Application of acetyl coenzyme A in preparation of medicine for treating pneumonia

Abstract

The invention belongs to the technical field of biological medicines, and particularly relates to application of acetyl coenzyme A in preparation of a medicine for treating pneumonia. In the invention, klebsiella pneumoniae is used as a representative, the improvement effect of the acetyl coenzyme A on mice with bacterial infectious pneumonia is researched, and the result shows that the acetyl coenzyme A can reduce the lung index of the mice with bacterial pneumonia to different degrees, can reduce the bacterial load in the lung tissue of the mice with bacterial pneumonia, and can improve the lung index of the mice with bacterial pneumonia. The injury of lung tissues of mice with bacterial pneumonia can be improved, and the expression level of inflammatory factors in the lung tissues of the mice with bacterial pneumonia and the oxidative stress level of the lung tissues can be reduced. In conclusion, the invention provides a new application of the acetyl coenzyme A, and a new thought and a theoretical basis are provided for treatment of the bacterial pneumonia.

Description

Technical Field

[0001] The invention belongs to the technical field of biomedicine, and in particular relates to the application of acetyl coenzyme A in preparing medicines for treating pneumonia. Background Art

[0002] Bacterial pneumonia is a lung infection caused by bacteria, usually affecting the alveoli (small air sacs in the lungs), causing inflammation of the lungs. It is one of the most common types of pneumonia and can be caused by a variety of different bacteria. Symptoms of bacterial pneumonia usually include cough, fever, shortness of breath, chest pain, fatigue, etc. In severe cases, respiratory failure and even death may occur, especially in the elderly, infants, and people with weak immune systems. Gram-negative bacteria such as Klebsiella pneumoniae and Pseudomonas aeruginosa are the most common types of bacterial pneumonia. Compared with patients with non-bacterial pneumonia, patients with bacterial pneumonia have significantly increased levels of C-reactive protein (CRP) and procalcitonin (PCT), and need to be treated with antibiotics. However, excessive use of antibiotics can easily lead to colonization of drug-resistant bacteria and repeated attacks of the disease, which affects the treatment efficacy of patients and greatly increases the economic burden on patients.

[0003] In recent years, studies have found that intestinal flora plays an important role in the body's defense. The metabolism of short-chain fatty acids (SCFAs) in the intestinal flora can regulate the lung immune response through the gut-lung axis, thereby responding to bacterial infections in the lungs. SCFAs in the body are mainly metabolized by intestinal flora. They are absorbed into the blood system through the intestine and can resist the invasion of pathogens by regulating the activation of immune cells. Targeting intestinal flora SCFAs may become a potential strategy for the treatment of bacterial pneumonia.

[0004] Acetyl-CoA is a very important molecule in cell metabolism, and is widely involved in multiple biochemical processes such as energy generation, fatty acid synthesis, and cholesterol synthesis. It is a compound formed by the combination of Coenzyme A (Coenzyme A) and an acetyl group (CH3CO), and plays a vital role in the metabolism of intestinal microorganisms. There are a large number of microbial communities in the intestines, which not only help digest food, but also participate in many metabolic processes, affecting the health and disease status of the host. As one of the core molecules of metabolism, acetyl-CoA is closely related to many aspects of intestinal microorganisms, such as energy metabolism, fermentation process, and synthesis of short-chain fatty acids (SCFAs).

[0005] Currently, there is no report on the use of acetyl-CoA in the preparation of drugs for treating bacterial pneumonia. Summary of the invention

[0006] Based on this, a new use of acetyl-CoA is provided, and specifically the use of acetyl-CoA in the preparation of drugs for treating pneumonia is proposed.

[0007] To achieve the above object, the present invention adopts the following technical solution.

[0008] The present invention provides application of acetyl-CoA in preparing medicine for treating bacterial infectious pneumonia.

[0009] In some embodiments, the pneumonia is bacterial pneumonia.

[0010] In some embodiments, the bacterial pneumonia includes bacterial pneumonia caused by Klebsiella pneumoniae, Pseudomonas aeruginosa, or Staphylococcus aureus.

[0011] In the present invention, Klebsiella pneumoniae is used as a representative to explore the improvement effect of acetyl-CoA on mice with bacterial infection pneumonia. Specifically, by constructing a mouse model of bacterial infection pneumonia, different concentrations of acetyl-CoA (low, medium and high doses are 100, 200, 400 mg / kg, respectively) are administered, and the administration is continued for 4 days. 12h after the last administration, the mouse lung tissue is obtained, and a part of the lung tissue is weighed and a part is taken for bacterial load detection, a part is HE stained, and a part is detected for lung tissue pathological indicators. It was found that acetyl-CoA can reduce the lung index of mice with bacterial pneumonia to varying degrees, can reduce the bacterial load in the lung tissue of mice with bacterial pneumonia, can also improve the damage of the lung tissue of mice with bacterial pneumonia, and can also reduce the expression level of inflammatory factors in the lung tissue of mice with bacterial pneumonia and the oxidative stress level of the lung tissue.

