CN105233298A - Paclitaxel phospholipid compound and drug combination and application thereof - Google Patents
Paclitaxel phospholipid compound and drug combination and application thereof Download PDFInfo
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- CN105233298A CN105233298A CN201510598962.6A CN201510598962A CN105233298A CN 105233298 A CN105233298 A CN 105233298A CN 201510598962 A CN201510598962 A CN 201510598962A CN 105233298 A CN105233298 A CN 105233298A
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Abstract
The invention discloses a paclitaxel phospholipid compound and a drug combination and application thereof. The drug combination comprises the paclitaxel phospholipid compound or a medicinal combination of the paclitaxel phospholipid compound and a pharmacodynamically-acceptable carrier, and can be a liquid preparation, a solid preparation, a semisolid preparation, capsules, granules, gel and an injection. The drug combination is liposome nanoparticles prepared from the paclitaxel phospholipid compound or the paclitaxel phospholipid compound and an auxiliary, and grain diameter of the liposome nanoparticles is 10-1000 nanometers. The paclitaxel phospholipid compound and the liposome nanoparticles utilize quickly-breakable spacer arms in molecular structure to quickly release paclitaxel drug, can be used as liquid preparations, solid preparations, semisolid preparations, sterilizing preparations and sterile preparations, are low in toxicity and can be used for efficient treatment of various tumors.
Description
Technical field
The present invention is a kind of taxanes phosphatide cpd and its pharmaceutical composition and purposes with antitumor action, relates to medical art.
Background technology
Paclitaxel is a kind of alkaloid, can extract from plant, also can carry out chemosynthesis.Paclitaxel is the secondary metabolite of a kind of complexity in Chinese yew genus plants, is also that understand at present uniquely a kind of can promote microtubule polymerization and the stable medicine being polymerized microtubule.Research shows, paclitaxel is attached on the microtubule of polymerization, does not react with unpolymerized tubulin dimer.Understand after cells contacting paclitaxel at a large amount of microtubule of thin intracellular accumulation, the accumulation of these microtubules disturbs the various functions of cell, particularly makes cell division stop at mitotic phase, has blocked the proper splitting of cell.By II-III phase clinical research, paclitaxel is mainly applicable to ovarian cancer and breast carcinoma, also has certain curative effect to pulmonary carcinoma, colorectal cancer, melanoma, incidence cancer, lymphoma, cerebroma.
Although paclitaxel has good antitumor action, poorly water-soluble, polyoxyethylene castor oil and ethanol hydrotropy need be adopted, and polyoxyethylene castor oil may cause anaphylaxis, needs to take Claritin in advance before patient medication.There are some researches show, the application of polyoxyethylene castor oil reduces the anti-tumor activity of paclitaxel, and paclitaxel is due to poor selectivity simultaneously, has stronger toxic action, easily produces drug resistance, cannot through shortcomings such as blood brain barrier.Anaphylaxis light symptoms is flushed face in the afternoon, dermoreaction, heart rate are slightly fast, blood pressure slightly falls, and severe reaction is that blood pressure is low, vasodilation, dyspnea, whole body urticaria etc.Side effect have refer to that toe is numb, transient tachycardia and hypotension, joint and myalgia, digestive tract reaction, slight alopecia and bilirubin, alkali phosphatase, glutamic oxaloacetic transaminase, GOT rising etc.
Paclitaxel and analog thereof can extract from plant, also can carry out chemosynthesis or semi-synthetic.The greatest drawback that paclitaxel or its analog are used for oncotherapy is poorly water-soluble, is difficult to the solution obtaining high concentration.In order to improve the defect of paclitaxel, many research institutions and enterprise have carried out modification to the chemical constitution of paclitaxel, or make liposome etc.On the other hand, there is the defects such as side reaction is large for oncotherapy in paclitaxel and analog thereof.Therefore it may be necessary and structural modification is carried out to paclitaxel and analog thereof, or adopt efficient carrier system, improve drug solubility, targeting to reach, improve curative effect, reduce toxicity, overcome the object of drug resistance.
Liposome is a kind of new medicinal preparation, is the important carrier of cancer therapy drug.Phospholipid is the main chemical compositions forming liposome, and when being scattered in aqueous phase, the hydrophobic tail in molecule is tended to flock together, and avoids aqueous phase, and hydrophilic head is exposed in aqueous phase, forms the vesicle with bilayer structure, i.e. liposome.During as carrier, hydrophobic drug usually in two lipid layers of liposome, in the aqueous phase of hydrophilic medicament in liposome.Due to the mobility of liposome membrane, cause medicine to be easy to leak out, be difficult to play good drug effect.Drug molecule being formed prodrug by being covalently bound in phospholipid molecule structure, can drug leakage be avoided.By making the breaking of covalent bonds between taxanes pharmaceutically active group and phospholipid, taxanes phospholipid prodrugs or liposome slow releasing medicine.Because the speed of prodrugs release medicine is too slow, taxanes phospholipid prodrug or liposome drug effect is caused to decline.
The present invention utilizes the paclitaxel of 2 molecules or its analog reactive moiety to be connected by covalent bond with phospholipid hydrophilic head as the spacerarm of hydrophobic tail by easily degraded, prepares taxanes phosphatide cpd, medicine can be made to discharge fast; The dissolubility of taxanes phosphatide cpd of the present invention is significantly better than paclitaxel and derivant thereof; Taxanes phosphatide cpd is prepared into nano-particle by the present invention, has the characteristic of liposome, has targeting feature, for oncotherapy.This taxanes phosphatide cpd nano-particle is not only a kind of prodrug, is also the drug release carrier that taxanes medicine is brand-new, has target function; Taxanes phosphatide cpd of the present invention and liposome thereof, through degradeds such as enzyme, glutathion effects, rapid delivery of pharmaceuticals, plays drug effect.
Summary of the invention
Technical problem: the object of the present invention is to provide one to discharge paclitaxel analog compound fast, improve the taxanes phosphatide cpd of paclitaxel analog compound dissolubility and drug effect, there is provided a kind of pharmaceutical composition based on this taxanes phosphatide cpd, and this taxanes phosphatide cpd is preparing the application in antitumor drug simultaneously.
Technical scheme: in order to Compound nomenclature and location, the position of each atom of definition paclitaxel is as follows:
The female ring of all paclitaxels that the present invention relates to and analog or its phosphatide cpd all defines according to the position in formula (A), and being configured as of the main carbon atom of parent nucleus: 1S, 2S, 3R, 4S, 5R, 7S, 8S, 10R, 13S, 2 ' R, 3 ' S.
Taxanes phosphatide cpd of the present invention is the pharmaceutically acceptable salt that the compound of following general formula (1) or the compound of described general formula (1) and counter ion counterionsl gegenions are formed:
In formula (1), R
1and R
2for spacerarm, to be carbon number be 2 ~ 20 the alkylene alkyl containing ehter bond, the carbon number oxygen base alkylene alkyl that is 1 ~ 20, the carbon number alkylene alkyl containing cystine linkage-S-S-that is 2 ~ 20, the carbon number alkylene alkyl containing peptide bond that is 2 ~ 60, the carbon number alkylene alkyl containing ester bond that is 2 ~ 20, carbon number be 2 ~ 20 the alkylene alkyl containing hydrazone key or carbon number be 2 ~ 20 not containing heteroatomic alkylene alkyl/sub-alkylene; L is 2-amino-2-carboxy ethyl, 2-amino-ethyl, 2-trimethyl amido ethyl cation, 2, the end group of 3-dihydroxypropyl, molecular weight to be the Polyethylene Glycol-amino-ethyl of the N-without targeting end group of 200-4000 or molecular weight be 200-4000 is the N-Polyethylene Glycol-amino-ethyl of targeting group;
R
xand R
ypaclitaxel analogs 10 bit substituent of to be structure be the paclitaxel analogs 2 ' bit substituent of following formula (2), paclitaxel analogs 7 bit substituent of formula (3) or formula (4), wherein the configuration of each carbon atom is: 1S, 2S, 3R, 4S, 5R, 7S, 8S, 10R, 13S, 2 ' R, 3 ' S:
In formula (2), (3), (4), R
3hydrogen; R
4any one in following group: benzoyl, C
1-C
6alkyl-carbonyl, C
1-C
6halogenated alkyl carbonyl, the C of straight or branched
1-C
6three substituted silane bases, the C of straight or branched alkyl
1-C
6straight or branched alkylene carbonyl; R
5any one in following group: acetyl group, C
1-C
6alkyl-carbonyl, C
1-C
6halogenated alkyl carbonyl, the C of straight or branched
1-C
6three substituted silane bases, the C of straight or branched alkyl
1-C
6straight or branched alkylene carbonyl; R
6hydrogen, C
1-C
6alkyl, the C of straight or branched
1-C
6alkyl-carbonyl, C
1-C
6haloalkyl, the C of straight or branched
1-C
6halogenated alkyl carbonyl, the C of straight or branched
1-C
6alkyl/alkyl-carbonyl, the C of hydroxyl of straight or branched
1-C
6three substituted silane bases, the C of straight or branched alkyl
1-C
6straight or branched alkylene carbonyl, C
1-C
6straight or branched alkoxymethylene, phenyoxymethylene or benzene methyl methylene; R7 is hydrogen, acetyl group, C
1-C
6alkyl, the C of straight or branched
1-C
6alkyl-carbonyl, C
1-C
6haloalkyl, the C of straight or branched
1-C
6halogenated alkyl carbonyl, the C of straight or branched
1-C
6three substituted silane bases, the C of straight or branched alkyl
1-C
6straight or branched alkylene carbonyl, C
1-C
6straight or branched alkoxymethylene, phenyoxymethylene or benzene methyl methylene; R
8hydrogen, C
1-C
6alkyl-carbonyl, C
1-C
6halogenated alkyl carbonyl, the C of straight or branched
1-C
6three substituted silane bases, the C of straight or branched alkyl
1-C
6straight or branched alkylene carbonyl, C
1-C
6the alkyl-carbonyl of hydroxyl of straight or branched or two dodecahexaene carbonyls; R
9benzoyl, t-butyloxycarbonyl, tert-butyl carbonyl, hydrogen, C
1-C
18alkyl-carbonyl, the C of straight or branched
1-C
6halogenated alkyl carbonyl, the C of straight or branched
1-C
6straight or branched hydroxyl/alkyl-carbonyl, C
1-C
6straight or branched alkylene carbonyl or two dodecahexaene carbonyls.
