Dipyridyl poison degradation method, device and application

Abstract

The invention belongs to the technical field of medical treatment, and particularly relates to a dipyridyl poison degradation method and device and application. According to the method, plasma, blood or water and other in-vitro matrixes containing toxicants are irradiated with 190-310 nm ultraviolet light, and efficient degradation of toxicant molecules is achieved; the device comprises an ultraviolet light generation module, the purpose of emitting ultraviolet light is achieved, the wavelength of the ultraviolet light is accurately controlled to be 190-310 nm, and the method and the device provide an efficient and safe technical scheme for in-vitro degradation of dipyridyl toxicants (such as diquat and paraquat). When the device is applied to preparation of a dipyridyl poison detoxification system, an in-vitro poison removal system can be formed, the problem that an existing blood purification technology is low in removal efficiency is solved, the poison removal efficiency is improved, and the poison removal effect after poisoning is improved.

Description

Technical Field

[0001] This invention relates to the field of medical technology, specifically to methods, apparatus, and applications for the degradation of bipyridine toxins. Background Technology

[0002] Bipyridine compounds (such as diquat and paraquat) are widely used in agriculture as highly effective herbicides, but their acute poisoning has become a global public health problem. Statistics show that these toxins cause approximately 85% of herbicide-related deaths, and even after my country banned the sale of paraquat aqueous solution in 2016, diquat poisoning cases have shown a significant upward trend. Current clinical treatment for bipyridine poisoning mainly relies on blood purification techniques (such as hemoperfusion and plasma exchange), but these techniques have significant limitations: firstly, the need for specialized equipment and technical teams makes early toxin removal difficult in rural and other grassroots areas; secondly, poisoned patients often ingest large doses of the toxin for various reasons, and the clearance efficiency of a single blood purification session is limited, resulting in unsatisfactory prognoses even after multiple treatments, with an overall mortality rate far higher than that of ordinary acute poisoning.

[0003] Phototherapy, as a non-invasive physical intervention, has demonstrated unique advantages in fields such as psoriasis and cancer, achieving target substance degradation through the synergistic effect of specific wavelengths of light and photosensitizers. However, current research on the application of phototherapy for toxin removal remains lacking, particularly in the in vitro photodegradation of bipyridine-based toxins. Furthermore, while blood purification devices can partially remove circulating toxins, they cannot address the problem of continuous release of toxins after they bind to tissues, nor can they rapidly reduce blood drug concentrations in the early stages of poisoning.

[0004] Therefore, developing an in vitro ultraviolet light degradation technology suitable for bipyridine toxins, along with a corresponding toxin degradation device, has become a key breakthrough in improving toxin removal efficiency. Summary of the Invention

[0005] The purpose of this invention is to provide a method, apparatus, and application for the degradation of bipyridine toxins. The method utilizes 190-310 nm ultraviolet light to irradiate plasma, blood, or water containing the toxins into an in vitro matrix, achieving efficient degradation of the toxin molecules. The apparatus includes an ultraviolet light generation module to generate ultraviolet light and precisely control the wavelength. The method and apparatus provide an efficient and safe technical solution for the in vitro degradation of bipyridine toxins (such as diquat and paraquat). When applied to the in vitro treatment of bipyridine toxins, the apparatus can form an in vitro toxin removal system, solving the problem of low removal efficiency in existing blood purification technologies, improving toxin removal efficiency, and enhancing the effect of toxin removal after poisoning.

[0006] The objective of this invention is achieved through the following technical solution:

[0007] First technical solution:

[0008] A method for in vitro degradation of bipyridine-type toxins involves irradiating an isolated matrix containing the toxin with 190–310 nm ultraviolet light, wherein the isolated matrix is ​​plasma, blood, or water.

[0009] Preferably, the duration of a single irradiation session is ≤4 hours.

[0010] Preferably, a photocatalyst, such as titanium dioxide, is added to the isolated matrix before irradiation to improve the degradation effect of bipyridine poisons.

[0011] Preferably, the ultraviolet light wavelength is 254nm.

[0012] Second technical solution:

[0013] A device for degrading bipyridine-type poisons includes an ultraviolet light generating module for producing 190–310 nm ultraviolet light. The ultraviolet light generating module can be a low-pressure mercury lamp, an LED ultraviolet light source, or the like.

[0014] Preferably, the toxic substance degradation device further includes a time control module for controlling the ultraviolet light irradiation time.

[0015] Preferably, the toxic substance degradation device further includes a photocatalyst dosing module.

[0016] Preferably, the toxic substance degradation device further includes a blood purification module.

