Application of enoxaparin combined with sivelestat in preparation of medicines and systems for preventing or treating decompression sickness

Abstract

The invention relates to the technical field of medicines, provides novel application of enoxaparin combined with sivelestat, and particularly relates to application of enoxaparin combined with sivelestat in preparation of medicines and systems for preventing or treating decompression sickness. The application of enoxaparin combined with sivelestat preventive administration in combination with hyperbaric oxygen therapy in prevention and treatment of decompression sickness lung injury caused by rapid buoyant escape can significantly reduce the death rate of decompression sickness caused by rapid buoyant escape, alleviate lung tissue pathological conditions and inflammatory cell infiltration, reduce pulmonary vascular leakage caused by rapid buoyant escape, and improve the clinical application value of decompression sickness lung injury caused by rapid buoyant escape and decompression sickness, and the application of enoxaparin combined with sivelestat in prevention and treatment of decompression sickness lung injury caused by rapid buoyant escape. The levels of lung tissue inflammatory factors and chemotactic factors are reduced.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology. Specifically, it is the application of enoxaparin combined with sivelestat in the preparation of a drug for preventing or treating decompression sickness, and a system for preventing or treating decompression sickness. Background Art

[0002] Rapid ascent escape is one of the main escape methods for submarine crew to get out of a disabled submarine. It has the advantages of simple equipment, easy method, large escape depth, and simple operation, and has become a conventional technology in the navies of developed countries and is widely used. However, with the increase of the escape depth, the allowable high-pressure exposure time for crew is quite limited. If the exposure time is too long due to improper pressure regulation or other reasons, serious decompression sickness will occur. At present, the conventional decompression procedures cannot completely avoid the occurrence of decompression sickness. How to take effective measures to reduce the damage of decompression sickness has become the focus of research on submarine rescue.

[0003] Previous studies found that rats that died of decompression sickness after rapid ascent escape showed obvious damage to the lung tissue, mainly manifested as lung tissue hemorrhage, edema, and alveolar structure destruction.

[0004] Hyperbaric oxygen therapy (HBO) is a method of breathing pure oxygen or high-concentration oxygen in a high-pressure (above normal pressure) environment to treat hypoxic diseases and related disorders. HBO treatment is the preferred treatment method for air embolism and decompression sickness, which can reduce the bubble volume, accelerate the bubble absorption, allow excessive gas to safely leave the human body within sufficient time, and enable the affected tissues to restore normal blood circulation and oxygen supply.

[0005] Enoxaparin is a low-molecular-weight heparin preparation, which can make the ratio of the activities of anticoagulant factors Ⅹa and Ⅱa greater than 4, thus exerting a strong anti-thrombotic function and a certain thrombolytic effect. It is mainly used for: 1. Preventing the formation of venous thrombosis after plastic surgery and general surgery and the coagulation during extracorporeal circulation in hemodialysis; 2. Preventing deep vein thrombosis and pulmonary embolism, and treating the already formed deep vein thrombosis.

[0006] Sivelestat is an inhibitor of neutrophil elastase and is an effective drug for treating acute lung injury or acute respiratory distress syndrome accompanied by systemic inflammatory response syndrome. Clinical studies have shown that sivelestat can effectively reduce the lung injury score of ARDS patients, improve lung function, shorten the mechanical ventilation time and the length of stay in the intensive care unit, and reduce the mortality rate.

[0007] So far, there is no study to confirm that the above two drugs can be applied to decompression sickness, and there is no relevant report on the combined use with HBO treatment to prevent and treat decompression sickness lung injury caused by rapid ascent escape. Summary of the invention

[0008] The purpose of the present invention is to provide the use of enoxaparin combined with sivelestat in the preparation of a drug for preventing or treating decompression sickness, especially the use of enoxaparin combined with sivelestat for preventive administration combined with hyperbaric oxygen therapy in the prevention and treatment of decompression sickness-induced lung injury caused by rapid ascent and descent, which can significantly reduce the mortality rate of decompression sickness caused by rapid ascent and descent, alleviate lung tissue pathology and inflammatory cell infiltration, reduce pulmonary vascular leakage caused by decompression sickness caused by rapid ascent and descent, and reduce the levels of inflammatory factors and chemokines in lung tissue.

