CN118021805A - Novel radiation damage protective agent and application thereof - Google Patents

Abstract

The invention provides a novel radiation injury protective agent and application thereof, wherein the novel radiation injury protective agent is SB 4, and further, the invention provides application of SB 4 or pharmaceutically acceptable salt thereof in preparation of medicines for treating or preventing diseases related to radiation injury of a subject. The inventor finds that SB 4 can effectively improve the survival rate of animals after being irradiated by a large dose of radiation.

Description

A new type of radiation damage protective agent and its application

Technical Field

The present invention relates to the field of biomedicine, and in particular to a novel radiation damage protective agent and application thereof.

Background technique

In recent years, with the progress and rapid development of science and technology, nuclear energy has been widely used in many fields such as national defense, medical care, industrial and agricultural production. However, while nuclear energy brings great benefits to today's society, it also poses a potential threat to public health. From previous nuclear power plant accidents, it can be seen that the leaked nuclear material will quickly release a large amount of highly penetrating gamma rays into the surrounding space, causing large-scale personnel injuries and environmental damage that lasts for many years. When the human body is irradiated by gamma rays, a variety of radiation-sensitive tissues and organs including the hematopoietic system, digestive tract, skin, and reproductive system will be severely damaged, causing patients to have a series of symptoms such as bleeding, infection, diarrhea, and skin ulcers. When the radiation dose accumulates to a certain level, the human body cannot achieve in situ repair and functional reconstruction of damaged tissues and organs through the regeneration of tissue stem cells, causing patients to die from severe radiation sickness. Therefore, the study of efficient and low-toxic radiation damage protection agents and radiation damage repair agents has very important application value and has become one of the core contents of radiation damage treatment research. In recent years, a number of studies have discovered and verified new drug targets for multiple nuclear radiation-induced tissue and organ damage, such as the PIKfyve inhibitor Apilimod, recombinant human thrombopoietin (rhTPO), and amifostine. However, for patients with severe or above bone marrow-type acute radiation sickness, due to combined damage to multiple tissues and organs such as the gastrointestinal tract, skin, and mucous membranes, the current conventional radiation damage drug treatment has very limited effects. Therefore, the development of drugs for the treatment of severe radiation sickness with multiple organ damage caused by high-dose radiation is still a very challenging but significant key research link.

Summary of the invention

The present invention aims to solve at least one of the technical problems existing in the prior art. To this end, one object of the present invention is to provide a compound that can be used to prepare an anti-radiation damage drug. Specifically, the present invention provides a new use of SB 4 - use in the preparation of an anti-radiation damage drug.

SB 4 (C ₁₄ H ₁₀ BrNOS, 2-((4-bromobenzyl)thio)benzo[D]oxazole, CAS: 100874-08-6) is a selective agonist of the BMP4 signaling pathway, with an EC50 value of 74nM. The compound has been shown to significantly enhance classical BMP signaling, activate SMAD-1/5/9 protein phosphorylation, and activate the expression of its downstream classical target genes such as Id1 and Id3. According to an embodiment of the present invention, the inventors unexpectedly found in the process of screening anti-radiation drugs that the administration of SB 4 has a significant effect on the survival rate of model mice subjected to lethal doses of radiation, and the compound has a good effect in preventing and treating severe radiation damage.

In view of this, the present invention proposes the use of SB 4 or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating or preventing radiation damage-related diseases in a subject. The inventor unexpectedly discovered that SB 4 can effectively prevent and resist radiation damage to mice. According to an embodiment of the present application, the radiation damage-related diseases include hematopoietic dysfunction, gastrointestinal damage, etc.

According to an embodiment of the present application, the above-mentioned use may also include at least one of the following additional technical features:

As used herein, a "pharmaceutically acceptable" ingredient is a substance that is suitable for use in humans and/or mammals without excessive adverse side effects (such as toxicity, irritation, and allergic response), ie, at a reasonable benefit/risk ratio.

