FGFR4 targeting affinity peptide and application thereof

Abstract

The invention discloses an FGFR4 targeting affinity peptide and application thereof. A tumor targeting peptide comprises six polypeptides such as YQGF-1: Ac-IIe-Asp-Pro-Asp-Gly-Tyr-Asn-His-NH2, and the like. The invention develops a series of novel high-affinity polypeptides simulating FGF19 (fibroblast growth factor 19), and the polypeptides can be used for targeting a fibroblast growth factor receptor 4 (FGFR4). Based on the principle that the YQGF-X (X = 1-6) polypeptide is specifically combined with the FGFR4 receptor, the YQGF-X polypeptide can be used for early diagnosis of FGFR4 high-expression tumors by utilizing high expression of the FGFR4 receptor in the tumors. The YQGF-X (X = 1-6) series polypeptide can be specifically combined with tumor cells, and in-vivo optical imaging and results prove that the YQGF-X (X = 1-6) series polypeptide has excellent imaging effects on various tumors including liver cancer, breast cancer, cervical cancer, colorectal cancer and the like.

Description

technical field

[0001] The invention belongs to the field of biomedical engineering, and in particular relates to an affinity peptide targeting FGFR4 and application thereof. Background technique

[0002] Malignant tumors seriously threaten human health. Early diagnosis and treatment of tumors is the key to improving the cure rate and quality of life of patients. For tumors, the current conventional imaging diagnostic techniques are mainly B-ultrasound, CT and MRI. These imaging diagnostic techniques achieve diagnostic results by showing the functional changes of tissues, and have good application value. With the continuous improvement of requirements, these traditional detection methods are gradually unable to meet the detection needs in differential diagnosis, systemic staging and early efficacy evaluation. Among many detection methods, molecular probes, as powerful tools for analytical sensing and optical imaging, can directly visualize biological analytes at the molecu...

Examples

Embodiment 1

[0048] Example 1 takes the polypeptide YQGF-1 as an example, including the following steps:

[0049] (1) Resin swelling

[0050] Add a certain amount of Rink Amide MBHA resin into the reaction column, then add an appropriate amount of dichloromethane (DCM), and blow nitrogen slightly for 10-30 minutes to fully swell the resin. The dichloromethane solution was sucked dry, washed 3 times with dimethylformamide (DMF) and drained.

[0051] (2) Remove Fmoc

[0052] Add 20% hexahydropyridine in DMF to the reaction column, and deprotect once every 5 minutes and once every 8 minutes. After the reaction, the resin was washed three times with DMF, DCM, and DMF respectively.

[0053] (3) Coupling

[0054] Accurately weigh Fmoc-IIe-OH and O-benzotriazole-tetramethyluronium hexafluorophosphate (HBTU) which is 3 times the mole number of the feed resin, completely dissolve in DMF, add N,N-diisopropyl After diethylamine (DIPEA) activates the carboxyl group, add the solution to the reacti...

Embodiment 2

[0059] Example 2 Preparation of polypeptide YQGF-X (X=2-6)

[0060] Prepare the tumor-targeting peptide YQGF-2 according to the method of Example 1, the sequence is the polypeptide shown in SEQ ID NO: 2, confirmed by mass spectrometry [M-H] - =933. Tumor targeting peptide YQGF-3, the sequence is the polypeptide shown in SEQ ID NO:3, confirmed by mass spectrometry [M-H] - =1953. Tumor targeting peptide YQGF-4, the sequence is the polypeptide shown in SEQ ID NO:4, confirmed by mass spectrometry [M-H] - =943. Tumor targeting peptide YQGF-5, the sequence is the polypeptide shown in SEQ ID NO:5, [M-H] - =933. Tumor targeting peptide YQGF-6, the sequence is the polypeptide shown in SEQ ID NO: 6, [M-H] - =933.

Embodiment 3

[0061] Example 3 Preparation of fluorescent targeting compound MPA-YQGF-1

[0062] MPA comes from an invention patent previously applied by our research group, authorized patent number: CN101440282.

[0063] (1) Dissolve 0.02mmol MPA in 200μL ultra-dry DMSO, add 3.7mg 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and 2.2mg N-hydroxysuccinyl Amine (EDCI / NHS) (molar ratio MPA:EDCI:NHS=1:1.5:1.5), react in the dark for 4 hours, and carry out carboxyl activation reaction.

