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Method and medicament for inhibiting generation of neonatal lymphatic vessel

An inhibitor, selected technology, applied in the direction of drug combination, chemical equipment and methods, antipyretics, etc., can solve ambiguous problems

Inactive Publication Date: 2014-04-23
TSINGHUA UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Whether chemokines, which are abundantly expressed in tumor tissues, especially under hypoxic conditions, can recruit lymphatic endothelial cells activated by growth factors, and thus participate in the regulation of neoplastic lymphangiogenesis is still unclear.

Method used

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  • Method and medicament for inhibiting generation of neonatal lymphatic vessel
  • Method and medicament for inhibiting generation of neonatal lymphatic vessel
  • Method and medicament for inhibiting generation of neonatal lymphatic vessel

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Lymphatic endothelial cells express multiple chemokine receptors

[0073] experimental method

[0074] 1. Total RNA extraction and RT-PCR detection of chemokine receptor expression on lymphatic endothelial cells

[0075] The isolation and extraction of total cellular RNA was performed using TRIZOL reagent (purchased from Invitrogen) and performed according to the standard operation of the reagent manual. Primary lymphatic endothelial cells collected by centrifugation (approximately 1×10 6 ) into 1mL TRIZOL, blow and suck repeatedly 30 times with the tip of the pipette, and let stand at room temperature for 5 minutes. Centrifuge at 10,000 g at 4°C for 10 minutes, and gently aspirate the supernatant. Add 0.2 mL of chloroform, shake vigorously for about 15 seconds, and let stand at room temperature for 3 minutes. Centrifuge at 10,000g at 4°C for 15 minutes, and the sample is divided into three layers: a yellow organic phase, an intermediate layer, and a colorless aque...

Embodiment 2

[0119] Chemokine receptor CXCR4 is highly expressed specifically on VEGF-C activated lymphatic endothelial cells

[0120] experimental method

[0121] 1. RT-PCR detection of chemokine receptor expression on lymphatic endothelial cells

[0122] Fluorescence quantitative Real-Time PCR using Stratagene kit (Brilliant II QRT-PCR Master Mix), the fluorescent quantitative PCR instrument is MX3000P (purchased from Stratagene), the fluorescent dye is SYBR Green, the reaction system is 20 μL, and the number of reaction cycles is 40. The internal reference is GAPDH, and the ΔCt value is obtained according to the fluorescence image given by the instrument, and the relative Δ(ΔCt) value is calculated to calculate the relative change of the corresponding gene level.

[0123] 2. Flow cytometry verification of the expression of chemokine receptor CXCR4 on the surface of lymphatic endothelial cells induced by vascular endothelial growth factor C (VEGF-C)

[0124] The mouse primary lympha...

Embodiment 3

[0142] CXCR4 specifically and highly expressed on the surface of neonatal lymphatic vessels in vivo

[0143] experimental method

[0144] 1. Detection of the distribution of chemokine CXCR4 in lymphatic vessels in vivo by tissue immunofluorescence

[0145] Take 8 healthy C57BL / 6 mice (6-8 weeks old, female, purchased from Beijing Weitong Lihua Co., Ltd.), and divide them into two groups. Inoculate 5×10 intradermally 6 Five tumor-bearing mice of B16 / F10 mouse melanoma cells (American Type Culture Collection, ATCC). After 14 days of inoculation, the tumor tissues and paratumor axillary lymph node tissues of tumor-bearing mice, as well as the colon and lymph node tissues of normal mice were removed.

[0146] Fixed embedding. Fix the removed tissue with 4% formaldehyde solution overnight, and then wash the tissue with tap water overnight to remove formaldehyde (you can wrap the tissue block with gauze, put it in a beaker, and wash it overnight with dripping water under the ta...

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Abstract

The invention provides a method for inhibiting generation of a neonatal lymphatic vessel of a subject, comprising: application of a CXCR4 inhibitor and / or a CXCL12 inhibitor with an effective amount for treating the subject. The invention further provides a method for inhibiting transfer of tumour lymphocyte in a tumour patient, comprising: combined application of (a) a CXCR4 inhibitor and / or CXCL12 inhibitor with an effective amount for treatment, and (b) a VEGF-C inhibitor, and / or a VEGF-D, and / or a VEGFR-3 inhibitor with an effective amount for treatment.

Description

technical field [0001] The invention relates to the field of biopharmaceuticals, in particular to a method and medicine for inhibiting the generation of new lymphatic vessels. Background technique [0002] In the body, blood vessels are responsible for transporting oxygen, nutrients and other substances to various tissues, and capillaries exchange substances with surrounding tissues. The existence of blood pressure causes plasma to continuously leak from capillaries to the interstitial space, which is called interstitial fluid. The main function of the lymphatic vessels is to collect this protein-rich fluid back into the blood circulation. The lymphatic capillary at the blind end of the lymphatic vessel can absorb water, macromolecules and cells to form lymph fluid, which passes through the collecting lymphatic vessel (Collecting Lymphatic Vessel) and finally returns to the blood at the junction of the lymphatic vessel and the vein, thereby maintaining Fluid balance. Durin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00A61K45/06A61P29/00A61P35/00A61P37/06A61P35/04
CPCC07K16/22A61K2039/507C07K16/2866A61K39/3955C07K2317/76A61K2039/505C07K16/2863A61K38/179A61K38/19A61P29/00A61P35/00A61P35/04A61P37/06A61K2300/00
Inventor 罗永章卓巍贾琳付彦常国栋
Owner TSINGHUA UNIV
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