WO2014094539A1 - Copolymère triséquencé du type aba à base de colle moléculaire, synthèse de celui-ci et utilisation - Google Patents

Copolymère triséquencé du type aba à base de colle moléculaire, synthèse de celui-ci et utilisation Download PDF

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WO2014094539A1
WO2014094539A1 PCT/CN2013/088478 CN2013088478W WO2014094539A1 WO 2014094539 A1 WO2014094539 A1 WO 2014094539A1 CN 2013088478 W CN2013088478 W CN 2013088478W WO 2014094539 A1 WO2014094539 A1 WO 2014094539A1
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peg
pla
compound
pcl
drug
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PCT/CN2013/088478
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Chinese (zh)
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沈玉梅
龚兵
邵志峰
杨晴来
徐宇虹
侯永泰
朱方霞
肖刚
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上海交通大学
上海其胜生物制剂有限公司
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/34—Macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyesters, polyamino acids, polysiloxanes, polyphosphazines, copolymers of polyalkylene glycol or poloxamers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/10—Dispersions; Emulsions
    • A61K9/107—Emulsions ; Emulsion preconcentrates; Micelles
    • A61K9/1075—Microemulsions or submicron emulsions; Preconcentrates or solids thereof; Micelles, e.g. made of phospholipids or block copolymers
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J3/00—Processes of treating or compounding macromolecular substances
    • C08J3/02—Making solutions, dispersions, lattices or gels by other methods than by solution, emulsion or suspension polymerisation techniques
    • C08J3/03—Making solutions, dispersions, lattices or gels by other methods than by solution, emulsion or suspension polymerisation techniques in aqueous media
    • C08J3/07—Making solutions, dispersions, lattices or gels by other methods than by solution, emulsion or suspension polymerisation techniques in aqueous media from polymer solutions
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J2353/00—Characterised by the use of block copolymers containing at least one sequence of a polymer obtained by reactions only involving carbon-to-carbon unsaturated bonds; Derivatives of such polymers

Definitions

  • the invention relates to a kind of molecular glue-based ABA type triblock copolymer and its synthesis and use, belonging to the fields of chemical synthesis, biochemistry, pharmaceutical science and nano material, and self-assembling micelles can be applied to improve the water solubility of poorly soluble drugs. And has a slow release, controlled release and other effects.
  • An amphiphilic block copolymer refers to a polymer compound formed by polymerizing two or more polymer fragments of different structures, which contains one or more hydrophobic segments, one or more hydrophilic segments, and an amphiphilic block.
  • the hydrophilic and hydrophobic fragments in the copolymer have large differences in solubility, and spontaneously form (self-assembled) nano-sized polymer micelles in water.
  • the micelles formed by the self-assembly of the amphiphilic block copolymer in aqueous solution have a unique core-shell structure.
  • the hydrophobic fragments are aggregated into the inner core in an aqueous environment, and the hydrophilic fragments are surrounded by the core, and the hydrophobic core can be solubilized.
  • Chitosan is a partially deacetylated product of chitin. It is a hydrophilic polysaccharide rich in natural sources. It has biocompatibility, biodegradability, bioadhesion and penetration enhancement. Its physical and chemical properties And its biological properties make it very suitable as a carrier material for drug controlled release systems (Jiang Tingda. Chitin [M]. Beijing: Chemical Industry Press, 2003). Low molecular weight chitosan (Mn ⁇ 10000) is characterized by low viscosity, water solubility, no hemolysis and low cytotoxicity with respect to high molecular weight chitosan (Mn > 50000) (Chme. Phann. Bull. 33 ( 1985) 3986-3922.
  • low molecular weight chitosan also has antitumor activity, enhances immunity, is resistant to microbial infections and promotes hemostasis (Chme. Phann. Bull. 29 (1981) 3067-3069.),
  • the use of low-quantity chitosan as a nano drug controlled release carrier has attracted the attention of many pharmacologists.
