阅读说明：本技术 铁死亡抑制剂Ferrostatin-1及其衍生物在制备药物中的应用 (Application of iron death inhibitor Ferrostatin-1 and derivatives thereof in preparation of medicines ) 是由 张晓红 李欣 邢雪双 秦双立 于 2019-10-08 设计创作，主要内容包括：本发明公开了铁死亡抑制剂Ferrostation-1及其衍生物在制备药物中的应用,属于医药技术领域,提供了Ferrostation-1及其衍生物在制备治疗骨髓型急性放射病和放疗诱导血细胞降低症药物中的应用的新用途。采用Ferrostatin-1腹腔注射致死剂量辐照的小鼠可使辐照小鼠存活率提高到60%(150天),外周血白细胞数量明显升高；骨髓粒-巨系造血祖细胞数量明显恢复；骨髓有核细胞明显恢复(HE切片)；脾脏淋巴细胞存活率明显恢复；脾脏淋巴细胞数量明显回升(HE切片),Ferrostation-1及其衍生物对治疗骨髓型急性放射病和放疗诱导血细胞降低症具有很好的效果。(The invention discloses an application of an iron death inhibitor Ferrostation-1 and derivatives thereof in preparing medicines, belongs to the technical field of medicines, and provides a new application of Ferrostation-1 and derivatives thereof in preparing medicines for treating bone marrow type acute radiation diseases and radiotherapy-induced cytopenia. The survival rate of the irradiated mice can be improved to 60 percent (150 days) by injecting the mice with lethal dose into the abdominal cavity of Ferrostatin-1, and the number of peripheral blood leucocytes is obviously increased; the number of myeloid granule-megalineage hematopoietic progenitor cells is obviously recovered; bone marrow nucleated cells were significantly restored (HE sections); the spleen lymphocyte survival rate is obviously recovered; the spleen lymphocyte number is obviously increased (HE slice), and the ferrostan-1 and the derivatives thereof have good effects on treating bone marrow type acute radiation diseases and radiotherapy-induced cytopenia.)

1. Application of iron death inhibitor Ferrostation-1 in preparing medicine for treating bone marrow type acute radiation diseases is provided.

2. Application of iron death inhibitor Ferrostation-1 in preparing medicine for treating radiotherapy induced cytopenia.

3. Application of iron death inhibitor Ferrostation-1 derivative in preparing medicine for treating bone marrow type acute radiation diseases is provided.

4. Application of iron death inhibitor Ferrostation-1 derivative in preparation of medicine for treating radiotherapy-induced cytopenia.

The invention belongs to the technical field of medicines, and particularly relates to an iron death inhibitor Ferrostatin-1 and application of a derivative thereof in preparation of a medicine.

Background artthe widespread use of nuclear technologies in the military and energy fields has greatly pushed the development of human society, but the use of these nuclear technologies also poses a serious threat to the survival of human society. According to the inquired record, at least 1054 nuclear test, 2 original bullet explosion, 7 submarine nuclear reactor accident and more than 100 nuclear power accidents occur worldwide. The most common consequence of ionizing radiation exposure in humans from nuclear weapons/radiation accidents is the development of acute radiation sickness of the bone marrow type. Acute radiation sickness of the bone marrow type is systemic radiation damage mainly caused by bone marrow tissue damage induced by acute exposure of ionizing radiation, and is characterized in that on the basis of hematopoietic tissue inhibition and destruction, pancytopenia is mainly manifested by bleeding and infection, and severe cases can cause death. The bone marrow type acute radiation diseases are divided into four types of mild (1-2 Gy), moderate (2-4 Gy), severe (4-6 Gy) and extreme severe (6-10 Gy) according to the difference of the irradiated dose. In the current state of the art of medical treatment, interventions are required and can be applied to moderate to very severe acute radiation diseases of the myeloid type. The more than moderate bone marrow type acute radiation sickness occurs in the mechanism that after the bone marrow hematopoietic stem cells are irradiated and damaged, the number of peripheral blood cells is reduced, and symptoms such as bleeding and infection are caused.

