CN111346235A - Drug-loaded nanoparticle based on elastic targeting polypeptide and preparation method and application thereof - Google Patents

Drug-loaded nanoparticle based on elastic targeting polypeptide and preparation method and application thereof Download PDF

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CN111346235A
CN111346235A CN201811598471.1A CN201811598471A CN111346235A CN 111346235 A CN111346235 A CN 111346235A CN 201811598471 A CN201811598471 A CN 201811598471A CN 111346235 A CN111346235 A CN 111346235A
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许铁峰
陈明南
董书芸
李丽
李光磊
张立明
吕卓璇
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First Affiliated Hospital Of Hainan Medical College
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Abstract

The invention discloses a drug-loaded nanoparticle based on elastic targeting polypeptide, which is prepared from the elastic targeting polypeptide and a modified drug, wherein the modified drug is modified paclitaxel PTX-LEV-MECH or modified salinomycin Sail-ABA-MPBH. The drug-loaded nanoparticle based on the elastic targeting polypeptide provided by the invention has the advantages of particle size of below 100nm, small dispersion degree and high drug loading rate, can be combined with in-vivo albumin to deliver drug loading, is combined with acidic cysteine rich in tumor cell specificity secretion, and can release drugs in a centralized manner in an acidic environment to kill breast cancer in situ in a targeted manner, and simultaneously reduce the toxic and side effects of the drugs on the whole body.

Description

Drug-loaded nanoparticle based on elastic targeting polypeptide and preparation method and application thereof
Technical Field
The invention relates to the technical field of medicines, in particular to a medicine-carrying nanoparticle based on elastic targeting polypeptide and a preparation method and application thereof.
Background
Tumors are one of diseases seriously harming human health, and the incidence and mortality of tumors in China are increased year by year. The first three of the malignant tumors are lung cancer, breast cancer and gastric cancer. Statistics show that the incidence rate of female cancer is obviously increased in the last two years, and about 21 ten thousand cases are newly transmitted every year, particularly the breast cancer which is the first malignant tumor of a female. The cancer incidence rate of China is close to the world level, but the death rate is higher than the world level, and the continuously rising incidence rate of malignant tumors becomes a public health problem and even a social problem which must be highly emphasized.
At present, Paclitaxel (Paclitaxel, PTX) and Salinomycin (Salinomycin, Sali) are clinically used as special anticancer drugs, but Paclitaxel and Salinomycin have advantages and disadvantages in prevention and treatment of breast cancer, Paclitaxel has a good antitumor effect, but cannot kill breast cancer stem cells at high selectivity, and the existence of breast cancer stem cells easily causes poor drug resistance, local recurrence, distant metastasis and other metastatic breast cancer chemotherapy failures. Salinomycin has selective killing power on human and mouse breast cancer stem cells, has the efficacy 100 times higher than that of taxol, can inhibit the generation of new tumor cells and simultaneously can slow down the growth rate of existing tumors, but has certain toxic and side effects on organisms, so the application of salinomycin in anticancer is limited.
How to accurately and efficiently deliver drugs to cancer cells and kill the cancer cells is a problem, and the prior report, for example, in patent CN201410838596.2, discloses a pegylated paclitaxel nanocrystal, which is composed of pegylated paclitaxel and paclitaxel, firstly, hydrophilic polymers polyethylene glycol (PEG), Paclitaxel (PTX) and small molecule linking agents are used as starting materials to prepare pegylated paclitaxel (PEG-PTX), and then, the pegylated paclitaxel nanocrystal (PEG-PTX-NCs) is prepared by using the hydrophilic polymers polyethylene glycol (PEG), paclitaxel and small molecule linking agents as modifiers, wherein the prepared pegylated paclitaxel nanocrystal has a small particle size, the drug loading capacity can reach more than 80%, but the targeting capability is poor.
Recently, there is a related literature (MacEWan SR, Hassouneh W, Chilkoti A. non-chromatographic purification of Recombinant Elastin-like Polypeptides and their Fusions with peptides and Proteins from Escherichia coli. journal of visualized experiments: journal. 2014: 51583.) reported that an immunocompatible lymph node targeting elastic Polypeptide (ABD-iTEP) has temperature sensitive properties, i.e., precipitates above the phase transition temperature, dissolves below the phase transition temperature, and can be applied to targeted tumor therapy.