  • low molecular weight chitosan does not have amphiphilic properties, introduces hydrophobic groups to make it amphiphilic, and acts as a nano drug controlled release carrier, while the amphiphilic chitosan block copolymer introduces sensitivity ( Active shells of PH, temperature, light, sound, etc.) Glycan block copolymers have become a hot spot in current research.
  • PCL Polycaprolactone
  • PCL has five non-polar methylene-CH 2 and one polar ester group -C00- on its structural repeating unit, and its ester structure is easily decomposed by microorganisms or enzymes, and the final product is 0) 2 and 0.
  • PCL has been widely used in the microcapsule nano drug controlled release system, which has the effects of reducing drug side effects, preventing drug inactivation, reducing the number of times of administration, and targeted drug delivery (Wang Jianguo. Functional Polymer) Material [M]. East China University of Science Press, 2006.).
  • hydrophobicity is to modify the above polymers by organic synthesis, and to introduce molecular chains with hydrogen bond sequence selectivity into known macromolecular polymers, respectively, under specific conditions. It can effectively form molecular glue, which can selectively and efficiently synthesize a series of amphiphilic triblock copolymers with different molecular weights. In the previous work, the applicant synthesized a series of diblock copolymers and multi-block copolymers by molecular glue.
  • the present invention is based on the hydrophilicity of polyethylene glycol (PEG) and low molecular weight chitosan (CS) and polylactic acid (PLA), polycaprolactone (PCL), lauric acid (C 12 ), stearic acid ( C 18 )
  • PEG polyethylene glycol
  • CS low molecular weight chitosan
  • PLA polylactic acid
  • PCL polycaprolactone
  • lauric acid C 12
  • stearic acid C 18
  • hydrophobicity is to introduce a molecular glue single chain with hydrogen bond matching into known macromolecular polymers, so that it can form molecular glue under specific conditions, so that it can be selectively and efficiently synthesized.
  • the invention relates to a molecular glue-based ABA type amphiphilic triblock copolymer PEG-PLA-PEG, the knot thereof
  • the copolymer is prepared by the following steps:
  • PLA Esterification with XT of succinic anhydride to form a modified carboxyl group The molar ratio of PLA, DMAP, succinic anhydride is 1:2.0:3; continue to use DMF as solvent, NMM (N-methylmorpholine), HATU (2-(7-azobenzotriazine) Oxazole) N, N' , N' -
  • the molar ratio of PLA, painting, HATU and Z1 is 1: 4.0: 4.0: (2.5 ⁇ 3.0);
  • the present invention relates to the use of the above molecular gel-based ABA type amphiphilic triblock copolymer for the preparation of drug-loaded micelles.
  • the ABA-type amphiphilic block copolymer and the poorly water-soluble drug are dissolved in a polar solvent to obtain a mixed solution, and the mixed liquid droplets are added to the stirred aqueous phase solution to form a micro-lactate, which will contain
  • the mixture of microemulsions is placed in a dialysis bag to permeate the polar solvent, that is, the drug-loaded micelles are obtained.
  • the polar solvent is DMS0, DMF or THF;
  • the poorly water-soluble drug is doxorubicin, paclitaxel, elemene, camptothecin or retinoic acid;
  • the aqueous phase solution is PBS buffer;
  • the dialysis bag has a molecular weight of 0.2 KD.
  • the present invention relates to an ABA-type amphiphilic triblock copolymer PLA-PEG-PLA based on a molecular gel, the structural formula of which is represented by the formula ( ⁇ )-
  • the copolymer is prepared by the following steps:
  • the molar ratio of N-PEG-NH 2 is 2.0: 4.0: 4.0: (0.3 ⁇ 0.5);
  • Segment compound PLA-Z1 The molar ratio of the PLA, the painting, the HATU and the Z1 is 1: (1.0 ⁇ 2.0): (1.5 ⁇ 3): (1.0 ⁇ 2); C, and composite amphiphilic triblock copolymer PLA-PEG-PLA: In a solvent of dichloromethane, under the effect of 12, the hydrophilic segment compound A4-PEG-A4 and the hydrophobic segment is oxidized compound PLA-Z1 the reaction to produce the amphiphilic triblock copolymer PLA-PEG-PLA, i.e. the amphiphilic ABA type triblock copolymer; the A4_PEG_A4, PLA-Z1 and a molar ratio of 12 to 1: 3:24 .