[0012] The present invention also provides a medicine for treating bacterial infectious pneumonia, wherein the medicine uses acetyl-CoA as the only effective ingredient.

[0013] In some embodiments, the content of acetyl-CoA in the drug is 1 wt% to 99 wt%.

[0014] In some embodiments, the medicament is a tablet, granules, capsule or injection.

[0015] In some embodiments, the formulation further includes other pharmaceutically acceptable excipients.

[0016] In some embodiments, the other pharmaceutically acceptable excipients include solvents, propellants, solubilizers, co-solvents, emulsifiers, colorants, binders, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, anti-adhesive agents, penetration enhancers, pH adjusters, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, inclusion agents, humectants, absorbents, diluents, flocculants and deflocculating agents, filter aids and release retardants.

[0017] In some embodiments, the filler is selected from starch, sucrose, lactose or microcrystalline cellulose; the binder is selected from cellulose derivatives, alginate, gelatin or polyvinyl pyrrolidone; the wetting agent is selected from water, glycerol, ethanol, methylcellulose, sodium carboxymethyl cellulose, low-substituted hydroxypropyl cellulose or hydroxypropyl methylcellulose; the disintegrant is selected from sodium carboxymethyl starch, hydroxypropyl cellulose, cross-linked carboxymethyl cellulose, agar, calcium carbonate or sodium bicarbonate; the absorption promoter is a quaternary ammonium compound; the surfactant is selected from cetyl alcohol or sodium lauryl sulfate; the adsorption carrier is selected from kaolin or bentonite; the lubricant is selected from talc, calcium and magnesium stearate, micropowdered silica gel or polyethylene glycol; the flavoring agent is selected from sucrose, simple syrup, aromatic syrup, glycerol, sorbitol or mannitol.

[0018] In some embodiments, the medicament further comprises a pharmaceutical carrier.

[0019] In some embodiments, the drug carrier comprises microcapsules, microspheres, nanoparticles and liposomes.

[0020] Based on the technical solution of the present invention, compared with the prior art, the present invention has the following beneficial effects:

[0021] The present invention provides a new use of acetyl-CoA, specifically provides the use of acetyl-CoA in the preparation of a drug for treating bacterial pneumonia, and provides a new idea for the treatment of bacterial pneumonia.

[0022] In the present invention, Klebsiella pneumoniae was used as a representative to explore the improvement effect of acetyl-CoA on mice with bacterial pneumonia. The results showed that acetyl-CoA could reduce the lung index of mice with bacterial pneumonia to varying degrees, reduce the bacterial load in the lung tissue of mice with bacterial pneumonia, improve the damage of the lung tissue of mice with bacterial pneumonia, and reduce the expression level of inflammatory factors in the lung tissue of mice with bacterial pneumonia and the oxidative stress level of the lung tissue.

[0023] The present invention also provides a drug for treating pneumonia. By administering the drug to mice suffering from bacterial pneumonia, the lung index of the mice with bacterial pneumonia can be reduced, the bacterial load in the lung tissue of the mice with bacterial pneumonia can be reduced, and the damage to the lung tissue of the mice with bacterial pneumonia can be improved. At the same time, the expression level of inflammatory factors in the lung tissue of the mice with bacterial pneumonia and the oxidative stress level of the lung tissue can be reduced, thereby achieving a good therapeutic effect on bacterial pneumonia and providing a theoretical basis for the treatment of bacterial pneumonia. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 The bar graph is a statistical result of the lung index of mice in different treatment groups.

[0025] Figure 2 The bar graph shows the statistical results of bacterial load in mouse lung tissue in different treatment groups.

[0026] Figure 3 Figure 2 shows the staining results of mouse lung tissue in different treatment groups.

[0027] Figure 4 This is a bar graph showing the statistical results of IL-1β mRNA expression levels in mouse lung tissues in different treatment groups.

[0028] Figure 5 The bar graph shows the statistical results of IL-6 mRNA expression levels in mouse lung tissues in different treatment groups.

[0029] Figure 6 This is a bar graph showing the statistical results of TNF-α mRNA expression levels in mouse lung tissues in different treatment groups.

[0030] Figure 7 The bar graph shows the results of measuring SOD levels in mouse lung tissues in different treatment groups.

[0031] Figure 8 The bar graph shows the results of measuring GSH-Px levels in mouse lung tissues in different treatment groups.

[0032] Fig. 9 The bar graph shows the results of measuring MDA levels in lung tissue of mice in different treatment groups. DETAILED DESCRIPTION

[0033] The experimental methods in the following examples of the present invention without specifying specific conditions are usually carried out under conventional conditions or under conditions recommended by the manufacturers.