Therefore, there is a need to prepare a drug-loaded nanoparticle based on an elastic targeting polypeptide, which solves the problems of poor targeting selectivity or large side effect of the existing anticancer drugs.
Disclosure of Invention
In view of the above, the invention provides a drug-loaded nanoparticle based on an elastic targeting polypeptide, which is prepared from the elastic targeting polypeptide and a modified drug, wherein the modified drug is convenient to be combined with a sulfhydryl group of cysteine and is released only in an acidic environment, and the drug is modified paclitaxel PTX-LEV-MECH or modified salinomycin Sail-ABA-MPBH. Furthermore, the elastic targeting polypeptide is prepared from ABD and iTEP, wherein the amino acid sequence of ABD is LAEAKVLANRELDKYGVSDFYKRLINKAKTVEGVEALKLHILAALP, iTEP, and the amino acid sequence of ABD is GAGVPG.
On the other hand, the preparation method of the drug-loaded nanoparticle based on the elastic targeting polypeptide comprises the following steps: adding elastic targeting polypeptide into a buffer solution for dissolving, adding tris (2-carboxyethyl) phosphine TCEP dissolved in the buffer solution, adjusting the pH value to 7, stirring and reacting for 1h at room temperature, introducing argon for protection, adding a modified drug dissolved in a solvent, reacting overnight, introducing argon for protection, after the reaction is finished, performing suction filtration and centrifugation, and reserving supernatant for ultrafiltration, washing and dispersing to obtain the nano-particles.
On the other hand, the drug-loaded nanoparticle based on the elastic targeting polypeptide can be used for targeted killing of tumor cells, and particularly treatment of breast cancer and lymph node metastatic carcinoma thereof.
The drug-loaded nanoparticle based on the elastic targeting polypeptide provided by the invention has the advantages of particle size of below 100nm, small dispersion degree and high drug loading rate, can be efficiently combined with in-vivo albumin to deliver drug loading, is combined with the acidic cysteine rich in tumor cell specificity secretion, and can intensively release the drug in an acidic environment to kill breast cancer in situ in a targeted manner, and simultaneously reduce the toxic and side effects of the drug on the whole body.
Drawings
FIG. 1 is a mass spectrum of modified paclitaxel PTX-LEV-MECH of example three of the present invention;
FIG. 2 is a nuclear magnetic spectrum of modified salinomycin Sail-ABA-MPBH in example IV of the invention;
FIG. 3 is the polypeptide-modified paclitaxel ABD-iTEP70- (G4C)8-PTX of example five
-laser scattering pattern of LEV-MECH nanoparticles;
FIG. 4 is a laser scattering diagram of the polypeptide-modified salinomycin ABD-iTEP-Sail-MPBH nanoparticle of example six;
FIG. 5 is a schematic diagram of two nanoparticles of the present invention for preventing and treating tumors in vivo.
Detailed Description
The principles and features of this invention are described below in conjunction with examples which are set forth to illustrate, but are not to be construed to limit the scope of the invention.
The first embodiment is as follows: preparation of elastic targeting polypeptide ABD-iTEP70- (GGGGC)8
Firstly designing amino acid sequences ABD and iTEP, and then preparing by adopting the existing polypeptide synthesis method
ABD- (iTEP)70- (GGGGC)8, wherein
ABD amino acid sequence: LAEAKVLANRELDKYGVSDFYKRLINKAKTVEGVEALKLHILAALP
iTEP amino acid sequence: GAGVPG
After the coding gene of the elastic targeting polypeptide ABD- (iTEP)70- (GGGGC)8 is synthesized, enzyme digestion is carried out to insert the coding gene into a modified pET25b (+) vector to construct a recombinant expression plasmid; the gene sequence is verified to be successfully prepared after sequencing.
Example two: preparation of elastic targeting polypeptide ABD- (iTEP)70- (GGGGGGC)8
Designing amino acid sequences ABD and iTEP, and preparing ABD- (iTEP)70- (GGGGC)8 by using the conventional polypeptide synthesis method, wherein
ABD amino acid sequence: LAEAKVLANRELDKYGVSDFYKRLINKAKTVEGVEALKLHILAALP
iTEP amino acid sequence: GAGVPG
After the coding gene of the elastic targeting polypeptide ABD- (iTEP)70- (GGGGGGC)8 is synthesized, enzyme digestion is carried out to insert the coding gene into a modified pET25b (+) vector to construct recombinant expression plasmid; the gene sequence is verified to be successfully prepared after sequencing.