At present, no very effective medicine exists for the bone marrow type acute radiation disease and the radiotherapy-induced cytopenia, the medicines such as granulocyte colony stimulating factor (G-CSF, filgrastim), polyethylene glycol granulocyte colony stimulating factor (samustine) and antoin capsule are mainly used for auxiliary treatment clinically to recover the hematopoietic function of bone marrow hematopoietic stem cells, but the medicines cannot completely prevent the disease from progressing and curing the disease, so that the development of more efficient medicines for treating the bone marrow type acute radiation disease and the radiotherapy-induced cytopenia is very important.

Ferrostation-1 is an iron death inhibitor based on inhibition of lipid peroxidation, and it has been reported that Ferrostatin-1 can inhibit iron-dependent cancer cell death by blocking cystine transport and glutathione production, and has the structural formula:

to date, no report about the use of Ferrostation-1 in preparing medicines for treating acute radiation diseases of bone marrow type and radiotherapy-induced cytopenia exists.

Disclosure of Invention

The invention provides application of an iron death inhibitor Ferrostation-1 and derivatives thereof in preparation of medicaments, and aims to provide new application of Ferrostation-1 and derivatives thereof in preparation of medicaments for treating bone marrow type acute radiation diseases and radiotherapy-induced cytopenia.

In order to better understand the essence of the invention, the application of Ferrostation-1 and the pharmacological experiment and the result of the derivative thereof in the preparation of the medicine for treating acute radiation diseases of bone marrow type and radiation therapy induced cytopenia is explained below.

Firstly, injecting a lethal dose of irradiated mice into an abdominal cavity of Ferrostatin-1, observing the survival rate of the irradiated mice, and confirming the functions of treating bone marrow acute radiation diseases and radiotherapy-induced cytopenia of Ferrostatin-1 from the whole animal level;

then respectively observing the change of the quantity of peripheral blood leukocytes and granulocyte-megakaryocyte progenitor cells of the irradiated mice acted by the Ferrostatin-1, confirming the recovery action of the Ferrostatin-1 on the quantity of the granulocyte-megakaryocyte progenitor cells of the irradiated mice, and further confirming the result by a bone marrow HE slice;

finally, the change of the irradiated mouse spleen lymphocyte survival rate under the action of the Ferrostatin-1 is observed, and the recovery effect of the Ferrostatin-1 on the irradiated mouse spleen lymphocyte survival rate is confirmed, and the result is further confirmed by a spleen HE section.

Has the advantages that: the invention provides application of an iron death inhibitor Ferrostation-1 and derivatives thereof in preparation of medicaments, and aims to provide new application of Ferrostation-1 and derivatives thereof in preparation of medicaments for treating bone marrow type acute radiation diseases and radiotherapy-induced cytopenia. Ferrostation-1 as an iron death inhibitor can block the lipid peroxidation process mediated by iron, thereby achieving the purpose of inhibiting the occurrence of cell iron death. The survival rate of the irradiated mice can be improved to 60 percent (150 days) by injecting the mice with lethal dose into the abdominal cavity of Ferrostatin-1, and the number of peripheral blood leucocytes is obviously increased; the number of myeloid granule-megalineage hematopoietic progenitor cells is obviously recovered; bone marrow nucleated cells were significantly restored (HE sections); the spleen lymphocyte survival rate is obviously recovered; the spleen lymphocyte number is obviously increased (HE slice), and the ferrostan-1 and the derivatives thereof have good effects on treating bone marrow type acute radiation diseases and radiotherapy-induced cytopenia.

Drawings

FIG. 1 is a schematic diagram showing the effect of Ferrostatin-1 on the survival rate of gamma ray irradiated mice in the embodiment of the present invention, wherein Ferrostatin-1 is shown in the diagram;

FIG. 2 is a graph showing the effect of Ferrostation-1 on the number of peripheral blood leukocytes in mice (15 and 30 days) irradiated according to the present invention, where Ferrostatin-1;

FIG. 3 is a graph showing the effect of Ferrostation-1 on the number of peripheral blood leukocytes in an irradiated mouse (120 days) in the present example, where Ferrostatin-1;

FIG. 4 is a diagram illustrating the effect of Ferrostation-1 on the number of granulocyte-megakaryocyte progenitors in irradiated mice according to an embodiment of the present invention, wherein A is a graph of granulocyte-megakaryocyte progenitors colonies, B is a data statistics graph of granulocyte-megakaryocyte progenitors colonies, and Ferrostatin-1 is shown in the graph;

