CN104689331A - Gold nanoparticle-anthracene ring pharmaceutical composition and preparation method thereof - Google Patents
Gold nanoparticle-anthracene ring pharmaceutical composition and preparation method thereof Download PDFInfo
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Abstract
The invention relates to design and construction of a novel gold nanoparticle-anthracene ring pharmaceutical composition. The gold nanoparticle-anthracene ring pharmaceutical composition consists of gold nanoparticles serving as the core, dihydrolipoic acid serving as a connecting arm, an anthracene ring drug serving as a carried medicine and a hydrophilic polymer serving as a biocompatible molecule. The half-life period of the anthracene ring drug can be prolonged, the concentration of the drug in unit volume is increased, the drug is sensitive to acidic pH value, and the bioavailability and the drug metabolism kinetic properties of the drug are improved. The invention also discloses a method for preparing each component of the gold nanoparticle-anthracene ring pharmaceutical composition, a method for preparing the composition, in-vitro release characteristics of the drug and a drug delivery application.
Description
Technical field
The invention belongs to biological medicine technology and nanosecond science and technology field, be specifically related to a kind of design and structure of gold nano anthracycline drug combination of novelty.This gold nano anthracycline drug combination take gold nano as kernel, dihydrolipoic acid is linking arm, anthracene nucleus medicament is contained medicine, hydrophilic polymer is that biocompatiblity molecules is formed, it can improve the half-life of anthracene nucleus medicament, increases pharmaceutical units volumetric concentration, to acidic-pH sensitive, improve bioavailability and the pharmacokinetic profile of medicine, can be used for intravenous injection diagnosing tumor and Therapy study.
Background technology
Recent two decades comes, Nano medication transmission system has become a study hotspot of field of pharmacology, it can well solve the problems that some medicines exist in clinical practice, as low in poorly water-soluble, poor stability, bioavailability, Half-life in vivo and action time is short, distribution in vivo is poor and toxic and side effects is large etc.And the application of nano-carrier in drug delivery, provide new thinking to the targeted delivery of people's drugs.
Golden nanometer particle (gold nanoparticles, GNPs, represent with A in the present invention) be the fine particle of elemental gold, diameter is at 1-100nm, a kind of stablizing and biocompatible metal nanoparticle, it has special physical chemistry and optical property, becomes one of focus of biomedical sector pharmaceutical formulations materials application research nearly ten years.Because gold nano has special physics and chemical characteristic, it has certain advantage in transmission and carrying medicament: (one) gold nano kernel is that inert, non-toxic substances has good biocompatibility; (2) Jenner's grain of rice is easy to preparation, and under different conditions, can prepare particle diameter is 1 ~ 150nm Jenner grain of rice; (3) gold nano surface by multiple modified with functional group, and can have different application functions; (4) in view of physics, chemistry and biology character that gold nano is special, it optionally controls transmission and the release of medicine.When using gold nano as carrier, therapeutant release in vivo can pass through internal stimulus (as glutathion or pH), or outside stimulus (as illumination) realizes.(5) gold nano has special light scatter properties, and photoluminescent property and photic thermotherapy character, it has started there is more deep research at diagnosing tumor and orientation treatment.
Anthracene nucleus medicament (Anthracyclines, represent with D in the present invention), comprise amycin, epirubicin, daunorubicin, idarubicin etc. (structure is shown below), it is the chemotherapeutics that a class derives from the steel gray mutation of ripple match streptomycete, the cancer species that they can be treated is all more than the chemotherapeutics of any other type, and use their chemotherapy to be one of the most effective current anti-cancer therapies, the cancer that can be used for treating comprises leukemia, lymphoma, breast carcinoma, uterus carcinoma, ovarian cancer and pulmonary carcinoma etc.But the toxic and side effects of this kind of medicine is larger, the problems such as the half-life is short limit its application, and hydrophilic polymer is generally biocompatibility macromolecule compound (representing with P in the present invention), it can improve the water solublity of medicine, increases the advantages such as medicine circulation time in vivo.
