CN101935365A - Method for synthesizing brain targeting head modification cyclodextrin (CD) derivative - Google Patents
Method for synthesizing brain targeting head modification cyclodextrin (CD) derivative Download PDFInfo
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Abstract
The invention relates to a method for synthesizing a brain targeting head modification cyclodextrin (CD) derivative. The method comprises the following steps of: preparing an intermediate, namely, a mono-6-p-methylbenzene sulfonic acid-beta-cyclodextrin ester from beta-CD by an alkali aqueous solution method and introducing active amino into the reaction of the mono-6-p-methylbenzene sulfonic acid-beta-cyclodextrin ester to obtain amino-modified beta-cyclodextrin; and connecting the amino-modified beta-cyclodextrin with a mercapto brain targeting head through iso-functional group disubstituted polyethylene glycol derivative N-hydroxysuccinimide polyethylene glycol-maleimide maleinimide to prepare the brain targeting head modification cyclodextrin derivative. The method has the advantages that: the method for preparing the brain targeting head modification cyclodextrin derivative is simple and convenient, a reaction condition is mild and the derivative is taken as a medicament carrier and is easy for industrial production; a polyethylene glycol (PEG) long-chain structure is contained, so that long circulation effect is achieved and a carrier is prevented from being phagocytosed by a netlike endothelial system; and the structure is connected with a brain targeting head Tf or Lf, so that the brain transport rate of the structure serving as a medicament conveying carrier is increased.
Description
[technical field]
The present invention relates to the biological medicine technology field, specifically, is the synthetic method of a kind of brain target base head modification property cyclodextrin derivative.
[background technology]
Hemato encephalic barrier (blood-brain barrier, BBB) mainly be the structure that the projection by capillary endothelial cell, basement membrane and neurogliocyte constitutes jointly with defense function, useful nutritive substance of brain and meta-bolites can freely be passed through, and prevent that extraneous objectionable impurities from entering brain.Yet various medicines especially protein and peptide medicament are difficult to break through BBB and advance people's central nervous system and bring into play its therapeutic action.In the past by the cranium drug transport, or the method for utilizing chemical process that soluble small molecular fatization etc. is attempted to increase medicine brain transport efficacy will be abandoned because of all deficiencies.
In recent years, the research of brain target administration has obtained bigger progress, has realized that medicine effectively by BBB, reaches the purpose of treatment central nervous system disease.Its method mainly is by by the endogenous vehicle that is on the hemato encephalic barrier, by changing methods such as medicines structure or design pharmaceutical carrier, makes it pass hemato encephalic barrier by the endogenous movement system.Therefore, the brain targeting drug delivery system is just causing increasing concern, becomes one of field, forward position of targeting drug delivery system research.Discover TfR (TfR), lactoferrin receptor (LfR) has high expression level at brain cell, brain capillary endothelial cell, connect Transferrins,iron complexes (Tf) or lactoferrin (Lf) target molecule at medicine itself or drug-carrying nanometer particle, make it optionally pass hemato encephalic barrier and advance human brain tissue, can effectively promote drug-carrying nanometer particle to break through BBB, make curative drug be transported to performance drug effect in the brain specifically.Tf and Lf are ionic bond strand glycoprotein, belong to Transferrins,iron complexes family.The Tf endogenous material that exists in the blood can be discerned by the corresponding acceptor of expressing on the capillary endothelial cell chamber face in the brain.Tf or TfR monoclonal antibody (TfR-MAb) or TfR-Mab fragment (fab ') can mediate active transport by TfR.TfR has high expression level at brain cell, brain capillary endothelial cell, utilizes above-mentioned transporting pathway that drug-carrying nanometer particle is connected and goes up the Tf part, can effectively promote drug-carrying nanometer particle to break through BBB, makes curative drug be transported to performance drug effect in the brain specifically.Moreover, because the concentration of endogenous Lf is about 5nmol/L, be lower than dissociation constant (Kd) value of high binding site, the drug-loading system that can modify exogenous Lf because of occupying LfR does not in a large number form competitive inhibition, be Lf, LfR monoclonal antibody (LfR-MAb) or LfR-Mab fragment (fab ') are as one of advantage of brain target part.