In taxanes phosphatide cpd of the present invention, Rx and Ry preferred paclitaxel 2 ' bit substituent, paclitaxel 7 bit substituent, paclitaxel 10 bit substituent, Docetaxel 2 ' bit substituent, Docetaxel 7 bit substituent, Docetaxel 10 bit substituent.
In the preferred version of taxanes phosphatide cpd of the present invention, counter ion counterionsl gegenions are a kind of cationes, or the combination of a kind of cation and a kind of anion.
In the preferred version of taxanes phosphatide cpd of the present invention, end group is in the N-Polyethylene Glycol-amino-ethyl of targeting group, and targeting group is any one in folic acid, galactose, polypeptide, antibody, antibody fragment, hyaluronic acid, asialoglycoprotein, polysaccharide, nucleic acid and plan peptide.
Taxanes phosphatide cpd of the present invention can be prepared in accordance with the following methods, first by the hydroxyl of paclitaxel or paclitaxel analogs and dicarboxylic anhydride or carboxylic acid reaction, prepare the intermediate containing carboxyl, then further described intermediate and hydrophilic segment are carried out condensation reaction under condensation reagent effect, namely obtain taxanes phosphatide cpd.
Pharmaceutical composition of the present invention, comprises acceptable carrier on above-mentioned taxanes phosphatide cpd or described taxanes phosphatide cpd and pharmacodynamics.
Pharmaceutical composition of the present invention is liquid preparation, solid preparation, semi-solid preparation, capsule, granule, gel, injection, slow releasing preparation or controlled release preparation.
In the preferred version of pharmaceutical composition of the present invention, pharmaceutical composition is the elaioplast nanometer particle of particle diameter 10-1000 nanometer.
In the preferred version of pharmaceutical composition of the present invention, pharmaceutical composition is the elaioplast nanometer particle of particle diameter 10-1000 nanometer, also comprises auxiliary agent in this pharmaceutical composition.
In the above-mentioned preferred version of pharmaceutical composition of the present invention, auxiliary agent is phospholipid, one or both combination cholesteric.
Taxanes phosphatide cpd of the present invention is preparing the application in antitumor drug, by taxanes phosphatide cpd or its pharmaceutically acceptable salt, is prepared into medicament with acceptable carrier on pharmacodynamics.
The compounds of this invention can the form of isomer exist, and usually described " the compounds of this invention " comprises the isomer of this compound.Can be there is asymmetric center and have S configuration or R configuration in the compounds of this invention, the present invention includes the mixture of all possible stereoisomer and two or more isomers.
The invention still further relates to and contain as the compounds of this invention of active ingredient or the pharmaceutical composition of the compounds of this invention and customary pharmaceutical auxiliaries.Usual pharmaceutical composition of the present invention contains the compounds of this invention of 0.1-100 % by weight.
The pharmaceutical composition of the compounds of this invention can be prepared according to method well known in the art.During for this object, if needed, the compounds of this invention or the compounds of this invention and one or more solids or liquid pharmaceutical excipients and/or adjuvant can be combined, make the suitable administration form or dosage form that can be used as the use of people's medicine.
The compounds of this invention or the pharmaceutical composition containing it can administrations in a unit, and 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.Form of administration can be liquid dosage form, solid dosage forms.
The compounds of this invention can be made ordinary preparation, also can be slow releasing preparation, controlled release preparation, targeting preparation and various particulate delivery system.
In order to administration unit is made capsule, effective ingredient the compounds of this invention is mixed with above-mentioned various carriers, and the mixture obtained thus is placed in hard gelatine capsule or soft capsule.Also effective ingredient the compounds of this invention can be made microcapsule, be suspended in aqueous medium and form suspensoid, also can load in hard capsule or make injection application.
The compounds of this invention is made injection preparation, as solution, suspensoid solution, Emulsion, lyophilized injectable powder, this preparation can be moisture or non-water, can containing acceptable carrier, diluent, binding agent, lubricant, antiseptic, surfactant or dispersant on a kind of and/or multiple pharmacodynamics.
Elaioplast nanometer particle is made, grain diameter 10-1000 nanometer by the compounds of this invention.
Elaioplast nanometer particle is made by the compounds of this invention and auxiliary agent, particle diameter 10-1000 nanometer, the auxiliary agent phospholipid used, the phospholipid of bonding Polyethylene Glycol, end group is one or more in the phospholipid of the bonding Polyethylene Glycol of targeting group or cholesterol, and targeting group is folic acid, galactose, antibody, biotin, polypeptide, antibody, antibody fragment, hyaluronic acid, asialoglycoprotein, polysaccharide, nucleic acid and plan peptide.
Medicine compound liposome nano-particle of the present invention is liquid preparation, solid preparation, semi-solid preparation, capsule, granule, gel, injection, slow releasing preparation or controlled release preparation.
From anti tumor activity in vitro screening, the compounds of this invention shows good anti-tumor activity.Test shows the toxicity in vivo of the compounds of this invention, is less than paclitaxel, Docetaxel.
From anti tumor activity in vitro screening, the compounds of this invention elaioplast nanometer particle shows good anti-tumor activity.Test shows that the toxicity in vivo of the compounds of this invention is much smaller than paclitaxel, Docetaxel.
From external degradation speed, the compounds of this invention performance fast degradation.
The preparation method of taxanes phosphatide cpd elaioplast nanometer particle of the present invention, the mixture by the compounds of this invention taxanes phosphatide cpd or the compounds of this invention and auxiliary agent, by method preparations such as film dispersion method, reverse phase evaporation, freeze-drying, ultrasonic dispersion, spray drying method, film squeezing and pressing method or high pressure homogenization methods.
The present invention utilizes the paclitaxel of 2 molecules or paclitaxel analogs to be connected by spacerarm covalency capable of being fast degraded with phospholipid hydrophilic head as hydrophobic tail, prepares taxanes phosphatide cpd, and degradation speed is fast, and dissolubility is significantly better than paclitaxel or paclitaxel analogs; Taxanes phosphatide cpd is prepared into nano-particle by the present invention, has the characteristic of liposome, for oncotherapy.This taxanes phosphatide cpd nano-particle is not only a kind of prodrug, is also the drug release carrier that taxanes medicine is brand-new, has target function; Taxanes phosphatide cpd of the present invention and elaioplast nanometer particle thereof, through effect fast degradation release medicines such as enzymes, fast onset drug effect, has low toxic and side effects.
Beneficial effect: the present invention compared with prior art, has the following advantages:
Taxanes phosphatide cpd of the present invention has following general structure (1):
Wherein, R
1and R
2for spacerarm, to be carbon number be 2 ~ 20 the alkylene alkyl containing ehter bond, the carbon number oxygen base alkylene alkyl that is 1 ~ 20, the carbon number alkylene alkyl containing cystine linkage-S-S-that is 2 ~ 20, the carbon number alkylene alkyl containing peptide bond that is 2 ~ 60, the carbon number alkylene alkyl containing ester bond that is 2 ~ 20, carbon number be 2 ~ 20 the alkylene alkyl containing hydrazone key or carbon number be 2 ~ 20 not containing heteroatomic alkylene alkyl/sub-alkylene; L is 2-amino-2-carboxy ethyl, 2-amino-ethyl, 2-trimethyl amido ethyl cation, 2, the end group of 3-dihydroxypropyl, molecular weight to be the Polyethylene Glycol-amino-ethyl of the N-without targeting end group of 200-4000 or molecular weight be 200-4000 is the N-Polyethylene Glycol-amino-ethyl of targeting group, R
xand R
ybe the substituent group of paclitaxel or paclitaxel analogs, specifically see technical scheme.
Taxanes phosphatide cpd of the present invention, R in structural formula (1)
1and R
2for spacerarm, this spacerarm contains the chemical bond of easy fracture, under glutathion or polypeptidase etc. exist, can fast fracture, and discharge active medicine paclitaxel or paclitaxel analogs fast, in the short time, form the former medicine of high concentration, play drug effect; A part taxanes phosphatide cpd discharges paclitaxel or the paclitaxel analogs of two molecules, thus makes the antineoplastic agent effect of paclitaxel or paclitaxel analogs phosphatide cpd reach paclitaxel or paclitaxel analogs more than 2 times;
Containing hydrophilic head part in taxanes phosphatide cpd structural formula (1) of the present invention, make phosphatide cpd water solublity significantly be better than paclitaxel, Docetaxel, there is hypotoxicity and excellent anti-tumor activity;
R in taxanes phosphatide cpd of the present invention
xand R
ymedicine substituent group has hydrophobic interaction, with hydrophobic R in formula (1)
1and R
2spacerarm is collaborative strengthens hydrophobicity, makes taxanes phosphatide cpd be assembled into stable elaioplast nanometer particle, and when overcoming general liposome paclitaxel or its analog dewatering medicament, medicine is easy to the shortcoming of leaking, and improves the efficiency of medicine parcel;
Taxanes phosphatide cpd of the present invention, R
1and R
2spacerarm to be carbon number be 2 ~ 20 the alkylene alkyl containing ehter bond, carbon number is the oxygen base alkylene alkyl of 1 ~ 20, carbon number is the alkylene alkyl containing cystine linkage-S-S-of 2 ~ 20, carbon number is the alkylene alkyl containing peptide bond of 2 ~ 50, carbon number is the alkylene alkyl containing hydrazone key of 2 ~ 20, carbon number be 2 ~ 20 containing the alkylene alkyl of ester bond or carbon number be 2 ~ 20 not containing heteroatomic alkylene alkyl/sub-alkylene, this spacerarm is by the fast degradation such as glutathion or polypeptidase, discharge paclitaxel or paclitaxel analogs fast, strong killing action is had to breast cancer cell,
The compound system that taxanes phosphatide cpd of the present invention or itself and phospholipid auxiliary agent etc. form can adopt simple process, as membrane process etc. is assembled into elaioplast nanometer particle easily, and particle diameter 10-1000 nanometer;
Taxanes phosphatide cpd of the present invention is assembled into stable elaioplast nanometer particle, there is the membrane structure similar with cell, be easy to by cellular uptake, make taxanes phosphatide cpd enter in cell by liposomal form, and discharge medicine, play stronger drug effect;
Taxanes phosphatide cpd elaioplast nanometer particle of the present invention has passive target effect, is easy to assemble in tumor cell, play drug effect, and the paclitaxel analogs such as paclitaxel, Docetaxel does not all possess this characteristic;
Taxanes phosphatide cpd of the present invention and elaioplast nanometer particle thereof are a kind of release vehicles of brand-new taxanes medicine, are also a kind of prodrugs;
The pharmaceutical composition of compound of the present invention or compound of the present invention and Conventional pharmaceutical carriers, the compounds of this invention containing 0.1-100 % by weight, has excellent anti-tumor activity, has strong killing action to breast cancer cell;
Taxanes phosphatide cpd of the present invention and elaioplast nanometer particle thereof, can be used as liquid preparation, solid preparation, semi-solid preparation, sterilization preparation and sterile preparation, can form liposome under aqueous phase system;
Taxanes phosphatide cpd of the present invention or taxanes phosphatide cpd liposome, in its structural formula (1), the end group of L to be molecular weight be 200-4000 is the N-Polyethylene Glycol-amino-ethyl of targeting group, the surface of liposome that this taxanes phosphatide cpd is assembled into has targeting group, has active targeting effect;
The surface of liposome that taxanes phosphatide cpd of the present invention is assembled into altogether with the phospholipid containing targeting group has targeting group, has active targeting effect.