Consider the character of golden nanometer particle (A), anthracene nucleus medicament (D) and hydrophilic polymer (P), anthracene nucleus medicament (D) is introduced by reasonably modifying biocompatibility part (P) is connected to gold nano (A) again surface through S-Au covalent bond, prepare golden nanometer particle-anthracycline drug combination A-D-P, drug delivery system can be made to reach " real " nanometer level (being less than 100nm), thus realize medicine analysis and control more accurately.
Summary of the invention
The invention discloses a kind of novel golden nanometer particle anthracycline drug combination A-D-P, the preparation method of each component part of such conjugate A-D-P, the preparation method of conjugate, drug release in vitro characteristic, and drug delivery applications.Gold nano anthracycline drug combination prepared by the present invention take golden nanometer particle as kernel, dihydrolipoic acid is linking arm, anthracene nucleus medicament is contained medicine, hydrophilic polymer is biocompatiblity molecules, gold nano anthracycline drug combination is formed by S-Au covalent bond and golden nanometer particle surface covalent bond, its size is less than 50nm, anthracene nucleus medicament circulation time in vivo can be improved, increase unit volume drug level, to acidic-pH sensitive, improve bioavailability and the pharmacokinetic profile of medicine.
In the present invention, anthracene nucleus medicament (D) comprises amycin, epirubicin, daunorubicin, idarubicin etc. have modifiable carbonyl and amino anthracene nucleus medicament simultaneously, hydrophilic polymer biocompatibility part (P) comprises the Polyethylene Glycol of different molecular weight, mono methoxy polyethylene glycol, polyvinyl alcohol, polyacrylic acid, the hydrophilic polymer biocompatibility parts such as polyacrylamide.
The present invention relates to a kind of gold nano anthracycline drug combination A-D-P had shown in following structure:
Wherein A is golden nanometer particle, and D is anthracene nucleus medicament mercapto derivatives, and P is hydrophilic polymer biocompatibility part;
It is characterized in that taking golden nanometer particle as kernel, anthracene nucleus medicament mercapto derivatives and hydrophilic polymer biocompatibility part are shell, to be combined with golden nanometer particle kernel by S-Au covalent bond and to form gold nano anthracycline drug combination, its particle diameter is less than 50nm;
Described gold nano anthracycline drug combination, is characterized in that taking golden nanometer particle as kernel;
Described gold nano anthracycline drug combination, is characterized in that anthracene nucleus medicament mercapto derivatives and hydrophilic polymer biocompatibility part are shell;
Described gold nano anthracycline drug combination, is characterized in that anthracene nucleus medicament mercapto derivatives is combined with golden nanometer particle kernel by S-Au covalent bond;
Described anthracene nucleus medicament comprises amycin, epirubicin, daunorubicin, idarubicin etc. have modifiable carbonyl and amino anthracene nucleus medicament simultaneously, hydrophilic polymer biocompatibility part comprises the Polyethylene Glycol of different molecular weight, mono methoxy polyethylene glycol, polyvinyl alcohol, polyacrylic acid, the hydrophilic polymer biocompatibility parts such as polyacrylamide;
Described anthracene nucleus medicament mercapto derivatives, is characterized in that the hydrazone key forming pH sensitivity at the carbonyl of anthracene nucleus medicament and thioctic acid hydrazides, connects sulfydryl functional group;
Described hydrophilic polymer biocompatibility part, is characterized in that possessing modifiable group as hydroxyl, carboxyl etc., and hydrophilic polymer biocompatibility part is combined with anthracene nucleus medicament mercapto derivatives by amide covalent bond;
Described gold nano anthracycline drug combination, is characterized in that its particle size is less than 50nm;
The preparation method of described gold nano anthracycline drug combination, it is characterized in that golden nanometer particle, anthracene nucleus medicament mercapto derivatives hydrophilic polymer biocompatibility part are in aqueous phase or organic facies single_phase system or the diphasic system that dissolves each other, stirring at room temperature is after 1 ~ 5 hour, obtain conjugate solution through deionized water dialysis, then obtain conjugate powder through lyophilization;
Described gold nano anthracycline drug combination is as drug delivery system application and pharmaceutically acceptable carrier or excipient.