Tf or Lf target molecule directly are connected on the drug molecule by covalent linkage, change the structural performance of medicine own easily, influence the medicine pharmacological action.Make up novel nano-carrier and aspect drug conveying, have many superiority: (1) slow releasing pharmaceutical, prolong drug action time; (2) guaranteeing to reduce dosage under the pharmaceutically-active prerequisite, alleviating or avoid toxic side effects; (3) improve stability of drug, help storing; (4) it is carried out structural modification, can reach the purpose of targeted.The nano-carrier that present stage adopts mainly concentrates on liposome, nanoparticle, and micellar research, the preparation of these nano-carriers is repeatable poor, and the aftertreatment complexity is unfavorable for suitability for industrialized production.
Cyclodextrin is a kind of good inclusion material, and its cylinder interior has-CH-glucoside bonded O atom, so be hydrophobicity; And 2 of glucose and 3-the OH base is positioned at an end opening place of cylinder, 6-the OH base is then at the other end opening part of cylinder, so its cylindrical outer presents wetting ability.Such hollow tube-shape structure makes a lot of molecules to be wrapped up by cyclodextrin molecular, forms supramolecular structure.After utilizing cyclodextrin that medicine is made inclusion compound, liquid drug is solidified, improve stability of drug, increase the solubleness of medicine, improve bioavailability of medicament.Yet, do not see as yet the relevant report of brain targeting molecular modification in the cyclodextrin carrier.The present invention, makes up novel controllability nanometer medicine-carried system, thereby is applied to field of medicaments as special delivery vehicles further combined with the target molecule by cyclodextrin is carried out structural modification.
[summary of the invention]
The objective of the invention is to overcome the deficiencies in the prior art, a kind of brain target base head modification property cyclodextrin derivative and synthetic method thereof are provided.
Of the present invention being contemplated that: adopt the alkaline aqueous solution method that β-CD is carried out the preparation of intermediate list 6-to Methyl benzenesulfonyl-beta-cyclodextrin ester, continue by single 6-is introduced active amino, obtains amido modified property-beta-cyclodextrin Methyl benzenesulfonyl-beta-cyclodextrin ester reaction; By the two PEG derivative NHS-PEG-MAL that replace of different functional group amido modified property-beta-cyclodextrin is connected with sulfhydrylation brain target base head again, makes brain target base head modification property cyclodextrin derivative.Advantage of the present invention: it is easy that preparation brain target base head is modified property cyclodextrin derivative method, and the reaction conditions gentleness is prepared into useful as drug carrier behind the cyclodextrin inclusion compound, is easy to suitability for industrialized production; Its structure has long circulation effect, effectively avoids carrier by reticuloendothelial system phagocytic; Structure is connected with the brain targeting group, effectively improves its brain transhipment rate as drug conveying carrier.
The objective of the invention is to be achieved through the following technical solutions:
A kind of brain target base head modification property cyclodextrin derivative, its structural formula is:
The synthetic method of a kind of brain target base head modification property cyclodextrin derivative, its concrete steps are: be raw material with the beta-cyclodextrin, adopt the alkaline aqueous solution method that β-CD is carried out the preparation of intermediate list 6-to Methyl benzenesulfonyl-beta-cyclodextrin ester earlier, continue by single 6-is introduced active amino, obtains amido modified property-beta-cyclodextrin Methyl benzenesulfonyl-beta-cyclodextrin ester reaction; By the two polyethyleneglycol derivatives that replace of different functional group amido modified property-beta-cyclodextrin is connected with sulfhydrylation brain target base head, makes brain target base head modification property cyclodextrin derivative; Wherein, Transferrins,iron complexes, lactoferrin, the monoclonal antibody of its corresponding acceptor and fragment thereof, below general designation brain target base head;
Detailed processing step is:
(1) beta-cyclodextrin is dispersed in the water, under 10～20 ℃, adds the sodium hydroxide solution of excessive 8.25mol/L, solution becomes must be clarified in the dropping process; Reaction continues to reduce after 1 hour temperature of reaction to 0～6 ℃, adds the Tosyl chloride acetonitrile solution, produces white precipitate; Under 4 ℃, stirring reaction 2h under the reaction solution filters, washing, and it is 8～9 that 10%HCl transfers to pH with filtrate, separates out white solid, places down at 4 ℃ and spends the night.Filter, collect precipitation twice, recrystallization 3 times in 50 ℃ of vacuum-dryings 6 hours, gets 6-tolysulfonyl chloro-beta-cyclodextrin;
Described cyclodextrin aqueous solution volume is directly proportional with the sodium hydroxide solution volume of dropping, is controlled at 30: 1～20: 1; And Tosyl chloride adds molar weight and is controlled at 0.5～1 times of excessive cyclodextrin all the time;
(2) 6-tolysulfonyl chloro-beta-cyclodextrin is dissolved in the anhydrous ethylenediamine, mixes back reaction 4～5h under 80 ℃, reaction finishes, and is cooled to room temperature, and reaction solution is dispersed in acetone soln, and precipitation is separated out product, and vacuum filtration gets crude product; Crude product is dissolved in the hot water, separates out with acetone precipitation, high speed centrifugation takes off layer precipitation, this process 5～6 times repeatedly, and 50 ℃ of vacuum-drying 7h get single 6-quadrol-beta-cyclodextrin (EDA-CD);
(3) it is 8 o'clock at the pH of borate buffer solution, to mix under brain target base head and 2-imino-sulfane hydrochloride (Traut ' s reagent) nitrogen protection, vibration, 4 ℃, reacted 1～1.5 hour, after reaction finished, solution washed with ultrafiltration membrance filter, remove unnecessary 2-imino-sulfane hydrochloride, obtain sulfhydrylation brain target base head;
The add-on mol ratio of the add-on of described 2-imino-sulfane hydrochloride and brain target base head is 35: 1～45: 1, is preferably 40: 1;
Described brain target base head comprises Tf, TfR monoclonal antibody (TfR-MAb), the TfR-Mab fragment (fab '), Lf, LfR monoclonal antibody (LfR-MAb) and LfR-Mab fragment (fab ');
(4) it is 7 o'clock at the pH of phosphoric acid buffer, the two polyethyleneglycol derivative N-hydroxy-succinamide-polyoxyethylene glycol-maleimide maleimides that replace of single 6-quadrol-beta-cyclodextrin and different functional group are mixed, jolting, normal-temperature reaction 15～30 minutes obtains maleimide maleimide-polyethyleneglycol modified property cyclodextrin derivative (CD-PEG-MAL);
Described single 6-quadrol-beta-cyclodextrin, the molar ratio scope of N-hydroxy-succinamide-polyoxyethylene glycol-maleimide maleimide and sulfhydrylation brain target base head is 900: 160: 1～20: 10: 1;
The two polyethyleneglycol derivative NHS-PEG-MAL that replace of described different functional group, its PEG long-chain comprises PEG3400, PEG5000;
(5) it is 7～8 o'clock at the pH of phosphoric acid buffer; mix under sulfhydrylation brain target base head and the maleimide maleimide-polyethyleneglycol modified property cyclodextrin derivative nitrogen protection; vibration; 4 ℃ were reacted 18～24 hours; after reaction finishes; the solution ultrafiltration membrance filter is removed unnecessary small-molecule substance, obtains brain target base head modification property cyclodextrin derivative.
Compared with prior art, positively effect of the present invention is:
The present invention prepares brain target base head, and to modify property cyclodextrin derivative method easy, and the reaction conditions gentleness is prepared into useful as drug carrier behind the cyclodextrin derivative, is easy to suitability for industrialized production; It contains the PEG backbone and has long circulation effect, effectively avoids carrier by reticuloendothelial system phagocytic; Its structure connects brain targeting base head Tf (comprise its TfR monoclonal antibody (MAb) and TfR-Mab fragment (fab ') or Lf (comprise LfR monoclonal antibody (MAb) or LfR-Mab fragment (fab ')), effectively improve its brain transhipment rate as drug conveying carrier.
[description of drawings]
Three groups of samples of Fig. 1 (CD-PEG5000-MAL, the mixture of CD-PEG5000-Tf and CD-PEG5000-MAL and free Tf) are at the UV at 200-600nm place scanning spectrogram;
The FTIR scanning spectrogram of Fig. 2 EDA-CD and CD-PEG5000-MAL;
The NMR spectrogram of Fig. 3 A NHS-PEG5000-MAL;
The NMR spectrogram of Fig. 3 B CD-PEG5000-Lf.
[embodiment]
The embodiment of a kind of brain target of the present invention base head modification property cyclodextrin derivative and synthetic method thereof below is provided.
