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a containing 131 i-labeled caerin1.1 polypeptide and its application

A technology of labeling and polypeptide solution, applied in the biological field, can solve the problems of progression-free survival, patient difficulty, etc., and achieve the effects of low cost, high labeling rate, and simple labeling process

Active Publication Date: 2020-10-02
ZHONG AO BIOMEDICAL TECH (GUANGDONG) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the current benefits of targeted drugs only improve the progression free survival (PFS) of patients, which can shrink tumors to a certain extent, but no studies of targeted drugs have proven that they can prolong the overall survival of patients At the same time, the above methods all have different degrees of adverse reactions. Once patients are enrolled in clinical trials, how to weigh the risks and benefits will bring difficulties to the selection of patients and the grasp of indications.

Method used

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  • a containing <sup>131</sup> i-labeled caerin1.1 polypeptide and its application
  • a containing <sup>131</sup> i-labeled caerin1.1 polypeptide and its application
  • a containing <sup>131</sup> i-labeled caerin1.1 polypeptide and its application

Examples

Experimental program
Comparison scheme
Effect test

experiment example 1

[0033] Experimental example 1 cell line and cell culture

[0034] B-CPAP human thyroid cancer cell B-CPAP (papillary) and CAL-62 human thyroid cancer cell CAL-62 (undifferentiated), these two types of cells are provided by the Chinese Academy of Sciences Stem Cell Bank, and the two types of cells are cultured according to the method in the product manual Tie. Wherein, B-CPAP cell culture fluid formula: 87% RPMI Medium 1640 (GIBCO), 10% heat-inactivated fetal bovine serum from New Zealand, 0.1% Penicillin-Streptomycin, Liquid (GIBCO), 1% MEM Non-Essential Amino Acids (NEAA, GIBCO), 1% Glutamax (GIBCO), 1% Sodium Pyruvate (GIBCO); CAL-62 cell culture medium formula: 90% DMEM Mediun (GIBCO), 10% from New Zealand % heat-inactivated fetal bovine serum, 0.1% Penicillin-Streptomycin, Liquid (GIBCO); both cells were stored at 37°C, 5% CO 2 cultured in an incubator.

experiment example 2

[0035] Experimental Example 2 Caerin1.1 Polypeptide Cell Proliferation Experiment

[0036] The activity of B-CPAP cells and CAL-62 cells was induced by 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfonic acid Benzene)-2H-tetrazolium monosodium salt (CCK-8, DOJINDO) detection, operate according to the instructions: B-CPAP cells in the logarithmic growth phase, CAL-62 cells in 5 × 10 per well 3 / 100μL cells were seeded in 96-well cell culture plates, placed at 37°C, 5% CO 2 Under culture for 24 hours, until the cell aggregation rate is about 60-70%. The Caerin1.1 polypeptide and control peptide P3 were added to each well in groups according to different concentrations so that the final concentration was in the range of 0-17.5 μg / mL (0, 1.25, 2.5, 5, 7.5, 10, 12.5, 15, 17.5 μg / mL), at the same time set the zero hole, each group of 4 auxiliary holes, and at 37 ℃, 5% CO 2 Conditioned for 24h. Add 10 μL CCK-8 to each well and incubate for 4-6 hours to stop the exper...

experiment example 3

[0038] Experimental Example 3 Caerin1.1 Peptide IC 50 determination

[0039] Logarithmically grown B-CPAP cells and CAL-62 cells were divided into 5×10 3 / 100μL cells were seeded in 96-well cell culture plates at 37°C, 5% CO 2 The cells were cultured for 24 h under the conditions of the above conditions, and they were allowed to adhere to the wall. Add the Caerin1.1 polypeptide to each well according to the concentration gradient of the multiple relationship (0, 1.25, 2.5, 5, 10, 20 μg / mL), and set the zero adjustment well at the same time. %CO 2 Conditioned for 24h. Add 10 μL CCK-8 to each well and incubate for 4-6 hours to stop the experiment. Cell viability was determined by measuring the absorbance (OD) at 450 nm using an enzyme-linked immunosorbent assay. Calculate IC with GraphPad software 50 (half inhibitory concentration).

[0040] see results image 3 , it can be seen that the IC of B-CPAP 50 = 4.038 μg / mL, IC of CAL-62 50 =9.856 μg / mL. Caerin1.1 polypepti...

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Abstract

The invention belongs to the technical field of biology, and particularly relates to a Caierin 1.1 polypeptide containing a 131I label and an application of the Caierin 1.1 polypeptide. The 131I-labeled Caerin 1.1 polypeptide provided by the invention is labeled by adopting a chloramine-T iodine direct labeling method, and the method has high labeling rate and strong stability. According to the 131I-labeled Caerin 1.1 polypeptide provided by the invention, the characteristic that a Caerin 1.1 polypeptide has specificity, enters a tumor cell through membrane receptor binding, and can inhibit growth and proliferation of the tumor cell is adopted; and after 131I-Caerin 1.1 specifically enters the tumor cell, the 131I-labeled Caerin 1.1 polypeptide can be used for emitting gamma rays and for tumor imaging, and the 131I can be used to emit beta rays for intra-tumor irradiation treatment, so that integration of tumor diagnosis and treatment and visualization of a tumor treatment process arerealized, the tumor treatment effect is improved, and tumor diagnosis and treatment cost are reduced.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a 131 I-labeled Caerin1.1 polypeptide and its application. Background technique [0002] Caerin polypeptides are a series of biologically active peptides extracted and identified from the glandular secretions of the back skin of tree frogs (Litoria genus) in coastal rainforests in Australia. Unified named: Caerin1.1 ~ Caerin1.9). The Caerin1.1 polypeptide (F1 polypeptide) used in this study for the first time is a biologically active targeting polypeptide that does not depend on NIS and can specifically bind to tumor cell membrane receptors and enter tumor cells. The growth and proliferation of various tumor cells (such as: thyroid cancer, lung cancer, esophageal cancer, etc.), and Caerin1.1 has more significant advantages than Caerin1.9 in the in vitro inhibition test of various tumor cells (see figure 1 ), figure 1 Among them, A represents thyroid papilloma, B represents anapla...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/46C07K1/13A61K51/08A61P35/00
CPCA61K51/08A61P35/00C07K14/463
Inventor 王天放袁建伟
Owner ZHONG AO BIOMEDICAL TECH (GUANGDONG) CO LTD
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