Example three: synthesis of modified paclitaxel PTX-LEV-MECH
Figure BDA0001920524400000041
Synthesis of Compound 1: hydrazine hydrate (20.0g, 0.40mmol) is dissolved in 80mL of isopropanol at 0 deg.C, di-tert-butyl dicarbonate (43.6g, 0.20mmol) is added to the solution, the reaction solution is warmed to room temperature, stirred for 1 hour, the solvent is removed, the residue is dissolved in dichloromethane, the insoluble material is removed by filtration, and the organic phase is concentrated to give a white solid, a combined compoundProduct 1(19.4g, 74% yield).1H NMR(400MHz，CDCl3)δ1.46(s，9H)
Synthesis of Compound 2: maleic anhydride (5.00g, 51.0mmol) was dissolved in 100mL of acetic acid at 0 deg.C, and 6-aminocaproic acid (6.69g, 51.0mmol) was then slowly added to the above solution. The reaction solution was warmed to room temperature and stirred for 4 hours. Subsequently, the reaction was heated to reflux overnight. Acetic acid was removed under reduced pressure. The residue was purified by alumina column chromatography to give compound 2(6.1g, 63% yield).1H NMR(400MHz，CDCl3)δ1.21-1.45(m，2H)，1.52-1.78(m，4H)，2.30-2.33(m，2H)，3.52(t，J＝6.0Hz，2H)，6.69(s，2H)
Synthesis of Compound 3: under the protection of argon, compound 2(5.92g, 28.0mmol) and 1-hydroxy-benzo-triazole (4.54g, 33.6mmol) are dissolved in 100mL of DMF and cooled to 0 ℃. Then, N' -diisopropylcarbodiimide (4.24g, 33.6mmol) was added to the reaction solution, and the mixture was stirred for 20 minutes. Compound 1 was dissolved in 60mL of DMF, added to the reaction and stirred at room temperature overnight. After the reaction was completed, the reaction solution was quenched with water and then extracted with ethyl acetate. The organic phases were combined, dried, concentrated and purified on a column to give 3 as a colorless oil (5.9g, 65% yield).1H NMR(400MHz，CDCl3)δ1.30-1.41(m，2H)，1.48(s，9H)，1.56-1.66(m，2H)，1.67-1.78(m，2H)，2.24(t，J＝6.0Hz，2H)，3.53(t，J＝8.0Hz，2H)，6.70(s，2H)
Synthesis of compound EMCH: compound 3(3.90g, 12.0mmol) and anisole (1.94g, 1.9mL, 18.0mmol) were dissolved in 30mL dioxane at room temperature, followed by addition of 48mL saturated HCl in ethyl acetate and stirring overnight. After compound 3 had reacted completely, it was filtered and the filter residue was washed with diethyl ether to give EMCH as a white solid (3.0g, 96% yield).1HNMR(400MHz，CD3OD)δ1.26-1.40(m，2H)，1.56-1.66(m，2H)，1.66-1.77(m，2H)，2.25-2.40(m，2H)，3.52(t，J＝6.0Hz，2H)，6.83(s，2H)
Figure BDA0001920524400000051
Synthesis of the Compound PTX-LEV: dicyclohexylcarbodiimide (1.35g, 6.55mmol) and levulinic acid (0.707g, 6.09mmol) were dissolved in 100mL anhydrous DMF under argon protection at-18 ℃. PTX (4.0g, 4.68mmol) and 4-dimethylaminopyridine (0.572g, 4.68mmol) were dissolved in 50mL of anhydrous DMF and added to the reaction mixture. Then, the reaction solution was heated to 4 ℃, stirred for 4 hours, and after the completion of PTX reaction, the reaction solution was filtered, and the residue was washed with water, and the filtrate was extracted with ethyl acetate. The combined organic phases were dried, concentrated and purified by column chromatography to give the compound PTX-LEV (4.2g, 94% yield) as a white solid, 1H NMR (400MHz, CD3OD) δ 1.13(s, 3H), 1.23(s, 3H), 1.67(s, 3H), 1.84(s, 2H), 1.91(s, 3H), 2.13(s, 3H), 2.23(s, 3H), 2.43(s, 3H), 2.48-2.58(m, 2H), 2.64-2.72(m, 2H), 2.72-2.78(m, 2H), 3.75-3.85(m, 1H), 4.16-4.26(m, 1H), 4.27-4.36(m, 1H), 4.38-4.50(m, 1H), 4.92-5.03(m, 1H), 4.46.8-4.8H, 1H), 89-6H (1H), 1H, 6-8H, 6 Hz, 6H, 8H, 6H, 1H, 1H) 6.97(d, J ═ 8.0Hz, 1H), 7.31-7.49(m, 7H), 7.48-7.56(m, 3H), 7.58-7.65(m, 1H), 7.75-7.82(m, 2H), 8.09-8.17(m, 2H)