Embodiment 1
The 60g beta-cyclodextrin is dispersed in the water, under 20 ℃, adds the 8.25mol/L sodium hydroxide solution of 20ml, 6min drips off, and solution becomes must be clarified in the process.Reaction continues to reduce after 1 hour temperature of reaction to 2 ℃, adds 15g Tosyl chloride acetonitrile solution, produces white precipitate.Stirring reaction 2h under the reaction solution normal temperature filters, washing, and 10%HCl transfers to pH 8 with filtrate, separates out white solid, places down at 4 ℃ and spends the night.Filter, collect precipitation twice, recrystallization 3 times in 50 ℃ of vacuum-dryings 6 hours, gets 6-tolysulfonyl chloro-beta-cyclodextrin, yield 10%.
6.5g 6-tolysulfonyl chloro-beta-cyclodextrin is dissolved in the 30ml anhydrous ethylenediamine, mixes the back and react 4h down in 80 ℃ of following nitrogen protections, reaction finishes; be cooled to room temperature; reaction solution is dispersed in a large amount of acetone solns, and precipitation is separated out product, and vacuum filtration gets crude product.Crude product is dissolved in an amount of hot water, separates out with a large amount of acetone precipitations, high speed centrifugation takes off layer precipitation, this process 5 times repeatedly, and 50 ℃ of vacuum-drying 7h get single 6-quadrol-beta-cyclodextrin (EDA-CD), yield 70%.
Under borate buffer solution (pH 8); the 2-imino-sulfane hydrochloride of 12.5nmol Transferrins,iron complexes and 0.5 μ mol (Traut ' s reagent) nitrogen protection is mixed down; vibration, was reacted 1 hour by 4 ℃; after reaction finishes; solution washs with ultrafiltration membrance filter, removes unnecessary 2-imino-sulfane hydrochloride, obtains the sulfhydrylation Transferrins,iron complexes; sulfydryl and Transferrins,iron complexes mol ratio are 0.8: 1, Transferrins,iron complexes yield 70%.
Under phosphoric acid buffer (pH 7), the single 6-quadrol-beta-cyclodextrin of 4mg and the two PEG of replacement of the different functional group of 10mg derivative N-hydroxy-succinamide-polyoxyethylene glycol 5000-maleimide maleimide (NHS-PEG5000-MAL) are mixed, jolting, normal-temperature reaction 30 minutes obtains maleimide maleimide-polyoxyethylene glycol 5000 modification property cyclodextrin derivative (CD-PEG5000-MAL).
At last; under phosphoric acid buffer (pH 7); above-mentioned sulfhydrylation Transferrins,iron complexes mixes vibration, 4 ℃ with maleimide maleimide-polyoxyethylene glycol 5000 modification property cyclodextrin derivative under nitrogen protection; reacted 24 hours; after reaction finished, the solution ultrafiltration membrance filter was removed unnecessary small-molecule substance; obtain Transferrins,iron complexes modification property cyclodextrin derivative (CD-PEG5000-Tf), Transferrins,iron complexes yield 50% (comparing) with the Transferrins,iron complexes input amount.
Embodiment 2
The 60g beta-cyclodextrin is dispersed in the water, under 20 ℃, adds the 8.25mol/L sodium hydroxide solution of 20ml, 6min drips off, and solution becomes must be clarified in the process.Reaction continues to reduce after 1 hour temperature of reaction to 2 ℃, adds 20g Tosyl chloride acetonitrile solution, produces white precipitate.Stirring reaction 2h under the reaction solution normal temperature filters, washing, and 10%HCl transfers to pH 8 with filtrate, separates out white solid, places down at 4 ℃ and spends the night.Filter, collect precipitation twice, recrystallization 3 times in 50 ℃ of vacuum-dryings 6 hours, gets 6-tolysulfonyl chloro-beta-cyclodextrin, and there is a small amount of 2-tolysulfonyl chloro-beta-cyclodextrin by product in yield 7.8%.
6.5g 6-tolysulfonyl chloro-beta-cyclodextrin is dissolved in the 30ml anhydrous ethylenediamine, mixes the back and react 4h down in 80 ℃ of following nitrogen protections, reaction finishes; be cooled to room temperature, reaction solution is dispersed in a large amount of acetone solns, high speed centrifugation; take off layer precipitation, vacuum filtration gets crude product.Crude product is dissolved in an amount of hot water, separates out with a large amount of acetone precipitations, high speed centrifugation takes off layer precipitation, this process 5 times repeatedly, and 50 ℃ of vacuum-drying 7h get single 6-quadrol-beta-cyclodextrin (EDA-CD), yield 80%.