[0009] In a first aspect of the present invention, there is provided an application of enoxaparin combined with sivelestat in the preparation of a drug for preventing or treating decompression sickness, wherein the enoxaparin comprises enoxaparin itself or a pharmaceutically acceptable salt or ester thereof, and the sivelestat comprises sivelestat itself or a pharmaceutically acceptable salt or ester thereof. The drug for preventing or treating decompression sickness is particularly suitable for use in combination with hyperbaric oxygen therapy in preventing and treating decompression sickness caused by rapid ascent and escape.

[0010] The pharmaceutically acceptable salt may be a sodium salt, a potassium salt or a calcium salt. Enoxaparin sodium and sivelestat sodium commonly used in clinical practice are preferably used.

[0011] The pharmaceutically acceptable ester may be methyl ester, ethyl ester, isopropyl ester, triglyceride, or the like.

[0012] The drug for preventing or treating decompression sickness is a pharmaceutical composition with enoxaparin and sivelestat as the only two active ingredients, or a pharmaceutical composition containing enoxaparin and sivelestat.

[0013] In some embodiments, the pharmaceutical composition further comprises a pharmaceutical excipient.

[0014] Furthermore, the auxiliary material is at least one of a solvent, a solubilizer, a diluent, a filler, a binder, a disintegrant, a flavoring agent, a preservative, a surfactant, and an excipient.

[0015] Preferably, the rat dose of enoxaparin is 1-3 mg / kg, and the rat dose of sivelestat is 10-30 mg / kg. Within the above range, the two doses can be arbitrarily matched. For example, the dose of enoxaparin can be formulated as 1 mg / kg, 1.3 mg / kg, 1.5 mg / kg, 1.8 mg / kg, 2 mg / kg, 2.2 mg / kg, 2.5 mg / kg, 2.8 mg / kg, 3 mg / kg or any other point value within the above range; the dose of sivelestat can be formulated as 10 mg / kg, 13 mg / kg, 15 mg / kg, 18 mg / kg, 20 mg / kg, 22 mg / kg, 25 mg / kg, 28 mg / kg, 30 mg / kg or any other point value within the above range.

[0016] Preferably, the rat dose of enoxaparin is 2 mg / kg, and the rat dose of sivelestat is 20 mg / kg.

[0017] The dose is for a single use. When used for humans or other animal species, it needs to be adjusted according to pharmacological regulations, body weight, and purpose of use, etc.

[0018] In some embodiments, the drug for preventing or treating decompression sickness is a combination of the drug preparations made from the enoxaparin and the sivelestat independently and conventional pharmaceutical excipients, or the drug preparation made from the combination of the enoxaparin and the sivelestat and conventional pharmaceutical excipients.

[0019] The drug preparation is at least one of a capsule, a suspension, an emulsion, a solution, a syrup, an injection, a tablet, a pill, and a granule.

[0020] The administration method of the drug preparation is oral administration and / or injection.

[0021] In some embodiments, the decompression sickness is decompression sickness caused by rapid ascent and escape.

[0022] In the second aspect of the present invention, a system for preventing or treating decompression sickness is provided, which includes the drug for preventing or treating decompression sickness and also includes a hyperbaric oxygen therapy device.

[0023] The hyperbaric oxygen therapy device can be an existing hyperbaric chamber or a pressurized oxygen therapy instrument. The hyperbaric chamber can be for single or multiple persons, and can also be large-sized or portable.

[0024] The system is particularly suitable for preventing or treating decompression sickness caused by rapid ascent and escape. The using time of enoxaparin and sivelestat is 13 - 17 minutes, preferably 15 minutes, before rapid ascent and escape, and the using time of the hyperbaric oxygen therapy device is immediately after rapid ascent and escape.