[0034] Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as those commonly understood by those skilled in the art of the present invention. The terms used in the specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention.

[0035] The terms "including" and "having" and any variations thereof of the present invention are intended to cover non-exclusive inclusions. For example, a process, method, device, product or equipment comprising a series of steps is not limited to the listed steps or modules, but may optionally include steps not listed, or may optionally include other steps inherent to these processes, methods, products or equipment.

[0036] In the present invention, the term "multiple" refers to two or more than two. "And / or" describes the association relationship of associated objects, indicating that three relationships may exist. For example, A and / or B can represent the following three situations: A exists alone, A and B exist at the same time, and B exists alone. The character " / " generally indicates that the associated objects are in an "or" relationship.

[0037] The following describes the invention in conjunction with specific embodiments.

[0038] Example 1

[0039] This example uses Klebsiella pneumoniae as a representative to explore the improving effect of acetyl-CoA on mice with bacterial pneumonia.

[0040] 1. Experimental animals: 60 SPF male BALB / c mice, 6-8 weeks old, weighing 20-22g, provided by Zhuhai Baishitong Biotechnology Co., Ltd., experimental animal production license number: SCXK (Guangdong) 2020-0051. All animals were kept in the SPF animal laboratory of Shenzhen Beijing University of Chinese Medicine Research Institute, experimental animal use license number: SYXK (Guangdong) 2024-0374, room temperature 22-25℃, relative humidity 40%-60%, follow 12h light / dark cycle, free food and water. All experiments designed for mice were approved by the Animal Ethics Committee of Shenzhen Beijing University of Chinese Medicine Research Institute.

[0041] 2. Animal grouping, modeling and drug administration: 60 mice were randomly divided into blank group, model group, ceftriaxone sodium group and acetyl-CoA (purchased from Shanghai Aladdin Biochemical Technology Co., Ltd., catalog number A463299) low-, medium- and high-dose groups using the random number table method, with 10 mice in each group.

[0042] The blank group was given an equal volume of sterile saline, and the mice in the other groups were anesthetized by intraperitoneal injection of 4 mg / mL sodium pentobarbital solution at a dose of 0.014 mL / g. The anesthetized mice were fixed on the operating table in supine position, and the mouth was opened with a mouse mouth opener. The epiglottal cartilage was found with the operating light, and a micro-syringe was inserted into the mouse trachea through the epiglottal cartilage. 9CFU of Klebsiella pneumoniae (purchased from China Industrial Microbiological Culture Collection Center, strain number CICC10870) was injected into the lungs of mice at a rate of 50 μL per mouse.

[0043] After modeling, the ceftriaxone sodium group was given a single dose of 200 mg / mL ceftriaxone sodium solution by tail vein injection; the low, medium and high dose groups of acetyl-CoA were gavaged with 100, 200 and 400 mg / kg of acetyl-CoA, respectively, and the blank group and model group were gavaged with normal saline. The gavage volume of each group was 10 mL / kg. Each group started to be administrated on the first day after modeling and continued to be administrated for 4 days.

[0044] 3. Sample collection: 12 hours after the last administration, weigh the mice in each group, then kill and dissect to obtain lung tissue. After the lung tissue is weighed, a portion of the lung tissue is taken for bacterial load detection, a portion is taken for HE staining, and a portion is taken for lung tissue pathology index detection.

[0045] 4. Index detection method

[0046] (1) Lung index detection: Lung tissues of mice in each group were collected and the whole lung weight was weighed. The lung index of mice in each group was calculated according to the formula “lung index (%) = lung wet weight (g) / body weight (g) × 100”.

[0047] (2) Lung bacterial load detection: About 8 mg of lung tissue was taken and homogenized with normal saline. After centrifugation, 2-3 mL was taken and placed in a cuvette. The lung tissue bacterial load was evaluated by detecting the OD600 value.

[0048] (3) Pathological examination of lung tissue: The lung tissue was embedded in paraffin and sliced ​​(4-5 μm thick) and stained with hematoxylin-eosin (H&E). Pathologists reviewed the slides and evaluated tissue inflammation based on the degree of lung tissue lesions: inflammatory cell infiltration, alveolar septum thickening, and fibrous nodular hyperplasia.

[0049] (4) Detection of inflammatory factor levels in lung tissue: Total RNA was extracted from lung tissue, and the expression of IL-1β, IL-6, and TNF-α mRNA was detected by real-time fluorescence quantitative PCR.

