WO2022199050A1 - Use of ulinastatin in preparation of medication for preventing and treating severe covid-19 - Google Patents

Abstract

Provided is the use of ulinastatin in preparation of a medication for preventing and treating severe COVID-19. The medication contains ulinastatin, polygonatum sibiricum polysaccharide, and carboxymethyl schizophyllan.

Description

Use of ulinastatin in the preparation of medicines for the prevention and treatment of severe novel coronavirus pneumonia technical field

The invention belongs to the technical field of medicine, and in particular relates to the use of ulinastatin in the preparation of a medicine for preventing and treating severe novel coronavirus pneumonia.

Background technique

Coronaviruses are a large class of viruses that exist widely in nature. Before 2019, a total of 6 coronaviruses that can infect humans were found, namely HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU1, SARS-CoV and MERS. -CoV, and the coronavirus that caused the outbreak was named the new coronavirus (SARS-CoV-2) after isolation and confirmation. According to existing studies, the common symptoms of patients with novel coronavirus infection are fever, cough and myalgia or fatigue; all patients suffered from pneumonia and abnormal chest CT examination findings; complications include acute respiratory distress syndrome , acute cardiac injury and secondary infection. The inflammatory factor storm of the new coronavirus will cause damage to multiple organs, mainly the lungs, and sepsis is a systemic inflammatory response syndrome caused by infection, which is one of the most prevalent critical illnesses in the world. Leukocyte and platelet recruitment, intravascular coagulation, endothelial injury, surfactant loss, and oxidative stress are all mechanisms of severe lung injury in sepsis. It is aimed at the injury caused by sepsis-like injury in the critical period of patients in the late stage of new coronavirus infection.

In addition, on the whole, the current treatment of the new coronavirus, while researching vaccines, is more of using existing drugs to play a corresponding therapeutic role. However, there is still no good drug in the existing technology to solve the new coronavirus and the complications caused by the new coronavirus. Therefore, further research on this topic is required.

SUMMARY OF THE INVENTION

The invention aims to provide the use of ulinastatin in the preparation of medicines for preventing and treating severe novel coronavirus pneumonia. Tests have shown that ulinastatin can play a therapeutic effect on the new coronavirus pneumonia, and the present invention further explores whether ulinastatin can play a therapeutic role on the complication sepsis caused by the new coronavirus pneumonia. Listatin can have a certain therapeutic effect on the complications of sepsis caused by the new coronavirus pneumonia, and when it is combined with Polygonatum polysaccharide, the therapeutic effect achieved is better than that of a single drug, and it has a significant effect. progress.

In order to achieve the above object, the present invention adopts the following technical solutions:

The use of ulinastatin in preparing a medicine for preventing and treating severe novel coronavirus pneumonia, the single administration dosage of ulinastatin is 300,000 to 10,000,000 units.

Further, the single dosage of ulinastatin is 1 to 9 million units.

Further, the novel coronavirus pneumonia is a disease caused by the novel coronavirus SARS-CoV-2.

Further, the medicine includes ulinastatin and polysaccharide polysaccharide.

Further, the medicine includes ulinastatin, polysaccharide polysaccharide and carboxymethyl schizophyllan.

Further, the medicine is freeze-dried powder injection, injection, spray preparation or atomized preparation.

In order to quickly apply the combination of ulinastatin and polysaccharide in medicine, the inventors further made ulinastatin and polysaccharide polysaccharide into an aerosol formulation, and found that the drug was easily unstable in the accelerated test. It may be caused by the inevitable need to adjust the pH in the preparation method and the inevitable exposure to the light environment. For this reason, the inventor found after a large number of experiments that adding carboxymethyl schizophyllan to the drug at the same time can significantly improve the stability of the drug. It is speculated that carboxymethyl schizophyllan has a protective effect on ulinastatin and polysaccharide polysaccharide. .