Under borate buffer solution (pH 8); the 2-imino-sulfane hydrochloride of 125nmol Transferrins,iron complexes and 5 μ mol (Traut ' s reagent) nitrogen protection is mixed down; vibration, was reacted 1.5 hours by 4 ℃; after reaction finishes; solution washs with ultrafiltration membrance filter, removes unnecessary 2-imino-sulfane hydrochloride, obtains the sulfhydrylation Transferrins,iron complexes; sulfydryl and Transferrins,iron complexes mol ratio are 1.22: 1, Transferrins,iron complexes yield 90%.
Under phosphoric acid buffer (pH 7), the single 6-quadrol-beta-cyclodextrin of 3mg and the two PEG of replacement of the different functional group of 6mg derivative N-hydroxy-succinamide-polyoxyethylene glycol 5000-maleimide maleimide (NHS-PEG5000-MAL) are mixed, jolting, normal-temperature reaction 30 minutes obtains maleimide maleimide-polyoxyethylene glycol 5000 modification property cyclodextrin derivative (CD-PEG5000-MAL).
At last; under phosphoric acid buffer (pH 7); above-mentioned sulfhydrylation Transferrins,iron complexes mixes vibration, 4 ℃ with maleimide maleimide-polyoxyethylene glycol 5000 modification property cyclodextrin derivative under nitrogen protection; reacted 24 hours; after reaction finished, the solution ultrafiltration membrance filter was removed unnecessary small-molecule substance; obtain Transferrins,iron complexes modification property cyclodextrin derivative (CD-PEG5000-Tf), Transferrins,iron complexes yield 80% (comparing) with the Transferrins,iron complexes input amount.
Embodiment 3
The 60g beta-cyclodextrin is dispersed in the water, under 20 ℃, adds the 8.25mol/L sodium hydroxide solution of 20ml, 6min drips off, and solution becomes must be clarified in the process.Reaction continues to reduce after 1 hour temperature of reaction to 2 ℃, adds 15g Tosyl chloride acetonitrile solution, produces white precipitate.Stirring reaction 2h under the reaction solution normal temperature filters, washing, and 10%HCl transfers to pH 8 with filtrate, separates out white solid, places down at 4 ℃ and spends the night.Filter, collect precipitation twice, recrystallization 3 times in 50 ℃ of vacuum-dryings 6 hours, gets 6-tolysulfonyl chloro-beta-cyclodextrin, yield 10%.
6.5g 6-tolysulfonyl chloro-beta-cyclodextrin is dissolved in the 30ml anhydrous ethylenediamine, mixes the back and react 4h down in 80 ℃ of following nitrogen protections, reaction finishes; be cooled to room temperature, reaction solution is dispersed in a large amount of acetone solns, high speed centrifugation; take off layer precipitation, vacuum filtration gets crude product.Crude product is dissolved in an amount of hot water, separates out with a large amount of acetone precipitations, high speed centrifugation takes off layer precipitation, this process 5 times repeatedly, and 50 ℃ of vacuum-drying 7h get single 6-quadrol-beta-cyclodextrin (EDA-CD), yield 80%.
Under borate buffer solution (pH 8); the 2-imino-sulfane hydrochloride of 250nmol lactoferrin and 10 μ mol (Traut ' s reagent) nitrogen protection is mixed down; vibration, was reacted 1.5 hours by 4 ℃; after reaction finishes; solution washs with ultrafiltration membrance filter, removes unnecessary 2-imino-sulfane hydrochloride, obtains the sulfhydrylation lactoferrin; sulfydryl and lactoferrin mol ratio are 1.22: 1, lactoferrin yield 96%.
Under phosphoric acid buffer (pH 7), the single 6-quadrol-beta-cyclodextrin of 3mg and the two PEG of replacement of the different functional group of 6mg derivative N-hydroxy-succinamide-polyoxyethylene glycol 5000-maleimide maleimide (NHS-PEG5000-MAL) are mixed, jolting, normal-temperature reaction 30 minutes obtains maleimide maleimide-polyoxyethylene glycol 5000 modification property cyclodextrin derivative (CD-PEG5000-MAL).