According to an embodiment of the present application, the radiation damage is caused by gamma rays.

According to an embodiment of the present application, the dose of the gamma ray is 1 to 10 Gy.

According to an embodiment of the present application, the subject is a mammal.

According to an embodiment of the present application, the subject is a human, a mouse, a rat, a rabbit, a monkey, a dog or a pig.

In addition, one or more pharmaceutically acceptable carriers can be added to the above-mentioned anti-radiation damage drugs according to actual needs. Specifically, according to an embodiment of the present invention, the carrier is at least one selected from conventional diluents, absorption enhancers and surfactants in the pharmaceutical field.

The pharmaceutically acceptable carriers of the present invention include (but are not limited to): water, saline, liposomes, lipids, proteins, protein-antibody conjugates, peptides, cellulose, nanogels, or combinations thereof. The choice of carrier should match the mode of administration, which are well known to those of ordinary skill in the art.

The effective amount of the active ingredient of the present invention may vary depending on the mode of administration and the severity of the disease to be treated. The selection of the preferred effective amount can be determined by a person of ordinary skill in the art based on various factors (e.g., through clinical trials). The factors include, but are not limited to: pharmacokinetic parameters of the active ingredient such as bioavailability, metabolism, half-life, etc.; the severity of the disease to be treated, the patient's weight, the patient's immune status, the route of administration, etc. For example, several divided doses may be administered daily, or the dose may be reduced proportionally, depending on the urgency of the treatment situation.

It should be noted that the present invention was discovered and completed unexpectedly based on the inventor's hard creative labor and optimization work. Experiments have shown that SB 4 can reduce the damage caused by lethal doses of radiation to various organs of mice and promote the survival of irradiated mice. The present invention provides a new idea for the research and development of anti-radiation damage drugs, has broad application prospects, and is of immeasurable significance.

In the second aspect of the present invention, the present invention provides a drug for treating or preventing radiation damage-related diseases in a subject. According to an embodiment of the present invention, the drug comprises: SB 4 or a pharmaceutically acceptable salt thereof as an active ingredient. As mentioned above, the inventors found that SB 4 can effectively prevent and resist radiation damage to mice, and can be used to prepare drugs for radiation damage-related diseases.

According to an embodiment of the present application, the above-mentioned use may also include at least one of the following additional technical features:

According to an embodiment of the present application, the pharmaceutically acceptable salt includes at least one of the following: sulfate, phosphate, citrate, tartrate, fumarate, benzoate, adipate, succinate, methanesulfonate and maleate.

According to an embodiment of the present application, the medicine comprises an injection.

According to an embodiment of the present application, the radiation damage-related disease includes at least one of hematopoietic dysfunction and gastrointestinal damage.

According to an embodiment of the present application, the drug is suitable for administration at 2.5-10 mg/kg human body weight.

In the third aspect of the present invention, a single-dose drug is provided for treating or preventing radiation damage-related diseases in a subject, the drug comprising: SB 4 or a pharmaceutically acceptable salt thereof suitable for administration at a dose of 2.5-10 mg/kg human body weight as an active ingredient.

In addition, one or more pharmaceutically acceptable carriers can be added to the above-mentioned single-dose drug according to actual needs. Specifically, according to an embodiment of the present invention, the carrier is at least one selected from conventional diluents, absorption enhancers and surfactants in the pharmaceutical field.

The pharmaceutically acceptable carriers of the present invention include (but are not limited to): water, saline, liposomes, lipids, proteins, protein-antibody conjugates, peptides, cellulose, nanogels, or combinations thereof. The choice of carrier should match the mode of administration, which are well known to those of ordinary skill in the art.

According to an embodiment of the present application, the above-mentioned use may also include at least one of the following additional technical features:

According to an embodiment of the present application, the radiation damage-related diseases include: at least one of hematopoietic dysfunction and gastrointestinal damage.

Additional aspects and advantages of the present invention will be given in part in the following description and in part will be obvious from the following description, or will be learned through practice of the present invention.