[0025] Experiments show that prophylactic administration of enoxaparin combined with sivelestat combined with hyperbaric oxygen therapy can significantly reduce pulmonary edema and injury caused by bubbles, thereby reducing the pathological changes caused by decompression sickness, significantly increasing the survival rate of decompression sickness caused by rapid ascent and escape, alleviating the symptoms of decompression sickness caused by rapid ascent and escape, and prolonging the survival time, which is beneficial to improving the rescue survival rate after escape.

[0026] The beneficial effects of the present invention are as follows:

[0027] The present invention provides the use of enoxaparin combined with sivelestat in the preparation of a drug for preventing or treating decompression sickness, and the drug for preventing or treating decompression sickness is particularly suitable for use in combination with hyperbaric oxygen therapy in the prevention and treatment of decompression sickness caused by rapid ascent and escape. The prophylactic administration of enoxaparin combined with sivelestat in combination with hyperbaric oxygen therapy can significantly reduce pulmonary edema and injury caused by bubbles, thereby reducing the pathological changes caused by decompression sickness, significantly increasing the survival rate of decompression sickness caused by rapid ascent and escape, alleviating the symptoms of decompression sickness caused by rapid ascent and escape, and prolonging the survival time, which is beneficial to improving the rescue survival rate after escape. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 It is the survival curve of rats after rapid ascent and escape with different prevention and treatment measures in Example 1.

[0029] Figure 2 It is the number of peripheral blood neutrophils in rats after rapid ascent and escape with different prevention and treatment measures in Example 1.

[0030] Figure 3 It is the total protein content in bronchoalveolar lavage fluid of rats after rapid ascent and escape with different prevention and treatment measures in Example 1.

[0031] Figure 4 It is the pathological changes of lung tissue in rats after rapid ascent and escape with different prevention and treatment measures in Example 1.

[0032] Figure 5 It is the immunohistochemical photograph of lung tissue in rats after rapid ascent and escape with different prevention and treatment measures in Example 1.

[0033] Figure 6 It is the levels of inflammatory factors and chemokines in lung tissue of rats after rapid ascent and escape with different prevention and treatment measures in Example 1.

[0034] The groups in the figure are: DCS: Vehicle control group; DCS + HBO: 1.8ATA 99% O2 group; Enoxaparin + DCS: 2mg / kg Enoxaparin group; Sivelestat + DCS: 20mg / kg Sivelestat group; Enoxaparin + Sivelestat + DCS + HBO: 2mg / kg Enoxaparin + 20mg / kg Sivelestat + 1.8ATA 99% O2 group. DETAILED DESCRIPTION OF THE INVENTION

[0035] The following provides a detailed description of the specific embodiments provided by the present invention in conjunction with the accompanying drawings. The following examples are implemented on the premise of the technical solution of the present invention, and detailed implementation manners are given, but the protection scope of the present invention is not limited to the following examples. The following reagents are all commercially available unless otherwise specified.

[0036] The enoxaparin sodium used in the experiments of the present invention was purchased from Shanghai Aladdin Biochemical Technology Co., Ltd., sivelestat sodium was purchased from MedChemExpress Biotechnology Co., Ltd. in the United States, and pure oxygen was purchased from Shanghai Shenwei Gas Canning Co., Ltd.

[0037] Example

[0038] Experimental injection drug: A Vehicle (solvent) prepared with 10% DMSO + 40% PEG300 + 5% Tween-80 + 45% Saline (normal saline) was used to prepare a mixed solution of 2 mg / mL enoxaparin sodium + 20 mg / mL sivelestat sodium into an injection.

[0039] Experimental animals and grouping: 80 male healthy SD rats, weighing 240 - 260 g, were purchased from Zhejiang Vital River Laboratory Animal Technology Co., Ltd. The SD rats were randomly divided into a DCS group, a DCS + HBO group, an Enoxaparin + DCS group, a Sivelestat + DCS group, and an Enoxaparin + Sivelestat + DCS + HBO group, with 20 rats in each group.

