A huashi baidu tang nanoparticle is derived from an assembled nanoparticle, and a preparation method and application thereof

Abstract

The application provides a Huashi Baidu Tang nanoparticle derived from self-assembled nanoparticles and a preparation method and application thereof, and belongs to the technical field of biological medicines. A monomer self-assembled nanoparticle (NPs) is constructed by using patchoulenone and rhein as raw materials, and the monomer self-assembled nanoparticle has excellent stability. In-vivo and in-vitro experiments show that the self-assembled nanoparticle can significantly improve the lung function of mice with staphylococcus aureus-induced pneumonia, reduce lung pathological damage, remove bacteria in the lung and inhibit the expression of inflammatory factors through a synergistic anti-inflammatory and antibacterial dual mechanism, has a good antibacterial pneumonia effect, and provides a new idea and method for the treatment of bacterial pneumonia.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and in particular to a self-assembled nanoparticle derived from Huashi Baidu Decoction nanoparticles, its preparation method, and its application. Background Technology

[0002] Bacterial pneumonia is the sixth leading cause of death worldwide and the only infectious disease among the top ten causes of death. Staphylococcus aureus (S. aureus) is the most common pathogen causing hospital-acquired bacterial pneumonia and one of the most common healthcare-associated infections. Staphylococcus aureus commonly exists as a human commensal bacterium in the nasal cavity, on the skin, and on mucous membranes, causing everything from minor skin infections to life-threatening conditions such as pneumonia, sepsis, and endocarditis. Notably, the multidrug resistance of Staphylococcus aureus poses a significant challenge to clinical treatment.

[0003] Traditional Chinese medicine (TCM) shows significant potential in treating bacterial pneumonia, with advantages including direct bactericidal activity, host immune modulation, symptom relief, and synergistic effects with antibiotics. Recent studies have revealed that TCM compound formulas commonly exhibit self-assembly during decoction, forming nanoscale self-assembled nanoparticles and supramolecular structures. These TCM self-assembled nanoparticles possess unique advantages, including utilizing molecular synergy, imparting novel biological activities (such as inducing protein conformational changes), and improving drug solubility and bioavailability.

[0004] Huashi Baidu Decoction (HSBD) is a compound preparation composed of 14 traditional Chinese medicines. Clinically, it is mainly used for the treatment of COVID-19, showing significant inhibitory effects on both the novel coronavirus and virus-induced inflammation. The components of Huashi Baidu Decoction include ephedra, apricot kernel, gypsum, licorice, patchouli, magnolia bark, atractylodes, rhubarb, pinellia, cardamom, poria, astragalus, lepidium seed, and red peony root. Current research on Huashi Baidu Decoction mainly focuses on its antiviral and anti-inflammatory effects. There are no reports in the literature regarding its activity against Staphylococcus aureus, nor are there any systematic studies on its antibacterial pneumonia activity. Furthermore, there are no reports of isolating natural self-assembled nanoparticles from Huashi Baidu Decoction and identifying its core active ingredient, let alone reports of constructing monomeric self-assembled nanoparticles using this core active ingredient and using them to treat Staphylococcus aureus pneumonia. Summary of the Invention

[0005] In view of this, the present invention provides self-assembled nanoparticles derived from Huashi Baidu Decoction nanoparticles, their preparation method, and applications. The present invention constructs monomeric self-assembled nanoparticles (NPs) using patchouli ketone and rhein as raw materials. These monomeric self-assembled nanoparticles exhibit excellent stability. In vivo experiments show that the self-assembled nanoparticles, through a synergistic anti-inflammatory and antibacterial dual mechanism, can significantly improve lung function in mice with Staphylococcus aureus-induced pneumonia, reduce lung pathological damage, clear lung bacteria, and inhibit the expression of inflammatory factors, demonstrating good antibacterial pneumonia effects.

[0006] To achieve the above objectives, the present invention adopts the following technical solution: In a first aspect, the present invention provides a self-assembled nanoparticle derived from Huashi Baidu Decoction nanoparticles, wherein the effective components of the self-assembled nanoparticles include patchouli ketone and rhein.

[0007] Furthermore, the molar ratio of patchouli ketone to rhein is 1:1.

[0008] Furthermore, the particle size of the self-assembled nanoparticles is 200-300 nm.

[0009] Furthermore, the zeta potential of the self-assembled nanoparticles is -8 to -12 mV.

