Nano-drug for reprogramming taurine metabolism of cancer cells by activating CDO1 and application of nano-drug in mediated tumor immune cell therapy

Abstract

The invention discloses a nano-drug for reprogramming taurine metabolism of cancer cells by activating CDO1 and application of the nano-drug in mediated tumor immune cell therapy. The CDO1 activating agent disclosed by the invention is a nano particle prepared by assembling miR-342-3p and / or MCPC2 and a biocompatible delivery carrier. Results of gene expression tests and extracellular taurine content determination experiments show that the miR-342-3p and MCPC2 nanoparticles have the capabilities of broad-spectrum activation of CDO1 and remodeling of taurine metabolism, cancer cells can be remodeled into a bioreactor to produce taurine in situ, and immune cells are activated in a tumor microenvironment; based on clinical data analysis, the nanoparticles have high medicinal values and good clinical application prospects for breast cancer, bile duct cancer, colon cancer, head and neck cancer, liver cancer, lung cancer, endometrial cancer, pancreatic cancer and the like.

Description

Technical Field

[0001] This application relates to nanomedicines that reprogram cellular taurine metabolism by activating CDO1 and their application in mediating tumor immune cell therapy, belonging to the field of biomedical technology. Background Technology

[0002] Cancer is a deadly and extremely difficult-to-treat disease, and traditional chemotherapy and surgery cannot change the current situation. Cellular immunotherapy, as a new technology, is receiving increasing attention. Currently, mRNA is expensive in cellular immunotherapy, and existing tumor cell therapies mainly rely on adoptive cell therapy, which has drawbacks such as short exogenous cell lifespan, difficulty in coping with the tumor-suppressive microenvironment, and cross-infection. Taurine is known to activate immune cells and mediate immune cell therapy in the tumor microenvironment. Taurine-related tumor immunomodulation technologies mainly revolve around direct supplementation strategies, but direct supplementation suffers from poor targeting and difficulty in concentration control. Delivering active molecules to reprogram immune cells is a recent international frontier, but existing targeted delivery strategies suffer from expensive vectors, poor practicality, and insufficient target specificity. Target regulation also suffers from insufficient pathway specificity and unknown long-term safety.

[0003] To address the shortcomings of the existing technologies, this invention develops nanomedicines that can activate the CDO1 gene in tumor cells and further reshape taurine metabolism, as well as taurine-related tumor immunomodulation technologies. Summary of the Invention

[0004] The objective of this invention is to develop a class of nanomedicines and their applications that utilize chemical reprogramming of cancer cells to produce taurine via a bioreactor, addressing the obstacles encountered in mRNA and immune cell delivery during cellular immunotherapy. These nanomedicines mediate immunotherapy within the tumor microenvironment. This invention uses MCPC2 (see Chinese Patent Application No. CN202410609485.8) and / or miR-342-3p as representative active molecules activating CDO1. The chemical structure of MCPC2 is as follows:

[0005]

[0006] To achieve the above objectives, the present invention adopts the following technical solution: This invention provides the use of miR-342-3p and / or MCPC2 nanoparticles, the uses including: 1) in the preparation of reagents or drugs that activate CDO1 to reprogram taurine metabolism in cancer cells; 2) in the preparation of drugs that mediate tumor immune cell therapy.

[0007] The miR-342-3p and / or MCPC2 nanoparticles use cancer cells or other somatic cells as a bioreactor to produce taurine in situ to activate immune cells for immunotherapy.

[0008] Preferably, the miR-342-3p nanoparticles are nanoparticles prepared by constructing miR-342-3p on a delivery carrier, wherein the delivery carrier includes a viral carrier or a non-viral carrier.

[0009] More preferably, the viral vector is selected from one or more combinations of retroviral vectors, adenovirus vectors, and adeno-associated virus vectors; the non-viral vector includes polymeric vectors (e.g., glycosylated peptides) and / or liposomes.

[0010] Preferably, the MCPC2 nanoparticles include nanoparticles obtained by self-assembly of MCPC2 or nanoparticles obtained by assembling MCPC2 with a biocompatible carrier.

[0011] More preferably, the biocompatible carrier is selected from at least one of Briji-35, dodecyl-β-D-maltodextrin, and DSPE-PEG-HA.