FIG. 5 is a graph of HE slices (20) of femoral bone marrow from irradiated mice (120 days) with Ferrostation-1 effect according to the example of the present invention, where Ferrostatin-1;

FIG. 6 is a graph showing the effect of Ferrostation-1 on the survival rate of splenic lymphocytes from irradiated mice (day 1) in the example of the present invention, where Ferrostatin-1;

FIG. 7 is a graph of HE sections (. times.20) of the spleen of irradiated mice (120 days) with Ferrostation-1 effect in the example of the present invention, where Ferrostatin-1 is shown.

Detailed Description

The invention is described in detail below with reference to the following figures and specific examples:

ferrostation-1 used in this example was purchased from APExBIO corporation (CAS number: 347174-05-4).

1. Effect of Ferrostatin-1 on survival of lethal dose irradiated mice

ICR mice were divided into four groups: 0Gy group, 0Gy + Ferrostatin-1 group, 10Gy group and 10Gy + Ferrostatin-1 group, wherein each group comprises 10 drugs; 0Gy and 10Gy groups contain solvent controls; co⁶⁰Mice were irradiated with gamma rays, and 3 days after irradiation, Ferrostatin-1(2 mg/kg, dissolved in 0.01% DMSO) was intraperitoneally injected, and then the survival of the irradiated mice was observed to investigate the effect of Ferrostatin-1 on the survival of mice irradiated with lethal dose.

As shown in figure 1, the survival rates of the mice in the 0Gy group and the 0Gy + Ferrostatin-1 group are not changed, which indicates that the toxic and side effects of Ferrostatin-1 are small, and as can be seen by comparing the 10Gy group with the 10Gy + Ferrostatin-1 group, the survival time of the mice in the 10Gy + Ferrostatin-1 group is prolonged, the survival rate of the mice in the 10Gy + Ferrostatin-1 group is already 0 after the 10Gy group is irradiated for 20 days, and the survival rate of the mice in the 10Gy + Ferrostatin-1 group is 60% after 150 days, so that the survival time and the survival rate of the mice can be obviously improved by the Ferrostatin-1.

2. Effect of Ferrostatin-1 on the number of peripheral blood leukocytes in mice irradiated with different doses

The ICR mice are divided into 0Gy group, 0Gy + Ferrostatin-1 group, irradiation group and irradiation + Ferrostatin-1 group, and each group comprises 6 mice; the 0Gy and irradiation groups contained solvent controls; co⁶⁰Mice were irradiated with gamma rays, and on the 3 rd day after irradiation, Ferrostatin-1(2 mg/kg, dissolved in 0.01% DMSO) was intraperitoneally injected, and the number of peripheral blood leukocytes of the mice was observed on 15 days, 30 days, and 120 days after intraperitoneal injection of the drug, respectively, to study the influence of Ferrostatin-1 on the number of peripheral blood leukocytes of the irradiated mice.

As shown in FIG. 2, the number of peripheral blood leukocytes of the mice was observed after the intraperitoneal injection of the drug for 15 days and 30 days, and it can be seen that the number of peripheral blood leukocytes of the mice irradiated 15 days and 30 days after the intraperitoneal injection of the drug was significantly increased.

As shown in FIG. 3, the number of peripheral blood leukocytes of the mice was observed 120 days after the intraperitoneal injection of the drug, and it can be seen from the figure that the number of peripheral blood leukocytes of the irradiated mice was significantly increased and approached to the number of peripheral blood leukocytes of the non-irradiated mice 120 days after the intraperitoneal injection of the drug.

3. Effect of Ferrostatin-1 on the number of myeloid-megakaryohematopoietic progenitors irradiated in mice

ICR mice were divided into four groups: 0Gy group, 0Gy + Ferrostatin-1 group, 4Gy + Ferrostatin-1 group, each group comprises 6; 0Gy and 4Gy groups contain solvent controls; co⁶⁰Irradiating a mouse by gamma rays, injecting Ferrostatin-1(2 mg/kg, dissolved in 0.01% DMSO) into the abdominal cavity on the 3 rd day after irradiation, taking the left femur of the mouse 24 hours after the intraperitoneal injection, preparing bone marrow tissue cell suspension, separating bone marrow nucleated cells from lymphocyte separating medium, inoculating the separated cells into a methyl cellulose culture medium containing granulocyte colony stimulating factors (10 ng/ml) for culturing for 14 days, observing the number of cell colonies by an inverted microscope, and researching the influence of the Ferrostatin-1 on the number of the irradiated bone marrow granulocyte-megalineage hematopoietic progenitor cells of the mouse.