[0010] Secondly, the present invention provides a method for preparing self-assembled nanoparticles derived from the aforementioned Huashi Baidu Decoction nanoparticles, comprising the following steps: (1) Dissolve patchouli and rhein in an organic solvent in proportion, mix well to obtain a mixture; (2) Add the mixture dropwise to preheated water and stir; (3) Dialyze the obtained solution and freeze-dry it to obtain the self-assembled nanoparticles derived from Huashi Baidu Decoction nanoparticles.

[0011] Thirdly, the present invention provides a Huashi Baidu Decoction nanoparticle, wherein the self-assembled nanoparticle is derived from the decoction of Huashi Baidu Decoction.

[0012] Furthermore, the particle size of the self-assembled nanoparticles is 300-400 nm.

[0013] Furthermore, the zeta potential of the self-assembled nanoparticles is -10 to -15 mV.

[0014] Furthermore, the self-assembled nanoparticles are spherical in shape.

[0015] Fourthly, the preparation method of the Huashi Baidu Decoction nanoparticles includes the following steps: (1) Boil the raw materials of Huashi Baidu Decoction with water and collect the decoction; (2) The decoction is subjected to gradient centrifugation and the precipitate is collected to obtain the Huashi Baidu Decoction nanoparticles.

[0016] Further, step (2) of gradient centrifugation includes centrifuging at 2000×g, 4000×g, 6000×g and 8000×g in sequence, collecting the supernatant; centrifuging the supernatant at 20000×g, collecting the precipitate.

[0017] Fifthly, the present invention also provides self-assembled nanoparticles derived from the Huashi Baidu Decoction nanoparticles, or the use of the Huashi Baidu Decoction nanoparticles in the preparation of medicaments for treating and / or preventing bacterial pneumonia.

[0018] Furthermore, the bacterial pneumonia is pneumonia caused by Staphylococcus aureus infection.

[0019] Furthermore, the drug has the following properties: improving lung function indicators in mice with bacterial pneumonia, reducing lung tissue pathological damage, decreasing the total number of white blood cells, the percentage of neutrophils and monocytes in the blood, increasing the percentage of lymphocytes in the blood, decreasing the level of inflammatory factors in serum and / or lung tissue, and inhibiting the expression of COX-2, IL-6 and IL-1β proteins in lung tissue.

[0020] Furthermore, the drug is administered via nebulized inhalation. Nebulized inhalation facilitates the deposition of nanomedicines at the site of lung lesions and prolongs their residence time at the site of infection.

[0021] Compared with the prior art, the present invention has the following beneficial effects: 1. This invention is the first to discover that Huashi Baidu Decoction can spontaneously form bioactive Huashi Baidu Decoction nanoparticles (HSBDN) during the decoction process, and successfully isolates and identifies these nanoparticles.

[0022] 2. This invention successfully constructed well-structured and stable monomer self-assembled nanoparticles (NPs) using pogostone (POG) and rhein (RHE) as raw materials. In vitro and in vivo experiments demonstrated that the monomer self-assembled nanoparticles (NPs) have a significant therapeutic effect on Staphylococcus aureus-induced pneumonia through a synergistic anti-inflammatory and antibacterial mechanism. Attached Figure Description

[0023] The above and / or additional aspects and advantages of the present invention will become apparent and readily understood from the description of the embodiments taken in conjunction with the following drawings, in which: Figure 1This invention presents the preparation process and characterization results of the Huashi Baidu Decoction nanoparticles (HSBDN). (A) Schematic diagram of the preparation process of HSBDN; (BC) Transmission electron microscopy images and particle size distribution of HSBDN; (DE) Zeta potential and polydispersity index (PDI) of HSBDN; (F) UPLC-Q-Orbitrap MS total ion chromatogram of HSBDN; (G) Screening results of antibacterial activity of 9 major compounds in HSBDN; (H) Screening results of anti-inflammatory activity of 9 major compounds in HSBDN.

[0024] Figure 2 The preparation process and characterization results of POG-RHE carrier-free self-assembled nanoparticles (NPs) are shown. (A) Schematic diagram of NP preparation process; (BC) Transmission electron microscopy images of NPs; (DE) Particle size and Zeta potential of NPs; (F) Stability of NPs within 7 days; (GH) Fourier transform infrared and ultraviolet-visible spectra of NPs; (I) Determination of patchouli ketone and rhein content in NPs; (J) Comparative evaluation of the NO inhibition effect of POG, RHE, NPs synthesized by both and their equal-proportion mixture; (KL) Comparative evaluation of the M1 polarization inhibition effect of POG, RHE, NPs synthesized by both and their equal-proportion mixture; (M) Comparative evaluation of the Staphylococcus aureus killing effect of POG, RHE, NPs synthesized by both and their equal-proportion mixture.