BRIEF DESCRIPTION OF THE DRAWINGS

The above and/or additional aspects and advantages of the present invention will become apparent and easily understood from the description of the embodiments in conjunction with the following drawings, in which:

FIG1 shows an example of an embodiment of the present invention in which mice were given different doses of SB 4 by intraperitoneal injection after receiving 8.0 Gy whole-body irradiation and the effects on the survival of the irradiated mice were observed (n=5).

Detailed ways

The embodiments of the present invention are described in detail below. The embodiments described below with reference to the accompanying drawings are exemplary and are only used to explain the present invention, and should not be construed as limiting the present invention.

In this article, the chemical formula of "SB 4, i.e. [2-((4-bromobenzyl)thio)benzo[D]oxazole" is C ₁₄ H ₁₀ BrNOS, CAS No.: 100874-08-6, and its chemical structure is:

In this document, the terms “include” or “comprising” are open expressions, that is, including the contents specified in the present invention but not excluding other contents.

As used herein, the terms "optionally", "optional" or "optionally" generally mean that the subsequently described event or circumstance may but need not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not.

As used herein, the term "treatment" refers to the use of drugs to obtain the desired pharmacological and/or physiological effects. The effect may be preventive in terms of completely or partially preventing a disease or its symptoms, and/or may be therapeutic in terms of partially or completely curing a disease and/or the adverse effects caused by the disease. "Treatment" as used herein covers diseases in mammals, especially humans, and includes: (a) preventing the occurrence of a disease or condition in individuals who are susceptible to the disease but have not yet been diagnosed with the disease; (b) inhibiting the disease, such as blocking the progression of the disease; or (c) alleviating the disease, such as alleviating symptoms associated with the disease. "Treatment" as used herein covers any medication that administers a drug or compound to an individual to treat, cure, alleviate, improve, reduce or inhibit an individual's disease, including but not limited to administering a drug or single-dose drug described herein for treating or preventing a disease related to radiation damage in a subject to an individual in need.

The scheme of the present invention will be explained below in conjunction with the embodiments. It will be appreciated by those skilled in the art that the following embodiments are only used to illustrate the present invention and should not be considered as limiting the scope of the present invention. Where specific techniques or conditions are not indicated in the embodiments, the techniques or conditions described in the literature in this area or the product specifications are used. The reagents or instruments used are not indicated by the manufacturer and are all conventional products that can be obtained commercially.

Example 1 Effect of intraperitoneal injection of SB 4 on the survival rate of radiation-induced model mice

1.1 Experimental methods

(1) Animal husbandry: C57BL/6N mice, 6-8 weeks old, weighing 20-22 g, male mice, were kept in an SPF-grade environment after purchase, with a temperature of 22±2°C and a humidity of 55±5%. The bedding was sterilized by gamma ray and changed twice a week. The light/dark hours were 12 hours each day, and 5 mice were placed in each cage. They were fed with sterile standard feed and acidified water. All mice were kept in the animal room for one week after purchase to adapt to the environment before the experiment was carried out.

(2) Experimental mouse grouping: 20 mice were randomly divided into 4 groups and ear-marked, with 5 mice in each group (control group, treatment groups 1-3). 2 hours before irradiation, different doses of SB 4 solution were injected intraperitoneally into the treatment groups 1-3, and the same volume of saline was injected intraperitoneally into the control group. The dosages of different groups are shown in Table 1.

(3) Before irradiation, the mice were placed in single-cell mouse boxes to restrict their free movement. The mouse boxes were placed 3 meters away from the cobalt 60 radiation source for irradiation, with a total irradiation dose of 8.0 Gy. After irradiation, the mice were immediately returned to the mouse cages in the SPF barrier environment to move freely and eat.

(4) The survival status of the mice was observed and recorded every day. The entire observation period lasted for 28 days. The schematic results are shown in the table below.

