Preparation method of breast cancer sentinel lymph node tracer agent indocyanine green-rituximab

Abstract

The invention discloses a preparation method of breast cancer sentinel lymph node tracer agent indocyanine green-rituximab. The method comprises the following steps: slowly dropping an indocyanine green solution into a rituximab solution, rapidly oscillating, mixing evenly and reacting; carrying out coupled reaction, carrying out centrifugal precipitation on a centrifuge tube with a coupled reactant, and observing presence and absence of sediments; if the sediments are generated, removing sediments at the bottom of the centrifuge tube, taking supernate and putting the supernate into a dialysis bag made of a semi-permeable membrane; immersing the dialysis bag into dialysate for sterilizing injection water to dialyze; and bonding the indocyanine green on rituximab through coupled reaction, so as to prepare a novel indocyanine green-rituximab combined tracer agent. The preparation method is simple, safe, and free of radiation; the prepared indocyanine green-rituximab can meet the requirements of developing and positioning of the sentinel lymph node.

Description

technical field [0001] The invention relates to a method for preparing a breast cancer sentinel lymph node specific tracer indocyanine green-rituximab. Background technique [0002] The sentinel lymph node is the first lymph node of breast cancer metastasis, and its pathological conditions have important guiding value for breast cancer surgery and postoperative comprehensive treatment. Sentinel lymph node biopsy is a technical method for lymph node staging of breast cancer. Accurate positioning of sentinel lymph nodes is a technical field that has been concerned in research on sentinel lymph node biopsy of breast cancer. Traditional tracers for sentinel lymph node localization include dyes, nuclides, and fluorescence, all of which are non-specific tracers. Among them, the dye method (methylene blue, patent blue and nano-carbon) locates the sentinel lymph node mainly through the preoperative dye around the primary breast tumor and (or) subcutaneously injecting the dye, and t...

AI Technical Summary

Problems solved by technology

Among them, the dye method (methylene blue, patent blue and nano-carbon) locates the sentinel lymph node mainly through the preoperative dye around the primary breast tumor and (or) subcutaneously injecting the dye, and then following the dye-stained lymphatic vessels in the axillary area for fine dissection Find the first stained lymph node to determine the specific location of the sentinel lymph node. However, this method requires superb surgical anatomy skills, and there is still a high false-negative rate of sentinel lymph node biopsy after the learning curve, which leads to sentinel lymph node biopsy. The accuracy of the method is difficult to grasp; while the nuclide method ( 99m Tc-sulfur colloid) is used to locate the sentinel lymph node by preoperatively injecting radionuclide into the glands around the primary breast tumor and (or) subcutaneously, and then use the gamma detector to detect the radioactive count during the operation, and find the lymph node with a high count according to the radioactive count , so as to determine the specific location of the sentinel lymph node (lymph node greater than 10% of the highest radioactive count), although this method of sentinel lymph node biopsy has high accuracy and good specificity, but there are risks of radionuclide contamination and radiation damage to medical staff during actual operation problems, and the sulfur colloid has not been approved by the China Food and Drug Administration, which limits its promotion and use in clinical practice; the method of locating sentinel lymph nodes by fluorescence method (indocyanine green) is similar to the dye method, mainly After the preoperative injection of a fluorescent tracer around the primary breast tumor and / or subcutaneously, the first lymph node that is positive for fluorescence imaging is found by using a near-infrared fluorescence imaging device to follow the bright lymph vessels of fluorescence imaging during surgery , so as to locate the specific location of the sentinel lymph node. However, since the fluorescent tracer molecules need to be excited by a near-infrared light source, the penetration of the fluorescent light released by the excited is only about 9mm, so it is difficult to find the deep lymphatic vessels during the biopsy , resulting in the missed detection of sentinel lymph nodes. Although the shallower lymphatic vessels can be clearly detected by fluorescence imaging, the fluorescent tracer in them leaks out together with the lymph fluid after they are disconnected, which can easily cause fluorescence in the tissues around the lymphatic vessels. This is not an ideal sentinel lymph node tracer.

In addition, current specific sentinel lymph node imaging agents 99m The preparation process of Tc-rituximab is relatively complicated, and it needs to go through steps such as antibody modification, separation after modification, antibody packaging, antibody labeling, separation and purification, etc. Exposure to radioactive substances can easily cause radiation damage to experimental technicians, surgeons, pathologists and other related personnel. In addition, the labeling rate of rituximab and radionuclides prepared by this method is sometimes only 90%, which is difficult to meet Sentinel lymph node imaging and positioning requirements may easily lead to inaccurate imaging

[0003] Indocyanine green is a traditional fluorescent tracer for sentinel lymph node biopsy of breast cancer. The light source (760nm) excites to generate fluorescence (820-830nm), the generated fluorescence can penetrate human tissue, and the imaging device can clearly and intuitively observe the drainage pathway of the lymphatic vessel and the imaging position of the sentinel lymph node, but it is in the sentinel lymph node The high imaging rate, high sensitivity and low accuracy of biopsy can easily lead to imaging of secondary lymph nodes other than sentinel lymph nodes

[0004] Currently there is a preparation method combining rituximab with indocyanine green, and there is no detailed operation steps and specific dosage instructions

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