[0025] Figure 3 The results of molecular dynamics simulations of the self-assembly of patchouli and rhein to form nanoparticles are shown. (A) Simulation snapshots every 5 ns; (B) Co-aggregation state of water molecules with POG-RHE at 30 ns; (C) π-π stacking and σ-π conjugation interactions; (D) RMSD over time; (E) SASA over time; (F) Number of hydrogen bonds over time; (G) Radius of gyration over time.

[0026] Figure 4 The therapeutic effects of HSBDT, HSBDN, and NPs on Staphylococcus aureus-induced pneumonia in mice are shown. (A) Schematic diagram of the animal experimental protocol; (BF) HSBDT, HSBDN, and NPs significantly improved the decline in lung function (peak inspiratory flow (B), respiratory rate (C), inspiratory volume (D), relaxation time (E), tidal volume (F)) in Staphylococcus aureus-induced pneumonia mice; (G) H&E staining of lung tissue; (HK) HSBDT, HSBDN, and NPs significantly improved the effects of Staphylococcus aureus-induced pneumonia on blood routine tests (white blood cells (H), neutrophils (I), lymphocytes (J), monocytes (K)).

[0027] Figure 5The study demonstrated the anti-inflammatory and bactericidal effects of HSBDT, HSBDN, and NPs on Staphylococcus aureus-induced pneumonia in mice. (A) HSBDT, HSBDN, and NPs significantly reduced the number of bacterial colonies in the lung tissue of mice with Staphylococcus aureus-induced pneumonia. (BE) HSBDT, HSBDN, and NPs significantly reduced the serum levels of IL-6, IL-1β, TNF-α, and NO in mice with Staphylococcus aureus-induced pneumonia. (FG) HSBDT, HSBDN, and NPs significantly reduced the protein expression of COX-2, IL-6, and IL-1β in the lung tissue of mice with Staphylococcus aureus-induced pneumonia.

[0028] Figure 6 HSBDT, HSBDN, and NPs were shown to have good biocompatibility. (A) H&E staining of heart, liver, spleen, and kidney of mice treated with HSBDT, HSBDN, and NPs; (B) Blood biochemical analysis of heart, liver, and kidney function and metabolic markers including ALT (B), AST (C), TBIL (D), CREA (E), BUN (F), CK (G), TG (H), and TCHO (I). Detailed Implementation

[0029] The embodiments of the present invention are described in detail below. The embodiments described below are exemplary and are only used to explain the present invention, and should not be construed as limiting the present invention.

[0030] It should be noted that the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of technical features indicated. Therefore, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. Furthermore, in the description of this invention, unless otherwise stated, "a plurality of" means two or more.

[0031] The endpoints and any values ​​of the ranges disclosed in this invention are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of the various ranges, the endpoint values ​​of the various ranges and individual point values, and individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed in this invention.

[0032] In this invention, the terms "comprising" or "including" are open-ended expressions, meaning they include the contents specified in this invention but do not exclude other aspects.

[0033] In this invention, the terms "optionally," "optionally," or "optionally" generally refer to events or conditions described below that may but may not occur, and the description includes both cases in which the event or condition occurs and cases in which the event or condition does not occur.

[0034] This invention proposes dehumidifying and detoxifying nanoparticles and their derived self-assembled nanoparticles, preparation methods, and applications, which will be described in detail below.

[0035] In this invention, the term "Huashi Baidu Tang" refers to a traditional Chinese medicine compound composed of ephedra, apricot kernel, gypsum, licorice, patchouli, magnolia bark, atractylodes, rhubarb, pinellia, cardamom, poria, astragalus, lepidium seed, and red peony root in a certain proportion. This formula has the effects of resolving dampness and detoxifying, and clearing heat from the lungs. Clinically, it is used to treat damp-heat accumulation in the lungs.

[0036] In this invention, the term "Huashi Baidu Decoction nanoparticles" refers to the nanoscale supramolecular structure formed by the spontaneous aggregation of active ingredients in Huashi Baidu Decoction through non-covalent interactions (such as hydrophobic interactions, hydrogen bonds, van der Waals forces, π-π stacking, etc.).

