrocamptothecin -20- phthalic acid ester-phosphatidyl choline compounds

9-nitrocamptothecin 0.5g, phthalic anhydride 0.5g, DMAP 0.5g, triethylamine 1g, add dichloromethane or three Chloromethanes are reaction dissolvent, add back flow reaction, react 12h；Dilute hydrochloric acid washs three times, filters dilute hydrochloric acid layer, takes filter cake；Plus first Alcohol dissolves, 4 DEG C of placement 12h, filters, takes filter cake, is dried, obtains intermediate product 9-nitrocamptothecin -20- phthalic monoester.

Take 9-nitrocamptothecin -20- phthalic monoester 0.3g, CDI 0.5g, add dichloromethane, chloroform or DMSO is reaction dissolvent, and room temperature reaction 2h adds 3- (4- methoxybenzyl epoxide) propane -1,2- glycol 0.3g, DBU 0.3g, room Temperature reaction 8h, revolves solvent evaporated；Add spirit of vinegar, room temperature reaction 2h, remove solvent；Add phosphorus oxychloride 0.5g, triethylamine 0.3g, adds dichloromethane, chloroform or THF to be reaction dissolvent, and 10 DEG C of reaction 24h revolve solvent evaporated；Add glycerol acetonide third Ketone 0.3g, triethylamine 0.3g, add dichloromethane, chloroform or DMSO to be reaction dissolvent, 10 DEG C of reaction 24h, and revolving is done molten Agent；Add isopropanol, aqueous acetic acid, reacting by heating 24h.Gained reactant liquor passes through column chromatography purification, obtains the happiness of product 9- nitro Tree alkali -20- phthalic acid ester-phosphatidyl choline compounds 0.18g.¹H NMR(500MHz,CD3OD:CDCl₃1:1):δ 8.31-6.94(18H,m),4.80(4H,s),4.61(4H,s),4.37-4.14(7H,m),3.79(2H,t),3.30(9H,s), 2.05(4H,m),1.08(6H,t).[M+H]⁺M/z, 1305.31.

Embodiment 16：

The synthesis (synthetic route is shown in Figure 14) of 9-aminocamptothecin -20- phosphatide cpd

Take the 9-aminocamptothecin 0.5g of Boc protection, succinic anhydride 2.5g, DMAP 0.1g, triethylamine 1.1g, add two Chloromethanes or chloroform are reaction dissolvent, heating reflux reaction 12h；Dilute hydrochloric acid washs three times, filters dilute hydrochloric acid layer, takes filter Cake, is dried, obtains intermediate product succinate monoester base amino camptothecin 0.4g.

Take intermediate product succinate monoester base amino camptothecin 0.3g, add CDI 0.4g, dichloromethane, chloroform or Person DMSO, as reaction dissolvent, adds GPC 0.2g and DBU 0.25g room temperature reaction 24h, solvent is dried；Addition trifluoroacetic acid/ Dichloromethane solution, room temperature reaction 2h.Gained reactant liquor passes through column chromatography purification, obtains product 9-aminocamptothecin phosphatide cpd 0.2g.

¹H NMR(500MHz,CD3OD:CDCl₃1:1):δ7.91-6.83(10H,m),4.82(4H,s),4.60(4H, s),4.49-4.18(7H,m),3.72(2H,t),3.26(9H,s),2.54(8H,tt),1.83(4H,m),1.02(6H,t).

[M+H]⁺M/z, 1149.16.

Embodiment 17：

The synthesis (synthetic route is shown in Figure 15) of DX-8951f-20- phosphatide cpd

Take the DX-8951f 0.4g of Boc protection, succinic anhydride 1g, DMAP 0.1g, triethylamine 0.5g, dichloromethane or three Chloromethanes are reaction dissolvent, heating reflux reaction 24h；Dilute hydrochloric acid washs three times, filters dilute hydrochloric acid layer, takes filter cake, is dried, in obtaining Between product succinate monoester base DX-8951f 0.27g.

Take intermediate product succinate monoester base DX-8951f 0.27g, add CDI 0.35g, add dichloromethane, trichlorine Methane or DMSO, as reaction dissolvent, add GPC 0.35g and DBU 0.25g room temperature reaction 12h, solvent are dried；Add three Fluoroethanoic acid/dichloromethane solution, room temperature reaction 2h.Gained reactant liquor passes through column chromatography purification, obtains product DX-8951f-20- phosphorus Compound 0.19g.

¹H NMR(500MHz,CD3OD:CDCl₃1:1):δ7.48(2H,s),6.83(2H,s),4.97(4H,s),4.65 (2H,t),4.48-4.21(7H,m),3.88(2H,t),3.41(13H,m),2.89(4H,t),2.56(8H,tt),2.21(6H, s),1.86(4H,m),0.97(6H,t).

[M+H]⁺M/z, 1294.66.