Figure BDA0001920524400000061
Synthesis of modified paclitaxel Compound PTX-LEV-MECH: to the compounds PTX-LEV (2.50g, 2.63mmol), EMCH (1.03g, 3.94mmol) and
Figure BDA0001920524400000062
to MS (2.50g) was added 50mL of methanol. Stirred at 45 ℃ for 36 hours in the dark. After the reaction was completed, methanol was removed, and the compound PTX-LEV-MECH (2.4g, 79% yield) was obtained as a white solid by column purification, and the structure thereof was confirmed by mass spectrometry (FIG. 1).
Example four: synthesis of modified salinomycin Sail-ABA-MPBH
Figure BDA0001920524400000071
Synthesis of Compound 1: hydrazine hydrate (8.20g, 163.80mmol) was dissolved in isopropanol at 0 ℃ and di-tert-butyl dicarbonate (15.78g, 72.39mmol) was added to the solution. The temperature of the reaction solution was raised to room temperature, and the mixture was stirred for 20 minutes. The solvent was removed, the residue was dissolved in dichloromethane, the insoluble material was removed by filtration, and the organic phase was concentrated to give compound 1 as a white solid. (7.48g, 78%).1H NMR(CDCl3，400MHz)δ1.46(d，J＝4.0Hz，9H)，3.68(s，2H)，7.27(s，1H)
Synthesis of Compound 2: maleic anhydride (13.63g, 139.00mmol) was dissolved in diethyl ether, and then 4- (4-aminophenyl) butyric acid (25.00g, 139.00mmol) and 2, 6-lutidine (16.25ml) dissolved in tetrahydrofuran were added to the solution, and the reaction solution was refluxed for 30 minutes. After the reaction was completed, the solid obtained by filtration was washed with diethyl ether. The resulting solid and potassium acetate (7.42g) were dissolved in acetic anhydride, followed by heating and refluxing for 30 minutes. Cooling the reaction solution to room temperature, quenching the reaction solution by using saturated saline solution, extracting by using ethyl acetate, combining organic phases, drying, concentrating, and purifying by a column to obtain a yellow solid compound 2. (15.22g, 42%).1H NMR(CDCl3，400MHz)δ1.86-2.07(m，2H)，2.22(d，J＝4.0Hz，1H)，2.40(t，J＝8.0Hz，1H)，2.44-2.52(m，1H)，2.72(q，J＝8.0Hz，2H)，6.85(s，2H)，7.21-7.34(m，4H)
Synthesis of Compound 3: compound 2(3.57g, 13.77mmol) and 1-hydroxybenzotriazole (2.04g, 15.15mmol) were dissolved in DMF, N' -diisopropylcarbodiimide (1.91g, 15.15mmol) was added, the reaction mixture was stirred at room temperature for 20 minutes, then Compound 1(2.00g, 15.15mmol) was added, and the reaction mixture was stirred at room temperature for 18 hours. After completion of the reaction, the reaction was quenched with saturated brine, followed by extraction with ethyl acetate, and the organic phases were combined, dried, concentrated, and purified by column chromatography to obtain compound 3(2.43g, 47%) as a yellow liquid.1H NMR(CDCl3，400MHz)δ1.47(s，9H)，2.01(d，J＝8.0Hz，2H)，2.23(t，J＝8.0Hz，2H)，2.70(t，J＝8.0Hz，2H)，6.85(s，2H)，7.19-7.37(m，4H)
