CN111228259A

CN111228259A - Medical application of 5-hydroxy-1-methylhydantoin

Info

Publication number: CN111228259A
Application number: CN202010250084.XA
Authority: CN
Inventors: 高利娜; 刘俊亭
Original assignee: China Medical University
Current assignee: China Medical University
Priority date: 2020-04-01
Filing date: 2020-04-01
Publication date: 2020-06-05

Abstract

The invention belongs to the technical field of biological medicines, relates to a new medical application of 5-hydroxy-1-methylhydantoin, and particularly relates to a medical application of 5-hydroxy-1-methylhydantoin in preventing lung injury caused by paraquat. The invention provides application of 5-hydroxy-1-methylhydantoin in preparing a medicament for treating lung injury caused by paraquat, which can be prepared into a medicament by taking the single 5-hydroxy-1-methylhydantoin as a raw material, and can also be prepared into a compound preparation by matching the 5-hydroxy-1-methylhydantoin with other medicaments. According to the invention, the research finds that the 5-hydroxy-1-methylhydantoin has a protection mechanism on paraquat-induced lung injury. The 5-hydroxy-1-methylhydantoin is a small molecule compound which can easily reach lung tissues through a diffusion mechanism to exert curative effects, including elimination of hydroxyl free radicals, antioxidation and anti-inflammatory mechanisms. The invention further explores the biological activity of the 5-hydroxy-1-methylhydantoin, and provides theoretical basis and experimental basis for preventing and treating lung injury diseases caused by paraquat.

Description

Medical application of 5-hydroxy-1-methylhydantoin

Technical Field

The invention belongs to the technical field of biological medicines, relates to a new medical application of 5-hydroxy-1-methylhydantoin, and particularly relates to a medical application of 5-hydroxy-1-methylhydantoin in preventing lung injury caused by paraquat.

Background

Paraquat was first synthesized in the nineteenth century, when used as a chemical indicator (redox indicator), and was used in agriculture as a herbicide in 1962. It can act on millions of growers and a hundred crops and is still widely used today by more than 120 countries worldwide. As an important non-selective contact herbicide, paraquat has the following characteristics: a) non-selective, that is, it kills most annual grassy and broadleaf weeds, as well as the top leaves of already-grown perennial weeds; b) the effect is very quick; c) the drug effect is not affected when the drug is applied for a few minutes and rains; d) the biological activity of the soil is passivated after the soil is contacted. Based on the characteristics, the paraquat brings many benefits to the farmers, the environment and the society. Nevertheless, paraquat is also claimed to cause serious acute and chronic health problems, such as: dermatitis, renal failure, respiratory failure, tachycardia, second degree burns, skin cancer, and parkinson's disease. In addition, paraquat is commonly used as a suicide agent in many countries (particularly developing countries). The use of paraquat has thus attracted attention from numerous organizations in the world, including the World Health Organization (WHO), the US environmental protection agency (US EPA) and the european chemical agency (ECB). Some non-governmental organizations from asia, america and europe have also launched the movement of "stopping Paraquat" in 2002 (Paraquat and Suicide, PAN Germany). Even so, due to the widespread use of paraquat, its widely available, low toxic dose (35 mg/kg for the lowest lethal dose in humans) (Pederson et al, 1981; Bismuth et al, 1982) and relatively inexpensive characteristics, there are a number of cases of accidental poisoning and suicide caused by paraquat which are continuously reported. The british medical scientist bulivant in 1966 first described 2 deaths due to accidental poisoning by paraquat, and subsequently, cases of paraquat poisoning were reported successively around the world. There are about ten thousand deaths reported in the united states, canada, europe, japan, singapore, hong kong, and the like. There are approximately thousands of suicide worldwide each year due to paraquat ingestion, although the actual figures are unknown due to incomplete reports and statistics in developing countries. The fundamental reason why paraquat causes such high mortality (total mortality is 25-75%, up to 95% in case of oral administration of 20% stock solution and survival reports in case of oral administration of more than 15 ml) is that paraquat poisoning has no specific antidote and effective therapeutic measure so far (A.L. Jones, R.Elton, R.Flanagan, QJM-int.J.Med.1999, 92: 573-.

Paraquat (PQ) is a widely used high-efficiency herbicide and has strong toxic effect on human. The death rate is high because of small toxic lethal dose, no specific antidote and extremely poor conventional symptomatic treatment effect. The characteristic change of paraquat poisoning is lung injury, which manifests as damage to alveolar epithelial cells in the early stage, hemorrhage and edema in the alveoli, and fibrosis in the alveoli and the lung interstitium in the later stage. At present, the mechanism of PQ poisoning is not clear and no effective antidote is clinically available, so that the death rate of PQ acute poisoning reaches 50-80%. The existing treatment method for paraquat poisoning is still in an exploration stage, so that a satisfactory treatment effect is difficult to obtain, and the death rate is still high. The treatment of acute PQ poisoning is mainly to reduce absorption, promote excretion, inhibit inflammatory response, and the like, and many studies at home and abroad include hemoperfusion combined with hemodialysis, gene therapy, antibody therapy, and the like. The clinical therapeutic drugs include anti-free radical and anti-oxidation drugs, hormones, and immunosuppressive drugs. At present, the treatment measures for PQ poisoning by scholars at home and abroad mainly focus on the application of exogenous drugs, but neglect the development and utilization of endogenous substances of organisms.