Further, each 1000ml of the medicine includes the following components:

Ulinastatin 300,000 to 10,000,000 units;

Povidone 0.3～1.5g;

Carboxymethyl Schizophyllan 0.1～0.5g;

Polysaccharide Polysaccharide 0.1～0.5g;

Isotonicity regulator 5～10g;

pH adjuster 0.01～15g;

Balance of water for injection.

Further, the isotonicity adjusting agent includes one of sodium chloride and glucose.

Further, the pH adjusting agent includes one or more of hydrochloric acid, citric acid-sodium citrate, and citric acid-disodium hydrogen phosphate.

Further, the preparation method of the medicine comprises the following steps:

S1) add ulinastatin, povidone, carboxymethyl schizophyllan, polysaccharide polysaccharide and isotonicity regulator to water for injection, mix and stir until completely dissolved, then add pH regulator, and adjust the pH to be 6.8～ 7.3, to obtain a mixed solution;

S2) adding water for injection to dilute the mixed solution of step S1 to 1000ml, sterilizing by filtration with a 0.22 μm filter membrane, sub-packaging, filling, and packaging, and that’s it.

Compared with the prior art, the present invention has the following beneficial effects:

(1) the present invention finds that ulinastatin can play a therapeutic effect on novel coronavirus, and the therapeutic effect of treatment group B is better than that of treatment group A, indicating that ulinastatin and polysaccharide polysaccharide combined can be used for the treatment of severe Complications of sepsis caused by novel coronavirus pneumonia and severe novel coronavirus pneumonia have important social significance.

(2) The present invention adjusts the composition and preparation method of the medicine. Experiment 1 shows that the medicine shows good stability in the 6-month accelerated test, reduces the risk of clinical medication, and is suitable for industrial production.

Detailed ways

The above-mentioned content of the present invention will be further described in detail below through the specific implementation in the form of examples. However, it should not be understood that the scope of the above-mentioned subject matter of the present invention is limited to the following examples.

Embodiment 1, a kind of medicine for preventing and treating severe novel coronavirus pneumonia

Take 100 million units of ulinastatin after filter sterilization, add 20g mannitol and 10g sodium chloride to dissolve, adjust the pH to 6.8, make up the volume to 1500mL with water for injection, sterile filter, and divide into 1000 vials. That's it.

Embodiment 2, a kind of medicine for preventing and treating severe novel coronavirus pneumonia

Take 100 million units of ulinastatin after filter sterilization, add 20 g of glucose and 5 g of sodium chloride to dissolve, adjust the pH to 7.1, make up the volume to 1500 mL with water for injection, sterile filter, and divide into 1000 vials. That's it.

Embodiment 3, a kind of medicine for preventing and treating severe novel coronavirus pneumonia

The preparation type of the medicine in this example is an atomized preparation, and each 1000ml of the medicine includes the following components:

Ulinastatin 2 million units;

Povidone 0.5g;

Carboxymethyl Schizophyllan 0.15g;

Polygonatum polysaccharide 0.15g;

Isotonicity regulator (sodium chloride) 6g;

pH adjuster (citric acid-sodium citrate) 0.01～15g;

Balance of water for injection.

The preparation method of the medicine comprises the following steps:

S1) adding ulinastatin, povidone, carboxymethyl schizophyllan, polysaccharide polysaccharide and isotonicity regulator to water for injection, mixing and stirring until completely dissolved, then adding pH regulator, and adjusting the pH to be 7.1, get the mixture;

S2) adding water for injection to dilute the mixed solution of step S1 to 1000ml, sterilizing by filtration with a 0.22 μm filter membrane, sub-packaging, filling, and packaging, and that’s it.

Embodiment 4. A kind of medicine for preventing and treating severe novel coronavirus pneumonia

The preparation type of the medicine in this example is an atomized preparation, and each 1000ml of the medicine includes the following components:

Ulinastatin 3 million units;

Povidone 0.75g;

Carboxymethyl Schizophyllan 0.3g;

Polygonatum polysaccharide 0.2g;

Isotonicity regulator (sodium chloride) 6g;

pH adjuster (citric acid-disodium hydrogen phosphate) 0.01～15g;

Balance of water for injection.