[0012] Preferably, the tumor includes any one of breast cancer, bile duct cancer, colon adenocarcinoma, head and neck cancer, liver cancer, lung cancer, endometrial cancer, and pancreatic cancer.

[0013] Compared with the prior art, the present invention has the following beneficial effects: This invention utilizes MCPC2 nanoparticles and / or miR-342-3p nanoparticles to activate CDO1 and reprogram the taurine metabolism pathway in cancer cells. Using cancer cells as a bioreactor, taurine is secreted into the tumor microenvironment to activate immune cells such as T cells, NK cells, macrophages, and neutrophils, further mediating cellular immunotherapy. These nanoparticles possess broad-spectrum anticancer activity (including against breast cancer, cholangiocarcinoma, colon adenocarcinoma, head and neck cancer, liver cancer, lung cancer, endometrial cancer, and pancreatic cancer). Furthermore, this invention is less expensive than mRNA delivery and safer and more convenient than cell injection or reinjection, facilitating the verification of clinical efficacy and clinical application. Attached Figure Description

[0014] Figure 1 Transmission electron microscopy (TEM) images (A) and (B) of MCPC2 assembled into nanoparticles in aqueous solution, and TEM images (C) and (D) of MCPC2 assembled with the biocompatible carrier Brij-35 in aqueous solution. Figure 2 Electron microscopy image of miR-342-3p mimic nanoparticles prepared by compression of glycosylated nanopeptides; Figure 3miR-342-3p mimic activates CDO1 in liver cancer cells, and MCPC2 activates CDO1 in liver cancer and endometrial cancer; Figure 4 Figure 1 shows the results of miR-342-3p mimic remodeling taurine metabolism in liver cancer cells and MCPC2 remodeling taurine metabolism in liver cancer and endometrial cancer. Figure 5 This is a graph showing the expression analysis of the CDO1 gene in breast cancer, bile duct cancer, colon cancer, head and neck cancer, liver cancer, lung cancer, and endometrial cancer in the TCGA database. Figure 6 This image shows the results of CDO1 overexpression activating tumor-associated immune cells such as NK cells, macrophages, T cells, and neutrophils in various cancers, based on the TIMER3 database. A represents changes in NK cell infiltration levels in pancreatic cancer; B represents changes in neutrophil infiltration levels in pancreatic cancer; C represents changes in NK cell infiltration levels in endometrial cancer; D represents changes in NK cell infiltration levels in liver cancer; E represents changes in M1 macrophage infiltration levels in colon cancer; F represents changes in neutrophil infiltration levels in colon cancer; and G represents changes in CD8+ in breast cancer. + Changes in T cell infiltration levels; H represents changes in macrophage M1 infiltration levels in breast cancer. Detailed Implementation

[0015] To make the present invention more apparent and understandable, preferred embodiments are described in detail below with reference to the accompanying drawings.

[0016] Unless otherwise specified, the experimental methods used in the following examples are conventional methods; the materials and reagents used are commercially available unless otherwise specified.

[0017] Example 1: Preparation of MCPC2 nanoparticles The preparation method of MCPC2 nanoparticles includes: dissolving MCPC2 in anhydrous ethanol to prepare a 30 mg / mL ethanol solution; adding 0.5 mL of this solution dropwise to approximately 10 mL of distilled water; dialysis to remove the organic solvent; and then adjusting the volume to 15 mL to obtain the MCPC2 nanoparticle solution. To achieve stability and targeting under specific conditions, it can be assembled with one of the following biocompatible carriers: Briji-35, dodecyl-β-D-maltose (liver-targeting), and DSPE-PEG-HA (tumor-targeting). This allows for the acquisition of MCPC2-encapsulated nanoparticles for CDO1 activation and taurine metabolism reprogramming.

[0018] By transmission electron microscopy (TEM) Figure 1 A) and visual inspection ( Figure 1 Characterization by B) revealed that MCPC2 nanoparticles were successfully prepared; transmission electron microscopy (TEM) confirmed that the nanoparticles were indeed successfully fabricated. Figure 1 C) and visual inspection Figure 1 Characterization of D) revealed that MCPC2 can assemble with biocompatible carriers (represented by Briji-35) into smaller nanoparticles, demonstrating its potential for assembly with other carriers.