  • the present invention relates to the use of the above molecular gel-based ABA type amphiphilic triblock copolymer for the preparation of drug-loaded micelles.
  • the ABA-type amphiphilic block copolymer and the poorly water-soluble drug are dissolved in a polar solvent to obtain a mixed solution, and the mixed liquid droplets are added to the stirred aqueous phase solution to form a micro-lactate, which will The mixed solution containing the microemulsions is placed in a dialysis bag to permeate the polar solvent, that is, the drug-loaded micelles are obtained.
  • the polar solvent is DMS0, DMF or THF;
  • the poorly water-soluble drug is doxorubicin, paclitaxel, elemene, camptothecin or retinoic acid;
  • the aqueous phase solution is PBS buffer; 2KD ⁇ The molecular weight of the dialysis bag is 0. 2KD.
  • the present invention relates to a molecular gel-based ABA type triblock copolymer having a structural formula as shown in formula (III) -
  • the present invention relates to a method for synthesizing the above-mentioned molecular gel-based ABA type triblock copolymer, which comprises the following steps: using dichloromethane/dimethyl sulfoxide as a mixed solvent, at 1 2 Hydrophilic fragment compound CS-A4 And hydrophilic fragment compound Z1-PEG- segment compound Z1-PLA- segment compound Z1-PCL-Z1 Generating the ABA type triblock
  • z/(w+z+l)iS15% 03 ⁇ 4 ⁇ 135, 23 ⁇ 4 ⁇ 3 ⁇ 4 ⁇ 225, l3 ⁇ 4St3 ⁇ 4S42; the molar ratio of the compound CS_A4, Zl-PEG-Zl, Zl-PLA-Zl or Zl-PCL-Zl and I 2 is (2 ⁇ 3) : 1 : (24 ⁇ 48).
  • the molar ratio of the PEG, PLA or PCL, DMAP and succinic anhydride is 1:2.0:3.0 ⁇ 4.0; continue to use DMF as solvent, NMM (N-methylmorpholine), HATU (2- (7-even) Nitrobenzotriazole) -N, N, N' , N' -tetra EG, terminal carboxyl PLA
  • the present invention relates to a molecular gel-based ABA type amphiphilic triblock copolymer having a structural formula as shown in formula (IV) - (IV) , where 23 ⁇ 4Sn3 ⁇ 4S
  • R 2 is a structural formula represented by formula (VE) or (IX):
  • the molar ratio of the A4_PEG_A4, PCL-Z1 or Cy_Zl and I 2 is 1:3:24.
  • the hydrophilic fragment compound A4-PEG-A4 is synthesized by the following steps: using DMF as a solvent, NMM (N-methylmorpholine), HATU (2-(7-azobenzotriene) Azole) - hydrazine, hydrazine, hydrazine, ⁇ '-tetramethylurea hexafluorophosphate. H
  • the hydrophobic fragment compound PCL-Z1 is synthesized by the following steps: using DMF as a solvent, NMM (N-methylmorpholine), HATU (2-(7-azobenzene-hydrazine, hydrazine, ⁇ ' , ⁇ ' - tetramethylurea hexafluorophosphate
  • the present invention relates to a molecular gel-based ABA type amphiphilic triblock copolymer having a structural formula (
  • the present invention relates to a method for synthesizing the above-mentioned molecular gel-based ABA type amphiphilic triblock copolymer, the method comprising the steps of: using dichloromethane as a solvent , under the action of 1 2 , hydrophilic fragment compound
  • Oxidation reaction occurs to form the ABA type amphiphilic triblock copolymer
  • hydrophilic fragment compound PEG-A4 The hydrophilic fragment compound PEG-A4; the molar ratio of the ⁇ 4, ⁇ , HATU, PEG-NH 2 is 1: (1.0 ⁇ 2.0): (1.5 ⁇ 3): (1.0 ⁇ 2);
  • the molar ratio of the anhydride is 1:2.0:3; continue with DMF as solvent, methylmorpholine), HATU (2-(7-azobenzotriazole)- ⁇ , ⁇ , ⁇ ', ⁇ ' - four Methylurea hexafluorophosphate) as an activator, the terminal carboxyl group PCL and
  • the amidation reaction of the compound Z1 gives the hydrophobic segment compound Z1-PCL-Z1; the molar ratio of the terminal carboxyl group PCL, the drawing, the HATU and the Z1 is 1: 4.0: 4.0: (2.5 to 3.0).