The compound MPBH-HCl: dissolving Compound 3To 1,4 dioxane was added 4N hydrochloric acid (22.65ml), and the reaction solution was stirred overnight at room temperature. After the reaction was completed, the solvent was removed to obtain MPBH-HCl (1.50g, 91%) as a yellow solid.1H NMR(CDCl3，400MHz)δ0.92-0.99(m，2H)，1.78-1.92(m，2H)，2.16-2.27(m，2H)，2.53-2.65(m，2H)，3.62(s，2H)，6.87(s，2H)，7.14(d，J＝4.0Hz，2H)，7.28(d，J＝4.0Hz，2H)
Figure BDA0001920524400000081
Compound Sail-ABA: 1-hydroxybenzotriazole (2.05g, 15.17mmol) was dissolved in anhydrous DMF at 0 ℃ followed by the addition of salinomycin (9.50g, 12.67mmol), (4- (1, 3-dioxolanyl) phenyl) methylamine (3.40g, 18.97mmol) and N, N' -diisopropylcarbodiimide (2.39g, 18.97mmol) in succession dissolved in anhydrous DCM, the reaction was stirred at 0 ℃ for 24 h, after which the reaction was warmed to room temperature and stirred for 24 h. After the reaction is finished, quenching the reaction liquid by using saturated saline solution, then extracting by using dichloromethane, combining organic phases, drying, concentrating, and purifying by using a column to obtain a white solid. The obtained white solid was dissolved in tetrahydrofuran, and then 2N hydrochloric acid (22.72ml) was added to stir the reaction solution at room temperature for 6 hours. After completion of the reaction, sodium hydrogencarbonate solution was added to the reaction mixture, followed by extraction with methylene chloride, and the organic phases were combined, dried, concentrated, and purified by column chromatography to obtain Sail-ABA (6.68g, 61%) as a white solid.1H NMR(CDCl3，400MHz)δ0.75-0.81(m，4H)，0.81-0.86(m，5H)，0.86-0.91(m，6H)，0.91-0.99(m，8H)，1.14(s，3H)，1.19-1.27(m，8H)，1.46-1.55(m，4H)，1.80-1.88(m，5H)，2.02-2.06(m，1H)，2.18-2.45(m，2H)，2.62-2.73(m，1H)，2.74-3.13(m，8H)，3.37-3.47(m，1H)，3.53(s，1H)，3.60-3.84(m，6H)，3.85-3.95(m，1H)，4.03-4.20(m，3H)，4.43-4.59(m，1H)，4.63-4.78(m，1H)，6.23-6.29(m，1H)，6.63-6.77(m，1H)，7.14(d，J＝4.0Hz，1H)，7.52(d，J＝8.0Hz，2H)，7.81(d，J＝8.0Hz，2H)，9.95(s，1H)
The compound Sail-ABA-MPBH: under the protection of argon gas, the method comprises the following steps of,a mixed solvent of isopropanol/propanol (v/v ═ 1:1) was added to a reaction flask containing Sail-ABA (compound 1.70g, 1.96mmol), MPBH-HCl (compound 0.91g, 2.97mmol) and molecular sieve (1g), and the reaction mixture was reacted at 40 ℃ overnight. After completion of the reaction, the reaction mixture was filtered, concentrated, and purified by column chromatography to obtain Sail-ABA-MPBH (0.90g, 41%) as a yellow solid.1H NMR(CDCl3，400MHz)δ0.76-0.82(m，4H)，0.82-0.87(m，6H)，0.87-0.93(m，8H)，0.93-1.00(m，7H)，1.14(s，3H)，1.19-1.27(m，11H)，2.01-2.07(m，5H)，2.17-2.36(m，3H)，2.63-2.77(m，6H)，2.77-3.13(m，8H)，3.37-3.53(m，2H)，3.53-3.63(m，1H)，3.64-3.74(m，2H)，3.75-3.84(m，2H)，3.86-3.97(m，1H)，4.05-4.19(m，3H)，4.22-4.32(m，1H)，4.36-4.51(m，1H)，4.56-4.73(m，1H)，6.20-6.32(m，1H)，6.45-6.60(m，1H)，6.81-6.90(m，2H)，7.05-7.13(m，1H)，7.20-7.28(m，2H)，7.30-7.41(m，4H)，7.45-7.54(m，2H)，7.69(s，1H)，9.38-9.62(m，1H)
Example five: preparation of polypeptide-modified paclitaxel ABD-iTEP70- (GGGGC)8-PTX-LEV-MECH nanoparticles
1. ABD-iTEP70- (GGGGC) 817 mg was added to 5ml of buffer (0.1M NaPO)41mM EDTA PH 7.0) dissolved with stirring;
2. taking 75mg of thiol reducing agent TCEP, using 500ul of buffer solution and adding NaHCO3Adjusting the pH value to 7.0;
3. adding the solution obtained in the step (2) into the solution obtained in the step (1), and stirring for reacting for 2 hours;