[0050] (5) Detection of oxidative stress indicators in lung tissue: The lung tissue of each group of mice was accurately weighed, and pre-cooled physiological saline was used as the homogenization medium to prepare a 10% tissue homogenate. The homogenate was then centrifuged at 4°C and 3000 rpm for 15 min. The supernatant was used for the malondialdehyde (MDA) assay kit, superoxide dismutase (SOD) assay kit, and glutathione peroxidase (GSH-Px) test kit to measure the levels of MDA, SOD, and GSH-Px.

[0051] 5. Experimental results

[0052] (1) Effect on lung index: The results are as follows Figure 1 As shown in the figure, compared with the blank group, the lung index of mice in the model group was significantly increased (P < 0.05), indicating that bacterial infection caused pathological damage to the lungs and the pneumonia mouse model was successfully established. Compared with the model group, the intervention of acetyl-CoA and ceftriaxone sodium can reduce the lung index of mice to varying degrees.

[0053] (2) Effect on bacterial load: The results are as follows Figure 2 As shown in the figure, compared with the blank group, the bacterial load in the lung tissue of the model group mice was significantly increased. Compared with the model group, the intervention of acetyl-CoA and ceftriaxone sodium decreased the bacterial load in the lung tissue of mice to varying degrees.

[0054] (3) Effects on lung tissue pathology: The results are as follows Figure 3 As shown in the figure, the alveolar and tracheal tissue structures of the blank group mice were clear, the alveolar wall had no congestion, edema, inflammatory cell infiltration, and no obvious lesions or necrosis of alveolar epithelial cells. Compared with the blank group, the alveolar wall of the model group mice was significantly thickened, and a large number of inflammatory cells (mainly macrophages and neutrophils) infiltrated the alveoli; compared with the model group, the degree of lung tissue damage of the mice in the acetyl-CoA and ceftriaxone groups was alleviated to varying degrees.

[0055] (4) Effects on inflammatory factors in lung tissue: The results are as follows Figure 4 , 5 , 6Compared with the blank group, the expression levels of IL-1β, IL-6 and TNF-α mRNA in the lung tissue of the mice in the model group were significantly increased (P<0.01), indicating that bacterial infection caused an inflammatory response in the lung tissue. Compared with the model group, the expression levels of inflammatory factor mRNA in the lung tissue of the mice in the acetyl-CoA and ceftriaxone groups decreased to varying degrees.

[0056] (5) Effects on oxidative stress in lung tissue: Results Figure 7 , 8As shown in Figures 9 and 10, compared with the blank group, the activities of SOD and GSH-Px in the lung tissue of mice in the model group were significantly decreased (P < 0.05), and the level of MDA was significantly increased (P < 0.05). Compared with the model group, the activities of SOD and GSH-Px in the lung tissue of mice in the acetyl-CoA and ceftriaxone groups increased to varying degrees, and the level of MDA decreased to varying degrees.

[0057] In summary, acetyl-CoA can reduce the lung index of mice with bacterial pneumonia to varying degrees, reduce the bacterial load in the lung tissue of mice with bacterial pneumonia, and improve the damage of the lung tissue of mice with bacterial pneumonia. It can also reduce the expression level of inflammatory factors in the lung tissue of mice with bacterial pneumonia and the level of oxidative stress in the lung tissue, and has a good therapeutic effect on bacterial pneumonia, providing a theoretical basis for the treatment of bacterial pneumonia.

[0058] The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0059] The above-mentioned embodiments only express several implementation methods of the present invention, and the description thereof is relatively specific and detailed, but it cannot be understood as limiting the scope of the patent of the present invention. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, which all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the attached claims.

Claims

1. Application of acetyl-CoA in the preparation of medicines for treating pneumonia.

2. The use according to claim 1, characterized in that The pneumonia is bacterial infection pneumonia.

3. The use according to claim 2, characterized in that The bacterial pneumonia includes bacterial pneumonia caused by Klebsiella pneumoniae, Pseudomonas aeruginosa or Staphylococcus aureus.

4. A drug for treating bacterial pneumonia, characterized in that: The drug contains the acetyl-CoA described in claim 1 as the only active ingredient.

5. The drug according to claim 4, characterized in that The content of acetyl-CoA in the medicine is 1wt% to 99wt%.

6. The drug according to claim 4, characterized in that The medicine is in the form of tablets, granules, capsules or injections.

7. The drug according to claim 4, characterized in that The medicine also includes other pharmaceutically acceptable excipients.

8. The drug according to claim 7, characterized in that The other pharmaceutically acceptable excipients include solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, adhesives, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, anti-adhesive agents, penetration enhancers, pH regulators, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, inclusion agents, humectants, absorbents, diluents, flocculants and deflocculating agents, filter aids and release retardants.

9. The drug according to claim 4, characterized in that The drug also includes a drug carrier.

10. The drug according to claim 9, characterized in that The drug carriers include microcapsules, microspheres, nanoparticles and liposomes.

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