Embodiment 18：

The preparation of camptothecine -20- phosphatide cpd liposome

Camptothecine -20- phosphatide cpd the 1mmol of Example 1, adds chloroform 20ml, 60 DEG C of rotation solvent evaporated； Add 20ml PBS (pH=7.4) 60 DEG C of skinnings, 200nm membrane filtration, obtain camptothecine phosphatide cpd elaioplast nanometer particle Solution.The following Figure 16 of granularmetric analyses result, mean diameter 35.40nm.Transmission electron microscope measures form such as Figure 17 of nano-particle.

Embodiment 19：

The preparation of camptothecine -20- acetass -10- phosphatide cpd liposome

Camptothecine -20- acetass -10- phosphatide cpd the 1mmol being obtained by embodiment 2, adds chloroform 20ml, 60 DEG C of rotation solvent evaporated；Add 20ml PBS (pH=7.4) 60 DEG C of skinnings, 200nm membrane filtration, obtain camptothecine -20- acetic acid Ester -10- phosphatide cpd elaioplast nanometer particle solution.Granularmetric analyses result shows mean diameter 166nm.By camptothecine -20- Acetass -10- phosphatide cpd elaioplast nanometer particle solution lyophilizing, obtains powdered nanoparticles granule.

Embodiment 20：

The preparation of 7- Ethyl-camptothecin -10- phosphatide cpd liposome

7- Ethyl-camptothecin -10- phosphatide cpd the 1mmol being obtained by embodiment 3, addition chloroform 20ml, 60 DEG C Rotation solvent evaporated；Add 20ml PBS (pH=7.4) 60 DEG C of skinnings, 200nm membrane filtration, obtain 7- Ethyl-camptothecin -10- phosphorus Compound elaioplast nanometer particle solution.Granularmetric analyses result shows, mean diameter 152nm.

Embodiment 21：

The preparation of irinotecan -20- phosphatide cpd liposome

Irinotecan -20- phosphatide cpd the 1mmol being obtained by embodiment 4, adds chloroform 20ml, 60 DEG C of revolvings Dry solvent；Add 20ml PBS (pH=7.4) 60 DEG C of skinnings, obtain irinotecan -20- phosphatide cpd elaioplast nanometer particle molten Liquid.Granularmetric analyses result shows, mean diameter 228nm.

Embodiment 22：

The preparation of topological Li Kang -10- phosphatide cpd liposome

Topological Li Kang -10- phosphatide cpd the 1mmol being obtained by embodiment 5, adds chloroform 20ml, 60 DEG C of revolvings Dry solvent；Add 20ml PBS (pH=7.4) 60 DEG C of skinnings, obtain topological Li Kang -10- phosphatide cpd elaioplast nanometer particle molten Liquid.Granularmetric analyses result display mean diameter 290nm (Figure 18).

Embodiment 23：

The preparation of camptothecine -20- phosphatide cpd liposome B

Take camptothecine -20- phosphatide cpd 0.5mmol, Distearoyl Phosphatidylcholine DSPC 0.5mmol, add trichlorine Methane 20ml, 60 DEG C of rotation solvent evaporated；Add 20ml PBS (pH=7.4) 60 DEG C of skinnings, obtain camptothecine phosphatide cpd lipid Body nanoparticles solution.Granularmetric analyses show mean diameter 380nm.

Embodiment 24：

The preparation of camptothecine -20- phosphatide cpd liposome C

Take camptothecine -20- phosphatide cpd 0.5mmol, Distearoyl Phosphatidylcholine DSPC 0.5mmol, distearyl Base PHOSPHATIDYL ETHANOLAMINE-Polyethylene Glycol DSPE-PEG- Folic Acid (molecular weight polyethylene glycol 400-3000) 0.1mmol, adds trichlorine Methane 20ml, 60 DEG C of rotation solvent evaporated；Add 20ml PBS (pH 7.4) 60 DEG C of skinnings, obtain camptothecine phosphatide cpd liposome Nanoparticles solution.Granularmetric analyses show, mean diameter 620nm.Lyophilization camptothecine -20- phosphatide cpd liposome is received Rice grain solution, obtains powder nano-particle.

Pharmacological evaluation

Experimental example 25：

Mtt assay human cancer cell fragmentation test

Medicine and reagent：Calf serum is Nanjing Sheng Xing Bioisystech Co., Ltd product；DMSO analysis is pure；RPMI1640 For GIBCO product.

Instrument：BIORAD 680 type microplate reader.

Collect well-grown tumor cell, be configured to 1 × 10-4/ with the RPMI1640 culture medium containing 10% calf serum ML cell suspension, inoculates, often empty 100 μ L (containing 1000 tumor cells), put 37 DEG C, in 5%CO2 incubator in 96 well culture plates Dosing after culture 24 hours, experiment sets blank and solvent control, and given the test agent sets 4 concentration, every 3 parallel holes of concentration, Put 37 DEG C, culture 4 days in 5%CO2 incubator.Discard culture fluid, every hole adds MTT solution (0.4mg/mL, RPMI1640 prepare) 100 μ L, 37 DEG C are incubated 4 hours.Abandoning supernatant, every hole adds MTT solution 150 μ L, dissolved particles, after gentle agitation, uses 680 Type microplate reader measures OD value under Detection wavelength 540nm, reference wavelength 450nm.