  • the ABA-type amphiphilic block copolymer and the poorly water-soluble drug are dissolved in a polar solvent to obtain a mixed solution, and the mixed liquid droplets are added to the stirred aqueous phase solution to form a micro-lactate, which will contain
  • the mixture of microemulsions is placed in a dialysis bag to permeate the polar solvent, that is, the drug-loaded micelles are obtained.
  • the present invention relates to a drug-loaded micelle obtained by dissolving the above-mentioned five ABA-type amphiphilic block copolymers and a poorly water-soluble drug in a polar solvent, and mixing the mixed liquid droplets
  • the microspheres are formed by adding the stirred aqueous phase solution, and the mixed solution containing the microemulsions is placed in a dialysis bag to permeate the polar solvent, thereby obtaining the drug-loaded micelles.
  • the polar solvent is DMS0, DMF or THF;
  • the poorly water-soluble drug is doxorubicin, paclitaxel, elemene, camptothecin or retinoic acid;
  • the aqueous phase solution is PBS buffer;
  • the dialysis bag has a molecular weight of 0.2 KD.
  • the block copolymer micelle prepared by the invention has good stability, and the blank micelle has no toxic effect on normal cells, and the drug-loaded micelle releases the packaged drug under the action of the reducing agent dithiothreitol, and the drug-loaded micelle can enter the cancer.
  • the cells release a drug to kill cancer cells under the action of glutathione.
  • the normal cells are NIH 313 cells, and the cancer cells are HeLa cells.
  • the drug contained therein is a poorly water-soluble drug such as azithromycin, paclitaxel, elemene, camptothecin and retinoic acid.
  • the present invention essentially relates to a class of molecular-based amphiphilic triblock copolymers having the structural formula shown below, the amphiphilic triblock copolymers being selectively controlled by hydrogen bond sequences. Formation of a double disulfide bond.
  • A1 is PLA, PCL or Cy
  • B1 is PEG
  • A2 is CS or PEG
  • B2 is PLA, PCL or PEG
  • the specific structural formula is as follows - Compared with the prior art, the present invention has the following beneficial effects:
  • organic polymer synthesis method was used to modify the polymers PEG, PLA, PCL, saturated fatty acid (C y ) and chitosan (CS) to introduce molecular chains with hydrogen bond sequence selectivity into polymer macromolecules. It can form molecular glue under certain conditions, so that the triblock copolymer can be selectively and efficiently synthesized.
  • the synthetic raw materials of the series of triblock copolymers are simple and easy to obtain, and the synthesis process is a conventional reaction, which is suitable for large-scale production. .
  • the block copolymer micelles prepared by the invention have good stability, and the blank micelles have no toxic effect on normal cells (NIH 313 cells), and the drug-loaded micelles are in the reducing agent dithiothreitol or reduced glutathione.
  • the drug can be effectively released by the action of a reducing agent, and the drug-loaded micelle can effectively enter the cancer cell and release the drug in the cancer cell to kill the cancer cell (for example, HeLa cells).
  • These compounds contain disulfide bonds and hydrogen bonds, which have the sensitivity of reducing agent. They can be used as sensitive drug carriers for fat-soluble drugs such as doxorubicin, paclitaxel, camptothecin, retinoic acid and elemene.