[0037] In this invention, the term "pogostone" (POG) is a natural active ingredient derived from patchouli (Agastache Herba) that has various pharmacological activities such as anti-inflammatory and antibacterial properties.

[0038] In this invention, the term "rhein" (RHE) refers to an anthraquinone active ingredient derived from rhubarb (Rheum palmatum) that possesses various pharmacological activities such as anti-inflammatory and antibacterial properties.

[0039] In this invention, the term "gradient centrifugation" refers to performing centrifugation operations at different centrifugal forces sequentially to gradually remove particles of different sizes. Specifically, large particles and micron-sized particles are removed first at a lower rotation speed, and then nanoscale particles are collected at a higher rotation speed.

[0040] In this invention, the term "treatment" refers to the attainment of a desired pharmacological and / or physiological effect. This effect may be preventative in terms of complete or partial prevention of the disease or its symptoms, and / or therapeutic in terms of partial or complete cure of the disease and / or adverse effects caused by the disease. As used herein, "treatment" encompasses diseases in mammals, particularly humans, including: (a) prevention of disease or the onset of symptoms in susceptible individuals who have not yet been diagnosed with the disease; (b) suppression of disease, such as inhibiting disease progression; or (c) alleviation of disease, such as reducing disease-related symptoms.

[0041] In a first aspect, this invention provides self-assembled nanoparticles derived from Huashi Baidu Decoction nanoparticles, wherein the effective components of the self-assembled nanoparticles include patchouli ketone and rhein. Preferably, the self-assembled nanoparticles are formed by the self-assembly of patchouli ketone and rhein. The inventors performed component analysis on Huashi Baidu Decoction nanoparticles (HSBDN) using UPLC-Q-Orbitrap MS, identifying nine major compounds (including safflowerin, pentamethoxyflavone, verrucoside-7-O-glucoside, rhein, norihesperidin, hesperidin, ligustrolactone M, rhein, and patchouli ketone). Through screening for anti-inflammatory (LPS-induced inhibition of NO release in RAW264.7 cells) and antibacterial (inhibition of Staphylococcus aureus) activities, patchouli ketone and rhein were found to be the two major components with the highest content and the best anti-inflammatory and antibacterial activities in Huashi Baidu Decoction nanoparticles (HSBDN). Based on this, the inventors successfully constructed monomer self-assembled nanoparticles (NPs) using patchouli ketone and rhein as assembly units.

[0042] In some embodiments of the present invention, the molar ratio of patchouli ketone to rhein is 1:1.

[0043] In some embodiments of the present invention, the self-assembled nanoparticles have a particle size of 200-300 nm. Transmission electron microscopy shows that the NPs have a uniform and stable spherical morphology with a hydration dynamic diameter of 273.8 nm.

[0044] In some embodiments of the present invention, the zeta potential of the self-assembled nanoparticles is -8 to -12 mV. The zeta potential of the NPs is -9.8 mV.

[0045] In some embodiments of the present invention, the self-assembled nanoparticles show no significant changes in particle size and polydispersity index over 7 days at room temperature, demonstrating good stability.

[0046] In this invention, patchouli and rhein self-assemble through non-covalent interactions such as π-π stacking, σ-π conjugation, and hydrogen bonding. Molecular dynamics simulations show that during a 30 ns simulation, patchouli and rhein molecules gradually aggregate through non-covalent interactions such as π-π stacking and σ-π conjugation to form a stable nanoparticle structure; the root mean square deviation (RMSD) of the system tends to equilibrium after 15 ns and stabilizes at approximately 3.256 ± 0.057 nm; the solvent accessible surface area (SASA) gradually decreases and eventually stabilizes at approximately 102.501 ± 3.977 Å. 2 The number of hydrogen bonds gradually increases and stabilizes in the range of 4-7; the radius of gyration decreases significantly and eventually reaches equilibrium in the range of 2.092±0.092 nm. The trends of the above parameters all demonstrate that POG and RHE can self-assemble into structurally stable nanoparticles.

[0047] Secondly, the present invention provides a method for preparing self-assembled nanoparticles derived from the aforementioned Huashi Baidu Decoction nanoparticles, comprising the following steps: (1) Dissolve patchouli and rhein in an organic solvent in proportion, mix well to obtain a mixture; (2) Add the mixture dropwise to preheated water and stir; (3) Dialyze the obtained solution and freeze-dry it to obtain the self-assembled nanoparticles derived from Huashi Baidu Decoction nanoparticles.