[0017] Preferably, the blood purification module is selected from hemodialysis, hemofiltration, hemoperfusion, continuous blood purification equipment, plasma exchange, and immunoadsorption equipment.

[0018] Third technical solution:

[0019] The application of a bipyridine-based toxin degradation device in the preparation of a bipyridine-based toxin detoxification system, wherein the bipyridine-based toxin is diquat or paraquat.

[0020] Preferably, when the ultraviolet light generating module directly irradiates the epidermis, the single irradiation time is less than 4 hours, which has the best effect on removing toxins in the early stage of poisoning.

[0021] Preferably, when the ultraviolet light generating module directly irradiates the epidermis, the patient first supplements with common vitamins or applies a photocatalyst to the skin surface, or both. Common vitamins include vitamins A, B, C, and D, while the photocatalyst is titanium dioxide. By supplementing with vitamins or applying a photocatalyst before ultraviolet light irradiation, excessive cell damage under ultraviolet light can be effectively prevented, and the degradation of bipyridine toxins can be further accelerated even under lower light intensity irradiation.

[0022] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0023] 1. This invention uses 190-310nm ultraviolet light to irradiate in vitro matrices such as plasma, blood, or aqueous solutions containing diquat and paraquat to verify the effect of different matrices and concentrations on the photodegradation of diquat and paraquat, and ultimately confirms that 190-310nm ultraviolet light can efficiently degrade pyridine toxins.

[0024] 2. This invention confirms that the 190-310 nm ultraviolet light emitted by the ultraviolet light emission module can significantly reduce serum creatinine and ALT levels in a mouse model of bipyridine poisoning, effectively reducing blood drug concentrations after bipyridine poisoning, significantly alleviating liver and kidney function damage in mice after bipyridine poisoning, and having a positive effect on improving physiological indicators in the poisoning model. It provides a verifiable technical solution for the efficient clearance of bipyridine poisons, and its effectiveness is fully supported by mouse poisoning model experiments and animal safety evaluations. It aims to solve the technical bottlenecks of the lack of efficient in vitro clearance methods for bipyridine poisoning and the limited efficiency of traditional blood purification, laying the foundation for the development of new treatments for bipyridine poisoning.

[0025] 3. By combining an ultraviolet light irradiation module and a blood purification module, this invention can effectively improve the clearance rate of bipyridine toxins and effectively solve the technical bottleneck of insufficient clearance efficiency of bipyridine toxins in blood or plasma when using a single blood purification module. Attached Figure Description

[0026] Figure 1 The effects of different light sources (190nm, 254nm, 310nm, 365nm and natural light) on the in vitro photodegradation of diquat;

[0027] Figure 2 Effects of different substrates (water, plasma) and concentrations on the in vitro photodegradation of diquat;

[0028] Figure 3 Effects of UV light generation module irradiation on blood drug concentrations in mice exposed to diquat;

[0029] Figure 4 Effects of ultraviolet light generation module irradiation on liver and kidney damage in mice exposed to dichlorvos;

[0030] Figure 5 The effects of different light sources (254nm, 365nm and natural light) on the in vitro photodegradation of paraquat;

[0031] Figure 6 Effects of different substrates (water, plasma) and concentrations on the in vitro photodegradation of paraquat. Detailed Implementation

[0032] Example 1: In vitro irradiation experiment on the rapid degradation of phytoalexin by ultraviolet light (emitted by an ultraviolet light generating module).

[0033] (1) Selection of ultraviolet light source

[0034] Light is a photon within a specific frequency band. Generally, visible light is the electromagnetic wave that humans can see, with a wavelength range typically between 360-400 nm and 760-830 nm, while ultraviolet light typically ranges from 100-400 nm. Here, we explore the effects of different light sources on the degradation of diquat and paraquat. A standard aqueous solution of diquat at a concentration of 1 μg / mL was irradiated with natural light, light sources with wavelengths of 365 nm, 310 nm, 254 nm, and 190 nm for 0.5, 1, 2, 4, 8, and 16 hours (duration measured by a time control module). Samples were collected and analyzed by chromatographic-mass spectrometry (LC-MS). The results are as follows: Figure 1 .