The preparation of gold nano anthracycline drug combination A-D-P of the present invention comprises following three steps:
(1) in the present invention, the preparation process of golden nanometer particle (A) kernel is as follows:
Glass apparatus used is cleaned with chloroazotic acid, distilled water, deionized water successively, prepares trisodium citrate aqueous solution, under ice-water bath (0 DEG C) condition, by gold chloride (HAuCl with deionized water
4) mother solution adds in above-mentioned citric acid three sodium solution, with vigorous stirring, by freezing sodium borohydride (NaBH
4) solution adds in mixture solution fast, reaction solution becomes orange to aubergine, keeps reaction temperature to be 0 DEG C, continues vigorous stirring 15 ~ 20min, till color no longer changes, naturally is warming up to room temperature, namely obtains and is of a size of 2 ~ 5nm golden nanometer particle.
(2) the thioctic acid hydrazides-anthracene nucleus medicament-hydrophilic polymer (D-P) in the present invention is the hydrazone key being formed pH sensitivity by the carbonyl of anthracene nucleus medicament and thioctic acid hydrazides, connect sulfydryl functional group, by amide covalent bond, hydrophilic polymer biocompatibility part is combined with anthracene nucleus medicament mercapto derivatives.
(3) preparation process of gold nano anthracycline drug combination A-D-P of the present invention is as follows:
Thioctic acid hydrazides-anthracene nucleus medicament-hydrophilic polymer first becomes free sulfhydryl groups by sodium borohydride reduction, it is in aqueous phase, organic facies single_phase system or the diphasic system that dissolves each other, stirring at room temperature is after 1 ~ 5 hour, obtain conjugate solution through deionized water dialysis, then obtain conjugate powder through lyophilization.
Accompanying drawing explanation
Accompanying drawing 1 is gold nano anthracycline drug combination structural representation of the present invention.
Accompanying drawing 2 is the nucleus magnetic hydrogen spectrum figure of thioctic acid hydrazides-amycin-mono methoxy polyethylene glycol.
Accompanying drawing 3 is the nucleus magnetic hydrogen spectrum figure of gold nano-amycin-mono methoxy polyethylene glycol conjugate.
Accompanying drawing 4 is the infrared absorpting light spectra of gold nano-amycin-mono methoxy polyethylene glycol conjugate.
Accompanying drawing 5 is the uv absorption spectra of golden nanometer particle and gold nano-amycin-mono methoxy polyethylene glycol conjugate.
Accompanying drawing 6 is the transmission electron microscope photo of golden nanometer particle (A) and gold nano-amycin-mono methoxy polyethylene glycol conjugate (B).
Accompanying drawing 7 is the gold nano-amycin-drug release patterns in vitro of mono methoxy polyethylene glycol conjugate under condition of different pH.
Detailed description of the invention
The present invention relates to a kind of novel gold nano anthracycline drug combination A-D-P to design, build, the preparation method of each component part of such conjugate, the preparation method of conjugate, drug release in vitro characteristic, and drug delivery applications.In the present invention, the preparation of gold nano anthracycline drug combination comprises: the preparation of (1) golden nanometer particle kernel; (2) synthesis of anthracene nucleus medicament sulfhydrylation biocompatibility part; (3) preparation of gold nano anthracycline drug combination.
The preparation method of gold nano-anthracene nucleus medicament-hydrophilic polymer conjugates A-D-P take gold nano as kernel, dihydrolipoic acid is linking arm, amycin is contained medicine, and mono methoxy polyethylene glycol is gold nano-amycin-mono methoxy polyethylene glycol conjugate that biocompatiblity molecules participates in preparing is that example is illustrated.
Wherein mono methoxy polyethylene glycol biocompatibility ligand molecular amount is 1000KD, and gold nanometer particle grain size is about 5nm, and reagent is analytical pure and chemical pure, and bag filter molecular cut off is 7,000 (MWCO7,000).