Result calculates：Suppression ratio mapping with the variable concentrations of medicine and to cell can get dose-effect curve, therefrom Obtain half-inhibition concentration (IC50).

Derivant of the present invention the results are shown in Table 1. from anti tumor activity in vitro screening to the anti-tumor activity of human tumor cell line From the point of view of, the half-inhibition concentration of the compounds of this invention is respectively less than irinotecan, illustrates that its activity is better than irinotecan.Of the present inventionization The half-inhibition concentration of compound is respectively less than camptothecine, the about half of camptothecine it may be possible to because camptothecin phospholipid chemical combination Thing contains two camptothecin molecular structure units.

The anti-tumor activity result to human tumor cell line for table 1. the compounds of this invention

MCF-7：Human breast cancer cell；Hep G2：Human liver cancer cell；A2780：Proliferation of Human Ovarian Cell；HCT-8：People's colon Cancerous cell；A549：Human lung adenocarcinoma cell；EC-304：Human vascular endothelial normal cell；ND：Not test (N.T.).

Experimental example 26

Camptothecin phosphatide cpd is tested in In vivotoxicity

Animal：ICR mice, male, 18-22g, purchased from laboratory animal technology Co., Ltd of dimension tonneau China.

The compounds of this invention camptothecine -20- phosphatide cpd, 10-hydroxycamptothecine -10- phosphatide cpd, 7- methylene Camptothecine phosphatide cpd, the result of the toxicity in vivo test of SN38-10- phosphatide cpd show, Its maximum tolerated dose is more than 100mg/kg, shows that its toxicity is much smaller than camptothecine, topotecan.

Table 2 camptothecin derivative is in In vivotoxicity result of the test

Above-described embodiment be only the preferred embodiment of the present invention it should be pointed out that：Ordinary skill for the art For personnel, under the premise without departing from the principles of the invention, some improvement and equivalent can also be made, these are to the present invention Claim improves and the technical scheme after equivalent, each falls within protection scope of the present invention.

Claims

1. a kind of camptothecin phosphatide cpd is it is characterised in that this phosphatide cpd is the compound of general formula (1) or institute State the pharmaceutically acceptable salt that the compound of formula (1) and counter ion counterionsl gegenions are formed：

In formula (1), R₁For spacerarm, it is alkylene, described alkylene is defined as the straight or branched that carbon number is 1～20 Alkylene alkyl/Asia alkylene or the aryl that carbon number is 6～10；L represent 2- amino -2- carboxy ethyl, 2- amino-ethyl, 2- trimethyl amido ethyl cation or 2,3- dihydroxypropyl；

R_xIt is that camptothecine compounds 20 bit substituent, the camptothecine compounds 10 of formula (3) that structure is following formula (2) take 7 methylene substituent of camptothecine compounds for base or formula (4)：

In formula (2) and (3) and (4), R₇For hydrogen, methyl, ethyl or methylol, R₉For hydrogen, amino, dimethylamino or nitro, R₁₀ For hydrogen, hydroxyl, methyl, halogen or connection piperazine carboxyl, R₁₁For hydrogen or halogen, R₁₂For hydrogen；

In formula (3) and (4), R represents methyl；

In formula (2) and (3) and (4), 20 carbon atoms are S configuration.

2. camptothecin phosphatide cpd according to claim 1 is it is characterised in that described counter ion counterionsl gegenions are cation matter Son, sodium ion, potassium ion, calcium ion, iron ion, magnesium ion, ammonium ion, zinc ion, anion chloride ion, sulfate ion, Sulfate ion, nitrate ion, carboxylic acid ion, carbanion, bromide ion, phosphate anion, formate, acetate, Citrate, lactate, fumaric acid radical, tartrate anion, any one or any two combination of gluconic acid radical ion.

3. a kind of pharmaceutical composition is it is characterised in that said composition includes the camptothecin phospholipid described in claim 1 or 2 Compound or described camptothecin phosphatide cpd and acceptable carrier on pharmacodynamicss.

4. pharmaceutical composition according to claim 3 is it is characterised in that described pharmaceutical composition is capsule, granule, coagulates Colloid, injection, slow releasing preparation or controlled release preparation.

5. pharmaceutical composition according to claim 3 is it is characterised in that described pharmaceutical composition is particle diameter 10-1000 nanometer Elaioplast nanometer particle, also include auxiliary agent in this pharmaceutical composition.

6. pharmaceutical composition according to claim 5 is it is characterised in that described auxiliary agent is phospholipid.

7. a kind of camptothecin phosphatide cpd described in claim 1 or 2 is being prepared the application in antitumor drug, its feature It is, this application is by described camptothecin phosphatide cpd or its pharmaceutically acceptable salt, acceptable with pharmacodynamicss Carrier is prepared into medicament.