[0035] The results showed that under sunlight, natural light, and light with a wavelength of 365 nm, diquat hardly underwent photodegradation, failing to achieve the purpose described in this patent. However, under ultraviolet light with wavelengths of 310 nm, 254 nm, and 190 nm, diquat aqueous solution was rapidly degraded, laying the foundation for the in vitro removal of diquat. Simultaneously, the results showed that the photodegradation capacity of wavelengths of 254 nm and 190 nm was superior to that of wavelength 310 nm. However, ultraviolet light with wavelengths of 180-190 nm reacts with air to produce ozone, a highly oxidizing substance harmful to humans. The 254 nm ultraviolet light source is the most readily available and commercially viable, already used in various fields such as environmental disinfection and sterilization, agriculture and food, and anti-counterfeiting technology. Considering all factors, subsequent ultraviolet irradiation experiments will utilize a 254 nm ultraviolet light source for further research.

[0036] (2) Effects of different substrates and different concentrations of diquat standard solutions on photodegradation

[0037] Using ultraviolet light with a wavelength of 254 nm as the irradiation source, the effects of different concentrations (0.5, 1.0, 2.0 μg / mL) of diquat standard solution on photodegradation under different matrices (water, plasma) were investigated. The results are as follows: Figure 2 The results showed that, under different substrates and concentrations, 254 nm light could achieve the in vitro degradation effect of diquat described in this invention.

[0038] Example 2: In vitro irradiation experiment of animal blood to accelerate the degradation of phytoalexin by ultraviolet light (emitted by ultraviolet light generation module).

[0039] (1) Effect of UV light generation module irradiation on blood drug concentration of diquat in mice

[0040] Monitoring blood drug concentrations of toxic substances helps in better understanding and managing poisoning cases, and more accurately determining the nature, severity, and prognosis of poisoning. Male BALB / c mice, SPF grade, 8-9 weeks old, weighing 22-25g, were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. (Beijing, China). Administration: Intravenous injection of dichlorvos at a toxic dose of 50 mg / kg. Intervention: 254nm irradiation. Groups: Blank group (irradiated only, no drug administration), drug administration without irradiation, and drug administration followed by irradiation, with 6 mice in each group. Blood was collected from the orbital venous plexus, centrifuged at 3500 rpm for 10 minutes, and the supernatant plasma sample was used for LC-MS analysis of blood drug concentration. Results are as follows: Figure 3 The results showed that at 0.5, 1, 2, and 4 hours after poisoning (duration measured by the time control module), the blood concentration of diquat in the irradiated group was lower than that in the unirradiated group (n=6), indicating that direct irradiation by the ultraviolet light generation module can significantly reduce the early blood concentration of diquat poisoning (P<0.001).

[0041] The above results demonstrate the safety of ultraviolet light generation module irradiation, which can effectively reduce the concentration of toxins after diquat poisoning and achieve the effect of toxin removal.

[0042] (2) Effects of UV light generation module irradiation on liver and kidney damage in mice exposed to dichlorvos

[0043] After poisoning in humans, the kidneys are the primary organ for excretion of diquat after absorption, and also the main target organ for damage. Furthermore, diquat can cause liver damage. Therefore, monitoring liver function (aspartate aminotransferase (AST), alanine aminotransferase (ALT)) and kidney function (serum creatinine (CrEA) and urea (U1REA)) are important clinical laboratory auxiliary diagnostic indicators for diquat poisoning (Expert Consensus on Diagnosis and Treatment of Acute Diquat Poisoning, 2020 Edition). Male BALB / c mice, SPF grade, 8-9 weeks old, weighing 22-25g, were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. (Beijing, China). Administration: Intravenous injection of dichlorvos at a toxic dose of 50 mg / kg. Treatment: 254nm irradiation. Groups: Blank group (irradiation treatment only, no drug administration), drug administration without irradiation, and drug administration followed by irradiation, 6 mice in each group. Blood was collected from the orbital venous plexus, centrifuged at 3500 rpm for 10 minutes, and the supernatant serum sample was collected for biochemical analysis. Results are as follows: Figure 4The results showed that after administration of diquat, liver and kidney markers aspartate aminotransferase (AST), alanine aminotransferase (ALT), serum creatinine (CrEA), and urea (UREA) significantly increased after 4 hours, indicating significant liver and kidney damage. Treatment with 254 nm ultraviolet light significantly reduced the degree of liver and kidney damage in mice (P < 0.001), demonstrating the therapeutic effect of ultraviolet light irradiation on diquat-exposed mice.

[0044] Example 3: In vitro irradiation experiment on the degradation of paraquat by ultraviolet light (emitted by an ultraviolet light generating module).

[0045] (1) Selection of ultraviolet light source

[0046] Paraquat, a bipyridine compound structurally similar to diquat, was used to irradiate standard blood samples with a paraquat concentration of 500 ng / mL under sunlight, natural light, and light sources at wavelengths of 365 nm and 254 nm for durations of 0.5, 1, 2, 4, and 8 hours (duration measured using a time control module). Samples were then collected and analyzed by LC-MS. The results are as follows: Figure 5 .