Gold nano-amycin-mono methoxy polyethylene glycol conjugate (be abbreviated as TA-NHN=DOX-mPEG, wherein mono methoxy polyethylene glycol molecular weight is 1000), general structure is:
Wherein the synthetic route of thioctic acid hydrazides-amycin-mono methoxy polyethylene glycol is as follows:
Its synthesis step is: first carry out ethyl esterifiedly obtaining thioctic acid ethyl ester (I) to thioctic acid (TA), thioctic acid ethyl ester (I) carries out hydrazinolysis reaction and forms thioctic acid hydrazides (II), II reacts the hydrazone key compound (III) forming pH sensitivity again with amycin, intermediate (IV) is activated to obtain to the terminal hydroxyl of mono methoxy polyethylene glycol, III and IV reaction forms the target compound of thioctic acid hydrazides-amycin-mono methoxy polyethylene glycol (being abbreviated as TA-NHN=DOX-mPEG), and wherein mono methoxy polyethylene glycol molecular weight is 1000.
Embodiment 1 gold nano-amycin-mono methoxy polyethylene glycol conjugate
1. the synthesis of golden nanometer particle (A):
Glass apparatus used is cleaned with chloroazotic acid, distilled water, deionized water successively, prepares 50mL (0.116mol/L) trisodium citrate aqueous solution, under ice-water bath (0 DEG C) condition, by 50 μ L (2.5 × 10 with deionized water
-6mol/L) gold chloride (HAuCl
4) mother solution adds in above-mentioned citric acid three sodium solution, with vigorous stirring, freezing sodium borohydride solution 5mL (1mg/mL) is added in mixture solution fast, reaction solution becomes orange to aubergine, keep reaction temperature to be 0 DEG C, continue vigorous stirring 15 ~ 20min, till color no longer changes, naturally be warming up to room temperature, namely obtain and be of a size of 5nm solution of gold nanoparticles.
2. the synthesis of thioctic acid hydrazides-amycin-mono methoxy polyethylene glycol:
The synthesis of thioctic acid ethyl ester (I): by thioctic acid (TA) (3.09g, 15mmol), ethanol (EtOH) (10.40g, 225mmol) to dimethylamino naphthyridine (DMAP) (0.92g, 7.5mmol) be dissolved in 120mL dichloromethane, be cooled to 0 DEG C, by N, N '-dicyclohexylcarbodiimide (DCC) (4.63g, 23mmol) be dissolved in 30mL dichloromethane, dropwise add in reaction system, after DCC dropwises, stir 1h and remove ice bath, rise to room temperature (25 DEG C), stirring reaction 24h.Sucking filtration, filtrate concentrates, through column chromatography (petroleum ether: ethyl acetate=15: 1) obtain yellow oil TA-COOEt2.96g, productive rate 84.6%.TLC Rf＝0.4(PE∶EA＝15∶1)；IR(KBr，v)：2493，1740，1448，1377，1183，1024cm
-1；
1H NMR(400MHz，CDCl
3)：δ1.47(t，3H)，1.53(m，2H)，1.66(m，4H)，1.92(m，1H)，2.35(m，2H)，2.49(m，1H)，3.14(m，2H)，3.59(m，1H)，4.13(q，2H)；
13C NMR(400MHz，CDCl
3)：δ24.46，28.54，33.78，34.43，38.32，40.05，56.16，61.52，63.30，69.01，70.15，70.41，72.48，173.25。
The synthesis of thioctic acid hydrazides (II): 2.96g I is dissolved in 50mL absolute methanol, 4.3mL hydrazine hydrate is dropwise added under 40 DEG C of stirrings, reaction stirring is spent the night, this reactant mixture of 50mL diluted ethyl acetate, secondary with saturated sodium-chloride extraction (20ml × 3), organic layer anhydrous magnesium sulfate drying spends the night.Sucking filtration, concentrated filtrate, through column chromatography, (dichloromethane: methanol=15: 1) obtain yellow solid 2.41g, productive rate is 86%, TLC (CH
2cl
2/ CH
3oH=15: 1), Rf=0.4, MS:m/z (%)=221 [M+H]+; IR (KBr, v): 3332,3182,3031,2908,2855,1643,1528,1439,1360,1263,1024,935,670,608,404cm
-1;
1h NMR (400MHz, DMSO-d
6): δ 1.34-1.36 (m, 2H), 1.48-1.56 (m, 3H), 1.61-1.68 (m, 1H), 1.83-1.88 (m, 1H), 2.00 (t, 2H), 2.38-2.43 (m, 1H), 3.08-3.20 (m, 2H), 3.58-3.62 (m, 1H), 4.14 (s, 2H), 8.92 (s, 1H);
13c NMR (400MHz, DMSO-d
6): δ 24.97,28.30,33.23,34.09,38.09,56.12,171.41.