At last; under phosphoric acid buffer (pH 7); above-mentioned sulfhydrylation lactoferrin mixes vibration, 4 ℃ with maleimide maleimide-polyoxyethylene glycol 5000 modification property cyclodextrin derivative under nitrogen protection; reacted 24 hours; after reaction finished, the solution ultrafiltration membrance filter was removed unnecessary small-molecule substance; obtain lactoferrin modification property cyclodextrin derivative (CD-PEG5000-Lf), lactoferrin yield 90% (comparing) with the lactoferrin input amount.
The present invention relates to and having synthesized is the brain target base head modification property cyclodextrin derivative of substrate with the beta-cyclodextrin, its synthesis step is simple, mild condition, repeatable high, its composite structure is as pharmaceutical carrier, effectively avoid being improved drug solubility and stability, have the rate of brain transhipment efficiently by the engulfing of reticuloendothelial cell in the body.
The structure of CD-PEG5000-Tf is identified:
(1) with maleimide maleimide-polyoxyethylene glycol 5000 modification property cyclodextrin derivative (CD-PEG5000-MAL), Transferrins,iron complexes modification property cyclodextrin derivative (CD-PEG5000-Tf) and maleimide maleimide-polyoxyethylene glycol 5000 modification property cyclodextrin derivative and Transferrins,iron complexes physical mixtures (CD-PEG5000-MAL+free Tf) scan respectively at ultraviolet-visible spectrophotometer 200-600nm.
As shown in Figure 1, CD-PEG5000-MAL the protein specificity absorption peak occurs at 280nm after adding free Tf, CD-PEG5000-Tf obviously reduces at the 280nm peak, and the uv-spectrogram of CD-PEG5000-MAL and CD-PEG5000-Tf has obvious difference, tentatively thinks synthetic CD-PEG5000-Tf.
(2) with CD-PEG5000-MAL and EDA-CD through the KBr compressing tablet, in the enterprising line scanning of Fourier transformation infrared spectrometer.
As shown in Figure 2, with the contrast of EDA-CD infrared spectra, CD-PEG5000-MAL is at 1670cm
-1(v
C=O) stronger acid amides characteristic absorbance cutting edge of a knife or a sword of appearance, 760cm simultaneously
-1, 710cm
-1(γ
-NH2) the primary amine charateristic avsorption band disappear, the NHS group that prove NHS-PEG5000-MAL success forms amido linkage with amino on the EDA-CD, synthesizes CD-PEG5000-MAL.
(3) will about 7mg CD-PEG5000-Lf and corresponding amount NHS-PEG5000-MAL freeze-drying after be dissolved in D
2O carries out spectral analysis of the nuclear magnetic resonance.
As Fig. 3 A, shown in the B, about 4.7ppm D
2The O solvent peak.There is NHS group feature hydrogen peak in NHS-PEG5000-MAL at the 2.8ppm place, have the feature overlapping peaks of PEG methylene radical repeating unit about 3.7ppm, has MAL group charateristic avsorption band (Fig. 3 A) at the 6.7ppm place.Also there is the charateristic avsorption band of methylene radical repeating unit among the PEG in CD-PEG-Lf about 3.7ppm, and with 1.30ppm, 3.63～3.90ppm, 5.07ppm the overlapping peaks at each amino acid hydrogen peak of the charateristic avsorption band of EDA-CD and the Lf of 2.5～3.5ppm, NHS (2.8ppm) and MAL (6.7ppm) characteristic peak disappearance (Fig. 3 B) simultaneously, illustrate NHS group and MAL group respectively with the mercapto groups effect of amino and the Lf of EDA-CD, successfully synthesize target product CD-PEG-Lf.
The above only is a preferred implementation of the present invention; should be pointed out that for those skilled in the art, without departing from the inventive concept of the premise; can also make some improvements and modifications, these improvements and modifications also should be considered within the scope of protection of the present invention.
Claims (9)
1. a brain target base head is modified the property cyclodextrin derivative, it is characterized in that its structural formula is:
2. a brain target base head is modified the synthetic method of property cyclodextrin derivative, it is characterized in that, concrete steps are: be raw material with the beta-cyclodextrin, adopt the alkaline aqueous solution method that β-CD is carried out the preparation of intermediate list 6-to Methyl benzenesulfonyl-beta-cyclodextrin ester earlier, continue by single 6-is introduced active amino, obtains amido modified property-beta-cyclodextrin Methyl benzenesulfonyl-beta-cyclodextrin ester reaction; By the two polyethyleneglycol derivatives that replace of different functional group amido modified property-beta-cyclodextrin is connected with sulfhydrylation brain target base head, makes brain target base head modification property cyclodextrin derivative.