[0040] Experimental method: 15 minutes before pressurization in the Enoxaparin + DCS group, the Sivelestat + DCS group, and the Enoxaparin + Sivelestat + DCS + HBO group, the corresponding solutions were injected through the tail vein (the injection drugs were 2 mg / kg enoxaparin sodium, 20 mg / kg sivelestat sodium, and a mixture of 2 mg / kg enoxaparin sodium and 20 mg / kg sivelestat sodium respectively); 15 minutes before pressurization in the DCS group and the DCS + HBO group, an equal volume of Vehicle was injected through the tail vein. Then the experimental animals were placed in a rapid ascent escape pressure chamber, the chamber door was closed, and a self-written computer automated rapid ascent escape program was used. Compressed air was pressurized to 1.5 MPa at an exponential rate of 2 t / 7 and stayed for 242 s, then decompressed out of the chamber at a speed of 3 m / s. The highest temperature in the chamber during pressurization was 40 °C, and the temperature in the chamber during pressure stabilization was 26 °C. Immediately after the DCS + HBO group and the Enoxaparin + Sivelestat + DCS + HBO group exited the chamber, the experimental animals were placed in an animal hyperbaric oxygen chamber for HBO treatment, with 1.8 ATA 99% O2 treatment for 30 minutes; the DCS group, the Enoxaparin + DCS group, and the Sivelestat + DCS group were placed in normal pressure and normal oxygen air. The mortality and behavior of all groups of rats were observed within 30 minutes after exiting the chamber, and all rats were sacrificed under isoflurane anesthesia 4 hours after exiting the chamber to collect samples.

[0041] It was observed that rats in the DCS group showed restlessness and rapid chest and abdominal undulations after emerging from the rapid ascent escape decompression chamber. The survival rate of rats was 55% as seen from the survival curve. Rats in the prevention groups of Enoxaparin + DCS group and Sivelestat + DCS group also showed restlessness and rapid chest and abdominal undulations after emerging from the rapid ascent escape decompression chamber, with survival rates of 60% and 55% respectively. Rats in the DCS + HBO group were in good condition after emerging from the rapid ascent escape decompression chamber, and the survival rate increased significantly to 85% compared with that of the DCS group, with a statistically significant difference (P < 0.05). Rats in the Enoxaparin + Sivelestat + DCS + HBO group were in good condition after leaving the chamber, and the survival rate increased to 95%, with a statistically significant difference compared with DCS (P < 0.01). The above experimental results indicate that independent administration of Enoxaparin and Sivelestat did not affect the survival time and survival rate of rats with rapid ascent escape-induced decompression sickness. However, after combined administration, it further increased the increase in the survival rate of HBO treatment for rats with rapid ascent escape-induced decompression sickness (see Figure 1 ).

[0042] Number of neutrophils in rat peripheral blood: There was no difference in the white blood cell (WBC) count in the peripheral blood of the five groups of rats (DCS: 8.068 ± 3.048, DCS + HBO: 8.470 ± 3.029, Enoxaparin + DCS: 8.541 ± 2.495, Sivelestat + DCS: 7.828 ± 2.721, Enoxaparin + Sivelestat + DCS + HBO: 7.523 ± 4.012), as shown in Figure 2 Figure A. Compared with the DCS group (DCS: 3.135 ± 1.483), the neutrophil (Neut) count in the peripheral blood of rats in the DCS + HBO group, Enoxaparin + DCS group, and Sivelestat + DCS group (DCS + HBO: 2.283 ± 1.259, Enoxaparin + DCS: 2.189 ± 0.7814, Sivelestat + DCS: 1.924 ± 0.7040) showed a downward trend, but there was no statistical difference. The neutrophil count in the peripheral blood of the Enoxaparin + Sivelestat + DCS + HBO group (1.189 ± 0.7063) decreased significantly, P < 0.01, as shown in Figure 2as shown in Figure B in []. Compared with the DCS group (DCS: 38.76 ± 10.35), the percentage of neutrophils (Neut%) in the peripheral blood of rats in the other groups decreased significantly (DCS + HBO: 25.97 ± 6.649, Enoxaparin + DCS: 25.60 ± 5.704, Sivelestat + DCS: 24.61 ± 3.318, Enoxaparin + Sivelestat + DCS + HBO: 15.85 ± 3.568). The P values of each group were less than 0.01, as Figure 2 shown in Figure C in []. Prophylactic administration of enoxaparin sodium combined with sivelestat sodium and hyperbaric oxygen therapy significantly reduced the neutrophil count and neutrophil ratio in the peripheral blood of decompression sickness rats, indicating that it alleviated the systemic inflammatory response in decompression sickness rats.