[0019] Example 2: Preparation of miR-342-3p mimic nanoparticles miR-342-3p is an active molecule that can broadly activate CDO1. Due to its instability in vivo, it requires a gene vector. Glycosylated synthetic peptides have good biocompatibility and designability. This example uses a glycosylated peptide, Poly(L-ASP(TET-GDL)). 20 - b -Poly(L-Phenylalanine)7 was used as a representative to compress miR-342-3p mimic to prepare nanoparticles. After optimizing the ratio, nanoparticles were successfully prepared. The preparation method is based on the electrostatic self-assembly of cationic glycosylated peptides and anionic miR-342-3p mimics. The specific preparation method is as follows: 1) Preparation of glycosylated peptide solution: Weigh 5 mg of glycosylated peptide, dissolve it in DEPC water to prepare a 1 mg / mL stock solution, filter through a 0.22 μm filter membrane, and aliquot at -20℃ for storage; 2) Preparation of miR-342-3p mimic solution: Dissolve miR-342-3p mimic in DEPC water to prepare a 5 μM stock solution, and aliquot at -20℃ for storage; 3) Mix the glycosylated peptide solution and miR-342-3p mimic solution at a ratio of 10:1 and incubate at room temperature for 30 min to form nanoparticles with uniform particle size. Figure 2 As shown.

[0020] Example 3: Broad-spectrum activation of the CDO1 gene by miR-342-3p or MCPC2 nanoparticles. Nanoparticles based on miR-342-3p or MCPC2 were prepared and used to treat BEL-7402, Hepa1-6, and Ishikawa cells. These studies showed significant activation of the CDO1 gene, such as... Figure 3 As shown.

[0021] Example 4: miR-342-3p or MCPC2 nanoparticles remodel taurine metabolism via CDO1 activation. After CDO1 was activated by nanoparticles based on miR-342-3p or MCPC2, taurine levels in the supernatant of treated BEL-7402, Hepa1-6, and KLE cells were significantly increased by ELISA. Figure 4 As shown.

[0022] Example 5: Analysis of the clinical significance of CDO1 activation A thorough analysis of the TCGA database revealed that CDO1 is silent or poorly expressed in cancers such as breast cancer, cholangiocarcinoma, colon cancer, head and neck cancer, liver cancer, lung cancer, and endometrial cancer. Therefore, activating CDO1 and remodeling taurine metabolism has broad therapeutic implications. Figure 5 As shown.

[0023] Example 6: Analysis of the regulatory effect on immune cells in the tumor environment and its clinical significance. A thorough analysis of the TIMER3 database reveals that CDO1 activation and taurine metabolic remodeling can activate T cells, NK cells, macrophages, and neutrophils in the tumor microenvironment. Therefore, this invention has broad clinical significance in immunocellular therapy. Figure 6 As shown.

[0024] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any form or substance. It should be noted that those skilled in the art can make several improvements and additions without departing from the present invention, and these improvements and additions should also be considered within the scope of protection of the present invention.

Claims

1. Uses of miR-342-3p and / or MCPC2 nanoparticles, the uses including: 1) Application in the preparation of reagents or drugs that activate CDO1 to reprogram taurine metabolism in cancer cells; 2) Application in the preparation of drugs that mediate tumor immune cell therapy. The miR-342-3p and / or MCPC2 nanoparticles use cancer cells or other somatic cells as a bioreactor to produce taurine in situ to activate immune cells for immunotherapy.

2. The use as described in claim 1, characterized in that, The miR-342-3p nanoparticles are nanoparticles constructed with miR-342-3p as the active molecule on a delivery carrier, which may include a viral carrier or a non-viral carrier.

3. The use as described in claim 2, characterized in that, The viral vector is selected from one or more combinations of retroviral vectors, adenovirus vectors, and adeno-associated virus vectors; the non-viral vectors include polymeric vectors and / or liposomes.

4. The use as described in claim 1, characterized in that, The MCPC2 nanoparticles include nanoparticles obtained by self-assembly of MCPC2 or nanoparticles obtained by assembling MCPC2 with a biocompatible carrier.

5. The use as described in claim 1, characterized in that, The biocompatible carrier is selected from at least one of Briji-35, dodecyl-β-D-maltodextrin, and DSPE-PEG-HA.

6. The use as described in any one of claims 1-5, characterized in that, The tumors include any one of the following: breast cancer, bile duct cancer, colon adenocarcinoma, head and neck cancer, liver cancer, lung cancer, endometrial cancer, and pancreatic cancer.

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