[0048] In some embodiments of the present invention, the organic solvent in step (1) includes dimethyl sulfoxide and / or methanol.

[0049] In some embodiments of the present invention, the temperature of the preheated water in step (2) is 70°C.

[0050] In some embodiments of the present invention, the molecular weight cutoff for dialysis in step (3) is 3.5 kDa; and the dialysis time is 8-12 h.

[0051] In some embodiments of the present invention, the dialysis is performed using a dialysis bag to remove organic solvents and small molecule impurities.

[0052] Thirdly, this invention provides nanoparticles derived from the decoction of Huashi Baidu Tang (a traditional Chinese medicine formula). During their research, the inventors discovered that Huashi Baidu Tang can spontaneously form bioactive self-assembled nanoparticles (HSBDNs) during the decoction process. These self-assembled nanoparticles can be separated from the decoction using gradient centrifugation. These nanoparticles are an important pharmacodynamic material basis for Huashi Baidu Tang.

[0053] In some embodiments of the present invention, the particle size of the Huashi Baidu Decoction nanoparticles is 300-400 nm.

[0054] In some embodiments of the present invention, the Huashi Baidu Decoction nanoparticles have a negatively charged surface with a Zeta potential of -10 to -15 mV, indicating that their surface is negatively charged.

[0055] In some embodiments of the present invention, the Huashi Baidu Decoction nanoparticles are spherical in shape with a hydration dynamic diameter of 379.2 nm.

[0056] In some embodiments of the present invention, the polydispersity index (PDI) of the Huashi Baidu Decoction nanoparticles is 0.22, indicating that the nanoparticles are uniform in size and stable.

[0057] Fourthly, the preparation method of the Huashi Baidu Decoction nanoparticles includes the following steps: (1) Boil the raw materials of Huashi Baidu Decoction with water and collect the decoction; (2) The decoction is subjected to gradient centrifugation and the precipitate is collected to obtain the Huashi Baidu Decoction nanoparticles.

[0058] In some embodiments of the present invention, the decoction includes a first decoction and a second decoction, and the decoction liquids from the two decoctions are combined.

[0059] In some embodiments of the present invention, the gradient centrifugation includes centrifugation at 2000 g, 4000 g, 6000 g, and 8000 g sequentially. Preferably, step (2) of the gradient centrifugation includes centrifugation at 2000×g, 4000×g, 6000×g, and 8000×g sequentially, collecting the supernatant; centrifuging the supernatant at 20000×g, and collecting the precipitate. The present invention, by gradually increasing the centrifugal force, can successively remove large particulate impurities and micron-sized aggregates, and finally collect nanoscale dehumidifying and detoxifying nanoparticles by high-speed centrifugation at 20000×g.

[0060] Preferably, the preparation method of the Huashi Baidu Decoction nanoparticles includes the following steps: (1) Soak the raw materials of Huashi Baidu Decoction in water and then decoct them. Filter and collect the decoction. (2) The decoction is subjected to gradient centrifugation, and the supernatant is collected; (3) The supernatant is centrifuged at high speed and the precipitate is collected to obtain self-assembled nanoparticles.

[0061] In some embodiments of the present invention, the high-speed centrifugation in step (3) is 20,000 g centrifugation for 30 minutes.

[0062] In some embodiments of the present invention, the dampness-resolving and detoxifying nanoparticles contain multiple components such as patchouli ketone and rhein. Patchouli ketone is derived from patchouli, and rhein is derived from rhubarb; both are effective components of the dampness-resolving and detoxifying decoction.

[0063] Fifthly, the present invention also provides self-assembled nanoparticles derived from the Huashi Baidu Decoction nanoparticles, or the use of the Huashi Baidu Decoction nanoparticles in the preparation of medicaments for treating and / or preventing bacterial pneumonia.

[0064] In some embodiments of the present invention, the bacterial pneumonia is pneumonia caused by Staphylococcus aureus infection.