The preparation method of the medicine comprises the following steps:

S1) adding ulinastatin, povidone, carboxymethyl schizophyllan, polysaccharide polysaccharide and isotonicity regulator to water for injection, mixing and stirring until completely dissolved, then adding pH regulator, and adjusting the pH to be 7.2, get the mixture;

S2) adding water for injection to dilute the mixed solution of step S1 to 1000ml, sterilizing by filtration with a 0.22 μm filter membrane, sub-packaging, filling, and packaging, and that’s it.

Comparative Example 1. A drug for the prevention and treatment of severe novel coronavirus pneumonia

The difference from Example 1 is that no carboxymethyl schizophyllan was added.

Comparative Example 2. A drug for the prevention and treatment of severe novel coronavirus pneumonia

The difference from Example 1 is that Polygonatum polysaccharide is not added.

Experiment 1. Clinical observation experiment

2.1 Treatment methods: 75 patients diagnosed with severe novel coronavirus pneumonia were randomly divided into treatment group A, treatment group B, and control group with 25 cases in each. The patient was treated with oxygen inhalation or non-invasive ventilator. The patients in the control group were treated with the standard treatment plan (conventional antiviral, anti-inflammatory, immune enhancer and supportive treatment); the treatment group A was combined with the example on the basis of the control group (conventional antiviral, anti-inflammatory, immune enhancer and supportive treatment). 1. Nebulized preparation for treatment; treatment group B was treated with the aerosolized preparation of Example 3 on the basis of the control group (conventional antiviral, anti-inflammatory, immune enhancer and supportive treatment); once every 8 hours, the treatment continued for 5 days.

2.2 Observation indicators

The valid criteria are the following indicators, and those not meeting the following indicators are considered invalid. Indicators include:

(1) CDC throat swabs were negative for novel coronavirus nucleic acid;

(2) Chest CT showed that the inflammation was absorbed earlier than before;

(3) The blood index returns to the health reference value, and the specific items include:

Cytokines (gamma-interferon, alpha-tumor necrosis factor, interleukin-10, interleukin-6, interleukin-4, interleukin-2 and interleukin-17A);

Cardiac three (myoglobin, troponin and creatine kinase isoenzymes);

Detection of oxidative stress levels (ROS, MDA and SOD enzyme expression levels) in plasma and blood cells;

Levels of neutrophils, macrophages and total apoptosis.

Table 1 Observations

As can be seen from Table 1, treatment group A and treatment group B showed that CDC throat swabs were negative for new coronavirus nucleic acid, chest CT showed that the inflammation was absorbed earlier, and blood indicators returned to healthy reference values. Compared with the control group, the effective number of treatment group A after treatment was lower than that of treatment group B. The specific reason was that the blood treatment of treatment group A did not return to the healthy reference value, so it was ineffective.

Experiment 2. The protective effect of drugs on mice with septic lung injury

Experimental method: 60 C57 mice of 240g～260g, half male and half male, were adaptively fed with standard diet for 3 days. A mouse model of septic lung injury was established by injecting 5 mg/kg LPS into the tail vein. The mice were randomly divided into 4 groups, including the model group, the treatment group, the treatment group 2, and the treatment group 3. The control group was injected with the same amount of normal saline and fed normally. . One group of treatment was injected with 0.4ml (20,000 units) of ulinastatin + 0.4ml of normal saline, the second group of treatment was injected with 0.4ml (0.2mg) of Polygonatum polysaccharide + 0.4ml of normal saline, and the third group of treatment was injected with 0.4ml (20,000 units) of Ulinastatin + 0.4ml (0.2mg) Polygonatum polysaccharide, model group and control group were injected with 0.8ml normal saline. Once every 24h, blood was collected after 72h, TNF-α, IL-1β and IL-6 were measured, and compared with the control group at 0h; the mortality after 72h was also measured.