The synthesis of thioctic acid hydrazides-amycin (III): by II (132mg, 0.6mmol), doxorubicin hydrochloride (116mg, 0.2mmol) be dissolved in 30mL absolute methanol, drip 50 μ l trifluoroacetic acids, nitrogen and the lower stirred overnight at room temperature of lucifuge protection, be concentrated into 1 ~ 2ml, add 25mL acetonitrile by reactant liquor, ultrasonic 5min, 3h is left standstill, sucking filtration, filter cake 10ml acetonitrile wash three times at 4 DEG C, filter cake is dry under vacuum, obtain red powder 140mg, productive rate 89%, MS:m/z (%)=746 [M+H]
+, IR (KBr, v): 3412,3244,2935,1670,1589,1413,1263,1112,997,820,581,448cm
-1,
1h NMR (400MHz, DMSO-d
6): δ 1.05-1.22 (m, 5H), 1.23-1.38 (m, 3H), 1.39-1.50 (m, 1H), 1.54-1.61 (m, 1H), 1.70-1.76 (m, 2H), 1.87-1.93 (m, 1H), 2.14-2.18 (m, 2H), 2.25-2.32 (m, 2H), 2.75 (d, 1H), 3.05-3.10 (m, 2H), 3.25-3.29 (m, 1H), 3.62 (m, 1H), 3.96 (s, 3H), 4.04-4.06 (m, 1H), 4.42 (m, 2H), 4.93 (m, 1H), 5, 31 (s, 1H), 5.46-5.54 (m, 2H), 5, 84 (m, 1H), 7.60 (m, 1H), 7.86 (m, 2H), 8.01 (m, 3H), 10.33 (s, 1H), 13.29 (s, 1H), 14.08 (s, 1H),
13c NMR (400MHz, DMSO-d
6): δ 19.55,25.60,28.91,28.96,31.72,34.23,34.67,38.50,47.12,56.43,56.57,57.07,66.44,66.81,72.44,72.95,99.42,111.02,119.44,120.20,120.44,135.11,136.28,136.68,137.02,153.12,154.76,157.01,161.30,174.31,186.87.
The synthesis of mono methoxy polyethylene glycol paranitrophenol formic acid esters (IV): by mono methoxy polyethylene glycol (molecular weight is 1000) (2.50g, 2.5mmol), triethylamine (TEA) (0.61g, 6.0mmol) be dissolved in 20mL dichloromethane, be cooled to 0 DEG C, will to nitroxyl chloride phenyl formate (NPC) (1.25g, 6.3mmol) be dissolved in 10mL dichloromethane, dropwise add in reaction system, after NPC dropwises, stir 1h and remove ice bath, rise to room temperature (25 DEG C), stirring reaction 24h, stop stirring, 40mL dchloromethane reactant liquor, saturated sodium-chloride (20ml × 3) washs, organic layer anhydrous magnesium sulfate drying spends the night.Sucking filtration, concentrated filtrate, through column chromatography (dichloromethane: methanol=30: 1) obtain yellow oil 1.77g, productive rate 61%, R
f=0.6, TLC (CH
2cl
2/ MeOH=15: 1) .IR (KBr, v): 3474,2890,1767,1643,1528,1448,1342,1218,1093,944,846cm
-1;
1h NMR (400MHz, CDCl
3): δ 1.11-1.40 (m, 2H), 3.38 (s, 3H), 3.40-3.98 (m, ~ 133H), 4.44 (t, 2H), 7.40 (d, 2H), 8.28 (d, 2H);
13c NMR (400MHz, CDCl
3): δ 29.67,42.73,59.04,68.31,68.60,70.56,70.63,70.70,71.92,121.81,125.31,145.36,152.47,155.50.