[0065] In some embodiments of the present invention, the drug includes one or more of the following properties: 1) Improve lung function indicators in mice with bacterial pneumonia, including peak inspiratory flow, peak expiratory flow, inspiratory volume, tidal volume, and respiratory rate; 2) Reduce pathological damage to lung tissues, including alveolar hemorrhage, edema, alveolar septal thickening, and inflammatory cell infiltration; 3) Reduces the total white blood cell count, percentage of neutrophils, and percentage of monocytes in the blood; 4) Increase the percentage of lymphocytes in the blood; 5) Reduce the bacterial load in the lungs; 6) Reduce the levels of inflammatory factors in serum and / or lung tissue, including IL-6, IL-1β, TNF-α, and NO; 7) Inhibits the expression of COX-2, IL-6 and IL-1β proteins in lung tissue.

[0066] In some embodiments of the present invention, the drug is administered via nebulized inhalation. Nebulized inhalation allows the nanomedicine to be delivered directly to the lung lesion site, which facilitates drug accumulation at the infection site and prolongs its residence time, thereby improving therapeutic efficacy and reducing systemic side effects.

[0067] In this invention, the term "object" is not limited and refers to a human or non-human mammal that is the target of administration of the drug of this invention, such as mammals including cattle, horses, dogs, sheep or cats. Humans are preferred as the target.

[0068] It should be noted that the characteristics and advantages described above for self-assembled nanoparticles also apply to this application, and will not be repeated here.

[0069] The present invention will be explained below with reference to embodiments. Those skilled in the art will understand that the following embodiments are for illustrative purposes only and should not be considered as limiting the scope of the invention. Where specific techniques or conditions are not specified in the embodiments, they are performed according to the techniques or conditions described in the literature in the field or according to the product instructions. Reagents or instruments whose manufacturers are not specified are all conventional products that can be obtained commercially.

[0070] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions of this invention will be clearly and completely described below. Obviously, the described embodiments are only a part of the embodiments of this invention, and not all of them. Based on the embodiments of this invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this invention.

[0071] Unless otherwise specified, the test methods or experimental methods described in the following examples are all conventional methods; unless otherwise specified, the raw materials and additives are obtained from conventional commercial sources or prepared by conventional methods.

[0072] Example 1 Preparation and characterization of self-assembled nanoparticles of Huashi Baidu Decoction.

[0073] 1.1 Drugs and reagents The 14 raw materials of Huashi Baidu Decoction (Ephedra 18g, Apricot Kernel 27g, Gypsum 45g, Licorice 9g, Patchouli 30g, Magnolia Bark 30g, Atractylodes 45g, Rhubarb 15g, Pinellia 27g, Amomum Fruit 18g, Poria 45g, Astragalus 30g, Lepidium Seed 30g, Red Peony Root 30g) were purchased from Dongzhimen Hospital.

[0074] Patchouli and rhein (purity >98%) were purchased from Shanghai Yuanye Biotechnology Co., Ltd. Reagent replenishment: Mangosteenin, p-methoxyflavone, verbascoside-7-O-glucoside, emodin, nobiletin, hesperidin, ligustrolactone M, rhein, and patchouli (purity >98%) were purchased from Shanghai Yeyuan Biotechnology Co., Ltd. LC / MS reagents, including TBTA, biotin-alkynyl, CuSO4, TAMRA-alkynyl, and TCEP (C4706), were purchased from Sigma-Aldrich (USA).

[0075] 1.2 Preparation of HSBDN Add the 14 raw materials of Huashi Baidu Decoction to 1L of water (the liquid level is 1 cm above the herbs) and soak for 30 minutes. Bring to a boil over high heat and then simmer over low heat for 60 minutes. Filter and collect the decoction. Add 1L of water again for a second extraction (bring to a boil over high heat and then simmer over low heat for 60 minutes). Combine the two decoctions and filter. Centrifuge the filtrate sequentially at 2000 g, 4000 g, 6000 g and 8000 g and collect the supernatant. Then centrifuge the supernatant at 20000 g for 30 minutes and collect the precipitate (HSBDN) and the supernatant (HSBDT). Freeze-dry and store.

[0076] 1.3 Hydrodynamic diameter and ζ-potential The lyophilized dehumidifying and detoxifying nanoparticles and self-assembled nanoparticles were reconstituted with double-distilled water to a concentration of 20 μg / mL. The hydrodynamic size distribution, polydispersity index (PDI), and zeta potential of the nanoparticles were then measured using dynamic light scattering (DLS) on a Zetasizer advance instrument. The reported values ​​represent the average of three consecutive measurements.

