Application of acetyl-CoA in preparing medicine for treating pneumonia

Abstract

The present invention belongs to the field of biomedicine technology, and in particular to the application of acetyl-CoA in the preparation of a medicine for treating pneumonia. In the present invention, taking Klebsiella pneumoniae as a representative, the improvement effect of acetyl-CoA on bacterial pneumonia mice was explored. The results showed that acetyl-CoA can reduce the lung index of bacterial pneumonia mice to varying degrees, can reduce the bacterial load in the lung tissue of bacterial pneumonia mice, can also improve the damage of bacterial pneumonia mouse lung tissue, and can also reduce the expression level of inflammatory factors in the lung tissue of bacterial pneumonia mice and the oxidative stress level of the lung tissue. In summary, the present invention provides a new use of acetyl-CoA, which provides a new idea and theoretical basis for the treatment of bacterial pneumonia.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and particularly relates to the application of acetyl-CoA in the preparation of medicines for treating pneumonia. Background Art

[0002] Bacterial pneumonia is a lung infection caused by bacteria, typically affecting the alveoli (small air sacs in the lungs), leading to 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 typically include cough, fever, shortness of breath, chest pain, and fatigue. In severe cases, it can lead to respiratory failure and even death, and the elderly, infants, and those with weakened immune systems are particularly susceptible. 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 elevated levels of C-reactive protein (CRP) and procalcitonin (PCT), requiring antibiotic treatment. However, overuse of antibiotics can easily lead to the colonization of drug-resistant bacteria and recurrent attacks of the disease, which can affect the efficacy of treatment and significantly increase the financial burden on patients.

[0003] Recent studies have revealed that the gut microbiota plays a crucial role in the body's defenses. The metabolism of short-chain fatty acids (SCFAs) by the gut microbiota can regulate the lung's immune response through the gut-lung axis, thereby combating bacterial lung infections. SCFAs in the body are primarily metabolized by the gut microbiota. Absorbed through the intestine and entering the bloodstream, they can protect against pathogens by regulating the activation of immune cells. Targeting gut microbiota SCFAs may be a potential strategy for treating bacterial pneumonia.

[0004] Acetyl-CoA is a crucial molecule in cellular metabolism, widely involved in multiple biochemical processes, including energy production, fatty acid synthesis, and cholesterol synthesis. It is a compound formed by the combination of Coenzyme A and an acetyl group (CH3CO), and plays a vital role in the metabolism of intestinal microorganisms. The intestines are home to a large number of microbial communities that not only help digest food but also participate in many metabolic processes, influencing the host's health and disease status. As one of the core metabolic molecules, Acetyl-CoA is closely related to multiple aspects of intestinal microbial energy metabolism, fermentation processes, and the 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-mentioned purpose, the present invention adopts the following technical solutions.

[0008] The present invention provides the use of acetyl-CoA in preparing a 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, with Klebsiella pneumonia as a representative, the improvement effect of acetyl-CoA on bacterial infection pneumonia mice is explored. Specifically by constructing a bacterial infection pneumonia mouse model, different concentrations of acetyl-CoA (low, medium and high doses are 100, 200, 400 mg / kg respectively) are administered afterwards, and the drug is administered continuously for 4 days. 12h after the last administration, the mouse lung tissue is obtained, and a part of the lung tissue is taken for bacterial load detection after the lung tissue is weighed, a part is stained with HE, and a part is detected by lung tissue pathology index. The results showed that acetyl-CoA can reduce the lung index of bacterial pneumonia mice to varying degrees, can reduce the bacterial load in the lung tissue of bacterial pneumonia mice, can also improve the damage of bacterial pneumonia mouse lung tissue, and can also reduce the expression level of inflammatory factors in the lung tissue of bacterial pneumonia mice 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 drug is a tablet, granule, capsule or injection.

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

[0016] In some embodiments, the other pharmaceutically acceptable excipients include solvents, propellants, solubilizers, cosolvents, 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 regulators, buffers, plasticizers, surfactants, foaming agents, defoaming agents, 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 carboxymethylcellulose, 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 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, providing 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, and improve the damage of the lung tissue of mice with bacterial pneumonia. At the same time, it could 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.

[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 shows the statistical results 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 The bar graph shows 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 The bar graph shows 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 the lung tissues of mice in different treatment groups.

[0031] Figure 8 The bar graph shows the results of determination of GSH-Px levels in lung tissue of mice in different treatment groups.

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

[0033] The experimental methods in the following examples of the present invention, where specific conditions are not specified, are generally carried out under conventional conditions or 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. 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 "comprising" and "having" in the present invention and any variations thereof are intended to cover non-exclusive inclusion. For example, a process, method, apparatus, product or device that includes a series of steps is not limited to the listed steps or modules, but optionally further includes steps not listed, or optionally further includes other steps inherent to these processes, methods, products or devices.

[0036] The "plurality" mentioned in the present invention means two or more. "And / or" describes the association relationship of associated objects and indicates that three relationships can exist. For example, A and / or B can represent: A exists alone, A and B exist simultaneously, and B exists alone. The character " / " generally represents an "or" relationship between the associated objects before and after.