The synthesis of thioctic acid hydrazides-amycin-mono methoxy polyethylene glycol (V): by III (208mg, 0.28mmol), IV (1.14g, 0.98mmol) is dissolved in 20mL dry DMF, add triethylamine (TEA, 136mg, 1.34mmol), nitrogen and the lower stirred overnight at room temperature of lucifuge protection, reactant liquor is concentrated, through column chromatography (dichloromethane: methanol=15: 1) obtain red oil 410mg, productive rate 83%, TLC (CH
2cl
2/ CH
3oH=15: 1), R
f=0.3.IR (KBr, v): 3440,2909,1653,1459,1299,1104,946,848cm
-1,
1h NMR (400MHz, DMSO-d
6): δ: 1.05-1.22 (m, 5H), 1.23-1.45 (m, 4H), 1.48-1.62 (m, 2H), 1.64-1.76 (m, 1H), 1.78-1.95 (m, 1H), 1.96-2.10 (m, 1H), 2.12-2.38 (m, 3H), 2.74-2.80 (m, 1H), 3.03-3.12 (m, 2H), 3.24-3.26 (m, 5H), 3.38-3.62 (m, ~ 119H), 3.65-3.80 (m, 4H), 3.90-4.10 (m, 6H), 4.52-4.76 (m, 2H), 4.80-4.96 (m, 2H), 5.23 (s, 1H), 5, 73 (s, 1H), 6, 83 (d, 1H), 7.61 (s, 1H), 7.84-7.88 (m, 2H), 10.60 (s, 1H), 13.33 (brs, 1H), 14.11 (brs, 1H),
13c NMR (400MHz, DMSO-d
6): δ 13.90, 15.12, 16.90, 22.07, 23.69, 23.76, 28.36, 28.43, 28.68, 28.99, 29.52, 31.27, 31.74, 33.66, 34.17, 37.97, 47.15, 51, 35, 54.84, 55.83, 56.51, 57.31, 58.01, 60.19, 63.09, 64.89, 66.91, 67.92, 68.77, 69.56, 69.76, 71.25, 71.86, 72.32, 72.64, 88.10, 99.87, 110.04, 118.93, 119.68, 119.92, 134.54, 136.17, 136.70, 145.68, 154.21, 154.50, 155.30, 156.07, 160.80, 174.77, 186.30, 186.38.
3. the preparation of gold nano-amycin-mono methoxy polyethylene glycol conjugate (A-D-P)
83mg V (TA-NHN=DOX-mPEG1000) is dissolved in 10ml methanol, be cooled to 0 DEG C, 4.4mg sodium borohydride is dissolved in 4mL deionized water, add in reaction system, 0 DEG C of stirring reaction 1h, reaction system about pH to 7 is adjusted with 1mol/ml HCL, add the gold nano solution reaction 1h of 0.5mL, be 7 with molecular cut off, the bag filter dialysis 24h of 000 also repeatedly changes fresh deionized water, in bag filter, liquid is through lyophilization, obtains gold nano-amycin-mono methoxy polyethylene glycol conjugate darkviolet powder.Redissolve in water, solution is homogeneous without precipitation.Transmission electron microscope observing golden nanometer particle is particle diameter is 4 ~ 7nm, gold nano-amycin-mono methoxy polyethylene glycol conjugate particle diameter increases to 5 ~ 8nm to some extent, and more disperse between particle, through nuclear-magnetism, infrared, ultraviolet characterizes proof mono methoxy polyethylene glycol and amycin is all connected to golden nanometer particle surface, this gold nano-amycin-mono methoxy polyethylene glycol conjugate is sustained drug release in 120h, and show the feature more responsive to acid pH.