[0047] The results showed that the photodegradation behavior of paraquat was similar to that of diquat. Under natural light and ultraviolet light of wavelength 365nm for 8 hours, the concentration of paraquat did not change significantly and almost no photodegradation occurred, which could not achieve the purpose described in this patent. However, under ultraviolet light of wavelength 254nm, the concentration of paraquat in the blood sample continued to decrease, proving that ultraviolet light of a certain wavelength can promote the rapid degradation of paraquat in the blood, laying the foundation for the in vitro clearance of paraquat.

[0048] (2) Effects of different substrates and different concentrations of diquat standard solutions on photodegradation

[0049] Using ultraviolet light with a wavelength of 254 nm as the irradiation source, the effects of different concentrations (100, 500, 1000 ng / mL) of diquat standard solution on photodegradation under different matrices (water, plasma) were investigated. The results are as follows: Figure 6 The results showed that, under different substrates and concentrations, 254nm light could achieve the in vitro degradation effect of paraquat described in this invention.

[0050] Example 4

[0051] In Example 2, the inventors discovered that prolonged direct irradiation of the skin (externally exposed area) using an ultraviolet light generating module can cause skin burns and related inflammatory reactions. Based on this, the inventors found that the related problems can be solved by any of the following methods:

[0052] 1) Supplement with vitamins (such as vitamin C, vitamin A, B vitamins, and vitamin D) in advance, with a minimum supplementation of vitamins B, C, and A. The dosage for each vitamin should be the maximum daily supplemental dose for that vitamin. This method can reduce skin damage and allergic reactions caused by ultraviolet radiation and free radicals.

[0053] 2) Apply the photocatalyst (titanium dioxide) to the animal's skin surface with a coating thickness of 1 mm (the coating thickness can be adjusted appropriately during actual application, with a range of 0 mm < coating thickness ≤ 2 mm). Under lower light intensity, this further accelerates the degradation of diquat, thereby reducing skin damage. If the conventional 25W 254nm wavelength ultraviolet light source can achieve the desired effect, after applying the photocatalyst (titanium dioxide), only 10-20W of ultraviolet light is needed to achieve the same desired effect as the conventional 25W.

[0054] 3) Using both methods 1) and 2) simultaneously yields better results.

[0055] Furthermore, in the in vitro irradiation studies of Examples 1 and 3, the inventors found that adding the photocatalyst (by manual means or by adding the photocatalyst via a photocatalyst dosing module) to the plasma in advance could also achieve better results and improve the degradation ability of diquat under the same light intensity.

[0056] Example 5

[0057] Based on the ultraviolet light (emitted by the ultraviolet light generating module) conditions optimized and verified in Example 1, it is further combined with common blood purification modules (hemoperfusion equipment, hemodialysis equipment, and continuous blood drug purification equipment selected in this example) to accelerate the removal rate of toxins. That is, in this example, the ultraviolet light generating module and the blood purification module are used together to achieve the purpose of accelerating the removal of toxins.

[0058] The experiment used male Landrace pigs aged 12–18 months, fed daily with free access to water, and kept at an ambient temperature of 22–25°C. The administration method was intraperitoneal injection of diquat solution at a toxic dose of 40 mg / kg.

[0059] Three blood purification methods for toxin removal: 1) Blood perfusion module: Blood perfusion was performed on Landrace pigs, with each perfusion lasting 2 hours, for a total of 3 times;

[0060] 2) Hemodialysis module: Hemodialysis is used on Landrace pigs, with each dialysis session lasting 2 hours, for a total of 3 sessions.

[0061] 3) Continuous Renal Purification (CRRT) module of continuous blood drug purification equipment: CRRT is used for Landrace pigs, with each purification session lasting 4 hours, for a total of 2 sessions.

[0062] Combined ultraviolet light therapy: The ultraviolet light generating module is used in combination with the above-mentioned blood perfusion module, hemodialysis module and CRRT module to treat Landrace pigs. The purification time and number of times are the same as above.

[0063] The experiment included a blank group (no toxin removal was performed, resulting in death), a control group (toxins were removed using three different blood purification modules individually), and an experimental group (toxins were removed using a combination of an ultraviolet light generation module and a blood purification module). Two hours after administration, the toxin removal intervention began. Blood samples were collected before and after toxin removal, denoted as Cs and Ce. After blood collection, the samples were centrifuged at 3500 rpm for 10 minutes, and the supernatant serum sample was collected for drug concentration analysis. The toxin clearance rate was calculated using the formula "Toxin clearance rate = (Cs - Ce) / Cs × 100%", and the prognostic results were recorded. The results are shown in Table 1.