[0043] Total protein content in bronchoalveolar lavage fluid (Total Protein in BALF) of rats: As Figure 3 shown, the total protein content in the bronchoalveolar lavage fluid of the DCS group was 1.203 ± 0.4490, that of the DCS + HBO group was 0.7225 ± 0.3022, showing a statistically significant difference compared with the DCS group (P < 0.05); that of the Enoxaparin + DCS group was 0.7313 ± 0.3342, showing a statistically significant difference compared with the DCS group (P < 0.05); that of the Sivelestat + DCS group was 0.7225 ± 0.3050, showing a statistically significant difference compared with the DCS group (P < 0.05); that of the Enoxaparin + Sivelestat + DCS + HBO group was 0.5425 ± 0.1773, showing a significant difference compared with the DCS group (P < 0.01). Prophylaxis with enoxaparin sodium, prophylaxis with sivelestat sodium, and HBO treatment all reduced pulmonary vascular leakage in rats and alleviated lung tissue damage in rats; combined prevention and treatment further improved pulmonary vascular leakage in rats.

[0044] Pathological changes in rat lung tissue: The pathological results of rat lung tissue showed that in the DCS group, the alveolar structure of rats was extensively damaged and fused, and eosinophilic fluid and red blood cells were visible exudating in the alveolar cavity, as Figure 4 shown in Figure A in []. Compared with the DCS group, the thickening degree of the alveolar wall and the degree of interstitial hemorrhage and exudation in the DCS + HBO group, Enoxaparin + DCS group, and Sivelestat + DCS group of rats were alleviated, as Figure 4 shown in Figures B - D in []. While in the Enoxaparin + Sivelestat + DCS + HBO group of rats, the alveolar structure was intact, the alveolar cavity was clear, and the thickening of the alveolar wall was not obvious, as Figure 4 shown in Figure E in [].

[0045] Changes in inflammatory cell infiltration in rat lung tissue: The results of immunohistochemical staining of CD11b in inflammatory cells in rat lung tissue showed that there was an increase in inflammatory cell infiltration in the lungs of rats in the DCS group, and some inflammatory cells died, as Figure 5 shown in Figure A in the middle; compared with the DCS group, there was less death of inflammatory cells in the lungs of rats in the DCS + HBO group, while in the Enoxaparin + DCS group and the Sivelestat + DCS group, the infiltration of inflammatory cells in the lung tissue decreased, and the morphology of inflammatory cells was better maintained, as Figure 5 shown in Figures B - D in the middle; the number and morphology of inflammatory cells in the lung tissue of rats in the Enoxaparin + Sivelestat + DCS + HBO group were normal, as Figure 5 shown in E.