Table 1 Dosage regimen for investigating the effects of different doses of SB 4 on the survival rate of irradiated mice

1.2 Experimental Results

C57BL/6N mice were given different concentrations of SB 4 intraperitoneally 2 hours before irradiation, and then received 8.0Gy gamma ray whole body irradiation, and the changes in the state and survival time of the mice within 28 days were observed. The results showed that, as shown in Figure 1, the mice in the control group died within 14 days after receiving 8.0Gy whole body irradiation (0/5); while the mice in the SB 4 administration group were in good condition after irradiation, and all the model mice survived after 5.0mg/kg SB 4 administration, with a survival rate of 100% (5/5), 2.5mg/kg SB 4 administration model mice survival rate 80% (4/5), 1.0mg/kg SB4 administration model mice survival rate 60% (3/5), which shows that SB 4 has a significant radiation protection effect on mice, and its radiation protection effect shows a dose-dependent effect.

Example 2 Effect of intraperitoneal injection of SB 4 on peripheral blood recovery in radiation-induced mice

2.1 Experimental methods

(1) Animal husbandry: C57BL/6N mice, male, 6-8 weeks old, weighing 20-22 g, were housed in an SPF barrier environment at a temperature of 22±2°C and a humidity of 55±5%. The bedding was sterilized by γ-rays and changed twice a week. The light/dark hours were 12 hours each day. Five mice were placed in each cage and fed with sterile standard feed and acidified water. All mice were housed in the animal room for one week to adapt to the environment before the experiment was conducted.

(2) The mice were randomly divided into two groups and ear-marked. Two hours before irradiation, the experimental group was given an intraperitoneal injection of SB 4 (5.0 mg/kg), while the control group was given an intraperitoneal injection of an equal volume of saline.

(3) Before irradiation, the mice were placed in a single-cell mouse box, which was placed 3 meters away from the γ-ray radiation source for irradiation, with a total irradiation dose of 6.5 Gy. After irradiation, the mice were immediately returned to the mouse cage in the SPF barrier environment to move freely and eat.

(4) Blood was collected from the tail vein of the mice at 0, 7, 14, 21 and 28 days after irradiation for blood test. The blood collection method was as follows: after the mice were fixed, the distal tail vein was cut with a blade, and 10 μl of blood was collected using a capillary glass tube. The blood was quickly mixed with 2 ml of diluent and then placed on a Nihon Kohden blood cell counter for detection.

2.2 Results Analysis

After irradiation with a sublethal dose of 6.5Gy, the peripheral blood red blood cells, white blood cells, and platelets of mice in the control group were significantly lower than the normal physiological state, reaching the lowest point 10-14 days after irradiation, and then the number of red blood cells, white blood cells, and platelets gradually recovered, reaching normal levels 28 days after irradiation. Compared with the irradiated control group, the peripheral blood blood count recovery time of irradiated mice after intraperitoneal administration of 5mg/kg SB 4 was significantly earlier than that of the control group, and the number of white blood cells, platelets, and red blood cells was significantly higher than that of the control group, indicating that the administration of SB 4 can effectively reduce the damage caused by radiation to mice, and this small molecule compound can promote the repair ability of hematopoietic damage in irradiated mice, and has the potential to treat severe bone marrow radiation sickness.

Example 3 Effect of a single injection of SB 4 on hematopoietic stem/progenitor cells in the bone marrow of mice after 6.5 Gy whole-body irradiation

3.1 Experimental methods

(1) Animal husbandry: C57BL/6N mice, male, 6-8 weeks old, weighing 20-22 g, were housed in an SPF barrier environment after purchase, with a temperature of 22±2°C and a humidity of 55±5%. The bedding was sterilized by γ-rays and changed twice a week. The light/dark hours were 12 hours each day, and 5 mice were placed in each cage. They were fed with sterilized standard feed and acidified water. All mice were housed in the animal room for one week after purchase to adapt to the environment before the experiment was carried out.