FIG. 4 shows Panel A showing the granulocyte-macrophage hematopoietic progenitor cell colony, which is a colony in the circle, wherein the number of 1 colony is 30 or more cells; b, the graph is the statistics of granulocyte-megakaryocyte progenitor cell colony data, and the comparison between a 4Gy group and a 4Gy + Ferrostatin-1 group shows that the number of the bone marrow granulocyte-megakaryocyte progenitor cells of the irradiated mice is obviously recovered after the Ferrostatin-1 is injected into the abdominal cavity.

4. Effect of Ferrostatin-1 on bone marrow histomorphology of irradiated mice

ICR mice are divided into three groups: 0Gy group, 0Gy + Ferrostatin-1 group and 10Gy + Ferrostatin-1 group; group 0Gy includes solvent control; co⁶⁰Gamma-ray irradiating the mouse, and injecting Ferrostatin-1(2 mg/kg, dissolved in 0.01% DMSO) into the abdominal cavity on the 3 rd day after irradiation; after 120 days of intraperitoneal injection of the medicine, bone marrow of the mouse is taken to prepare an HE staining section, morphological change of bone marrow tissue of the mouse is observed, and the effect of Ferrostatin-1 on the irradiation of the bone marrow tissue of the mouse is researched. As shown in FIG. 5, after 120 days of Ferrostatin-1 irradiation, the bone marrow nucleated cell number of the mice is obviously recovered and is close to the level of the control group.

5. Effect of Ferrostatin-1 on spleen lymphocyte survival Rate in irradiated mice

The ICR mice are divided into a 0Gy group, a 0Gy + Ferrostatin-1 group, an irradiation group and an irradiation + Ferrostatin-1 group, and each group comprises 6 mice; 0Gy and irradiation group contain solvent control; co⁶⁰Gamma ray irradiating mouse, irradiatingAnd 3, injecting Ferrostatin-1(2 mg/kg, dissolved in 0.01% DMSO) into the abdominal cavity, taking the spleen of the mouse 24 hours after the intraperitoneal injection to prepare a spleen cell suspension, separating lymphocytes from a lymphocyte separation solution, detecting the survival rate of the lymphocytes by a CCK method, and researching the influence of the Ferrostatin-1 on the survival rate of irradiated spleen lymphocytes of the mouse.

As shown in FIG. 6, spleen lymphocyte survival rate of irradiated mice injected with Ferrostatin-1 in the intraperitoneal direction was significantly restored by comparing the 4Gy group with the 4Gy + Ferrostatin-1 group, the 8Gy group with the 8Gy + Ferrostatin-1 group.

6. Effect of Ferrostatin-1 on irradiated mouse spleen histomorphology

Male ICR mice were divided into three groups: 0Gy group, 0Gy + Ferrostatin-1 group and 10Gy + Ferrostatin-1 group; group 0Gy includes solvent control; co⁶⁰The mice are irradiated by gamma rays, the 3 rd day after irradiation is intraperitoneally injected with Ferrostatin-1(2 mg/kg, dissolved in 0.01% DMSO), the spleen of the mice is taken 120 days after the intraperitoneal injection of the drug to prepare HE stained sections, the histological change of the spleen tissue of the mice is observed, and the effect of the Ferrostatin-1 on the irradiated spleen tissue of the mice is researched. As shown in FIG. 7, the spleen myelogenous lymphocyte number of the mice is obviously recovered after the mice are irradiated by Ferrostatin-1 for 120 days, and is already close to the level of the control group.

The foregoing are only preferred embodiments of the present invention, which will aid those skilled in the art in further understanding the present invention, and are not intended to limit the invention in any way. It should be noted that various changes and modifications can be made by those skilled in the art without departing from the spirit and scope of the invention.