[0037] The following is illustrated with specific embodiments.

[0038] Example 1

[0039] In this example, Klebsiella pneumoniae was used as a representative to explore the improvement effect of acetyl coenzyme A on mice with bacterial infectious pneumonia.

[0040] 1. Experimental animals: 60 SPF-grade male BALB / c mice, 6 - 8 weeks old, with a body weight of 20 - 22 g, provided by Zhuhai Besttone Biotechnology Co., Ltd., experimental animal production license number: SCXK(Yue)2020 - 0051. All animals were housed in the SPF-grade animal laboratory of the Shenzhen Institute of Beijing University of Chinese Medicine, experimental animal use license number: SYXK(Yue)2024 - 0374, at a room temperature of 22 - 25°C and a relative humidity of 40% - 60%, following a 12h light / dark cycle, with free diet and water. All experiments designed for mice were approved by the Animal Ethics Committee of the Shenzhen Institute of Beijing University of Chinese Medicine.

[0041] 2. Animal grouping, model establishment and administration: 60 mice were divided into a blank group, a model group, a ceftriaxone sodium group, and low, medium, and high-dose groups of acetyl coenzyme A (purchased from Shanghai Aladdin Biochemical Technology Co., Ltd., product number A463299) by the random number table method, with 10 mice in each group.

[0042] The blank group was given an equal volume of sterile saline. The mice in the other groups were anesthetized by intraperitoneal injection of a 4mg / mL pentobarbital sodium solution at a dose of 0.014 mL / g. The anesthetized mice were fixed supine on the operating table. The oral cavity was opened with a mouse mouth opener, and the epiglottic cartilage was found under the illumination of a surgical lamp. A micro syringe was used to insert into the trachea of the mouse through the epiglottic cartilage, and the concentration was 1×10 9A bacterial solution of CFU 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 acetyl-CoA groups were gavaged with 100, 200 and 400 mg / kg 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 receive the drug on the first day after modeling and continued to receive the drug for 4 consecutive days.

[0044] 3. Sample collection: 12 hours after the last administration, weigh the mice in each group, then kill them and dissect to obtain lung tissue. After weighing the lung tissue, take a portion of the lung tissue for bacterial load detection, a portion for HE staining, and a portion 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) Lung tissue pathology: Lung tissue was embedded in paraffin and sectioned (4-5 μm thick) and stained with hematoxylin-eosin (H&E). Pathologists reviewed the sections and assessed the degree of lung lesions for tissue inflammation, including inflammatory cell infiltration, alveolar septum thickening, and fibrous nodular proliferation.

[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 using real-time fluorescence quantitative PCR.

[0050] (5) Detection of oxidative stress indicators in lung tissue: The lung tissues of each group of mice were accurately weighed and homogenized with pre-cooled physiological saline to prepare a 10% tissue homogenate. The homogenate was then centrifuged at 4°C and 3000 rpm for 15 min. The supernatant was collected and assayed using a malondialdehyde (MDA) assay kit, a superoxide dismutase (SOD) assay kit, and a glutathione peroxidase (GSH-Px) assay 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 reduced 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, the alveoli and trachea of ​​the mice in the blank group had clear histological structures, with no congestion, edema, or inflammatory cell infiltration in the alveolar walls, and no obvious lesions or necrosis in the alveolar epithelial cells. Compared with the blank group, the alveolar walls of the mice in the model group were significantly thickened, with a large number of inflammatory cells (mainly macrophages and neutrophils) infiltrating the alveoli. Compared with the model group, the degree of lung tissue damage in the mice in the acetyl-CoA and ceftriaxone sodium groups was alleviated to varying degrees.

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

[0056] (5) Effects on oxidative stress in lung tissue: The results are as follows Figure 7 、 8As shown in Figures 9 and 10, compared with the blank group, the activities of SOD and GSH-Px in the lung tissues of the 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 tissues of the mice in the acetyl-CoA and ceftriaxone sodium 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. At the same time, it can also reduce the expression level of inflammatory factors and the oxidative stress level of the lung tissue of mice with bacterial pneumonia, 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 can be combined arbitrarily. 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-described embodiments merely illustrate several implementations of the present invention, and while their descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the present invention. It should be noted that a person skilled in the art would be able to make numerous variations and improvements without departing from the spirit of the present invention, all of which fall within the scope of protection of the present invention. Therefore, the scope of protection of the present invention shall be determined by the appended claims.

Claims

1. Use of acetyl-CoA as the sole active ingredient in the preparation of a drug for treating bacterial pneumonia caused by Klebsiella pneumoniae.

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

3. The use according to claim 1, characterized in that The medicine is in the form of tablets, granules, capsules or injections.

4. The use according to claim 1, wherein The drug also includes other pharmaceutically acceptable excipients.

5. The use according to claim 1, characterized in that The drug also includes a drug carrier.

6. The use according to claim 5, characterized in that The drug carrier comprises one of microcapsules, microspheres, nanoparticles and liposomes.

Images