[0064] Table 1:

[0065]

[0066] As shown in Table 1, the combined use of the ultraviolet light generation module and the blood purification module significantly improved the toxin removal rate compared to the blood purification module alone, resulting in a higher survival rate and better toxin removal after combined treatment.

[0067] Example 6

[0068] Similar to Example 5, blood perfusion combined with continuous venous-venous hemofiltration (HP+CVVH) was used to treat Landrace pigs contaminated with paraquat. After a single treatment of 8 hours (duration measured by the time control module), the paraquat clearance rate was 53.5%, and after combined with ultraviolet irradiation for 8 hours, the paraquat clearance rate was 82.1%, which is better than the conventional single blood purification module for paraquat clearance.

[0069] Example 7

[0070] A device for degrading bipyridine-type poisons includes an ultraviolet light generating module for producing 190–310 nm ultraviolet light. The ultraviolet light generating module can be a low-pressure mercury lamp, an LED ultraviolet light source, etc. The ultraviolet light generating modules mentioned in Examples 1-6 all use LED ultraviolet light sources. As can be seen from Examples 1 or 3, the ultraviolet light generating module can achieve the purpose of degrading bipyridine-type poisons.

[0071] Furthermore, in order to more precisely control the irradiation time of the ultraviolet light generating module, the degradation device also includes a time control module in addition to the ultraviolet light generating module, which is used to control the light irradiation time of the ultraviolet light generating module. The time control module used in Examples 1-6 is an electronic timer.

[0072] Furthermore, in order to further improve the degradation efficiency of bipyridine toxins, the degradation device also includes a photocatalyst dosing module, which can be any device in the prior art capable of dispensing solid materials.

[0073] In practice, a photocatalyst is first added to the isolated matrix through a photocatalyst addition module, and then the isolated matrix is ​​irradiated through an ultraviolet light generation module. During the irradiation, the irradiation time of the ultraviolet light generation module is controlled by a time control module.

[0074] Furthermore, to further improve the degradation efficiency of bipyridine toxins, the degradation device, in addition to the ultraviolet light generating module, also includes a blood purification module. The blood purification module includes a hemodialysis device, a hemofiltration device, a hemoperfusion device, a continuous blood purification device, a plasma exchange device, and an immunoadsorption device. That is, in the actual degradation of toxin concentrations in blood and plasma, the ultraviolet light generating module and the blood purification module are used in combination, as in Examples 5 and 6.

[0075] Comparative Example 1

[0076] When other types of pesticides, including glyphosate, glufosinate, chlorfenapyr, and other non-bipyridine pesticides (non-diquat-like structures), are irradiated with natural light or light sources with wavelengths of 365nm, 310nm, 254nm, and 190nm, they cannot be degraded by ultraviolet light and thus cannot achieve the purpose described in this patent.

Claims

1. A method for in vitro degradation of bipyridine-type poisons, characterized in that, An in vitro matrix containing toxic substances is irradiated with 190–310 nm ultraviolet light, wherein the in vitro matrix is ​​plasma, blood, or water.

2. The in vitro degradation method for bipyridine poisons according to claim 1, characterized in that, Single irradiation time ≤ 4 hours.

3. The method for in vitro degradation of bipyridine-type poisons according to claim 1, characterized in that, A photocatalyst is added to the isolated matrix before irradiation.

4. The method for in vitro degradation of bipyridine poisons according to claim 1, characterized in that, The ultraviolet light wavelength is 254 nm.

5. A device for degrading bipyridine-type poisons, characterized in that, It includes an ultraviolet light generation module for generating 190–310 nm ultraviolet light.

6. The device for degrading bipyridine-type poisons according to claim 5, characterized in that, It also includes a timing module for controlling the duration of ultraviolet light irradiation.

7. The device for degrading bipyridine-type toxins according to claim 6, characterized in that, It also includes a photocatalyst dosing module.

8. A device for degrading bipyridine-type poisons according to any one of claims 5-7, characterized in that, It also includes a blood purification module, which is selected from hemodialysis, hemofiltration, hemoperfusion, continuous blood purification equipment, plasma exchange, and immunoadsorption equipment.

9. The application of the bipyridine-type poison degradation device according to claim 6 or 8 in the preparation of a bipyridine-type poison detoxification system, characterized in that, The bipyridine-type toxins mentioned are diquat or paraquat.

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