The physicochemical properties of embodiment 2 conjugate characterize
Nuclear-magnetism: the conjugate got after a certain amount of amycin part and lyophilizing is dissolved in deuterochloroform, and TMS is primary standard substance, Bruker (Avace) AV-300AV-500 type nmr determination measures hydrogen nuclear magnetic resonance spectrum;
Infrared: the conjugate got after a certain amount of lyophilizing is scattered in KBr, tabletting, Bruker TENSOR type determination of infrared spectroscopy infrared spectrum;
Ultraviolet: (1) gets the solution of gold nanoparticles of the sodium citrate protection of fresh preparation; (2) conjugate after precision takes a certain amount of lyophilizing is dissolved in ultra-pure water wiring solution-forming, is diluted to suitable concn, adopts UV-2401pc uv-vis (Japanese Shimadzu) ultraviolet spectrophotometer scanning 400 to the uv absorption of 800nm;
Transmission electron microscope: get the golden nanometer particle of certain volume and the solution of conjugate particle, drip respectively on the copper mesh with carbon film, room temperature is dried, and adopts JEM-200CX transmission electron microscope (Japanese JEOL Ltd.) to observe the form of particle, size and deployment conditions.
Embodiment 3 drug release experiment
Release in vitro:
Adopt the crack conditions of Bag filter method research gold nano doxorubicin conjugates hydrazone key under different pH solution.Take a certain amount of conjugate (containing amycin 4mg).Be dissolved in the pH7.4 of 3mL respectively, the phosphate buffer (pH7.4,0.2M PBS) of ion concentration 0.2M.Solution solution being divided into three parts of i.e. every part of 1mL is placed in the bag filter of 7KDa, tightens with jag two by bag filter.Bag filter immerses 9mLpH5.5 respectively, in the 0.2M phosphate buffer of 6.5,7.4.Take out solution 200 μ L at different time points, add the release liquid of 200 μ L.The 200 μ L taken out discharge liquid and enter efficient Liquid Detection.
Liquid-phase condition:
Chromatographic condition: HPLC (Agilent1290Infinity series, USA), is separated and adopts reverse chromatograms post to carry out analyzing (ZORBAX Eclipse Plus C18,2.1 × 150mm, 3.5 μm, Agilent Technologies, USA).
Column temperature is 25 DEG C.
Flow velocity is 0.8mL/min, and sampling volume is 20 μ L.
Mobile phase is acetonitrile: water (ammonium acetate containing millesimal formic acid and 10mmol)=40: 60 (v/v).Determined wavelength: excitation wavelength is 480nm, emission wavelength is 550nm.
Claims (8)
1. a gold nano anthracycline drug combination, it is characterized in that taking golden nanometer particle as kernel, anthracene nucleus medicament mercapto derivatives and hydrophilic compound biocompatibility part are shell, to be combined with golden nanometer particle kernel by S-Au covalent bond and to form gold nano anthracycline drug combination, its particle diameter is less than 50nm.
2. gold nano anthracycline drug combination according to claim 1, it is characterized in that the preparation of golden nanometer particle adopts sodium citrate to be the sodium borohydride reduction of stabilizing agent, the golden nanometer particle size of preparation is within the scope of 2 ~ 5 nm.
3. anthracycline drug combination according to claim 1, the derivatization of described anthracene nucleus medicament is selected from has modifiable carbonyl and amino anthracycline compound simultaneously, can be selected from amycin, epirubicin, daunorubicin, idarubicin.
4. anthracene nucleus medicament mercapto derivatives according to claim 1, it is characterized in that the hydrazone key forming pH sensitivity at the carbonyl of anthracene nucleus medicament and thioctic acid hydrazides, obtain the anthracene nucleus medicaments derivative of thioctic acid, the disulfide bond of thioctic acid generates two sulfydryl functional groups under the reduction of sodium borohydride.