  • Figure 1 is a schematic diagram showing the synthesis route of the hydrophilic fragment compound PEG-A4;
  • FIG. 2 is a schematic diagram of a synthetic route of a hydrophilic fragment compound A4-PEG-A4;
  • Figure 3 is a schematic view showing the synthesis route of the hydrophobic segment compound PLA-Z 1;
  • Figure 4 is a schematic view showing the synthesis route of the hydrophobic segment compound Z 1-PLA-Z 1 ;
  • Figure 5 is a schematic diagram showing the synthesis route of the amphiphilic triblock copolymer PEG-PLA-PEG;
  • Figure 6 is a schematic diagram of the synthetic route of the amphiphilic triblock copolymer PLA-PEG-PLA;
  • Figure 7 is a GPC molecular weight distribution diagram of the amphiphilic triblock copolymer of Example 1;
  • Figure 8 is a graph showing the relationship between the micelle concentration and the absorbance A
  • Figure 10 is PEG 5 . . . _PLA 5 . . . - PEG 5 . . . Particle size distribution map of drug-loaded micelles;
  • Figure 11 is PEG 5 . . . _PLA 5 . . . - PEG 5 . . . Micellar stability test results;
  • Figure 12 is PEG 5 . . . _PLA 5 . . . - PEG 5 . . . Particle size change diagram of micelles treated with reducing agent DTT;
  • Figure 13 is a drug-loaded micelle PEG 5 . . . _PLA 5 . . . - PEG 5 . . . Release rate map of D0X under different concentrations of DTT treatment conditions;
  • Figure 14 is a drug-loaded micelle PEG 5 . . . _PLA 5 . . . - PEG 5 . . . In vitro drug release profile;
  • Figure 15 is a blank micelle PEG 5 . . . _PLA 5 . . . - PEG 5 . . . MTT assay results for NIH 3T3 cytotoxicity;
  • Figure 16 is a blank micelle PEG 5 . . . -PLA 5 . . . - PEG 5 . . . A0/EB double staining experiment results;
  • Figure 17 is a drug-loaded micelle PEG 5 . . . _PLA 5 . . . - PEG 5 . . . Intracellular release profile;
  • Figure 18 is a drug-loaded micelle PEG 5 . . . _PLA 5 . . . - PEG 5 . . . Results of MTT assay on Hela cells;
  • Figure 19 is a triblock copolymer PEG 2 . . . _PLA 3 . . . - PEG 2 . . .
  • Figure 20 is a triblock copolymer PLA 3 . . . - PEG 2 . . . _PLA 3 . . . 3 ⁇ 4_NMR comparison of each polymer in the synthesis
  • Figure 21 shows PEG 5 . . . _PLA 5 . . . - PEG 5 . . . TEM image of blank micelles;
  • Figure 22 is PEG 5 . . . _PLA 5 . . . - PEG 5 . . . TEM image of drug-loaded micelles;
  • Figure 23 is a schematic view showing the synthesis route of the hydrophilic fragment compound Z 1-PEG-Z 1 ;
  • Figure 24 is a schematic view showing the synthesis route of the hydrophilic fragment compound CS-A4;
  • Figure 25 is a schematic view showing the synthesis route of the hydrophobic segment compound PCL-Z 1;
  • Figure 27 is a schematic view showing the synthesis route of the hydrophobic segment compound C 12 _Z 1 and C 18 _Z 1 ;
  • Figure 29 is a triblock copolymer PCL 47 . . - PEG 5 . . . -PCL 47 . . 1 H-NMR comparison chart of each polymer in the synthesis;
  • Fig. 30 is a schematic diagram of a synthetic route of a triblock copolymer (-PEG-;
  • Figure 33 is a triblock copolymer CS 25 . . - PEG 5 . . . -CS 25 . . 3 ⁇ 4-NMR comparison of each polymer in the synthesis;
  • Figure 34 is a schematic diagram of the synthesis route of the triblock copolymer CS-PLA-CS;
  • the step is specifically: weigh A4 (0.841 g, lmmol) in 15 ml of DMF in a 25 ml single-necked flask, and add NMM (N-methylmorpholine) (224 L, 2. Ommol) under stirring in an ice water bath, HATU (2_(7_Azobenzotriazole)-N, N, ⁇ ', ⁇ '-tetramethylurea hexafluorophosphate) (0.76g, 2.