3. the synthetic method of a kind of brain target base head modification property cyclodextrin derivative as claimed in claim 2 is characterized in that its concrete steps are:
(1) beta-cyclodextrin is dispersed in the water, under 10～20 ℃, adds the sodium hydroxide solution of excessive 8.25mol/L, solution becomes must be clarified in the dropping process; Reaction continues to reduce after 1 hour temperature of reaction to 0～6 ℃, adds the Tosyl chloride acetonitrile solution, produces white precipitate; Under 4 ℃, stirring reaction 2h under the reaction solution filters, washing, and it is 8～9 that 10%HCl transfers to pH with filtrate, separates out white solid, places down at 4 ℃ and spends the night; Filter, collect precipitation twice, recrystallization 3 times in 50 ℃ of vacuum-dryings 6 hours, gets 6-tolysulfonyl chloro-beta-cyclodextrin;
(2) 6-tolysulfonyl chloro-beta-cyclodextrin is dissolved in the anhydrous ethylenediamine, mixes back reaction 4～5h under 80 ℃, reaction finishes, and is cooled to room temperature, and reaction solution is dispersed in acetone soln, and precipitation is separated out product, and vacuum filtration gets crude product; Crude product is dissolved in the hot water, separates out with acetone precipitation, high speed centrifugation takes off layer precipitation, this process 5～6 times repeatedly, and 50 ℃ of vacuum-drying 7h get single 6-quadrol-beta-cyclodextrin;
(3) be 8 o'clock at the pH of borate buffer solution, brain target base head and the nitrogen protection of 2-imino-sulfane hydrochloride mixed vibration down, 4 ℃, reacted 1～1.5 hour, after reaction finishes, solution washs with ultrafiltration membrance filter, removes unnecessary 2-imino-sulfane hydrochloride, obtains sulfhydrylation brain target base head;
(4) it is 7 o'clock at the pH of phosphoric acid buffer, the two polyethyleneglycol derivative N-hydroxy-succinamide-polyoxyethylene glycol-maleimide maleimides that replace of single 6-quadrol-beta-cyclodextrin and different functional group are mixed, jolting, normal-temperature reaction 15～30 minutes obtains maleimide maleimide-polyethyleneglycol modified property cyclodextrin derivative;
(5) it is 7～8 o'clock at the pH of phosphoric acid buffer; mix under sulfhydrylation brain target base head and the maleimide maleimide-polyethyleneglycol modified property cyclodextrin derivative nitrogen protection; vibration; 4 ℃; reacted 18～24 hours, after reaction finishes, the solution ultrafiltration membrance filter; remove unnecessary small-molecule substance, obtain brain target base head modification property cyclodextrin derivative.
4. the synthetic method of a kind of brain target base head modification property cyclodextrin derivative as claimed in claim 3, it is characterized in that, in described step (1), described cyclodextrin aqueous solution volume is directly proportional with the sodium hydroxide solution volume of dropping, is controlled at 30: 1～20: 1; And Tosyl chloride adds molar weight and is controlled at 0.5～1 times of excessive cyclodextrin all the time.
5. the synthetic method of a kind of brain target base head modification property cyclodextrin derivative as claimed in claim 3, it is characterized in that, in described step (3), described brain target base head comprises Tf, TfR monoclonal antibody (TfR-MAb), the TfR-Mab fragment (fab '), Lf, LfR monoclonal antibody (LfR-MAb) and LfR-Mab fragment (fab ').
6. the synthetic method of a kind of brain target base head modification property cyclodextrin derivative as claimed in claim 3, it is characterized in that, in described step (3), the add-on mol ratio of the add-on of described 2-imino-sulfane hydrochloride and brain target base head is 35: 1～45: 1.
7. the synthetic method of a kind of brain target base head modification property cyclodextrin derivative as claimed in claim 5 is characterized in that the add-on mol ratio of the add-on of described 2-imino-sulfane hydrochloride and brain target base head is 40: 1.
8. the synthetic method of a kind of brain target base head modification property cyclodextrin derivative as claimed in claim 3, it is characterized in that, in described step (4), the two polyethyleneglycol derivative NHS-PEG-MAL that replace of described different functional group, its PEG long-chain comprises PEG3400, PEG5000.