5. anthracene nucleus medicament mercapto derivatives according to claim 1, it is characterized in that being combined with the hydrophilic polymer molecules of functionalization at the amino of anthracene nucleus medicament, the functional group of hydrophilic polymer can form covalent bond with amino, as carboxyl, acyl chlorides etc.
6. anthracene nucleus medicament mercapto derivatives according to claim 6, is characterized in that hydrophilic polymer biocompatibility part is selected from the Polyethylene Glycol of different molecular weight, mono methoxy polyethylene glycol, polyvinyl alcohol, polyacrylic acid, polyacrylamide.
7. the preparation method of gold nano anthracycline drug combination according to claim 1, it is characterized in that golden nanometer particle, anthracene nucleus medicament mercapto derivatives, hydrophilic polymer biocompatibility part are in aqueous phase or organic facies single_phase system or the diphasic system that dissolves each other, stirring at room temperature is after 1 ~ 5 hour, obtain conjugate solution through deionized water dialysis, then obtain conjugate powder through lyophilization.
8. gold nano anthracycline drug combination according to claim 1, is characterized in that also containing pharmaceutically acceptable carrier or excipient.
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| CN110256402A (en) * | 2019-07-02 | 2019-09-20 | 江苏第二师范学院（江苏省教育科学研究院） | A kind of lipoic acid hydroxylamine derivatives and its preparation method and application |
| CN113004536A (en) * | 2021-03-19 | 2021-06-22 | 中国药科大学 | Metal-amino acid/peptide coordination polymer and application thereof |
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| CN102727905A (en) * | 2011-04-14 | 2012-10-17 | 中国药科大学 | Gold nano-paclitaxel conjugate, and preparation method and application thereof |
| CN103230596A (en) * | 2013-04-12 | 2013-08-07 | 山东大学 | Gold nanometer medicament carrier with synergistic antineoplastic effect |
| CN103768080A (en) * | 2013-12-30 | 2014-05-07 | 浙江工业大学 | Targeting preparation for resisting drug-resistant tumor, as well as preparation method and application thereof |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102727905A (en) * | 2011-04-14 | 2012-10-17 | 中国药科大学 | Gold nano-paclitaxel conjugate, and preparation method and application thereof |
| CN103230596A (en) * | 2013-04-12 | 2013-08-07 | 山东大学 | Gold nanometer medicament carrier with synergistic antineoplastic effect |
| CN103768080A (en) * | 2013-12-30 | 2014-05-07 | 浙江工业大学 | Targeting preparation for resisting drug-resistant tumor, as well as preparation method and application thereof |
Non-Patent Citations (4)
| Title |
|---|
| M.PRABAHARAN,ET AL: "Gold nanoparticles with a monolayer of doxorubicin-conjugated amphiphilic block copolymer for tumor-targeted drug delivery", 《BIOMATERIALS》 * |
| SANTOSH ARYAL,ET AL: "Doxorubicin conjugated gold nanoparticles as water-soluble and pH-responsive anticancer drug nanocarriers", 《JOURNAL OF MATERIALS CHEMISTRY》 * |
| 王峰: "响应性金纳米药物载体系统用于克服肿瘤细胞耐药性的研究", 《 中国博士学位论文全文数据库 医药卫生科技辑》 * |
| 陆晨熠等: "纳米金的药用研究进展", 《中国药学杂志》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110256402A (en) * | 2019-07-02 | 2019-09-20 | 江苏第二师范学院（江苏省教育科学研究院） | A kind of lipoic acid hydroxylamine derivatives and its preparation method and application |
| CN110256402B (en) * | 2019-07-02 | 2021-10-22 | 江苏第二师范学院（江苏省教育科学研究院） | Lipoic acid hydroxylamine derivatives and preparation method and application thereof |
| CN113004536A (en) * | 2021-03-19 | 2021-06-22 | 中国药科大学 | Metal-amino acid/peptide coordination polymer and application thereof |
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