Table 2 Experimental results

Note: The units of TNF-α, IL-1β and IL-6 in Table 2 are multiples.

As can be seen from Table 2, ulinastatin can inhibit TNFα, IL-1β and IL-6-induced inflammatory cytokines, play an active anti-inflammatory effect, and can reduce the mortality of mice with septic lung injury. At the same time, it can also be seen from Table 2 that the effect of treatment group 3 is better than that of treatment group 1 and treatment group 2, indicating that the combination of ulinastatin and polysaccharide polysaccharide can play a more positive protective effect.

Experiment 3. Stability test

Accelerated experimental method: According to the 2015 edition of "Chinese Pharmacopoeia" "Guidelines for Stability Experiments of Raw Materials and Preparations". The examples/comparative examples were placed in a test chamber with a temperature of 30±2°C and a relative humidity of 60±5% for 6 months, and samples were taken at the end of the 0th, 1st, 2nd, 3rd, and 6th months to investigate the appearance, main drug and related substances.

Table 3 Accelerated experimental results

It can be seen from Table 3 that the contents of ulinastatin and polysaccharide polysaccharides and the contents of related substances in the medicines of Examples 1 to 3 of the present invention did not change significantly in the 6-month accelerated test, and the performance was stable. Compared with Example 1, Comparative Example 1 is unstable in the 6-month accelerated experiment, and it can be seen that carboxymethyl schizophyllan can play a synergistic effect with Polygonatum polysaccharide and Ulinastatin to jointly improve the drug's efficacy. stability.

The above-mentioned embodiments merely illustrate the principles and effects of the present invention, but are not intended to limit the present invention. Anyone skilled in the art can modify or change the above embodiments without departing from the spirit and scope of the present invention. Therefore, all equivalent modifications or changes made by those with ordinary knowledge in the technical field without departing from the spirit and technical idea disclosed in the present invention should still be covered by the claims of the present invention.

Claims (7)

The use of ulinastatin in preparing a medicine for preventing and treating severe novel coronavirus pneumonia, characterized in that the single administration dosage of ulinastatin is 300,000 to 10,000,000 units; The medicament includes ulinastatin, polysaccharide polysaccharide and carboxymethyl schizophyllan; The medicine is freeze-dried powder injection, injection, spray preparation or atomized preparation. The use according to claim 1, wherein the single dosage of ulinastatin is 1 million to 9 million units. The use according to claim 1 or 2, wherein the novel coronavirus pneumonia is a disease caused by the novel coronavirus SARS-CoV-2. The use according to claim 1, wherein each 1000ml of the medicine comprises the following components: Ulinastatin 300,000 to 10,000,000 units; Povidone 0.3～1.5g; Carboxymethyl Schizophyllan 0.1～0.5g; Polysaccharide Polysaccharide 0.1～0.5g; Isotonicity regulator 5～10g; pH adjuster 0.01～15g; Balance of water for injection. The use according to claim 4, wherein the isotonicity adjusting agent comprises one of sodium chloride and glucose. The use according to claim 4, wherein the pH adjusting agent comprises one or more of hydrochloric acid, citric acid-sodium citrate, and citric acid-disodium hydrogen phosphate. The use according to any one of claims 4 to 6, wherein the preparation method of the medicine comprises the following steps: S1) add ulinastatin, povidone, carboxymethyl schizophyllan, polysaccharide polysaccharide and isotonicity regulator to water for injection, mix and stir until completely dissolved, then add pH regulator, and adjust the pH to be 6.8～ 7.3, to obtain a mixed solution; S2) adding water for injection to dilute the mixed solution of step S1 to 1000ml, filter and sterilize with a 0.22 μm filter membrane, subpackage, fill, and package, and that’s it.