9. the synthetic method of a kind of brain target base head modification property cyclodextrin derivative as claimed in claim 3, it is characterized in that, in described step (4), described single 6-quadrol-beta-cyclodextrin, the molar ratio scope of NHS-PEG-MAL and sulfhydrylation brain target base head is 900: 160: 1～20: 10: 1.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103083652A (en) * | 2013-02-06 | 2013-05-08 | 中国科学院过程工程研究所 | Meningococcal polysaccharide conjugate vaccine treating heterobifunctional reagent as conjugation bridge, and its preparation method |
| CN105999296A (en) * | 2016-06-07 | 2016-10-12 | 上海纳米技术及应用国家工程研究中心有限公司 | Preparation method for fluorescent blood-brain-barrier-membrane-crossing silica nanoparticles with different particle sizes |
| CN110755638A (en) * | 2019-10-30 | 2020-02-07 | 西南交通大学 | Bone-targeting drug carrier and preparation method and application thereof |
| CN111253505A (en) * | 2020-03-06 | 2020-06-09 | 西南交通大学 | Water-soluble cyclodextrin drug carrier with cell targeting and preparation method thereof |
| CN111393543A (en) * | 2020-05-20 | 2020-07-10 | 黑龙江八一农垦大学 | Cyclodextrin derivative, herbicide coated by cyclodextrin derivative, preparation method and application |
| CN111505140A (en) * | 2020-04-24 | 2020-08-07 | 厦门大学 | Chemical signal amplification multiplier based on virus capsid protein nanostructure, preparation method and application |
| CN113461836A (en) * | 2021-06-16 | 2021-10-01 | 华南农业大学 | Application of amino acid cyclodextrin derivative as pesticide synergist |
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| US20030017972A1 (en) * | 2000-12-19 | 2003-01-23 | California Institute Of Technology And Insert Therapeutics, Inc | Complexing agents for compositions containing inclusion complexes |
| WO2009001364A2 (en) * | 2007-06-28 | 2008-12-31 | Capsutech Ltd | Targeting conjugates comprising active agents encapsulated in cyclodextrin-containing polymers |
| CN101357990A (en) * | 2008-08-21 | 2009-02-04 | 上海交通大学 | Tadpole polymer with controllable molecular weight and good biocompatibility and preparation method thereof |
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Patent Citations (3)
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| US20030017972A1 (en) * | 2000-12-19 | 2003-01-23 | California Institute Of Technology And Insert Therapeutics, Inc | Complexing agents for compositions containing inclusion complexes |
| WO2009001364A2 (en) * | 2007-06-28 | 2008-12-31 | Capsutech Ltd | Targeting conjugates comprising active agents encapsulated in cyclodextrin-containing polymers |
| CN101357990A (en) * | 2008-08-21 | 2009-02-04 | 上海交通大学 | Tadpole polymer with controllable molecular weight and good biocompatibility and preparation method thereof |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103083652A (en) * | 2013-02-06 | 2013-05-08 | 中国科学院过程工程研究所 | Meningococcal polysaccharide conjugate vaccine treating heterobifunctional reagent as conjugation bridge, and its preparation method |
| CN105999296A (en) * | 2016-06-07 | 2016-10-12 | 上海纳米技术及应用国家工程研究中心有限公司 | Preparation method for fluorescent blood-brain-barrier-membrane-crossing silica nanoparticles with different particle sizes |
| CN110755638A (en) * | 2019-10-30 | 2020-02-07 | 西南交通大学 | Bone-targeting drug carrier and preparation method and application thereof |
| CN111253505A (en) * | 2020-03-06 | 2020-06-09 | 西南交通大学 | Water-soluble cyclodextrin drug carrier with cell targeting and preparation method thereof |
| CN111505140A (en) * | 2020-04-24 | 2020-08-07 | 厦门大学 | Chemical signal amplification multiplier based on virus capsid protein nanostructure, preparation method and application |
| CN111393543A (en) * | 2020-05-20 | 2020-07-10 | 黑龙江八一农垦大学 | Cyclodextrin derivative, herbicide coated by cyclodextrin derivative, preparation method and application |
| CN113461836A (en) * | 2021-06-16 | 2021-10-01 | 华南农业大学 | Application of amino acid cyclodextrin derivative as pesticide synergist |
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| CN101935365B (en) | 2012-03-21 |
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