4. putting the solution obtained in the step (3) into an ultrafiltration tube (with molecular weight of 1W) for ultrafiltration (at 4 ℃, 5500rpm, 30 minutes), carrying out ultrafiltration with 8ml of reaction liquid each time for 5 times, finally dissolving with 2ml of buffer solution, and putting into a reaction bottle;
5. dissolving PTX-LEV-MECH 2mg in 2ml DMF (dimethylformamide);
6. slowly adding the solution obtained in the step 5 into the solution obtained in the step 4, reacting overnight, centrifuging (10 ℃, 12000rpm, 10 minutes), and taking a supernatant; adding PBS buffer solution to total volume of 8ml, adding 2ml of acetonitrile, and ultrafiltering in ultrafiltration tube (tube size 1W) (4 deg.C, 5500rpm, 30 min); ultrafiltering with (PBS: acetonitrile: 8: 2) 8ml each time for 2 times; then using PBS for ultrafiltration, using 8ml each time, 3 times, finally using 2ml PBS for dissolution and dispersion to obtain ABD-iTEP70- (G4C)8-PTX-LEV-MECH nano particles, wherein the laser scattering diagram is shown in figure 3, and the average particle size is 86 nm.
Example six: the preparation process of the polypeptide-modified salinomycin ABD-iTEP-Sail-MPBH nanoparticle comprises the following steps:
1. taking polypeptide 60mgABD-iTEP- (GGGGGGC)8, dissolving with 9mL PBS, and adding EDTA (10mg/mL)167 uL;
2. dissolving TCEP 75mg in 500uL PBS buffer solution;
3. adding the step 2 into the solution of the step 1, and then adding NaHCO380mg, the pH value is adjusted to 7, the mixture is stirred and reacted for 1 hour at room temperature, argon is introduced for protection, and 16.7mL of acetonitrile is added;
4. dissolving 20mg of Sail-ABA-MPBH in 3.3mL of acetonitrile;
5. adding the step 4 into the step 3, continuing to react overnight, introducing argon for protection, after the reaction is finished, pumping out acetonitrile (about 30min), centrifuging (4 ℃, 16000rpm, 15min), and reserving supernatant;
6. the supernatant was ultrafiltered with 10000 ultrafilter tube and washed with PBS 5 times (4 deg.C, 5500rpm, 30min), each time adding PBS to 10mL, and mixing by blowing to obtain dispersed ABD-iTEP-Sail-MPBH nanoparticle solution with laser scattering pattern as shown in FIG. 4 and average particle diameter below 100 nm.
In conclusion, the drug-loaded nanoparticle based on the elastic targeting polypeptide provided by the invention has the advantages that the particle size is below 100nm, the dispersity is small, the drug-loading rate is high, the drug-loaded nanoparticle can be efficiently combined with in-vivo albumin after injection administration, the albumin receptor gp60 mediates the drug to enter vascular endothelial cells, the envelope transports the drug out of blood vessels and gathers around tumor cells through endocytosis, the drug is combined with acidic cysteine (SPARC) secreted by the tumor cells specifically, the drug is released in a concentrated manner in an acidic environment, breast cancer in-situ is killed in a targeted manner, and meanwhile, the toxic and side effects of the drug on the whole body are reduced; and simultaneously, the ABD-iTEP-SaliNPs and ABD-iTEP-PTX NPs linked by the albumin are easier to deliver drugs through a lymphatic system, and the drug concentration in lymph nodes is higher, so that the targeted killing of lymph metastasis cancer is facilitated, and the lymph node cancer metastasis is prevented and treated.