  • PLA 3 is weighed. . . (3.0 g, l mmol), HATU (2-(7-azobenzotriazole)-N, hydrazine, ⁇ ', ⁇ '-tetramethyluron hexafluorophosphate) (1.14 g, 3. Ommol) , Zl (1.566, 2 ⁇ ol) in a 25 ml dry single-necked flask, NMM (N-methylmorpholine) (223 ⁇ L, 2. Ommol) was added under ice-cooled water, and dried DMF 15 ml was stirred for 45 min.
  • PLA 5 is weighed. . . (5.0g, lmmol), HATU (2-(7-azobenzotriazole)-N, ⁇ , ⁇ ', ⁇ '-tetramethylurea hexafluorophosphate) (0.76g, 2. Ommol) , Zl (l.174, 1.5 mmol) was added to a 25 ml dry single-necked flask, NMM (N-methylmorpholine) (167 L, 1.5 mmol) was added under ice-cooling, and the dried DMF 15 ml was stirred for 45 min.
  • HATU 2-(7-azobenzotriazole)-N, ⁇ , ⁇ ', ⁇ '-tetramethylurea hexafluorophosphate
  • the reaction was heated to 35 ° C for 14 h, the reaction was stopped, the appropriate amount of dichloromethane was extracted, washed twice, washed twice with saturated NaCl solution, and the organic phase was distilled under reduced pressure to give 5.8 g of pale yellow solid. The yield was 71%.
  • the step is specifically as follows: weighing ⁇ 1 ⁇ . . . . (l.Og, 0. Immol), HATU (2-(7-azobenzotriazole)-N, N, ⁇ ', ⁇ '-tetramethylurea hexafluorophosphate) (0.057g, 0.15 Methyl), Z1 (0.0783, 0.1 mmol) in a 25 ml dry single-necked flask, NMM (N-methylmorpholine) (17 ⁇ L, 0.1 mmol), dry DMF 15 ml After stirring for 45 min, the temperature was raised to 35 ° C for 18 h, the reaction was stopped, the appropriate amount of dichloromethane was added, the water was washed twice, the saturated NaCl solution was washed twice, and the organic phase was distilled under reduced pressure to give a pale yellow solid.
  • the steps are specifically as follows: Weigh PLA 162 (L62g lOmmol) dissolved in 20 ml C C1 2 , add DMAP (2.44g 20 ol) once in ice water bath, succinic anhydride (3.0g 30 ol) and stir for 1 hour after room temperature After stirring for 18 hours, an appropriate amount of water was added, and the mixture was extracted with DCM. The organic phase was washed with saturated NaHCO ⁇ and brine, dried over anhydrous sodium sulfate,
  • PLA 3 is weighed. . . 3.0g lmmol) dissolved in 20 ml CH 2 C1 2 , added with DMAP (0.244g 2 ol), succinic anhydride (0.3g 3mmol), stirred for 1 hour, stirred at room temperature for 15h, and then added with water.
  • the organic phase was washed with a saturated NaHCO ⁇ solution and a saturated aqueous sodium chloride solution and dried over anhydrous sodium sulfate.
  • the steps are specifically as follows: Weigh PLA 5 (5.0 g lmmol) dissolved in 20 ml CH 2 C1 2 , add DMAP (0.244 g 20 mmol), succinic anhydride (0.3 g 30 mmol), and stir for 1 hour at room temperature under ice water bath. After stirring for 22 hours, the reaction was completed, and an appropriate amount of water was added thereto, and the mixture was extracted with DCM. The organic phase was washed with saturated NaHCO ⁇ and brine, dried over anhydrous sodium sulfate, and evaporated to remove solvent.
  • the steps are specifically as follows: weigh Z1-PLA 5 -Z1 (99.45mg 0.015mmol) and PEG 5 _A4 (116.46mg, 0.02mmol), respectively, into 20ml of dichloromethane in a 250ml single-mouth bottle, stir, dissolve and mix evenly after the dry distillation under reduced pressure, the residue was added 60 ml of dichloromethane a solution of 12 (where 12 is 6.0 mM) was dissolved, stirred for lh at room temperature, the reaction solution was cooled to 0V, was added N3 ⁇ 4S 2 0 3 (3. OmM) until The color of 1 2 disappears.
