Programmed multi-targeting dendrimer assembly drug delivery system and its preparation method and application

Abstract

The invention discloses a programmed multi-target dendrimer assembly drug delivery system, a preparation method and application thereof, and belongs to the field of biomedical materials. The drug delivery system of the present invention uses the amphiphilic dendrimer assembly as the carrier, realizes programmed multiple targeting by responding to the tumor microenvironment, thereby overcoming the main biological obstacles encountered by the drug carrier in the process of transporting the drug in vivo and finally The drug is delivered to its destination. The resulting self-assembled body has programmed multiple targeting effects through various terminal functionalizations. On the one hand, the advantages of good stability, high mechanical strength, multivalent end groups, high branched structure and virus-like intracellular delivery of dendrimers as drug carriers are used; on the other hand, multiple synergistic The self-assembly strategy integrates the advantages of each functional group on the self-assembly motif of dendrimers to facilitate the multi-functionalization of the assembly and the efficient delivery of drug systems. Especially suitable for the delivery of antitumor drugs.

Description

technical field

[0001] The invention belongs to the field of biomedical materials, in particular to a programmed multi-target drug carrier. technical background

[0002] At present, malignant tumors are a serious threat to human health and development, and are one of the common problems faced by all mankind. Chemotherapy, as one of the most commonly used treatment methods, has achieved a certain degree of tumor inhibition; however, current chemotherapy drugs face problems such as poor water solubility, unsatisfactory metabolism and distribution in the body, and large toxic and side effects that need to be resolved urgently. Nano-delivery systems have significant advantages in increasing drug solubility, improving drug metabolism, and reducing side effects. After research and development in recent years, some nano-drug delivery systems have been successfully used in clinical treatment (such as nano-liposomes loaded with doxorubicin Doxil ® , Nanomicelle Genexol-PM loaded wi...

Examples

Embodiment 1

[0073] Example 1: Preparation of programmed multi-targeted amphiphilic dendrimer self-assembly motif 1 (synthetic route as shown in figure 1 )

[0074] Second-generation lysine dendrimers (HO-Lys(G2)-Boc 4 )Synthesis

[0075] Weigh 4.0 g of H-Lys-OMe·2HCl, 14.8 g of Boc-Lys(Boc)-OH, 5.1 g of HOBT and 13.2 g of EDC·HCl in a 100 mL round-bottomed flask with a branch tube, vacuumize, fill with nitrogen, add 30 mL of redistilled DCM was stirred to dissolve, and 28.3 mL of DIEA was added to react for 0.5 h under ice-cooling, and then reacted at room temperature for 48 h. The solvent was removed by rotary evaporation with a water pump, dissolved in chloroform, and washed successively with 1 M HCl, saturated NaHCO 3 After washing with NaCl, anhydrous MgSO 4 After drying overnight, filter, spin off the solvent, and separate by 200-300 mesh silica gel column chromatography (eluent DCM / EA = 1:1) to obtain white powder MeO-Lys(G2)-Boc 4 (compound 1).

[0076] Demethylation protecti...

Embodiment 2

[0087] Example 2: Preparation of programmed multi-targeted amphiphilic dendrimer self-assembly motif II (synthetic route as shown in image 3 )

[0088] Functionalized hydrophilic end (PEG 1500 -Biotin) synthesis

[0089] Accurately weigh 1.0 g of biotin, 1.6 g of EDC·HCl and 0.6 g of HOBT in a 250 mL round bottom flask with a branch tube, vacuumize, fill with nitrogen protection, and add 20 mL of DMSO to dissolve. After biotin was activated for 2 h, add 12.2 g PEG 1500 and 3.4 mL of DIEA in DCM for 24 h at room temperature. After the oil pump was rotated to remove the solvent, the residue was dissolved in chloroform and washed with saturated NaHCO 3 After washing with NaCl, anhydrous MgSO 4 After drying overnight, filter, spin off the solvent, and separate by 200-300 mesh silica gel column chromatography to obtain white solid PEG 1500 -Biotin (compound 11).

[0090] Synthesis of Hydrophobic End of Oleic Acid-Lys-OH

[0091] Accurately weigh 5.0 g of H-Lys-OMe·2HCl, 16...