(2) The mice were randomly divided into two groups and ear-marked. Two hours before irradiation, the experimental group was given an intraperitoneal injection of SB 4 (5.0 mg/kg), while the control group was given an intraperitoneal injection of an equal volume of saline.

(3) Before irradiation, the mice were placed in a single-cell mouse box, which was placed 3 meters away from the γ-ray radiation source for irradiation, with a total irradiation dose of 6.5 Gy. After irradiation, the mice were immediately returned to the mouse cage in the SPF barrier environment to move freely and eat.

(4) On the 14th day after irradiation, the femur and tibia of the hind limbs of mice in each group were separated and all bone marrow cells were flushed out. The bone marrow samples were treated with red blood cell lysis buffer, and then the total nucleated cells in the femur and tibia of each mouse were counted. Each sample was counted twice and the average value was taken. After counting, the cells were labeled with flow cytometry antibodies Lineage, c-Kit, Sca-1, CD48 and CD150, and after incubation and washing, flow cytometry was performed.

3.2 Results Analysis

Under normal circumstances, mouse hematopoietic stem/progenitor cells only account for 0.01% of bone marrow cells, and are mainly enriched in the Lin-c-Kit+Sca-1+ (LSK) cell population. They are the core of maintaining the normal physiological function of the hematopoietic system and the regeneration and repair of hematopoietic damage. The number and proportion of bone marrow hematopoietic stem/progenitor cells after irradiation were significantly lower than those of normal mice, indicating that bone marrow hematopoietic stem/progenitor cells were damaged after irradiation. After SB 4 administration, the number of LSK and long-cycle hematopoietic stem cells (LT-HSC) in the bone marrow of the experimental group of mice and their proportion in bone marrow nucleated cells were significantly increased compared with those of the control group of mice, indicating that SB 4 administration can indeed act on hematopoietic stem/progenitor cells, reduce the damaging effects of sublethal doses of irradiation on these cells, and promote the regeneration and repair of the hematopoietic system of irradiated mice.

In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "examples", "specific examples", or "some examples" means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representation of the above terms does not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described may be combined in any one or more embodiments or examples in a suitable manner.

Although the embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the claims and their equivalents.

Claims (9)

1. Use of SB 4 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating or preventing a radiation damage-related disease in a subject. 2. The use according to claim 1, characterized in that the pharmaceutically acceptable salt comprises at least one of the following: sulfate, phosphate, citrate, tartrate, fumarate, benzoate, adipate, succinate, methanesulfonate and maleate. 3. The use according to claim 1, characterized in that the radiation damage-related disease includes at least one of hematopoietic dysfunction and gastrointestinal damage. 4. The use according to claim 1, characterized in that the radiation damage is caused by gamma rays; Optionally, the gamma ray dose is 1 to 10 Gy. 5. The use according to claim 1, characterized in that the subject is a mammal; Optionally, the subject is a human, mouse, rat, rabbit, monkey, dog or pig. 6. A drug for treating or preventing radiation damage-related diseases in a subject, comprising: SB 4 or a pharmaceutically acceptable salt thereof as an active ingredient. 7. The drug according to claim 6, characterized in that the pharmaceutically acceptable salt comprises at least one of the following: sulfate, phosphate, citrate, tartrate, fumarate, benzoate, adipate, succinate, methanesulfonate and maleate; Optionally, the medicament comprises an injection; Optionally, the radiation damage-related disease includes at least one of hematopoietic dysfunction and gastrointestinal tract damage. 8. The drug according to claim 6, characterized in that the drug is suitable for administration at 2.5-10 mg/kg human body weight. 9. A single-dose drug for treating or preventing radiation damage-related diseases in a subject, the drug comprising: SB 4 or a pharmaceutically acceptable salt thereof suitable for administration at a dosage of 2.5-10 mg/kg human body weight as an active ingredient; Optionally, the radiation damage-related disease includes at least one of hematopoietic dysfunction and gastrointestinal tract damage.