Further, as shown in fig. 5, the two nanoparticles of the present invention have synergistic effect in breast cancer treatment, expanding and improving the efficacy of breast cancer treatment. Thereby reducing the occurrence of breast cancer cell lymphatic metastasis and reducing the occurrence of serious complications such as iatrogenic lymphedema and the like caused by lymph node cleaning operation.
Furthermore, the nanoparticle has strong binding capacity with serum albumin, the targeted lymph node enrichment capacity is improved by 3 times, and the local CTL immune response capacity is improved; by using the ABD-iTEP carrier, on one hand, the 5' end of the ABD-iTEP carrier can have super strong binding capacity with albumin, so that the special function of the albumin carrier is realized, the albumin carrier becomes a nano-scale carrier, the EPR effect of nano-drugs is realized, and the tumor is treated in a targeted manner; on the other hand, the 3' end of the polypeptide can be designed to be combined with small molecular substances (anti-cancer drugs or immunomodulatory peptides) to realize the targeted delivery function.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like made within the spirit and principle of the present invention should be included in the scope of the present invention.

Claims (4)

1. A drug-loaded nanoparticle based on elastic targeting polypeptide is characterized in that: the modified taxol peptide is prepared from an elastic targeting polypeptide and a modified drug, wherein the modified drug is modified taxol PTX-LEV-MECH or modified salinomycin Sail-ABA-MPBH.
Figure FDA0001920524390000011
2. The drug-loaded nanoparticle based on the elastic targeting polypeptide of claim 1, wherein: the elastic targeting polypeptide is prepared from ABD and iTEP, and the amino acid sequence of ABD is LAEAKVLANRELDKYGVSDFYKRLINKAKTVEGVEALKLHILAALP, iTEP and the amino acid sequence of ABD is GAGVPG.
3. A preparation method of drug-loaded nanoparticles based on elastic targeting polypeptide is characterized by comprising the following steps: adding elastic targeting polypeptide into a buffer solution for dissolving, adding tris (2-carboxyethyl) phosphine TCEP dissolved in the buffer solution, adjusting the pH value to 7, stirring and reacting for 1h at room temperature, introducing argon for protection, adding a modified drug dissolved in a solvent, reacting overnight, introducing argon for protection, after the reaction is finished, performing suction filtration and centrifugation, and reserving supernatant for ultrafiltration, washing and dispersing to obtain the nano-particles.
4. The drug-loaded nanoparticle based on the elastic targeting polypeptide in claim 1 or 2 is used for preventing and treating breast cancer and lymph node metastatic cancer thereof.
CN201811598471.1A 2018-12-20 2018-12-20 Drug-loaded nanoparticle based on elastic targeting polypeptide and preparation method and application thereof Pending CN111346235A (en)

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CN112999160A (en) * 2021-01-12 2021-06-22 海南医学院第一附属医院 Nano-drug based on copper-doped metal hydroxide co-loaded 5-fluorouracil and albumin-bound paclitaxel, and method and application thereof
CN113521301A (en) * 2021-05-17 2021-10-22 海南医学院第一附属医院 LDH (layered double hydroxide) nanoparticle-based drug for loading salinomycin and albumin paclitaxel together as well as preparation method and application thereof

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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111944061A (en) * 2020-08-10 2020-11-17 广东药科大学 Glucagon-like peptide-1 analogue monomer, dimer and application thereof
CN111944061B (en) * 2020-08-10 2023-03-10 广东药科大学 Glucagon-like peptide-1 analogue monomer, dimer and application thereof
CN112999160A (en) * 2021-01-12 2021-06-22 海南医学院第一附属医院 Nano-drug based on copper-doped metal hydroxide co-loaded 5-fluorouracil and albumin-bound paclitaxel, and method and application thereof
CN112999160B (en) * 2021-01-12 2022-06-24 海南医学院第一附属医院 Nano-drug based on copper-doped metal hydroxide co-loaded 5-fluorouracil and albumin-bound paclitaxel, and method and application thereof
CN113521301A (en) * 2021-05-17 2021-10-22 海南医学院第一附属医院 LDH (layered double hydroxide) nanoparticle-based drug for loading salinomycin and albumin paclitaxel together as well as preparation method and application thereof

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