The hydantoin compounds are five-membered nitrogen heterocyclic compounds containing various substituents, and are also called hydantoin compounds. Because of the existence of various functional groups in the structure and the relatively high reactivity, various kinds of different substituted hydantoin and derivatives thereof are widely applied to the fields of medicines, pesticides, chemical industry, textiles and the like. The pharmacological actions of the hydantoin compounds are mainly represented by antibacterial, anti-inflammatory, antiepileptic, blood sugar lowering, sodium channel blockers, uremic toxin inhibiting and the like. The 5-hydroxy-1-methylhydantoin is an important derivative in hydantoin compounds, the structure is shown in figure 1, and research shows that 5-hydroxy-1-methylhydantoin is applicable to diseases such as intractable vasculitis, hyposerum albuminemia, renal failure, free radical and active oxygen elimination, has obvious inhibiting effect on guinea pig cough caused by citric acid, and 5-hydroxy-1-methylhydantoin has pharmacological activity such as protective effect on kidney injury caused by paraquat poisoning. However, reports on the application of 5-hydroxy-1-methylhydantoin in preparing medicaments for treating lung injury caused by paraquat are not found so far.

Disclosure of Invention

The target organ of paraquat poisoning is the lung, and no specific antidote is currently available for paraquat treatment. In view of the problems of the prior art, the invention aims to provide the medical application of 5-hydroxy-1-methylhydantoin in preventing lung injury caused by paraquat. According to the invention, the research finds that the 5-hydroxy-1-methylhydantoin has a protection mechanism on paraquat-induced lung injury. 5-hydroxy-1-methylhydantoin is an endogenous substance and is one of the creatinine metabolites. The 5-hydroxy-1-methylhydantoin is a small molecule compound which can easily reach lung tissues through a diffusion mechanism to exert curative effects, including elimination of hydroxyl free radicals, antioxidation and anti-inflammatory mechanisms.

In order to achieve the above object, the present invention provides the following technical solutions.

Use of 5-hydroxy-1-methylhydantoin in preparing medicine for treating lung injury caused by paraquat.

Furthermore, the application of the 5-hydroxy-1-methylhydantoin in preparing the medicine for treating the lung injury caused by the paraquat can be used for preparing a medicament by taking the single 5-hydroxy-1-methylhydantoin as a raw material, and can also be used for preparing a compound preparation by matching the 5-hydroxy-1-methylhydantoin with other medicines.

Compared with the prior art, the invention has the following beneficial effects.

(1) The 5-hydroxy-1-methylhydantoin is a small molecule compound which can easily reach lung tissues through a diffusion mechanism to exert curative effects, including elimination of hydroxyl free radicals, antioxidation and anti-inflammatory mechanisms.

(2) The invention researches the application of the gold 5-hydroxy-1-methylhydantoin in preparing the medicine for treating the lung injury caused by the paraquat, further explores the biological activity of the 5-hydroxy-1-methylhydantoin, and provides theoretical basis and experimental basis for preventing and treating the lung injury diseases caused by the paraquat.

Drawings

FIG. 1 is the structural formula of 5-hydroxy-1-methylhydrogen and a partial metabolite of creatinine (5-hydroxy-1 methylhydrogen is a degradation product in vivo from creatinine).

FIG. 2 is an image of a pathological section of a normal lung of a control group stained with H & E pathological sections showing intact alveolar structures, no edema in alveolar walls, and no inflammatory cell infiltration in lung parenchyma.

FIG. 3 is an H & E stained pathological section image of lung in paraquat (25mg/kg) poisoning group, which shows massive infiltration of inflammatory cells, significant intra-alveolar bleeding and significant film formation.

FIG. 4 is a H & E stained pathological section of lung after 5 consecutive days of treatment with 5-hydroxy-1-methylhydantoin (100mg/kg) after paraquat poisoning showing mild damage to alveolar structures and bleeding of small amounts of inflammatory cells.

FIG. 5 shows that Western Blot is used for detecting the HO-1 protein expression of lung tissues of various groups of mice, wherein Western Blot is used for a protein immunoblotting test, HO-1 is heme oxygenase-1, and β -actin is β -actin.

Figure 6 is a volcano plot of the differential metabolites obtained in negative ion mode. Red for up-regulation, green for down-regulation, grey for no change, VIP for the significance projection of the substance on the comparative OPLS-DA model.

Figure 7 volcano plots of the differential metabolites obtained in positive ion mode. Red for up-regulation, green for down-regulation, grey for no change, VIP for the significance projection of the substance on the comparative OPLS-DA model.

Fig. 8 is a Principal Component Analysis (PCA), an upper graph representing a principal component analysis obtained in the negative ion mode, and a lower graph representing a principal component analysis obtained in the positive ion mode. The abscissa t 1 and the ordinate t 2 in the figure represent the scores of the first and second ranked principal components, respectively, the blue point representing the PQ group sample and the red point representing the HMH group sample.

