CN-122010900-A - Organic ionic liquid formed by mixing double medicaments and application thereof

Abstract

The invention discloses an organic ionic liquid formed by mixing double medicaments and application thereof, wherein the organic ionic liquid with different molar ratios is prepared by using Imatinib (IM) and 3-Bromopyruvate (BP), the optimal molar ratio is obtained according to in-vitro toxicity experiments and theoretical calculation on tumor cells, and the organic ionic liquid with the optimal molar ratio is packaged into an engineering bacteria exosome-liposome heterozygous vesicle (IM-BP@HV-Pep-PEG) to construct a double medicament co-delivery system. The dosage is reduced, and simultaneously, better synergistic treatment effect is obtained. Meanwhile, the outer membrane vesicle part of the carrier can reverse tumor immunity microenvironment, promote tumor cell immunogenicity death, and realize the anti-tumor effect of the adjuvant double-drug synergistic treatment of immunotherapy.

Inventors

• LIU JIAN
• WANG ZHEXIANG
• LI XINYI

Assignees

• 苏州大学

Dates

Publication Date

20260512

Application Date

20260211

Claims (10)

1. 1. The organic ionic liquid formed by mixing double medicaments is characterized in that the double medicaments are respectively organic base imatinib and organic acid 3-bromopyruvate, and the hydrophobic organic base has four nitrogen-containing cationization sites and can be ionized with hydrophilic organic acids with different equivalent weights of 1:1-4 in aqueous solution.
2. 2. A preparation method of the organic ionic liquid according to claim 1 is characterized by comprising the steps of dispersing imatinib in an organic solvent, dissolving 1-4 times of 3-bromopyruvic acid in an equal volume of the organic solvent, fully mixing the two, dropwise adding ethyl acetate, stirring to obtain a clear and transparent solution, and rotationally evaporating the organic solvent to obtain a yellow solid 3-bromopyruvate-imatinib, namely the organic ionic liquid formed by mixing the two medicaments.
3. 3. The hybridized nano vesicle material of the targeted tumor drug-resistant cell strain serving as the efficient carrier of the organic ionic liquid as claimed in claim 2, which is characterized in that the nano vesicle material is formed by compounding an engineering bacterial outer membrane vesicle and a liposome, and the organic ionic liquid can be doped in the preparation process for encapsulation.
4. 4. The nano vesicle material according to claim 2, wherein the nano vesicle material is prepared by cloning a gene encoding transferrin T12 into a plasmid vector, introducing an engineering bacterium, feeding the engineering bacterium with 6-azido-galactose, adding isopropyl beta-D-1-thiogalactonucleoside to induce T12 expression, taking a bacterial solution, centrifuging to remove thalli, concentrating by using an ultrafiltration tube, ultracentrifugating to obtain bacterial outer membrane vesicle sediment, mixing 3-bromopyruvate-imatinib Bao Fengru liposome, repeatedly pushing the liposome loaded with 3-bromopyruvate-imatinib with the outer membrane vesicle by using a liposome extruder with a polycarbonate porous membrane, collecting the obtained hybrid membrane vesicle, mixing Pep-PEG dry powder with the obtained vesicle, oscillating, and modifying to the surface of the vesicle by a click reaction.
5. 5. The hybrid nanovesicle material of claim 4, wherein the bacterium is e.
6. 6. The hybridized nano vesicle material according to claim 5, wherein the specific preparation process of the outer membrane vesicle comprises the steps of inoculating ClyA-Flag-T12 BL21 escherichia coli into LB broth culture medium, respectively adding isopropyl beta-D-1-thiogalactose nucleoside and 6-azido-galactose when the bacterial liquid OD 600 value is 0.6-0.8, and obtaining the bacterial outer membrane vesicle through centrifugation and ultracentrifugation after the culture is performed for overnight.
7. 7. The hybridized nano vesicle material according to claim 5, wherein the Pep-PEG is synthesized by dissolving mPEG-COOH, N-hydroxysuccinimide and N, N' -dicyclohexylcarbodiimide in anhydrous N, N-dimethylformamide, reacting for 20-30h, filtering with a 0.1-0.3 mu m filter head after the reaction is finished, settling with anhydrous diethyl ether to obtain white solid, and vacuum drying to obtain mPEG-NHS; Fmoc-e8XPLGLAGr c, dibenzocyclooctyne DBCO-maleimide and methylmorpholine are dissolved in N, N-dimethylformamide for reaction, mPEG-NHS is dissolved in an organic solvent, then the mixture is added into the reaction system for overnight reaction, anhydrous diethyl ether is settled after the reaction is finished, white solid is obtained, and the white solid is dried in vacuum, thus obtaining the product.
8. 8. The application of the combination of the imatinib and the 3-bromopyruvate in preparing medicaments for treating malignant tumors, wherein the malignant tumors comprise breast cancer and brain glioma.
9. 9. Use of a nanovesicle material according to claim 3 for the preparation of a medicament in the field of bio-orthogonal catalytic therapy.
10. 10. Use of the nanovesicle material of claim 3 in the preparation of a medicament for the treatment of malignant neoplasms.