[0064] (2) Take 0.02mmol of the solid-phase synthesized polypeptide YQGF-1 (X=1-6), add 0.1mmol triethylamine and 200μL ultra-dry DMSO into a 5mL reaction bottle, and react for 10min under nitrogen protection; the above reaction (1 ) solution into the reaction solution of (2), stirred at room temperature for 12 hours;

[0065] (3) After the reaction, the reaction solution was concentrated by freeze-drying, then diluted with distilled water, and separated and purified with the preparative liquid ph...

technical field

[0001] The invention belongs to the field of biomedical engineering, and in particular relates to an affinity peptide targeting FGFR4 and application thereof. Background technique

[0002] Malignant tumors seriously threaten human health. Early diagnosis and treatment of tumors is the key to improving the cure rate and quality of life of patients. For tumors, the current conventional imaging diagnostic techniques are mainly B-ultrasound, CT and MRI. These imaging diagnostic techniques achieve diagnostic results by showing the functional changes of tissues, and have good application value. With the continuous improvement of requirements, these traditional detection methods are gradually unable to meet the detection needs in differential diagnosis, systemic staging and early efficacy evaluation. Among many detection methods, molecular probes, as powerful tools for analytical sensing and optical imaging, can directly visualize biological analytes at the molecu...

Examples

Embodiment 1

[0048] Example 1 takes the polypeptide YQGF-1 as an example, including the following steps:

[0049] (1) Resin swelling

[0050] Add a certain amount of Rink Amide MBHA resin into the reaction column, then add an appropriate amount of dichloromethane (DCM), and blow nitrogen slightly for 10-30 minutes to fully swell the resin. The dichloromethane solution was sucked dry, washed 3 times with dimethylformamide (DMF) and drained.

[0051] (2) Remove Fmoc

[0052] Add 20% hexahydropyridine in DMF to the reaction column, and deprotect once every 5 minutes and once every 8 minutes. After the reaction, the resin was washed three times with DMF, DCM, and DMF respectively.

[0053] (3) Coupling

[0054] Accurately weigh Fmoc-IIe-OH and O-benzotriazole-tetramethyluronium hexafluorophosphate (HBTU) which is 3 times the mole number of the feed resin, completely dissolve in DMF, add N,N-diisopropyl After diethylamine (DIPEA) activates the carboxyl group, add the solution to the reacti...

Embodiment 2

[0059] Example 2 Preparation of polypeptide YQGF-X (X=2-6)

[0060] Prepare the tumor-targeting peptide YQGF-2 according to the method of Example 1, the sequence is the polypeptide shown in SEQ ID NO: 2, confirmed by mass spectrometry [M-H] - =933. Tumor targeting peptide YQGF-3, the sequence is the polypeptide shown in SEQ ID NO:3, confirmed by mass spectrometry [M-H] - =1953. Tumor targeting peptide YQGF-4, the sequence is the polypeptide shown in SEQ ID NO:4, confirmed by mass spectrometry [M-H] - =943. Tumor targeting peptide YQGF-5, the sequence is the polypeptide shown in SEQ ID NO:5, [M-H] - =933. Tumor targeting peptide YQGF-6, the sequence is the polypeptide shown in SEQ ID NO: 6, [M-H] - =933.

Embodiment 3

[0061] Example 3 Preparation of fluorescent targeting compound MPA-YQGF-1

[0062] MPA comes from an invention patent previously applied by our research group, authorized patent number: CN101440282.

[0063] (1) Dissolve 0.02mmol MPA in 200μL ultra-dry DMSO, add 3.7mg 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and 2.2mg N-hydroxysuccinyl Amine (EDCI / NHS) (molar ratio MPA:EDCI:NHS=1:1.5:1.5), react in the dark for 4 hours, and carry out carboxyl activation reaction.

[0064] (2) Take 0.02mmol of the solid-phase synthesized polypeptide YQGF-1 (X=1-6), add 0.1mmol triethylamine and 200μL ultra-dry DMSO into a 5mL reaction bottle, and react for 10min under nitrogen protection; the above reaction (1 ) solution into the reaction solution of (2), stirred at room temperature for 12 hours;

[0065] (3) After the reaction, the reaction solution was concentrated by freeze-drying, then diluted with distilled water, and separated and purified with the preparative liquid ph...

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