Accompanying drawing explanation
Fig. 1 is two (paclitaxel-7-dithio diethyl alkyd) Phosphatidylcholine biosynthesis route maps.
Fig. 2 is two (Docetaxel-2 '-dithio diethyl alkyd) Phosphatidylcholine biosynthesis route map.
Fig. 3 is two (paclitaxel-7-diglycolic acid) Phosphatidylcholine biosynthesis route maps.
Fig. 4 is two (paclitaxel-7-succinyl-glycyl) Phosphatidylcholine biosynthesis route maps.
Fig. 5 is two (paclitaxel-7-succinyl-glycyl-phenylalanyl-leucyl-glycyl) Phosphatidylcholine biosynthesis route maps.
Fig. 6 is two (paclitaxel-7-succinyl-GFQGVQFAGF) Phosphatidylcholine biosynthesis route map (G is GFQGVQFAGF).
Fig. 7 is two (paclitaxel-7-carbonic ester-butyryl) Phosphatidylcholine biosynthesis route maps.
Fig. 8 is two (paclitaxel-7-dithio diethyl alkyd) PHOSPHATIDYL ETHANOLAMINE synthetic route charts.
Fig. 9 is two (paclitaxel-7-dithio diethyl alkyd) PHOSPHATIDYL ETHANOLAMINE-N-polyethylene glycol compound synthetic route charts.
Figure 10 is two (paclitaxel-7-dithio diethyl alkyd) PHOSPHATIDYL ETHANOLAMINE-N-Polyethylene Glycol-folic acid synthetic route charts.
Figure 11 is two (paclitaxel-7-dithio diethyl alkyd) phosphatidyl glycerol synthetic route charts.
Figure 12 is two (paclitaxel-2 '-succinic acid) Phosphatidylcholine biosynthesis route map.
Figure 13 is the synthetic route chart of two (paclitaxel-7-succinic acid) phosphatidylcholine
Figure 14 is the synthetic route chart of two (Docetaxel-2 '-adipic acid) phosphatidylcholine
Figure 15 is the particle size distribution of two (paclitaxel-7-succinic acid) phosphatidylcholine liposome.
Figure 16 is two (paclitaxel-7-succinic acid) phosphatidylcholine elaioplast nanometer particle transmission electron microscope pictures.
Figure 17 is two (paclitaxel-7-succinyl) phosphatidylcholine lipidosome antineoplastic activity and the weight of animals change (a) anti-tumor activity, and (b) the weight of animals changes.
Detailed description of the invention
Below in conjunction with Figure of description and embodiment, technical scheme of the present invention is described in further details.
A kind of taxanes phosphatide cpd of the present invention is the pharmaceutically acceptable salt that the compound of following general formula (1) or the compound of described general formula (1) and counter ion counterionsl gegenions are formed:
In formula (1), R
1and R
2for spacerarm, to be carbon number be 2 ~ 20 the alkylene alkyl containing ehter bond, carbon number be 1 ~ 20 oxygen base alkylene alkyl, carbon number be 2 ~ 20 the alkylene alkyl containing cystine linkage-S-S-or carbon number be 2 ~ 60 the alkylene alkyl containing peptide bond, carbon number be 2 ~ 20 the alkylene alkyl containing ester bond, carbon number be 2 ~ 20 the alkylene alkyl containing hydrazone key or carbon number be 2 ~ 20 not containing heteroatomic alkylene alkyl/sub-alkylene; L is 2-amino-2-carboxy ethyl, 2-amino-ethyl, 2-trimethyl amido ethyl cation, 2, the end group of 3-dihydroxypropyl, molecular weight to be the Polyethylene Glycol-amino-ethyl of the N-without targeting end group of 200-4000 or molecular weight be 200-4000 is the N-Polyethylene Glycol-amino-ethyl of targeting group, targeting group is any one in folic acid, galactose, polypeptide, antibody, antibody fragment, hyaluronic acid, asialoglycoprotein, polysaccharide, nucleic acid and plan peptide;
R
xand R
ypaclitaxel analogs 10 bit substituent of to be structure be the paclitaxel analogs 2 ' bit substituent of following formula (2), paclitaxel analogs 7 bit substituent of formula (3) or formula (4), wherein the configuration of each carbon atom is: 1S, 2S, 3R, 4S, 5R, 7S, 8S, 10R, 13S, 2 ' R, 3 ' S:
In formula (2), (3), (4), R
3hydrogen; R
4benzoyl, C
1-C
6alkyl-carbonyl, C
1-C
6halogenated alkyl carbonyl, the C of straight or branched
1-C
6three substituted silane bases, the C of straight or branched alkyl
1-C
6straight or branched alkylene carbonyl; R
5acetyl group, C
1-C
6alkyl-carbonyl, C
1-C
6straight or branched halogenated alkyl carbonyl, C
1-C
6three substituted silane bases, the C of straight or branched alkyl
1-C
6straight or branched alkylene carbonyl; R
6hydrogen, C
1-C
6alkyl, the C of straight or branched
1-C
6alkyl-carbonyl, C
1-C
6haloalkyl, the C of straight or branched
1-C
6halogenated alkyl carbonyl, the C of straight or branched
1-C
6alkyl/alkyl-carbonyl, the C of hydroxyl of straight or branched
1-C
6three substituted silane bases, the C of straight or branched alkyl
1-C
6straight or branched alkylene carbonyl, C
1-C
6straight or branched alkoxymethylene, phenyoxymethylene or benzene methyl methylene; R
7hydrogen, acetyl group, C
1-C
6alkyl, the C of straight or branched
1-C
6alkyl-carbonyl, C
1-C
6haloalkyl, the C of straight or branched
1-C
6halogenated alkyl carbonyl, the C of straight or branched
1-C
6three substituted silane bases, the C of straight or branched alkyl
1-C
6straight or branched alkylene carbonyl, C
1-C
6straight or branched alkoxymethylene, phenyoxymethylene or benzene methyl methylene; R
8hydrogen, C
1-C
6alkyl-carbonyl, C
1-C
6halogenated alkyl carbonyl, the C of straight or branched
1-C
6three substituted silane bases, the C of straight or branched alkyl
1-C
6straight or branched alkylene carbonyl, C
1-C
6the alkyl-carbonyl of hydroxyl of straight or branched or two dodecahexaene carbonyls; R
9benzoyl, t-butyloxycarbonyl, tert-butyl carbonyl, hydrogen, C
1-C
18alkyl-carbonyl, the C of straight or branched
1-C
6halogenated alkyl carbonyl, the C of straight or branched
1-C
6straight or branched hydroxyl/alkyl-carbonyl, C
1-C
6straight or branched alkylene carbonyl or two dodecahexaene carbonyls.
In taxanes phosphatide cpd of the present invention, Rx and Ry preferred paclitaxel 2 ' bit substituent, paclitaxel 7 bit substituent, paclitaxel 10 bit substituent, Docetaxel 2 ' bit substituent, Docetaxel 7 bit substituent, Docetaxel 10 bit substituent.
Counter ion counterionsl gegenions are combinations of any one or any two of cation hydrion, 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, gluconic acid radical ion, preferred hydrion, sodium ion, chloride ion.
The method of the taxanes phosphatide cpd described in preparation, first the method by the hydroxyl of paclitaxel or paclitaxel analogs and anhydride or carboxylic acid reaction, prepares the intermediate containing carboxyl.Then, described intermediate and choline glycerophosphatide are carried out condensation reaction, namely obtains taxanes phosphatidyl choline compounds.
Pharmaceutical composition of the present invention is liquid preparation, solid preparation, semi-solid preparation, capsule, granule, gel, injection, slow releasing preparation or controlled release preparation.
Pharmaceutical composition of the present invention is the elaioplast nanometer particle of particle diameter 10-1000 nanometer, is made up of the taxanes phosphatide cpd self assembly with general formula (1) structure.
The elaioplast nanometer particle forming particle diameter 10-1000 nanometer is jointly assembled by the taxanes phosphatide cpd and phospholipid with general formula (1) structure.
Taxanes phosphatide cpd of the present invention is characterized in that preparing the application in antitumor drug, and the method, by described taxanes phospholipid or its pharmaceutically acceptable salt, is prepared into medicament with acceptable carrier on pharmacodynamics.The compounds of this invention or the compounds of this invention and one or more solids or liquid pharmaceutical excipients and/or adjuvant can be combined, make and can be used as the suitable administration form or dosage form that people's medicine uses.Usual pharmaceutical composition of the present invention contains the compounds of this invention of 0.1-100 % by weight.
The compounds of this invention or the pharmaceutical composition route of administration containing it can be intestinal or non-bowel, as oral, muscle, subcutaneous, nasal cavity, oral mucosa, skin, peritoneum or rectum etc.Can be drug administration by injection, comprise intravenous injection, intramuscular injection, subcutaneous injection, intradermal injection and acupoint injection therapy etc.Form of administration can be liquid dosage form, solid dosage forms.If liquid dosage form can be true solution class, colloidal type, particulate formulations, emulsion dosage form, mixed suspension form.Other dosage forms are tablet, capsule, drop pill, aerosol, pill, powder, solution, suspensoid, Emulsion, granule, suppository, lyophilized injectable powder etc. such as.
