Fluorescence-coupled specific sentinel node photographic developer and preparation method thereof

A sentinel lymph node and specific technology, applied in the field of fluorescence-coupled specific sentinel lymph node imaging agent and its preparation, can solve the problem that the total number of injected tracer particles is difficult to control, and the secondary lymphatic system has development, imaging and biopsy. The problem of high time is to achieve the effect of unlimited imaging and biopsy time, low injection point retention, and strong operability.

Inactive Publication Date: 2015-04-08
BEIJING CANCER HOSPITAL PEKING UNIV CANCER HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In many studies, the selected radiotracers are almost all radiocolloids that rely on lymph node phagocytosis for detection ( 99m Tc-sulfur colloid or 99m Tc-human serum albumin, etc.), the particle size is uneven, and the s...

Method used

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  • Fluorescence-coupled specific sentinel node photographic developer and preparation method thereof
  • Fluorescence-coupled specific sentinel node photographic developer and preparation method thereof

Examples

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Embodiment 1

[0025] Example 1 Preparation of Fluorescent-Coupled Specific Sentinel Lymph Node Imaging Agent (1)

[0026] 1) Purification of CD20-targeted monoclonal antibody (take rituximab as an example): Take 1.0mL of 10mg / ml CD20 antigen-targeted monoclonal antibody, add it to the pre-treated PD-10 column, and then add 0.01M pH 7.4 PBS solution, collect 1mL of monoclonal antibody. And quantify the protein by biological method, and add 0.01M PBS solution with pH 7.4 to it, dilute to 5mg / mL for use;

[0027] 2) Take 0.01M PBS solution with pH 7.4 as the buffer solution, and take the concentration as 1×10 -2 10 μL of mol / mL Cy5.5-NHS DMF solution was mixed with the above 0.1 mL rituximab solution, 1 mL of 0.1M PBS solution (pH=7.4) was added thereto, and 1 mol / L Na 2 The HPO4 solution adjusted the pH of the reaction system to 8.0, mixed well, and reacted at 4°C in the dark for 12 hours. Separation and purification by PD-10 column. A specific sentinel lymph node drug Cy5.5-SLN-F capable...

Embodiment 2

[0028] Example 2 Preparation of Fluorescent-Coupled Specific Sentinel Lymph Node Imaging Agent (2)

[0029] 1) Purification of CD20-targeting monoclonal antibody (taking rituximab as an example): Take 1.0mL of 50mg / ml CD20 antigen-targeting monoclonal antibody, add it to the pre-treated PD-10 column, and then add 0.01M PBS solution with pH 7.4, in which rituximab was collected. And quantify the protein by biological method, and add 0.1M PBS solution with pH 7.4 to it, dilute to 10mg / mL for use;

[0030] 2) Take 0.1M PBS solution with pH 7.4 as the buffer solution, and take the concentration as 1×10 -2 mol / mL Cy5.5-NHS DMF solution 10μL was mixed with the above 1.0mL rituximab solution, 1mL 0.1M PBS solution (pH=7.4) was added to it, and 1mol / L Na 2 The HPO4 solution adjusted the pH of the reaction system to 9.5, mixed well, and reacted at 4°C in the dark for 24 hours. Separation and purification by PD-10 column. A specific sentinel lymph node drug Cy5.5-SLN-F capable of in...

Embodiment 3

[0031] Example 3 Preparation of Fluorescent-Coupled Specific Sentinel Lymph Node Imaging Agent (3)

[0032] 1) Purification of CD20-targeted monoclonal antibody (take rituximab as an example): Take 1.0mL 20mg / ml CD20 antigen-targeted monoclonal antibody, add it to the pre-treated PD-10 column, and then add 0.01M PBS solution with pH 7.4, collect 0.5-1.5ml of monoclonal antibody. And carry out protein quantification by biological method, and add PBS solution to it, dilute to 7mg / mL for use;

[0033] 2) Take 0.05M PBS solution with pH 7.4 as the buffer solution, and take the concentration as 1×10 -3 mol / mL Cy5.5-NNHS DMF solution 10 μL was mixed with 1.0 mL of the above-mentioned rituximab solution, 1 mL of 0.1M PBS solution (pH=7.4) was added thereto, and 1 mol / L Na 2 HPO 4 The solution adjusted the pH of the reaction system to 8.5, mixed well, and reacted at 4°C in the dark for 18 hours. Separation and purification by PD-10 column. A specific sentinel lymph node drug Cy5....

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Abstract

The invention provides a fluorescence-coupled specific sentinel node photographic developer and a preparation method thereof and belongs to the field of preparation of biological medicines. The specific sentinel node photographic developer disclosed by the invention is formed by separating and purifying rituximab coupled with Cy5.5-N-hydroxyl pyrrolidine dione ester (Cy5.5-NHS) which is Cy5.5-SLN-F for short. The photographic developer is convenient to use, stable in performance, as long as 24 months in preservation time, above 95% in labeling rate, high in operability, applicable to noninvasive detection of an infrared-developed sentinel node, low in ingestion by a secondary lymph node, low in retention at an injection point, not limited in development and biopsy time, good in development within 30min to 24 hours after injection, as high as 100% in development rate, higher in safety, low in cost and relatively good in clinical application prospect.

Description

technical field [0001] The invention relates to the field of biopharmaceuticals, in particular to a fluorescence-coupled specific sentinel lymph node imaging agent and a preparation method thereof. Background technique [0002] Surgery is the main method for the treatment of breast cancer. Since the status of axillary lymph nodes is an important independent prognostic factor affecting the survival of breast cancer patients, axillary lymph node dissection is often used clinically as a means of diagnosis and treatment of axillary lymph node status. Axillary lymph node dissection (ALND) is the most commonly used surgical method. With the screening and early diagnosis of breast cancer, 50% to 70% of newly discovered breast cancer patients have negative postoperative pathological axillary lymph nodes, and ALND for this part of patients is overtreatment; therefore, ALND is performed for all breast cancer patients. The rationality has been questioned, and an accurate and minimally...

Claims

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Application Information

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IPC IPC(8): A61K49/00
Inventor 杨志朱华刘菲
Owner BEIJING CANCER HOSPITAL PEKING UNIV CANCER HOSPITAL
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