  • PLA 5 -Z1 (173 mg, 0.03 mmol) and A4_PEG 5 _A4 (66.46 mg 0. Olmmol) are respectively weighed and dissolved in 20 ml of dichloromethane in a 250 ml single-mouth bottle, stirred, dissolved and uniformly mixed. dry pressure distillation, the residue was added 60 ml dichloro-1 2 A solution of the embankment (wherein 1 2 is 6. OmM) was dissolved, was stirred lh at room temperature, the reaction solution was cooled to 0 ° C, was added N3 ⁇ 4S 2 0 3 (3. OmM) 12 until the color disappears.
  • the molecular weight of the polymer obtained in Example 1 was determined by the GPC method; Apparatus: Agilent Model 260 gel permeation chromatograph, GPC column: 7. 5 X 300 10 ⁇ m gel column, solvent: tetrahydrofuran, flow rate: 1 OmL/min, column temperature: 35 ° C, standard: polystyrene.
  • the GPC molecular weight distribution of each polymer is shown in Fig. 7.
  • Example 3 Preparation and characterization of micelles (with PEG 5 -PLA 5 -PEG 5 ... as an example)
  • 1,6-diphenyl-1,3,5-hexatriene (DF3 ⁇ 4) as an ultraviolet molecular probe (maximum absorption wavelength of 313 nm)
  • Determine the CMC value take 10 10ml EP tubes, add 20 ⁇ 1 ImM DPH acetone solution to each tube, after the acetone is evaporated, add different concentrations to each tube (concentration range: 0. 5 mg/mL, 0. 25 mg / mL, 0. 1 mg / mL, 0. 05mg / mL, 0. 025 mg / mL, 5 X 10- 3 mg / mL, 1 X 10- 3 mg / mL, 0. 25 X 10- 3 mg / mL, 0.
  • DTT treatment of blank micelles preparation of 0. 5mg / ml of micellar solution 10ml, adding 15. 4mg of DTT (dithiothreitol) DTT solution concentration of 10 mM, constant temperature 37 ° C stirring, at lh, 3h, Particle size change at 6h, 8h.
  • the particle size change chart is shown in Fig. 12; it can be seen from Fig. 12 that under the treatment of the reducing agent DTT, the disulfide bond in the micelle is destroyed, so that the particle size of the micelle changes, and it can be seen that the particle size is slightly smaller. Because of the hydrogen bonding force in the fragment after the fracture, the smaller particle size particles will accumulate again and the partial particle size becomes larger.
  • the drug-loaded micelles prepared above were separately prepared into 3 mL of drug-loaded micelle solution containing DTT concentration of 0.1 mM, 1.0 mM, 10 mM, and placed in a water bath at 37 ° C for a period of time.
  • the fluorescence absorption value of the emission wavelength of 590 nm was measured on the fluorescence spectrophotometer at an excitation wavelength of 485 nm, and the fluorescence absorbance change curve was plotted as shown in Fig. 13.
  • the DTT concentration was increased, and the micelle solution was increased.
  • the fluorescence absorption value is increased, and it can be seen that under the treatment condition of the reducing agent DTT, the micelle disulfide bond is broken, and the micelle-encapsulated D0X is released.
  • the encapsulation ratio refers to the ratio of the drug encapsulated in the nanoparticles to the total amount of the drug in the nanoparticle suspension. Freeze-dried to upload the drug micelles, add 5ml DMF to dissolve the dry powder, determine the drug encapsulation rate by ultraviolet spectrophotometry, and calculate the formula as follows -
  • the experimental results are shown in Fig. 18.
  • the survival rate of cancer cells is increased under the action of BS0, and the survival rate of cancer cells is reduced by the action of GSH-OEt, wherein the role of BS0 is to inhibit cancer cells.