The compounds of this invention can be made ordinary preparation, also can be slow releasing preparation, controlled release preparation, targeting preparation and various particulate delivery system.
The carrier that pharmaceutical composition of the present invention adopts is, such as diluent, absorbent, wetting agent, binding agent, disintegrate inhibitor, lubricant etc.Tablet can also be made coated tablet further, such as sugar coated tablet, thin membrane coated tablet, ECT, or double-layer tablet and multilayer tablet.
The compounds of this invention is made injection preparation, as solution, suspensoid solution, Emulsion, lyophilized injectable powder, this preparation can be moisture or non-water, can containing acceptable carrier, diluent, binding agent, lubricant, antiseptic, surfactant or dispersant on a kind of and/or multiple pharmacodynamics.
Make elaioplast nanometer particle by the compounds of this invention and auxiliary agent, particle diameter 10-1000 nanometer, the auxiliary agent of use is one or more in phospholipid, targeting phospholipid or cholesterol.Targeting group in targeting phospholipid is any one in folic acid, galactose, polypeptide, antibody, antibody fragment, hyaluronic acid, asialoglycoprotein, polysaccharide, nucleic acid and plan peptide.
From anti tumor activity in vitro screening, the compounds of this invention shows good anti-tumor activity.Test shows the toxicity in vivo of the compounds of this invention, is less than paclitaxel.From anti tumor activity in vitro screening, the compounds of this invention elaioplast nanometer particle shows good anti-tumor activity.Test shows that the toxicity in vivo of the compounds of this invention is much smaller than paclitaxel.From external degradation speed, the compounds of this invention shows fast degradation.
The preparation method of taxanes phosphatide cpd elaioplast nanometer particle of the present invention, the mixture by the compounds of this invention taxanes phosphatide cpd or the compounds of this invention and auxiliary agent, by method preparations such as film dispersion method, reverse phase evaporation, freeze-drying, ultrasonic dispersion, spray drying method, film squeezing and pressing method or high pressure homogenization methods.
Taxanes phosphatide cpd of the present invention, its spacerarm is under the effect such as glutathion or polypeptidase, and fast fracture, discharges active medicine paclitaxel or paclitaxel analogs fast, forms the former medicine of paclitaxel analog compound high concentration in the short time, plays drug effect.A part taxanes phosphatide cpd fast degradation of the present invention discharges paclitaxel or the paclitaxel analogs of two molecules, thus makes the antitumor drug effect of paclitaxel or paclitaxel analogs phosphatide cpd reach paclitaxel or paclitaxel analogs more than 2 times.The present invention utilizes the paclitaxel of 2 molecules or paclitaxel analogs to be connected by covalent bond with phospholipid hydrophilic head as hydrophobic tail, and prepare taxanes phosphatide cpd, dissolubility is significantly better than paclitaxel or paclitaxel analogs, has lower toxic and side effects.Taxanes phosphatide cpd is prepared into nano-particle by the present invention, has the characteristic of liposome, can form liquid preparation, solid preparation, semi-solid preparation, sterilization preparation and sterile preparation, for oncotherapy.This taxanes phosphatide cpd elaioplast nanometer particle is not only a kind of prodrug, is also the drug release carrier that taxanes medicine is brand-new.Taxanes phosphatide cpd liposome of the present invention, in structural formula (1), the end group of L to be molecular weight be 200-4000 is the N-Polyethylene Glycol-amino-ethyl of targeting group, and the surface of liposome that this taxanes phosphatide cpd is assembled into has targeting group, has active targeting effect; The surface of liposome that taxanes phosphatide cpd of the present invention is assembled into altogether with the phospholipid containing targeting group has targeting group, has active targeting effect.
Portion of reagent code name below for using in preparation process:
DMAP4-dimethylamino naphthyridine
CDIN, N '-carbonyl dimidazoles
GPC Phosphorylcholine glycerol
DBU1,5-diazabicylo [5.4.0] 11-5-alkene
EDC1-ethyl-(3-dimethylaminopropyl) carbodiimide
NHSN-N-Hydroxysuccinimide
TFA trifluoroacetic acid
DMF dimethyl formamide
DMSO dimethyl sulfoxine
BNF tetrabutyl ammonium fluoride
(BOC)
2o Bis(tert-butoxycarbonyl)oxide
BOC tertbutyloxycarbonyl
TEA triethylamine
Gly glycine
GFLG glycyl-phenylalanyl-leucyl-glycine
GFQGVQFAGF glycyl-Phenylalanyl-glutamyl-glycyl-a word used in person's names aminoacyl-glutamy-phenylalanyl-alanyl-glycyl-phenylalanine
Followingly further illustrate the present invention by embodiment, but the invention is not restricted to following examples.
Embodiment 1
The synthesis (route is shown in Fig. 1) of two (paclitaxel-7-dithio diethyl alkyd) phosphatidylcholine
Paclitaxel 1.5g (1mmol) is dissolved in 30mL chloroform, add triethylamine 1mL, add 1.5g tert-butyl chloro-silicane (TBDMSCl), at 0 DEG C, react 3h, add ice ether, separate out white sedimentation, centrifugalize, further column chromatography for separation (eluant, chloroform/methanol: 7/1, v/v) obtaining paclitaxel 2 ' position end-cap product 2 '-tertiary butyl dimethyl Si base-paclitaxel 1.24g, is white powdery solids.
2 '-tertiary butyl dimethyl Si base-paclitaxel 0.20g, is dissolved in 10ml chloroform, adds DMAP0.12g (1mmol), dithio diglycolic anhydride 0.6g, stirred at ambient temperature reaction 24h.Then, with the above-mentioned reactant liquor of 2%HCl solution washing, removing DMAP, column chromatography is separated (eluant, chloroform/methanol: 5/1, v/v) obtain white powder 2 '-tertiary butyl dimethyl Si base-paclitaxel-7-dithio diethanol acid monoester 0.13g.2 '-tertiary butyl dimethyl Si base-paclitaxel-7-dithio diethanol acid monoester is dissolved in 5mL chloroform, add CDI0.2g, activate 40 minutes, then GPC0.05g and DBU0.2g is added, room temperature reaction 24h, gained reactant liquor uses tetrabutyl ammonium fluoride process further, removes TBDMS, by column chromatography purification after cold diethyl ether precipitation, obtain two (paclitaxel-7-dithio diethyl alkyd) the phosphatidylcholine 0.08g of product.
1HNMR(500MHz，CD3OD∶CDCl
31∶1)：δ8.0-7.08(32H，m)，5.59-5.51(4H，m)，5.06-4.93(6H，m)，4.64(1H，m)，4.43-4.32(4H，m)，4.12-3.43(10H，m)，3.52-3.37(10H，m)，3.30(9H，s)，2.81(2H，d)，2.01-1.71(30H，m)，1.26-1.21(18H，s)。[M+H]
+m/z，2258.40。
Two (paclitaxel-7-dithio diethyl alkyd) phosphatidylcholine is dissolved in 0.1M sodium-chloride water solution, and lyophilizing obtains the phosphocholine compound solid powder containing sodium ion and chloride ion.
Embodiment 2:
The synthesis (synthetic route is shown in Fig. 2) of two (Docetaxel-2 '-dithio diethyl alkyd) phosphatidylcholine
Docetaxel (Docetaxel) 0.6g, adds 30ml chloroform and dissolves, add DMAP0.3g, dithio diglycolic anhydride 0.64g (6mmol), stirred at ambient temperature reaction 24h.Then, with the above-mentioned reactant liquor of 2%HCl solution washing, removing DMAP, column chromatography is separated (eluant, chloroform/methanol: 5/1, v/v) and obtains white powder Docetaxel-2 '-dithio diethanol acid monoester 0.43g.
0.20g Docetaxel-2 '-dithio diethanol acid monoester is dissolved in 20ml pyridine, add 0.4g tert-butyl chloro-silicane, 3h is reacted under room temperature, reactant liquor is added drop-wise in 50ml ice ether, separate out white precipitate, centrifugalize, further column chromatography is separated (eluant, chloroform/methanol: 7/1, v/v) end-cap product 7 is obtained, 10-bis-(tertiary butyl dimethyl Si base)-Docetaxel-2 '-dithio diethanol acid monoester 0.15g is white powdery solids.
Above-mentioned end-cap product 0.10g is dissolved in 10mL chloroform, add CDI0.1g, activation 1h, adds GPC0.05g and DBU0.1g, room temperature reaction 24h, reactant liquor uses tetrabutyl ammonium fluoride process further, remove TBDMS, reactant liquor is by column chromatography purification (eluant, chloroform/methanol: 5/1, v/v), product two (Docetaxel-2 '-dithio diethyl alkyd) phosphatidylcholine 0.06g is obtained.
1HNMR(500MHz，CD3OD∶CDCl
31∶1)：δ8.0-7.08(20H，m)，6.10-5.8(4H，dd)，4.93(4H，s)，4.77(2H，s)，4.64(1H，m)，4.43-3.37(24H，m)，3.30(9H，s)，2.81(2H，d)，2.01-2.00(16H，m)，1.72-1.71(10H，m)，1.41(18H，s)，1.26-1.21(18H，d)。[M+H]
+m/z，2166.35。
Two (Docetaxel-2 '-dithio diethyl alkyd) phosphocholine compound is dissolved in 0.1M sodium phosphate aqueous solution, and lyophilizing obtains the phosphocholine compound solid powder containing sodium ion and phosphoric acid acid group.
Embodiment 3:
The synthesis (route is shown in Fig. 3) of two (paclitaxel-7-diglycolic acid) phosphatidylcholine
Paclitaxel 2 ' position the end-cap product 2 '-tertiary butyl dimethyl Si base-paclitaxel 0.20g of embodiment 1 is dissolved in 10ml chloroform, adds DMAP0.5g, diglycolic anhydride 0.5g, stirred at ambient temperature reaction 24h.Then, with the above-mentioned reactant liquor of 2%HCl solution washing, removing DMAP, column chromatography is separated (eluant, chloroform/methanol: 5/1, v/v), obtains white powder 2 '-tertiary butyl dimethyl Si base-paclitaxel-7-diglycolic acid monoesters.2 '-tertiary butyl dimethyl Si base-paclitaxel-7-diglycolic acid monoesters is dissolved in 5mL chloroform, add CDI0.1g, activation 1h, add GPC0.05g and DBU0.1g, room temperature reaction 24h, reactant liquor 0.5g tetrabutyl ammonium fluoride process, removes TBDMS, reactant liquor, by column chromatography purification, obtains two (paclitaxel-7-diglycolic acid) the phosphatidylcholine 0.09g of product.