[0077] The self-assembled nanoparticles were stabilized over 7 days. The self-assembled nanoparticles were reconstituted with double-distilled water to 20 μg / mL, and then divided into 7 aliquots of 1 mL each. The hydrodynamic size distribution and PDI of the nanoparticles were measured on days 1, 2, 3, 4, 5, 6, and 7, respectively.

[0078] 1.4 Transmission Electron Microscopy The nanoparticles were resuspended in double-distilled water to a concentration of 20 μg / mL. Ten μL aliquots of the suspension were cast onto a glow discharge-coated carbon-coated copper grid and air-dried for 30 minutes before TEM imaging. Transmission electron microscopy (TEM) imaging was then performed.

[0079] 1.5 UHPLC-LTQ-orbitt trap mass spectrometry Prior to analysis, the nanoparticles were dissolved in methanol, and the composition of the HSBDN lyophilized powder was analyzed using ultra-high performance liquid chromatography coupled with high resolution mass spectrometry (UHPLC-MS / MS) (Phase A: 0.1% formic acid water, Phase B: acetonitrile). Data acquisition and processing were performed using Metworks, Xcalibur, and Mass Frontier 7.0 software.

[0080] 1.6 Screening for antibacterial and anti-inflammatory effects of small molecules derived from HSBDN RAW264.7 cells were incubated with LPS (100 ng / mL) and 20 μg / mL of gentianin, pentamethoxyflavonoid glycoside, verbascoside-7-O-glucoside, emodin, nobiletin, hesperidin, ligustrolactone M, rhein, and patchouli for 24 hours. Subsequently, the culture supernatant was collected, and nitric oxide (NO) levels were measured using a nitric oxide assay kit (S0021S).

[0081] A Staphylococcus aureus suspension (5 × 10⁵ CFU / 100 μL / well) was added to 96-well plates and treated for 24 hours with 20 μg / mL of gentianin, pentamethoxyflavone, verbascoside-7-O-glucoside, emodin, norimethanil, ligustilide M, rhein, and patchouli. A bacterial control group (untreated) and a blank control group (culture medium only) were included. Absorbance was measured at 600 nm.

[0082] 1.7 Ultraviolet-Visible Spectroscopy and Fourier Transform Infrared Spectroscopy (FTIR) Rhein, patchouli ketone, and NPs were dispersed and dissolved in methanol. The UV-Vis spectra were then recorded using a micro-ultraviolet spectrophotometer, and the background signal of methanol was subtracted to obtain the baseline.

[0083] Powder (1 mg) of rhein, patchouli ketone, and NPs was analyzed by Fourier transform infrared (FTIR) spectroscopy. For each measurement, the air background spectrum was subtracted for baseline correction.

[0084] 1.8 The contents of patchouli ketone and rhein in the nanoparticles were determined by HPLC.

[0085] HPLC analysis was performed on an ACQUITY UPLC BEH C18 column (100 mm × 2.1 mm, 1.7 μm) on an UltiMate 3000 UHPLC (Thermo Fisher Scientific, USA). The mobile phase consisted of water containing 0.1% formic acid (A) and acetonitrile (B), delivered at a flow rate of 1.00 mL / min according to the following gradient program: 0 to 5 min 5%–30% B, 5 to 15 min 30%–80% B, 15 to 20 min 80%–95% B, 20 to 21 min held at 95% B, and returned to 5% B after 21 min. The injection volume was 10 μL, and the detection wavelength was 254 nm.

[0086] 1.9 Preparation of NPs Patchouli ketone and rhein self-assembled nanoparticles were prepared using reverse osmosis, and the specific preparation method is as follows: Patchouli and rhein were dissolved in DMSO and methanol, respectively, and mixed in a 1:1 molar ratio. The mixture was stirred at room temperature for 30 minutes. The mixture was then added dropwise to double-distilled water preheated to 70 °C and stirred for another 30 minutes. The resulting solution was transferred to a dialysis bag and dialyzed overnight. The molecular weight cutoff for the dialysis was 3.5 kDa. The self-assembled nanoparticles (NPs) were obtained by freeze-drying.

[0087] 1.10 Evaluation of the anti-inflammatory and antibacterial efficacy of Pog, Rhe, Pog+Rhe, and NPs RAW264.7 cells were then co-treated with LPS (100 ng / mL) and Pog, Rhe, Pog+Rhe, and NPs (20 μg / mL) for 24 hours. A nitric oxide (NO) assay kit was purchased from Beijing Beyotime Biotechnology Co., Ltd. (S0021S). The levels of NO inflammatory factors were detected in the culture supernatant. For polarization analysis, cells were stained with FITC-CD86 and PE-F4 / 80, and the results were analyzed by flow cytometry to assess the effects of the drugs on macrophage polarization.