  • the production of reduced glutathione, the role of GSH-OEt is to promote the increase of reduced glutathione content in cells, which indicates that the drug-loaded micelles have reduced sensitivity, which is synergistic under the synergy of GSH-OEt.
  • the killing effect of drug micelles on cancer cells Strong, wherein the results showed that when the D0X concentration in the drug-loaded micelles was 0.0001 ⁇ ⁇ . mL -1 or more, the drug-loaded micelles inhibited cancer cells by more than 50%.

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Abstract

L'invention concerne un copolymère triséquencé du type ABA à base d'une colle moléculaire, la synthèse de celui-ci et son utilisation. La formule de structure du copolymère est choisie parmi la Formule (I). Le copolymère selon l'invention peut être utilisé pour préparer une micelle auto-assemblée portant un médicament. La présente invention est fondée sur le caractère hydrophile du polyéthylène glycol et du chitosane de faible poids moléculaire, et sur le caractère hydrophobe de l'acide polylactique, du polycaprolactone, de l'acide laurique, et du stéarate. Des chaînes simples de colle moléculaire ayant une sélectivité de séquence de liaison hydrogène sont introduites séparément dans des polymères macromoléculaires connus, de telle sorte que dans certaines conditions, deux chaînes simples complémentaires peuvent former une colle moléculaire. Une série de copolymères triséquencés amphiphiles peut ainsi être synthétisée efficacement. En outre, la colle moléculaire introduite contient une liaison disulfure, et le copolymère est donc sensible à l'oxydoréduction. Les agents de matière première pour la synthèse selon l'invention peuvent être facilement obtenus, les conditions sont tempérées, et le procédé de synthèse ne comporte que des réactions chimiques habituelles et est adapté à une production à grande échelle.
PCT/CN2013/088478 2012-12-21 2013-12-04 Copolymère triséquencé du type aba à base de colle moléculaire, synthèse de celui-ci et utilisation WO2014094539A1 (fr)

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CN110997284A (zh) * 2017-08-18 2020-04-10 通用电气公司 用于粘合剂喷射增材制造的热塑性粘合剂

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CN103059311B (zh) * 2012-12-21 2014-11-19 上海交通大学 基于分子胶的aba型两亲性三嵌段共聚物及其用途
CN103396556B (zh) * 2013-07-25 2015-08-26 上海交通大学 基于壳聚糖的两亲性嵌段共聚物及其合成方法
CN104031268B (zh) * 2014-05-09 2017-01-11 上海交通大学 聚乙烯亚胺三嵌段共聚物及其在基因载体中的用途
CN106366264B (zh) * 2016-09-30 2018-12-11 东莞理工学院 一种三嵌段共聚物以及其调控方法
CN106967211B (zh) * 2017-04-05 2019-05-17 南京大学 紫杉醇-聚乳酸-紫杉醇共聚物及其制备方法与应用

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CN102153761A (zh) * 2011-01-21 2011-08-17 上海交通大学 用于合成两亲性嵌段共聚物的模块化合物及其制备方法
CN102558535A (zh) * 2011-12-14 2012-07-11 上海交通大学 通过分子胶连接的两亲性嵌段共聚物及其合成方法与应用
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WO2006071769A1 (fr) * 2004-12-28 2006-07-06 The Trustees Of The University Of Pennsylvania Liberation controlee a partir de micelles de type ver de copolymere sequence
CN102153761A (zh) * 2011-01-21 2011-08-17 上海交通大学 用于合成两亲性嵌段共聚物的模块化合物及其制备方法
CN102558535A (zh) * 2011-12-14 2012-07-11 上海交通大学 通过分子胶连接的两亲性嵌段共聚物及其合成方法与应用
CN103059311A (zh) * 2012-12-21 2013-04-24 上海交通大学 基于分子胶的aba型两亲性三嵌段共聚物及其用途

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110997284A (zh) * 2017-08-18 2020-04-10 通用电气公司 用于粘合剂喷射增材制造的热塑性粘合剂
CN110997284B (zh) * 2017-08-18 2022-04-05 通用电气公司 用于粘合剂喷射增材制造的热塑性粘合剂

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