1HNMR(500MHz，CD3OD∶CDCl
31∶1)：δ8.01-7.08(32H，m)，5.59-5.51(4H，m)，5.06-4.93(6H，m)，4.64-4.32(13H，m)，4.12-3.37(12H，m)，3.30(9H，s)，2.81(2H，d)，2.01-2.00(20H，m)，1.81-1.71(10H，m)，1.26-1.21(18H，s)。[M+H]
+m/z，2162.15。
Two (paclitaxel-7-diglycolic acid) phosphatidylcholine is dissolved in 0.1M sodium citrate aqueous solution, and lyophilizing obtains the phosphatidylcholine pressed powder containing citrate ion and sodium ion.
Embodiment 4:
The synthesis (synthetic route is shown in Fig. 4) of two (paclitaxel-7-succinyl-glycyl) phosphatidyl choline compounds
Paclitaxel 2 ' position the end-cap product 2 '-tertiary butyl dimethyl Si base-paclitaxel 0.20g of embodiment 1, succinic anhydrides 1.0g, is dissolved in 30mL pyridine, reacts 24h under room temperature; Revolve steaming to desolventize, separate out precipitation in cold diethyl ether, dilute hydrochloric acid washs, and obtains intermediate product 2 '-tertiary butyl dimethyl Si base-paclitaxel-7-monomester succinate 0.15g.Intermediate product 0.15g is dissolved in DMF, adds 0.2gCDI and 0.2gTEA, activation 1h, then add glycine 0.5g, room temperature reaction 24h, reactant liquor pillar layer separation, obtain 2 '-tertiary butyl dimethyl Si base-paclitaxel-7-succinyl-glycine 0.11g.Above-mentioned product is dissolved in 5mL dimethyl sulfoxine, add CDI0.1g, activation 1h, add GPC0.05g and DBU0.1g, react 24h under room temperature, use tetrabutyl ammonium fluoride process further, remove TBDMS, reactant liquor cold diethyl ether separates out precipitation, is separated obtains two (paclitaxel-7-succinyl-glycyl) phosphatidylcholine 0.06g by column chromatography purification.
1HNMR(500MHz，CD3OD∶CDCl31∶1)：δ8.01-7.08(32H，m)，5.59-5.51(4H，m)，5.06-4.93(6H，m)，4.64(1H，m)，4.43-4.32(4H，m)，4.16-3.43(14H，m)，3.37(2H，t)，3.30(9H，s)，2.81-2.64(10H，t)，2.01-2.00(20H，m)，1.81-1.71(10H，m)，1.26-1.21(18H，s)。[M+H]
+m/z，2243.25。
Embodiment 5:
The synthesis (synthetic route is shown in Fig. 5) of two (paclitaxel-7-succinyl-glycyl-phenylalanyl-leucyl-glycyl) phosphatidylcholine
The 2 '-tertiary butyl dimethyl Si base-paclitaxel 0.2g of embodiment 1 is dissolved in 10mL chloroform, add succinic anhydrides 0.5g, DMAP0.5g, 12h is reacted under room temperature, precipitation is separated out in cold diethyl ether, dilute hydrochloric acid washs three times, filters, obtains intermediate product 2 '-tertiary butyl dimethyl Si base-paclitaxel-7-monomester succinate with pillar layer separation.Intermediate product is dissolved in 10mLDMF, add 0.2gCDI and 0.2gTEA, activation 1h, then N-glycyl-phenylalanyl-leucyl-glycine (GFLG) 0.3g is added, room temperature reaction 24h, reactant liquor pillar layer separation, obtains 2 '-tertiary butyl dimethyl Si base-paclitaxel-7-succinyl-glycyl-phenylalanyl-leucyl-glycine 0.12g; This product is dissolved in 10mL dimethyl sulfoxine, add CDI0.1g and activate 1h, add GPC0.05g and DBU0.1g, room temperature reaction 24h, 0.5g tetrabutyl ammonium fluoride is added in reactant liquor, react 6h under room temperature, separate out precipitation in cold diethyl ether, column chromatography for separation obtains two (paclitaxel-7-succinyl-glycyl-phenylalanyl-leucyl-glycyl) phosphatidylcholine 0.06g.
1HNMR(500MHz，CD3OD∶CDCl
31∶1)：δ8.0-7.08(50H，m)，5.59-5.51(4H，m)，5.06-4.12(25H，m)，4.01-3.05(23H，m)，2.81-2.53(10H，m)，2.01-1.71(36H，m)，1.26-1.21(18H，m)，1.01(12H，d)。[M+H]
+m/z，2879.06。
Embodiment 6:
The synthesis (synthetic route is shown in Fig. 6) of two (paclitaxel-7-succinyl-GFQGVQFAGF) phosphatidyl choline compounds
The 2 '-tertiary butyl dimethyl Si base-paclitaxel 0.2g of embodiment 1 is dissolved in 10mL chloroform, add succinic anhydrides 0.5g, DMAP0.5g, 12h is reacted under room temperature, precipitation is separated out in cold diethyl ether, dilute hydrochloric acid washs three times, filters, obtains intermediate product 2 '-tertiary butyl dimethyl Si base-paclitaxel-7-monomester succinate 0.12g with pillar layer separation.Intermediate product 0.12g is dissolved in 10mLDMF, adds 0.1gCDI and 0.1gTEA, activation 1h, then add polypeptide N-glycyl-Phenylalanyl-glutamyl-glycyl-a word used in person's names aminoacyl-glutamy-phenylalanyl-alanyl-glycyl-phenylalanine (NH
2gFQGVQFAGFCOOH, represents with G in reaction equation) 0.3g, react 24h under room temperature, separate out precipitation in cold diethyl ether, column chromatography for separation obtains 2 '-tertiary butyl dimethyl Si base-paclitaxel-7-succinyl-GFQGVQFAGF0.06g.Above-mentioned product is dissolved in 10mL dimethyl sulfoxine, add CDI0.06g and activate 1h, add GPC0.03g and DBU0.06g, 24h is reacted under room temperature, 0.3g tetrabutyl ammonium fluoride is added in reactant liquor, react 2h under room temperature, separate out precipitation in cold diethyl ether, column chromatography for separation obtains two (paclitaxel-7-succinyl-GFQGVQFAGF) phosphatidylcholine 0.053g.
Embodiment 7:
The synthesis (synthetic route is shown in Fig. 7) of two (paclitaxel-7-carbonic ester-butyryl) phosphatidylcholine
4 hydroxybutyric acid 1g is dissolved in chloroform 20ml, adds triethylamine 0.5g, adds 0.5g tert-butyl chloro-silicane, and room temperature reaction 3h adds ice ether, separates out white sedimentation, is product 4-tertiary butyl dimethyl Si base-butanoic acid 0.7g.0.3g4-tertiary butyl dimethyl Si base-butanoic acid is dissolved in 10mL chloroform; add CDI0.2g; activate after 1 hour; revolve and steam except desolventizing, add DMSO10mL, DBU0.2g; add GPC0.10g; react 12h under room temperature, and with tetrabutyl ammonium fluoride process deprotection base 4-tertiary butyl dimethyl Si base, pillar layer separation obtains two (4 hydroxybutyric acid) phosphatidylcholine 0.22g.
Two (4 hydroxybutyric acid) phosphatidylcholine 0.2g is scattered in the anhydrous chloroform of 20mL; add 0.5g triethylamine; solid phosgene 0.3g; after reaction 6h; be spin-dried for solvent; add the anhydrous chloroform of 10mL; add triethylamine 0.3g; add the 2 '-tertiary butyl dimethyl Si base-paclitaxel 0.2g of embodiment 1, under room temperature, react 6h; add tetrabutyl ammonium fluoride reaction 6h; deprotection base TBDMS, adds cold diethyl ether and separates out precipitation in reactant liquor, precipitation uses column chromatography and obtains two (paclitaxel-7-carbonic ester-butyryl) phosphatidylcholine 0.13g.
1HNMR(500MHz，CD3OD∶CDCl31∶1)：δ8.01-7.08(32H，m)，5.59-5.51(4H，m)，5.06-4.93(6H，m)，4.64(1H，m)，4.43-4.32(4H，m)，4.16-3.43(14H，m)，3.37(2H，t)，3.30(9H，s)，2.25(4H，t)，2.81(2H，st)，2.01-2.00(20H，m)，1.96-1.71(14H，m)，1.26-1.21(18H，s)。[M+H]+m/z，2190.20。
Embodiment 8:
The synthesis (synthetic route is shown in Fig. 8) of two (paclitaxel-7-dithio diethyl alkyd) phosphatidyl ethanol amines
3-(4-methoxyl group benzyloxy) propane-1,2-glycol 0.2g, dithio diglycolic anhydride 1g, DMAP0.5g, triethylamine 0.5g are dissolved in 30mL chloroform, back flow reaction 6h, and reactant liquor dilute hydrochloric acid washs three times, filter, solid is dissolved in 15mLDMSO, adds CDI0.2g, activation 1h, then the 2 '-tertiary butyl dimethyl Si base-paclitaxel 0.2g of embodiment 1 is added, DBU0.2g, reacts 24h under room temperature, and reactant liquor pillar layer separation obtains intermediate product; Intermediate product is dissolved in spirit of vinegar, logical hydrogen under palladium/carbon catalyst, room temperature reaction 24h, and except desolventizing, pillar layer separation obtains two (2 '-TBDMS-paclitaxel-7-dithio diethyl alkyd) glyceride 0.16g.