Description

Organic ionic liquid formed by mixing double medicaments and application thereof Technical Field The invention relates to the design of organic ionic liquid, the preparation of bio-based vesicles, the synthesis of multifunctional PEG polymers and click chemistry, in particular to an organic ionic liquid formed by mixing double medicaments and application thereof. Background Tyrosine Kinase (TK) is one of important targets for tumor signal regulation, and can be classified into non-receptor tyrosine kinase and receptor tyrosine kinase according to the presence or absence of cell membrane receptors. Tyrosine kinase inhibitors (tyrosine kinase inhibitor, TKI) are capable of selectively blocking abnormal activation of tyrosine kinases, inhibiting a number of signal pathways associated with tumor cell growth, proliferation, differentiation, apoptosis, and have recently been playing a significant role in tumor therapy. The first generation of TKIs, imatinib (Imatinib, IM) was approved by the united states food and drug administration in 2001 for the treatment of chronic myeloid leukemia. Thereafter, TKI-type targeted drugs, such as gefitinib, erlotinib, afatinib, oxatinib, and the like, are increasingly being added to clinical use, most of which are used for cancer treatment. Although the current TKI-type targeted drugs have better therapeutic effects, drug resistance is a troublesome problem in clinical treatment, 10-40% of patients cannot use the drug due to primary drug resistance, and almost all patients acquire secondary drug resistance after long-term drug use for 1-2 years. The main action targets of imatinib are BCR-ABL fusion protein, stem cell factor receptor c-KIT and Platelet Derived Growth Factor Receptor (PDGFR), and can specifically identify the binding site of BCR-ABL protein and ATP, replace ATP and competitively inhibit BCR-ABL protein phosphorylation, thereby inhibiting phosphorylation of self and downstream Ras/Erk, PI3K/Akt, stat5, crkL, c-Myc and other channels. It has achieved a good effect in clinical treatment, however statistics show that about 25% of patients develop drug resistance after prolonged treatment with IM, withdrawal from drug resistance affects the effectiveness of subsequent treatments, and withdrawal syndrome exists. ATP binding cassette proteins (e.g., ABCB1, ABCC1, ABCG2, etc.) are a transporter located on the surface of a cell membrane that mediates increased efflux of a variety of chemotherapeutic agents. ABCB1 protein has higher ATP hydrolysis rate, and under the existence of a substrate, a plurality of inhibitors and regulators, the ATP hydrolysis rate is continuously increased to reach the level of 1 mu mol ATP/min/mg protein. In clinical conditions, ABCB1 inhibitors suffer from a lack of efficacy, significant adverse effects, probably because the drug efflux effects of ABCB1 are replaced by other transporters. Due to the abnormal growth, proliferation and invasion behavior of tumor cells, tumor microenvironments (tumor microenvironment, TME) show significant differences from normal cells, including endothelial cells, fibroblasts, immune cells and other cell types and extracellular components such as cytokines, growth factors, hormones, extracellular matrix and the like, with the most significant features including dynamic hypoxia, high concentration of redox components, elevated intracellular Adenosine Triphosphate (ATP) and lactate levels, extracellular acidosis. Transferrin receptor (TfR) is an important factor mediating iron uptake by cells by transporting iron to cells through binding to plasma glycoprotein transferrin, and transferrin receptor 1 (TfR 1) has been demonstrated to be highly expressed on the surface of various cells such as breast cancer, lung cancer, colon cancer, etc. TfR1 is clinically used as a tumor marker for early diagnosis and treatment of breast cancer. Matrix metalloproteinases (matrix metalloproteinases, MMPs) are key mediators of cell-cell and cell-extracellular matrix communication, acting to release bioactive peptides and growth factors, degrading cell surface and extracellular proteins, and can have a decisive influence on the growth and proliferation of cells. In malignant tumors, the expression and activity of MMPs vary significantly. Disclosure of Invention The invention aims to solve the key technical difficulty of the synergistic treatment of tumors by using double medicaments. There is an optimal dosage range between the drugs, but it is difficult to precisely control the molar ratio between the drugs during actual treatment. In the nano delivery system, the physical embedding method is difficult to control the drug loading rate of different drugs, and the preparation process of methods such as polymer prodrugs, host-object interactions and the like is complex and difficult to generalize. Aiming at the large-class medicine combination of hydrophobic medicines (organic alkali) and hydrophilic medicines (organic acid), the inven