[0046] Changes in inflammatory factors and chemokines in rat lung tissue: Interleukin 1β (IL-1β), tumor necrosis factor α (TNF-α), and interleukin 8 (CXCL8) are commonly used inflammatory factors and chemokines. When acute inflammation occurs, multiple cells are damaged and inflammatory cells are activated, and multiple cells will express and release these cytokines. Therefore, by measuring the mRNA changes of IL-1β, TNF-α, and CXCL8 in lung tissue, the level of lung inflammation and the degree of damage can be reflected. The results showed that several prevention and treatment methods could significantly reduce the levels of IL-1β (DCS: 8.163 ± 3.526, DCS + HBO: 4.220 ± 1.835, Enoxaparin + DCS: 4.041 ± 1.798, Sivelestat + DCS: 3.078 ± 1.403, Enoxaparin + Sivelestat + DCS + HBO: 1.723 ± 0.7544) and TNF-α (DCS: 10.31 ± 2.591, DCS + HBO: 5.658 ± 1.880, Enoxaparin + DCS: 5.814 ± 2.650, Sivelestat + DCS: 4.674 ± 1.791, Enoxaparin + Sivelestat + DCS + HBO: 3.564 ± 2.260) in lung tissue, as Figure 6As shown in Figure A-B. Compared with the DCS group (2.620±1.822), the CXCL8 level in the DCS+HBO group (1.474±0.7210) showed a downward trend, but there was no significant difference; the CXCL8 level in the Enoxaparin+DCS group (1.228±0.4822) was lower than that in the DCS group, P<0.05; in the Enoxaparin+DCS group and the Enoxaparin+Sivelestat+DCS+HBO group (Sivelestat+DCS: 0.8563±0.6059, Enoxaparin+Sivelestat+DCS+HBO: 0.4775±0.2692), the CXCL8 level decreased significantly, and the difference was significant compared with the DCS group (P<0.01).

[0047] In summary, through the above experiments, it can be confirmed that prophylactic administration of enoxaparin sodium combined with sivelestat sodium combined with hyperbaric oxygen therapy can significantly reduce pulmonary edema, inflammatory cell infiltration and injury caused by bubbles, thereby reducing the pathological changes caused by decompression sickness and significantly improving the survival rate of decompression sickness caused by rapid ascent and escape.

[0048] The preferred embodiments of the present invention have been specifically described above, but the present invention is not limited to the described embodiments. Those skilled in the art can also make various equivalent variations or substitutions without departing from the spirit of the present invention, and these equivalent variations or substitutions are all included within the scope defined by the claims of this application.

Claims

1. Use of enoxaparin combined with sivelestat in the preparation of a drug for preventing or treating decompression sickness, wherein the enoxaparin includes enoxaparin itself or its pharmaceutically acceptable salt or ester, and the sivelestat includes sivelestat itself or its pharmaceutically acceptable salt or ester.

2. The application according to claim 1, characterized in that: The drug for preventing or treating decompression sickness is a pharmaceutical composition with enoxaparin and sivelestat as the only two active ingredients, or a pharmaceutical composition containing enoxaparin and sivelestat.

3. The application according to claim 2, characterized in that: The pharmaceutical composition further includes pharmaceutical excipients.

4. The application according to claim 3, characterized in that: The excipients are at least one of a solvent, a solubilizer, a diluent, a filler, a binder, a disintegrant, a flavoring agent, a preservative, a surfactant, and an excipient.

5. The application according to claim 1, wherein: The drug for preventing or treating decompression sickness is a combination of drug preparations made from the enoxaparin and the sivelestat independently and conventional pharmaceutical excipients in pharmacy, or a drug preparation made from the combination of the enoxaparin and the sivelestat and conventional pharmaceutical excipients in pharmacy.

6. The application according to claim 5, wherein: The drug preparation is at least one of a capsule, a suspension, an emulsion, a solution, a syrup, an injection, a tablet, a pill, and a granule.

7. The application according to claim 5, characterized in that: The administration method of the drug preparation is oral administration and / or injection.

8. The application according to claim 1, characterized in that: The rat dose of the enoxaparin is 1 - 3 mg / kg, and the rat dose of the sivelestat is 10 - 30 mg / kg.

9. A system for preventing or treating decompression sickness, which includes the drug for preventing or treating decompression sickness according to any one of claims 1 - 8, and further includes a hyperbaric oxygen therapy device.

10. The system for preventing or treating decompression sickness according to claim 9, characterized in that: The decompression sickness is decompression sickness caused by rapid ascent and escape; the use time of the drug for preventing or treating decompression sickness is 13 - 17 minutes before rapid ascent and escape, and the use time of the hyperbaric oxygen therapy device is immediately after rapid ascent and escape.

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