Two (2 '-TBDMS-paclitaxel-7-dithio diethyl alkyd) glyceride 0.16g is dissolved in the anhydrous chloroform of 10mL, add phosphorus oxychloride 0.1g, triethylamine 0.3g, 4h is reacted at 10 DEG C, revolve solvent evaporated, then 5mL chloroform solvent is added, add ethylaminoethanol 0.1g, triethylamine 0.3g, react 4h at 10 DEG C, in reactant liquor, add 0.2g tetrabutyl ammonium fluoride, reaction 2h, reactant liquor, by column chromatography purification, obtains two (paclitaxel-7-dithio diethyl alkyd) the PHOSPHATIDYL ETHANOLAMINE 0.12g of product.
1HNMR(500MHz，CD3OD∶CDCl31∶1)：δ8.01-7.08(32H，m)，5.59-5.51(4H，m)，5.06-4.93(6H，m)，4.64(1H，m)，4.43-4.32(4H，m)，4.12-3.43(10H，m)，3.37(2H，t)，3.30(9H，s)，2.81-2.64(10H，t)，2.01-2.00(20H，m)，1.81-1.71(10H，m)，1.26-1.21(18H，s)。[M+H]
+m/z，2216.36。
Embodiment 9:
The synthesis (synthetic route is shown in Fig. 9) of two (paclitaxel-7-dithio diethyl alkyd) PHOSPHATIDYL ETHANOLAMINE-N-polyethylene glycol compound
One-ended hydroxy mPEG (mean molecule quantity 2000) 0.8g, CDI0.1g are dissolved in 20mL chloroform, room temperature reaction 1h, add two (paclitaxel-7-dithio diethyl alkyd) the PHOSPHATIDYL ETHANOLAMINE 0.06g in embodiment 8, DBU0.1g, 40 DEG C of reacting by heating 12h, reactant liquor, by column chromatography purification, obtains two (paclitaxel-7-dithio diethyl alkyd) PHOSPHATIDYL ETHANOLAMINE-N-Polyethylene Glycol 0.04g of product.
1HNMR(500MHz，CD3OD∶CDCl
31∶1)：δ8.01-7.12(32H，m)，6.10-5.51(6H，m)，4.93-4.64(5H，m)，4.43-4.25(6H，m)，4.01(2H，d)，3.79-3.37(？H，broad)，3.15-2.64(12H，m)，2.01-1.71(30H，m)，1.21(12H，s)。
Embodiment 10:
The synthesis (synthetic route is shown in Figure 10) of two (paclitaxel-7-dithio diethyl alkyd) PHOSPHATIDYL ETHANOLAMINE-N-Polyethylene Glycol-folic acid
Terminal hydroxy group-N-BOC-amino-polyethyleneglycols (mean molecule quantity 2000) 0.5g is dissolved in 15mL chloroform, add CDI0.2g, activate after 1 hour, revolve and steam except desolventizing, add DMSO10mL, DBU0.2g, add two (paclitaxel-7-dithio diglycolic anhydride) PHOSPHATIDYL ETHANOLAMINE 0.06g of embodiment 8, room temperature reaction 12 hours, adds cold diethyl ether and separates out precipitation; Precipitation joins in oxolane, removes end group protecting group BOC, carry out purification by column chromatography process with TFA process, and obtaining end group is amino two (paclitaxel-7-dithio diethyl alkyd) PHOSPHATIDYL ETHANOLAMINE-N-Polyethylene Glycol.Above-mentioned product joins in 5mLDMSO, adds EDC0.2g, NHS0.2g, add folic acid 0.2g, room temperature reaction 24 hours, reactant liquor, through column chromatography, obtains two (paclitaxel-7-dithio diethyl alkyd) PHOSPHATIDYL ETHANOLAMINE-N-Polyethylene Glycol-folic acid 0.035g of product.11.0(2H，s)，8.57(2H，s)，8.0-7.08(40H，m)，6.61(4H，d)，5.59-5.51(4H，m)，5.06-4.32(19H，m)，4.12(2H，t)，4.01(2H，d)，3.79-3.37(？H，broad)，3.15(2H，t)，2.81(2H，d)，2.64(8H，t)，2.18-2.00(28H，m)，1.81-1.71(10H，m)，1.26-1.21(18H，s)。
Embodiment 11:
The synthesis (synthetic route is shown in Figure 11) of two (paclitaxel-7-dithio diethyl alkyd) phosphatidyl glycerol compound
Two (2 '-TBDMS-paclitaxel-7-dithio diethyl alkyd) glyceride 0.1g of intermediate product of embodiment 8 is dissolved in 10mL chloroform, adds phosphorus oxychloride 0.1g, triethylamine 0.3g, and 10 DEG C of reaction 24h, revolve solvent evaporated; Solid is dissolved in 10mL chloroform, adds glycerol acetonide acetone 0.1g, triethylamine 0.2g, and 10 DEG C of reaction 3h, add proper amount of acetic acid in reactant liquor, remove acetone protecting group; Add cold diethyl ether and separate out precipitation, solid is dissolved in 10mLDMF, adds 0.2g tetrabutyl ammonium fluoride room temperature reaction 3h, and gained reactant liquor, by column chromatography purification, obtains two (paclitaxel-7-dithio diethyl alkyd) the phosphatidyl glycerol 0.06g of product.
1HNMR(500MHz，CD3OD∶CDCl
31∶1)：δ8.0-7.08(32H，m)，5.59-5.51(4H，m)，5.06-4.93(6H，s)，4.64(1H，m)，4.43-4.32(4H，m)，4.12-4.01(4H，m)，3.77-3.68(6H，m)，3.39-3.37(3H，m)，2.81-2.64(10H，t)，2.01-2.00(20H，m)，1.81-1.71(10H，m)，1.26-1.21(18H，s)。[M+H]
+m/z，2246.33。
Two (paclitaxel-7-dithio diethyl alkyd) phosphatidyl glycerol is dissolved in pure water, the sodium hydroxide adding 0.01M is dissolved in and regulates pH to be 7, and lyophilizing obtains two (paclitaxel-7-dithio diethyl alkyd) the phosphatidyl glycerol pressed powders containing sodium ion.
Embodiment 12:
The synthesis (route is shown in Figure 12) of two (paclitaxel-2 '-succinic acid) phosphatidylcholine
Paclitaxel (0.85g, 1mmol) is dissolved in 30mL chloroform, and constantly vibration makes it fully dissolve.Add DMAP (0.12g, 1mmol), succinic anhydrides (0.32g, 3mmol), stirring at room temperature reaction 24h.Then, cold diethyl ether precipitates, and adds 2%HCl solution washing, removing DMAP, and column chromatography is separated and obtains white powder paclitaxel-2 '-monomester succinate (eluant v/v, chloroform/methanol: 5/1) 0.56g.
Getting 0.5g paclitaxel monomester succinate (0.5mmol) is dissolved in 15mL pyridine, add 0.4g tert-butyl chloro-silicane (TBDMSCl, 2.6mmol), 3h is reacted under room temperature, add ice ether sedimentation, the white solid of separating out is separated with column chromatography further and obtains paclitaxel 7 end-cap product 7-tertiary butyl dimethyl Si alkane-paclitaxels-2 '-monomester succinate (eluant v/v, chloroform/methanol: 7/1) white solid 0.26g.
Above-mentioned end-cap product 0.22g, CDI0.3g, be dissolved in 10mL chloroform, adds GPC0.15g and DBU0.2g room temperature reaction 24h; Ice ether sedimentation, precipitation be fixedly dissolved in 5mLDMF, add tetrabutyl ammonium fluoride 0.5g, room temperature reaction 6h, reactant liquor by column chromatography purification, obtains product two (paclitaxel-2 '-succinic acid) phosphatidylcholine 0.16g.
1HNMR(500MHz，CD
3OD∶CDCl
31∶1)：δ7.99-7.12(32H，m)，6.10-5.8(4H，dd)，5.51(2H，s)，4.93(4H，s)，4.54-4.21(5H，m)，4.01-3.37(12H，m)，3.30(9H，s)，2.81-2.64(10H，m)，2.01-2.00(20H，m)，1.72-1.71(10H，m)，1.21(12H，s).[M+H]
+m/z，2130.15。
Embodiment 13:
The synthesis (route is shown in Figure 13) of two (paclitaxel-7-succinic acid) phosphatidylcholine
Paclitaxel (0.85g, 1mmol) is dissolved in 30mL chloroform, adds triethylamine 1mL, add 0.4g tert-butyl chloro-silicane (TBDMSCl, 2.6mmol), at-20 DEG C, react 3h, add ice ether, separate out white sedimentation, centrifugalize, further column chromatography is separated and obtains paclitaxel 2 ' position end-cap product 2 '-tertiary butyl dimethyl Si base-paclitaxel (eluant, chloroform/methanol: 7/1, v/v) 0.61g is white powdery solids.
Get end-cap product 0.20g, be dissolved in 30mL chloroform, add DMAP (0.36g, 3mmol), succinic anhydrides (0.32g, 3mmol), stirred at ambient temperature reaction 24h.Then, the above-mentioned reactant liquor of 2%HCl solution washing, removing DMAP, column chromatography is separated (eluant, chloroform/methanol: 5/1, v/v) and obtains white powder 2 '-tertiary butyl dimethyl Si base-paclitaxel-7-monomester succinate; Use tetrabutyl ammonium fluoride process further, remove TBDMS, reactant liquor, by column chromatography purification, obtains Product Taxol-7-monomester succinate 0.15g.
Paclitaxel-7-monomester succinate is dissolved in chloroform, adds CDI0.3g, GPC0.05g and DBU0.2g room temperature reaction 24h, and gained reactant liquor, by column chromatography purification, obtains two (paclitaxel-7-succinic acid) the phosphatidylcholine 0.10g of product.
1HNMR(500MHz，CD3OD∶CDCl31∶1)：δ8.01-7.08(32H，m)，5.59-5.51(4H，m)，5.06-4.93(6H，m)，4.64(1H，m)，4.43-4.32(4H，m)，4.12-3.43(10H，m)，3.37(2H，t)，3.30(9H，s)，2.81-2.64(10H，t)，2.01-2.00(20H，m)，1.81-1.71(10H，m)，1.26-1.21(18H，s)。[M+H]+m/z，2130.25。
3 execute example 14:
The synthesis (synthetic route is shown in Figure 14) of two (Docetaxel-2 '-adipic acid) phosphatidylcholine
Docetaxel (Docetaxel) 0.6g, adds 30ml chloroform and dissolves.Add DMAP (0.12g, 1mmol), adipic anhydride (0.64g, 6mmol), stirred at ambient temperature reaction 24h.Then, the above-mentioned reactant liquor of 2%HCl solution washing, removing DMAP, column chromatography is separated and obtains white powder paclitaxel-2 '-adipate monoester 0.43g (eluant, chloroform/methanol: 5/1, v/v).