Embodiment 3

[0095] Example 3: Determination of Amphiphilic Dendrimer Self-Assembly Motif-Critical Aggregation Concentration

[0096] First prepare a concentration of 6.02 × 10 -7 mol / L aqueous solution of pyrene, and then dilute the amphiphilic dendrimer with end group functionalization prepared in Example 1 with the above prepared pyrene water to a concentration of 1.0 mg / mL to 1.0 × 10 -7 mg / mL solutions with different concentrations. The emission wavelength was fixed at 395 nm, the excitation wavelength in the range of 300 nm - 380 nm was measured with a fluorescence spectrophotometer, and the fluorescence values ​​at 338 nm and 334 nm were recorded. with I 338 / I 334 The ratio of is the ordinate, and the concentration is the abscissa, and the result is as follows Figure 5 shown. The measured critical aggregation concentration is 4.85 μg / mL, and the smaller critical aggregation concentration value indicates that the assembly has better potential stability and prevents disassembl...

technical field

[0001] The invention belongs to the field of biomedical materials, in particular to a programmed multi-target drug carrier. technical background

[0002] At present, malignant tumors are a serious threat to human health and development, and are one of the common problems faced by all mankind. Chemotherapy, as one of the most commonly used treatment methods, has achieved a certain degree of tumor inhibition; however, current chemotherapy drugs face problems such as poor water solubility, unsatisfactory metabolism and distribution in the body, and large toxic and side effects that need to be resolved urgently. Nano-delivery systems have significant advantages in increasing drug solubility, improving drug metabolism, and reducing side effects. After research and development in recent years, some nano-drug delivery systems have been successfully used in clinical treatment (such as nano-liposomes loaded with doxorubicin Doxil ® , Nanomicelle Genexol-PM loaded wi...

Examples

Embodiment 1

[0073] Example 1: Preparation of programmed multi-targeted amphiphilic dendrimer self-assembly motif 1 (synthetic route as shown in figure 1 )

[0074] Second-generation lysine dendrimers (HO-Lys(G2)-Boc 4 )Synthesis

[0075] Weigh 4.0 g of H-Lys-OMe·2HCl, 14.8 g of Boc-Lys(Boc)-OH, 5.1 g of HOBT and 13.2 g of EDC·HCl in a 100 mL round-bottomed flask with a branch tube, vacuumize, fill with nitrogen, add 30 mL of redistilled DCM was stirred to dissolve, and 28.3 mL of DIEA was added to react for 0.5 h under ice-cooling, and then reacted at room temperature for 48 h. The solvent was removed by rotary evaporation with a water pump, dissolved in chloroform, and washed successively with 1 M HCl, saturated NaHCO 3 After washing with NaCl, anhydrous MgSO 4 After drying overnight, filter, spin off the solvent, and separate by 200-300 mesh silica gel column chromatography (eluent DCM / EA = 1:1) to obtain white powder MeO-Lys(G2)-Boc 4 (compound 1).

[0076] Demethylation protecti...

Embodiment 2

[0087] Example 2: Preparation of programmed multi-targeted amphiphilic dendrimer self-assembly motif II (synthetic route as shown in image 3 )

[0088] Functionalized hydrophilic end (PEG 1500 -Biotin) synthesis

[0089] Accurately weigh 1.0 g of biotin, 1.6 g of EDC·HCl and 0.6 g of HOBT in a 250 mL round bottom flask with a branch tube, vacuumize, fill with nitrogen protection, and add 20 mL of DMSO to dissolve. After biotin was activated for 2 h, add 12.2 g PEG 1500 and 3.4 mL of DIEA in DCM for 24 h at room temperature. After the oil pump was rotated to remove the solvent, the residue was dissolved in chloroform and washed with saturated NaHCO 3 After washing with NaCl, anhydrous MgSO 4 After drying overnight, filter, spin off the solvent, and separate by 200-300 mesh silica gel column chromatography to obtain white solid PEG 1500 -Biotin (compound 11).

[0090] Synthesis of Hydrophobic End of Oleic Acid-Lys-OH

[0091] Accurately weigh 5.0 g of H-Lys-OMe·2HCl, 16...

Embodiment 3

[0095] Example 3: Determination of Amphiphilic Dendrimer Self-Assembly Motif-Critical Aggregation Concentration

[0096] First prepare a concentration of 6.02 × 10 -7 mol / L aqueous solution of pyrene, and then dilute the amphiphilic dendrimer with end group functionalization prepared in Example 1 with the above prepared pyrene water to a concentration of 1.0 mg / mL to 1.0 × 10 -7 mg / mL solutions with different concentrations. The emission wavelength was fixed at 395 nm, the excitation wavelength in the range of 300 nm - 380 nm was measured with a fluorescence spectrophotometer, and the fluorescence values ​​at 338 nm and 334 nm were recorded. with I 338 / I 334 The ratio of is the ordinate, and the concentration is the abscissa, and the result is as follows Figure 5 shown. The measured critical aggregation concentration is 4.85 μg / mL, and the smaller critical aggregation concentration value indicates that the assembly has better potential stability and prevents disassembl...

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