阅读说明：本技术 喷他脒在制备抑制pd1和pd-l1蛋白之间相互作用药物中的应用 (Application of pentamidine in preparing medicine for inhibiting interaction between PD1 and PD-L1 protein ) 是由 董子钢 李美贤 顾庭轩 田雪利 于 2019-04-10 设计创作，主要内容包括：喷他脒在制备抑制PD1和PD-L1蛋白之间相互作用药物中的应用,属于医药领域。本发明经研究发现：Pentamidine可以作为PD1/PD-L1蛋白相互作用的抑制剂,其中,PD1浓度为10μg/ml,PD-L1浓度为10μg/ml,喷他脒的浓度为0.5-4μM。PD1/PD-L1信号通路与免疫应答、肿瘤免疫逃逸密切相关,抑制PD1/PD-L1信号通路的异常激活能够起到预防及治疗肿瘤的作用。本发明的Pentamidine能够抑制PD1/PD-L1蛋白质之间相互作用。(An application of pentamidine in preparing a medicament for inhibiting interaction between PD1 and PD-L1 protein, which belongs to the field of medicines. The research of the invention finds that: pentamidine can be used as an inhibitor of PD1/PD-L1 protein interaction, wherein the concentration of PD1 is 10 μ g/ml, the concentration of PD-L1 is 10 μ g/ml, and the concentration of Pentamidine is 0.5-4 μ M. The PD1/PD-L1 signal channel is closely related to immune response and tumor immune escape, and the effect of preventing and treating tumors can be achieved by inhibiting abnormal activation of the PD1/PD-L1 signal channel. The Pentamidine provided by the invention can inhibit the interaction between PD1/PD-L1 proteins.)

1. Use of pentamidine in the manufacture of an immunotherapeutic agent characterized as an inhibitor of the interaction between PD1 and PD-L1 protein.

2. The use of pentamidine in the preparation of an immunotherapeutic agent according to claim 1 where the PD1 concentration is 10 μ g/ml, the PD-L1 concentration is 10 μ g/ml and the pentamidine concentration is 0.5-4 μ M.

3. Use of pentamidine in the preparation of an immunotherapeutic agent according to claim 1 to exert an immune-enhancing action by enhancing the killing ability of T cells against cancer cells.

4. The use of pentamidine in the preparation of an immunotherapeutic agent according to claim 3, wherein the cancer cells are lung cancer cells.

5. Use of pentamidine in the manufacture of an immunotherapeutic agent according to claim 4, characterized by a number of cancer cells of 1 x 10⁵Pentamidine concentration was 0.5-4 μ M per ml.

Technical Field

The invention belongs to the field of medicines, and particularly relates to application of pentamidine in preparing a medicine for inhibiting interaction between PD1 and PD-L1 protein.

Background

Current approaches to the treatment of immunotherapy consist mainly of two major components, immunopotentiation, which is used to enhance processes thought to play a key role in the immune process of tumors, mainly active immunotherapy and passive immunotherapy. Active immunotherapy includes antibody-targeted therapy, tumor vaccines, cytokine therapy, adoptive immune cell therapy, and CAR-T cell therapy. The therapy directly kills or enhances the killing effect of tumor cells by using effector cells of the immune system and corresponding effector molecules. The immune response is improved to a higher and stronger level so as to achieve the aim of killing tumor cells.

The second step of enhancing immunity can be achieved by regulating endogenous cell signals or related mechanisms of immune activation and inhibition, and the corresponding specific steps of enhancing immune response are called passive immunotherapy, such as enhancing the process of uptake and processing of antigens and presenting to T cells by antigen presenting cells, and the drugs comprise tumor vaccines aiming at tumor antigens and adjuvants for enhancing immunity, and in addition, the corresponding purposes can be achieved by additional activation of certain immune enhancing pathways, and the drugs comprise type I interferon, agonists of Toll-like receptors and agonists of STING pathway, so as to enhance the activity of antigen presenting cells. Yet another class of drugs, including dendritic cell vaccines and monoclonal antibodies against the cell-effector T lymphocyte antigen-4 (CTLA-4), even adoptive immunotherapy, which expands and activates tumor-infiltrating T cells in vitro and is transfused, is immunopotentiating therapy that enhances the process of initial T cell activation and expansion and enhances the killing ability of effector T cells.

Disclosure of Invention

The invention aims to overcome the defects of the prior art and provides application of Pentamidine (Pentamidine) in a medicine for inhibiting interaction between PD1 and PD-L1 protein.

Pentamidine is a small molecule compound of the formula: c₁₉H₂₄N₄O₂Molecular weight: 340.427 Pentamidine, 100-33-4, 4' - (Pentane-1,5-diylbis (oxy)) dibenzimidamide. FDA approvalFor first-line treatment of visceral leishmaniasis (kala-azar) and pneumocystis carinii, commercially available products are available directly.

The invention provides a natural compound Pentamidine used as an inhibitor for the action between PD1 and PD-L1 protein.

Specifically, the pentamidine is applied to the preparation of the immunotherapy medicament, the concentration of PD1 is 10 mug/ml, the concentration of PD-L1 is 10 mug/ml, and the concentration of pentamidine is 0.5-4 muM.

Further, the immune enhancement effect is exerted by enhancing the killing ability of the T cells to cancer cells. The cancer cell is a lung cancer cell. Number of cancer cells 1 x 10⁵Pentamidine concentration was 0.5-4 μ M per ml.

The invention discovers that: the application of Pentamidine in preparing the medicament for inhibiting the PD1/PD-L1 signal channel is to find the inhibition effect of Pentamidine on the PD1/PD-L1 signal channel, and the suitable concentration of the Pentamidine for inhibiting the Pentamidine is as follows: 0.5-4 μ M.

The research of the invention finds that: pentamidine can be used as an inhibitor of the interaction of PD1/PD-L1 protein. The PD1/PD-L1 signal channel is closely related to immune response and tumor immune escape, and the effect of preventing and treating tumors can be achieved by inhibiting abnormal activation of the PD1/PD-L1 signal channel. The Pentamidine provided by the invention can inhibit the interaction between PD1/PD-L1 proteins.

Drawings

FIG. 1 and Table 1 show the ability of Pentamidine to bind human PD1 and PD-L1 in vitro, wherein Pentamidine is capable of binding human PD-L1 with a KD of 3.498 x 10^-4Can bind murine PD-L1 with KD of 1.588 x 10^-4And no significant binding to human and murine PD-1: in the figure, a commercial PD1/PD-L1 inhibitor is used as a control group, and the Pentamidine is tested for the capability of combining with human PD1 and PD-L1 in vitro under different concentration gradients;

FIGS. 2 and 3 show that Pentamidine enhances the ability of primary T cells to kill tumor cells, wherein Pentamidine in FIG. 2 promotes primary T cells to kill non-small cell lung cancer cells H1975 at a concentration of 1. mu.M; the graph shows the tumor cell death ratio at different time points with different concentrations of the drug;

FIG. 4 is a graph of cell viability of H1975 cells treated with different concentrations of Pentamidine for different times.

Detailed Description

The technical solution of the present invention is further described in detail with reference to the following examples, but the scope of the present invention is not limited thereto.