0.20g paclitaxel-2 '-adipate monoester is dissolved in 2ml pyridine, add 0.4g tert-butyl chloro-silicane, react 3h under room temperature, reactant liquor is added drop-wise among 100ml ice ether, separates out white sedimentation, centrifugalize, further column chromatography is separated and obtains paclitaxel end-cap product (eluant, chloroform/methanol: 7/1, v/v) 7,10-bis-(tertiary butyl dimethyl Si base)-paclitaxel-2 '-adipate monoester 0.12g is white powdery solids.
Above-mentioned end-cap product 0.12g, add chloroform to dissolve, add CDI0.3g, add GPC0.05g and DBU0.2g room temperature reaction, use tetrabutyl ammonium fluoride process further, remove TBDMS, reactant liquor by column chromatography purification, obtains product two (Docetaxel-2 '-adipic acid) phosphatidylcholine 0.08g.
1HNMR(500MHz，CD3OD∶CDCl
31∶1)：δToxal：8.0-7.12(22H，m)，6.10-5.8(4H，dd)，4.93(4H，s)，4.77(2H，s)，4.64(1H，m)，4.43-3.37(16H，m)，3.30(9H，s)，2.81(2H，d)，2.25(8H，t)，2.01-2.00(16H，m)，1.72-1.68(18H，m)，1.41(18H，s)，1.26-1.21(18H，s)。[M+H]
+m/z，2094.20。
Embodiment 15:
The preparation of two (paclitaxel-7-dithio diethyl alkyd) phosphatidylcholine liposome
Two (paclitaxel-7-dithio diethyl alkyd) the phosphatidylcholine 1mmol obtained by embodiment 1, add chloroform 20ml, 60 DEG C revolve solvent evaporated; Add 20mlPBS (pH=7.4) 60 DEG C of skinnings, 200nm membrane filtration, obtain two (paclitaxel-7-dithio diethyl alkyd) phosphatidylcholine elaioplast nanometer particle solution.The following Figure 15 of granularmetric analysis result, mean diameter 160nm.Transmission electron microscope measures the form of nano-particle as Figure 16.By two (paclitaxel-7-dithio diethyl alkyd) phosphatidylcholine elaioplast nanometer particle solution lyophilizing, obtain powdered nanoparticles granule.
Embodiment 16:
The preparation of two (paclitaxel-7-succinyl-glycyl-phenylalanyl-leucyl-glycyl) phosphatidylcholine liposome
Two (paclitaxel-7-succinyl-glycyl-phenylalanyl-leucyl-glycyl) the phosphatidylcholine 1mmol obtained by embodiment 5, add chloroform 20ml, 60 DEG C revolve solvent evaporated; Add 20mlPBS (pH=7.4) 60 DEG C of skinnings, obtain two (paclitaxel-7-succinyl-glycyl-phenylalanyl-leucyl-glycyl) phosphatidylcholine elaioplast nanometer particle solution.Granularmetric analysis result shows, mean diameter 260nm.
Embodiment 17:
The preparation of two (paclitaxel-7-succinyl-glycyl) phosphatidylcholine liposome
Two (paclitaxel-7-succinyl-glycyl) phosphatidyl choline compounds 1mmol that embodiment 4 obtains, add chloroform 20ml, 60 DEG C revolve solvent evaporated; Add 20mlPBS (pH=7.4) 60 DEG C of skinnings, obtain two (paclitaxel-7-succinyl-glycyl) phosphatidyl choline compounds elaioplast nanometer particle solution.Granularmetric analysis shows, mean diameter 210nm.
Embodiment 18:
The preparation of two (paclitaxel-7-dithio diethyl alkyd) phosphatidylcholine liposome B
Two (paclitaxel-7-dithio diethyl alkyd) phosphatidylcholine 0.5mmol that embodiment 1 obtains, distearoyl phosphatidylcholine DSPC0.5mmol, DSPE-PEG-folic acid (Polyethylene Glycol mean molecule quantity 1500) 0.1mmol, add chloroform 20ml, 60 DEG C revolve solvent evaporated; Add 20mlPBS (pH=7.4) 60 DEG C of skinnings, obtain two (paclitaxel-7-dithio diethyl alkyd) phosphatidylcholine elaioplast nanometer particle solution.Granularmetric analysis shows, mean diameter 560nm.Two (paclitaxel-7-dithio diethyl alkyd) the phosphatidylcholine liposome B nanoparticles solution of lyophilization, obtains Powdered nano-particle.
Embodiment 19:
The preparation of two (paclitaxel-7-succinic acid) phosphatidylcholine liposome
Two (paclitaxel-7-succinic acid) phosphatidylcholine 1mmol that embodiment 13 obtains, add chloroform 10ml, 60 DEG C revolve solvent evaporated; Add 20mlPBS (pH=7.4) 60 DEG C of skinnings, obtain two (paclitaxel-7-succinic acid) phosphatidylcholine elaioplast nanometer particle solution.Granularmetric analysis shows, mean diameter 190nm.Two (paclitaxel-7-succinic acid) the phosphatidylcholine elaioplast nanometer particle solution of lyophilization, obtains Powdered nano-particle.
Embodiment 20:
The preparation of two (paclitaxel-7-succinic acid) phosphatidylcholine liposome
Two (paclitaxel-7-succinic acid) phosphatidylcholine 1mmol that embodiment 13 obtains, add chloroform 20ml, 60 DEG C revolve solvent evaporated; Add 20mlPBS (pH=7.4) 60 DEG C of skinnings, obtain two (paclitaxel-7-succinic acid) phosphatidylcholine elaioplast nanometer particle solution.Granularmetric analysis shows, mean diameter 170nm.
Embodiment 21:
The Degrading experiment of two (paclitaxel-7-succinyl-glycyl-phenylalanyl-leucyl-glycyl (GFLG)) phosphatidyl choline compounds liposome
Two (paclitaxel-7-succinyl-glycyl-phenylalanyl-leucyl-glycyl (GFLG)) phosphatidyl choline compounds elaioplast nanometer particle 10mLPBS solution of 0.1mmol prepared by embodiment 16, two (paclitaxel-7-succinic acid) phosphatidylcholine elaioplast nanometer particle 10mLPBS solution prepared by embodiment 19, be divided into 2 parts respectively, a copy of it adds the PBS buffer 0.5mL (enzyme concentration 50mmol) of papain (papain), another part adds PBS buffer, place and hatch at 37 DEG C in incubator, content (the Agilent1100LC of paclitaxel is detected by high performance liquid chromatography, the anti-phase C18 post of Zorbax, 150 × 4.6mm, 5 μm, sample size 20 μ L, column temperature 25 DEG C, determined wavelength λ=254nm, gradient elution: 2-90% buffer B/A, flow velocity 1.0mL/min, the deionized water of buffer A: 0.1%TFA, the acetonitrile of buffer B: 0.1%TFA).
Result shows, the paclitaxel discharged after the taxol drug that two (paclitaxel-7-succinyl-glycyl-phenylalanyl-leucyl-glycyl (GFLG)) phosphatidyl choline compounds liposome solutions of Papain ferment treatment discharge after 6 hours reaches 35%, 15 hours of total amount reaches 65%; Liposome solutions without ferment treatment does not detect paclitaxel.Two (paclitaxel-7-succinic acid) phosphatidylcholine liposome solutions of Papain ferment treatment do not detect paclitaxel after 6 hours, the paclitaxel discharged after 15 hours only 2%; Liposome solutions without ferment treatment does not detect paclitaxel.Visible, two (paclitaxel-7-succinyl-glycyl-phenylalanyl-leucyl-glycyl (GFLG)) phosphatidyl choline compounds liposome has enzyme fast degradation, discharges the former medicine of paclitaxel fast.
Embodiment 22:
The external degradation test of two (paclitaxel-7-dithio diethyl alkyd) phosphatidylcholine liposome
Two (paclitaxel-7-dithio diethyl alkyd) phosphatidylcholine elaioplast nanometer particle 10mLPBS solution of 0.1mmol prepared by embodiment 15, two (paclitaxel-7-succinic acid) the phosphatidylcholine elaioplast nanometer particle 10mLPBS solution of 0.1mmol prepared by embodiment 20, be divided into 2 parts respectively, a copy of it adds the PBS buffer 0.5mL (GSH concentration 50mmol) of glutathion (GSH), another part adds PBS buffer, place and hatch at 37 DEG C in incubator, content (the Agilent1100LC of paclitaxel is detected by high performance liquid chromatography, the anti-phase C18 post of Zorbax, 150 × 4.6mm, 5 μm, sample size 20 μ L, column temperature 25 DEG C, determined wavelength λ=254nm, gradient elution: 2-90% buffer B/A, flow velocity 1.0mL/min, the deionized water of buffer A: 0.1%TFA, the acetonitrile of buffer B: 0.1%TFA).
Result shows, the taxol drug that two (paclitaxel-7-dithio diethyl alkyd) phosphatidylcholine liposome solutions of GSH process discharge after 3 hours reaches 94% of total amount; Two (paclitaxel-7-dithio diethyl alkyd) phosphatidylcholine liposome solutions without GSH process did not detect taxol drug after 3 hours; Two (paclitaxel-7-succinic acid) phosphatidylcholine liposome solutions of GSH process and do not detect taxol drug after 3 hours without pair (paclitaxel-7-succinic acid) phosphatidylcholine liposome solutions of GSH process.
So two (paclitaxel-7-dithio diethyl alkyd) phosphatidylcholine liposome has GSH sensitivity, the former medicine of release paclitaxel fast can be controlled by GSH.Two (paclitaxel-7-succinic acid) phosphatidylcholine liposome can not be degraded by GSH, is difficult to rapid delivery of pharmaceuticals.
Pharmacological evaluation
Experimental example 23:
Mtt assay human cancer cell fragmentation test
Medicine and reagent: calf serum is Nanjing Sheng Xing Bioisystech Co., Ltd product; DMSO analytical pure; RPMI1640 is GIBCO product.