FIG. 9 is a partial least squares-discriminant analysis (PLS-DA) diagram, in which the upper diagram represents the negative ion mode and the lower diagram represents the positive ion mode. The abscissa is the score of the sample in the first principal component; the ordinate is the score of the sample on the second principal component.

FIG. 10 is a pharmacological mechanism of 5-hydroxy-1-methylhydantoin in the protection against lung injury caused by paraquat.

Detailed Description

The present invention will be described in detail with reference to specific examples, wherein the starting materials and equipment used in the specific examples of the present invention are known products and can be obtained by purchasing commercially available and commonly known starting materials.

Example 5-hydroxy-1-methylhydantoin for the experimental study of the treatment of lung injury caused by paraquat.

5-30 Kunming mice (body weight about 30 g) were purchased from Liaoning Biotechnology Ltd. Animals were acclimatized in the animal department for 1 week from the date of procurement. 30 mice were randomly divided into a normal saline group (control group), a paraquat poisoning group (gavage 20mg/kg, completed at one time), and a 5-hydroxy-1-methylhydantoin treatment group (paraquat poisoning mouse model was prepared, 5-hydroxy-1-methylhydantoin (100mg/kg) was intraperitoneally injected, and drug was administered every 8 hours for 5 days, the whole experiment was continued for 5 days, during which the condition of the mice was observed, and the mice were sacrificed on the sixth day.

The 5-hydroxy-1-methylhydantoin has a protective mechanism for paraquat-induced lung injury.

Through observation, the following results are found: the NS group mice have normal appetite and respiration, good mental status and flexible action. Most of the PQ group mice have toxic symptoms within 2 hours after the infection, and the symptoms are manifested by lassitude, dysphoria, reduction of different degrees of intake and water intake, blood secretion in mouth and nose, loose hair, and easy capture of slow action. After administration for 12h, the mice in the HMH group showed toxic symptoms, manifested by lassitude, bradykinesia, and no bloody secretion in the mouth and nose. H & E staining experiments show that the damage of the lung is relatively reduced in the paraquat poisoning group through the administration of 5-hydroxy-1-methylhydantoin, as shown in figures 2-4. Observing the H & E stained pathological section image, wherein the image of the pathological section of the normal lung of the control group is shown in FIG. 2, and the result shows that the alveolar structure is complete, the alveolar wall has no edema, and the lung parenchyma has no inflammatory cell infiltration; FIG. 3 is a pathological section image of lung of paraquat (20mg/kg) poisoning group, which shows a large amount of inflammatory cell infiltration, significant intra-alveolar bleeding and significant film formation; FIG. 4 is a pathological section of lung after 5 days of continuous treatment with 5-hydroxy-1-methylhydantoin (100mg/kg) after paraquat poisoning, showing mild damage to alveolar structure and bleeding of a small amount of inflammatory cells. Through measuring SOD and MDA indexes of lung tissues, the result is shown in table 1, and 5-hydroxy-1-methylhydantoin can be found to have antioxidant effect.

TABLE 1 MDA content and SOD activity between groups

Further observation of the change in the expression of HO-1 protein in lung tissue of mice revealed that the lung tissue of the control group had low levels of the change in the expression of HO-1 protein, as shown in Table 1 and FIG. 5. Compared with a control group, the HO-1 expression level of lung tissues of the infected group is obviously increased, and the expression level of the HMH group is obviously increased compared with that of the PQ group.

Metabonomic studies on lung tissue extraction of mice of each group found that the metabolites were observed to change in a mass spectrometer in a positive and negative ion mode, and the results are shown in fig. 6-7, wherein red represents an up-regulated metabolite compared to the PQ group, green represents a down-regulated metabolite compared to the PQ group, and gray represents a metabolite having no difference between the PQ group and the HMH group. The VIP value represents the projected value of the significance of the metabolite in the OPLS-DA model of the group comparison.

To obtain meaningful statistical results, Principal Component Analysis (PCA) and partial least squares discriminant analysis (PLS-DA) were performed. The HMH and PQ groups were well separated within 95% confidence intervals, see fig. 8, and the HMH and PQ groups, see fig. 9.

Differential metabolites were analyzed for the HMH group and PQ group, see table 2. The differential metabolite metabolism pathways between the HMH group and the PQ group are mainly focused on the citric acid cycle (TCA cycle), alanine, aspartate and glutamate metabolism, taurine and hypotaurine metabolism, microbial metabolism, adipocyte lipolysis regulation, purine pyrimidine metabolism.

TABLE 2 differential metabolites between HMH and PQ groups

Claims

Use of 5-hydroxy-1-methylhydantoin in preparing medicine for treating lung injury caused by paraquat is provided.
2. The use of 5-hydroxy-1-methylhydantoin as claimed in claim 1 in the preparation of a medicament for the treatment of lung injury caused by paraquat, wherein the medicament is prepared from 5-hydroxy-1-methylhydantoin alone or 5-hydroxy-1-methylhydantoin in combination with other medicaments to form a compound preparation.