Instrument: BIORAD680 type microplate reader.Collect well-grown tumor cell MCF-7: human breast cancer cell, be mixed with 8 × 10 by the RPMI1640 culture medium containing 10% calf serum
3/ mL cell suspension, inoculate in 96 well culture plates, every empty 100 μ L (containing 1000 tumor cells), put 37 DEG C, cultivate dosing after 24 hours in 5%CO2 incubator, blank and solvent control are established in experiment, given the test agent establishes 4 concentration, every concentration 3 parallel holes, put 37 DEG C, cultivate 4 days in 5%CO2 incubator.Discard culture fluid, every hole adds MTT solution (0.4mg/mL, RPMI1640 prepare) 100 μ L, hatches 4 hours for 37 DEG C.Abandoning supernatant, every hole adds MTT solution 150 μ L, and dissolved particles, after gentle agitation, measures OD value by microplate reader under determined wavelength 540nm, reference wavelength 450nm.Result calculates: can obtain dose-effect curve with the variable concentrations of medicine and to the suppression ratio mapping of cell, therefrom obtain half-inhibition concentration (IC50).
Result: the anti-tumor activity of the compounds of this invention liposome to human tumor cell line the results are shown in Table 1. and screen from anti tumor activity in vitro, the half-inhibition concentration of two (paclitaxel-7-dithio diethyl alkyd) the phosphatidylcholine liposome of the compounds of this invention, two (paclitaxel-7-succinyl-glycyl) phosphatidylcholine liposome is all less than two (paclitaxel-7-succinic acid) phosphatidylcholine liposome, also be less than paclitaxel, illustrate that its activity is better than paclitaxel and two (paclitaxel-7-succinic acid) phosphatidylcholine liposome.This is because two (paclitaxel-7-dithio diethyl alkyd) phosphatidylcholines containing spacerarm of easily degrading, two (paclitaxel-7-succinyl-glycyl) phosphatidylcholine liposome are easy to by cellular uptake, and discharge former medicine fast.
Table 1. the compounds of this invention liposome is to the anti-tumor activity result of human tumor cell line MCF-7
Experimental example 24:
Taxanes phosphatide cpd is tested at In vivotoxicity
Animal: ICR mice, male, 18-22g, purchased from Nanjing KaiJi Biology Science Development Co., Ltd.
The toxicity in vivo test of two (paclitaxel-7-dithio diethyl alkyd) phosphatidylcholine of the compounds of this invention, two (paclitaxel-7-succinyl-glycyl) phosphatidylcholine the results are shown in Table 2.Result shows, the maximum tolerated dose of the compounds of this invention taxanes phosphatide cpd is all greater than 100mg/kg, and toxicity is much smaller than paclitaxel.
Table 2 taxanes phosphatide cpd is in In vivotoxicity result of the test
Experimental example 25:
Drug effect and toxicity test in two (paclitaxel-7-dithio diethyl alkyd) phosphatidyl choline compounds Via Liposomes
The preparation of nude mice model: collect the MCF-7 cell suspension cultivated, concentration is 1 × 10
7individual/ml, is inoculated in axillary fossa on the right side of nude mouse with every 0.1ml subcutaneous.Grouping and administration: transplanted tumor in nude mice vernier caliper measurement transplanted tumor diameter, tumor growth is to 75mm
3time by animal random packet.Meanwhile, each group nude mice starts administration, and dosage regimen is shown in group and dosage regimen, uses the method measuring tumor footpath, dynamically observes the Graft Versus Tumor of given the test agent.The computing formula of gross tumor volume (TV) is: TV=1/2 × a × b
2(formula 3-2), wherein a, b represent length and width respectively.
Group and dosage regimen: blank group: normal saline, intravenous injection, once, volume is 0.2ml, continuous 3 weeks in injection in every three days.
Matched group: paclitaxel is dissolved in normal saline (micro-DMSO hydrotropy, dosage 5mg/kg), intravenous injection, once, volume is 0.2ml, continuous 3 weeks in injection in every three days.
Medicine group: two (paclitaxel-7-dithio diethyl alkyd) phosphatidyl choline compounds liposome solutions (dosage is equivalent to paclitaxel 5mg/kg) of embodiment 15, intravenous injection, once, volume is 0.2ml, continuous 3 weeks in injection in every three days.Period, measure body weight change.
Anti-tumor activity and body weight change the results are shown in Figure 17.From anti-tumor activity (Figure 17 a), two (paclitaxel-7-dithio diethyl alkyd) the phosphatidylcholine liposome of the present invention has good Tumor suppression growth, and the weight of animals does not decline (Figure 17 b), display avirulence.
Above-described embodiment is only the preferred embodiment of the present invention; be noted that for those skilled in the art; under the premise without departing from the principles of the invention; some improvement and equivalent replacement can also be made; these improve the claims in the present invention and are equal to the technical scheme after replacing, and all fall into protection scope of the present invention.
Claims (8)
1. a taxanes phosphatide cpd, is characterized in that, the pharmaceutically acceptable salt that the compound that this phosphatide cpd is following general formula (1) or the compound of described general formula (1) and counter ion counterionsl gegenions are formed:
In formula (1), R
1and R
2for spacerarm, to be carbon number be 2 ~ 20 the alkylene alkyl containing ehter bond, the carbon number oxygen base alkylene alkyl that is 1 ~ 20, the carbon number alkylene alkyl containing cystine linkage-S-S-that is 2 ~ 20, the carbon number alkylene alkyl containing peptide bond that is 2 ~ 60, the carbon number alkylene alkyl containing ester bond that is 2 ~ 20, carbon number be 2 ~ 20 the alkylene alkyl containing hydrazone key or carbon number be 2 ~ 20 not containing heteroatomic alkylene alkyl/sub-alkylene; L is 2-amino-2-carboxy ethyl, 2-amino-ethyl, 2-trimethyl amido ethyl cation, 2, the end group of 3-dihydroxypropyl, molecular weight to be the Polyethylene Glycol-amino-ethyl of the N-without targeting end group of 200-4000 or molecular weight be 200-4000 is the N-Polyethylene Glycol-amino-ethyl of targeting group;
R
xand R
ypaclitaxel analogs 10 bit substituent of to be structure be the paclitaxel analogs 2 ' bit substituent of following formula (2), paclitaxel analogs 7 bit substituent of formula (3) or formula (4), wherein the configuration of each carbon atom is: 1S, 2S, 3R, 4S, 5R, 7S, 8S, 10R, 13S, 2 ' R, 3 ' S:
In formula (2), (3), (4), R
3hydrogen; R4 is any one in following group: benzoyl, C
1-C
6alkyl-carbonyl, C
1-C
6halogenated alkyl carbonyl, the C of straight or branched
1-C
6three substituted silane bases, the C of straight or branched alkyl
1-C
6straight or branched alkylene carbonyl; R
5any one in following group: acetyl group, C
1-C
6alkyl-carbonyl, C
1-C
6halogenated alkyl carbonyl, the C of straight or branched
1-C
6three substituted silane bases, the C of straight or branched alkyl
1-C
6straight or branched alkylene carbonyl; R
6hydrogen, C
1-C
6alkyl, the C of straight or branched
1-C
6alkyl-carbonyl, C
1-C
6haloalkyl, the C of straight or branched
1-C
6halogenated alkyl carbonyl, the C of straight or branched
1-C
6alkyl/alkyl-carbonyl, the C of hydroxyl of straight or branched
1-C
6three substituted silane bases, the C of straight or branched alkyl
1-C
6straight or branched alkylene carbonyl, C
1-C
6straight or branched alkoxymethylene, phenyoxymethylene or benzene methyl methylene; R7 is hydrogen, acetyl group, C
1-C
6alkyl, the C of straight or branched
1-C
6alkyl-carbonyl, C
1-C
6haloalkyl, the C of straight or branched
1-C
6halogenated alkyl carbonyl, the C of straight or branched
1-C
6three substituted silane bases, the C of straight or branched alkyl
1-C
6straight or branched alkylene carbonyl, C
1-C
6straight or branched alkoxymethylene, phenyoxymethylene or benzene methyl methylene; R
8hydrogen, C
1-C
6alkyl-carbonyl, C
1-C
6halogenated alkyl carbonyl, the C of straight or branched
1-C
6three substituted silane bases, the C of straight or branched alkyl
1-C
6straight or branched alkylene carbonyl, C
1-C
6the alkyl-carbonyl of hydroxyl of straight or branched or two dodecahexaene carbonyls; R
9hydrogen, benzoyl, t-butyloxycarbonyl, tert-butyl carbonyl, C
1-C
18alkyl-carbonyl, the C of straight or branched
1-C
6halogenated alkyl carbonyl, the C of straight or branched
1-C
6straight or branched hydroxyl/alkyl-carbonyl, C
1-C
6straight or branched alkylene carbonyl or two dodecahexaene carbonyls.
2. taxanes phosphatide cpd according to claim 1, is characterized in that, described counter ion counterionsl gegenions are a kind of cationes, or the combination of a kind of cation and a kind of anion.
3. taxanes phosphatide cpd according to claim 1, it is characterized in that, described end group is in the N-Polyethylene Glycol-amino-ethyl of targeting group, and targeting group is any one in folic acid, galactose, polypeptide, antibody, antibody fragment, hyaluronic acid, asialoglycoprotein, polysaccharide, nucleic acid and plan peptide.
4. a pharmaceutical composition, is characterized in that, said composition comprises acceptable carrier on taxanes phosphatide cpd described in claim 1,2 or 3 or described taxanes phosphatide cpd and pharmacodynamics.
5. pharmaceutical composition according to claim 4, is characterized in that, said composition is liquid preparation, solid preparation, semi-solid preparation, capsule, granule, gel, injection, slow releasing preparation or controlled release preparation.
6. pharmaceutical composition according to claim 4, is characterized in that, said composition is the elaioplast nanometer particle of particle diameter 10-1000 nanometer, also comprises auxiliary agent in this pharmaceutical composition.
7. pharmaceutical composition according to claim 6, is characterized in that, described auxiliary agent is phospholipid, cholesterol.
8. described in claim 1,2 or 3, taxanes phosphatide cpd is preparing the application in antitumor drug, it is characterized in that, this application, by described taxanes phosphatide cpd or its pharmaceutically acceptable salt, is prepared into medicament with acceptable carrier on pharmacodynamics.
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