[0088] 100 μL of a solution of Pog, Rhe, Pog+Rhe, and NPs (20 μg / mL) was added to each well of a 96-well cell culture plate. 10 μL of a 5 × 10⁻⁶ concentration was seeded into each well. 5 CFU of Staphylococcus aureus was incubated at 37 °C for 24 hours. After incubation, bacterial growth was quantified by measuring absorbance at 600 nm.

[0089] Example 2 Molecular dynamics simulation of the self-assembly mechanism of patchouli and rhein.

[0090] Molecular dynamics simulations were performed using the GROMACS 2021.3 software package with Amber ff99SB force fields. The molecular geometries of patchouli and rhein were optimized at the B3LYP / 6-311G level using Gaussian 09. The confined electrostatic potential (RESP) charge was calculated using Multifwn software, and the topology file was generated using the sobtop program. The simulation system was constructed by randomly filling 40 molecules each of patchouli and rhein into a 7 × 7 × 7 nm cubic box using PACKMOL. The system was dissolved in TIP3P water within a periodic boundary box, maintaining 10 layers between the solute and the box edge. After applying periodic boundary conditions (PBC), energy minimization was performed over 50,000 steps using the steepest descent algorithm. The system was then equilibrated for 1 ns under NVT and NPT ensembles, with temperature and pressure controlled by a V-rescale thermostat and a Berendsen barometer, respectively. Production runs of 30 ns were performed with an integration time step of 2.0 fs. Unbonded van der Waals interactions were handled with a cutoff of 10, while long-range electrostatics were handled using the Particle Mesh Ewald (PME) method. Bonds containing hydrogen atoms were constrained using the SHAKE algorithm.

[0091] Example 3 Evaluation of the therapeutic effects and safety of HSBDT, HSBDN, and NPs on Staphylococcus aureus-induced pneumonia in mice.

[0092] 3.1 Animal model establishment and dosing regimen This invention employs a combined prophylactic and therapeutic dosing regimen to evaluate the in vivo efficacy of HSBDT, HSBDN, and NPs against Staphylococcus aureus-induced bacterial pneumonia. The animal study protocol was approved by the Ethics Committee of the China Academy of Chinese Medical Sciences (Approval No. 2025B238). Male / female C57BL / 6J mice (8 weeks old) were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd., and housed under standard environmental conditions. After a 3-day acclimatization period, 36 mice were randomly divided into 6 groups (n=6 per group, half male and half female). These included: Normal control group: nebulized inhalation of 50 μL of physiological saline; Model control group: nebulized inhalation of 50 μL of physiological saline; Amoxicillin group: administered by gavage (180 mg / kg); The group receiving Huashi Baidu Decoction (HSBDT group): 50 μL HSBDT (5 mg / kg, 2.5 mg / mL) was nebulized and inhaled. The HSBDN nanoparticle group: 50 μL HSBDN (5 mg / kg, 2.5 mg / mL) was nebulized and inhaled. Self-assembled nanoparticle group (NPs group): 50 μL NPs (5 mg / kg, 2.5 mg / mL) by nebulization.

[0093] For the first three consecutive days, mice were treated with the corresponding drug daily. On day 4, a bacterial pneumonia model was established by intranasal inoculation with 50 μL of Staphylococcus aureus (OD600 = 1.0). Drug administration continued 1 hour and 24 hours post-infection. Twenty-four hours after the last administration, lung function was assessed using the Small Animal Lung Solution System (EMKA-WBP). The mice were then anesthetized, their eyes were enucleated to collect blood, and the animals were euthanized. Lung, liver, spleen, heart, kidney tissue, and bronchoalveolar lavage fluid (BALF) were collected for subsequent analysis.

[0094] 3.2 Histopathological observation Mouse lung tissue was fixed overnight in 4% paraformaldehyde, then dehydrated and embedded in paraffin. Subsequently, lung, liver, spleen, kidney, and heart tissue sections were prepared, dewaxed, hydrated, and stained with hematoxylin and eosin (H&E) for histopathological observation.

[0095] 3.3 Hematological parameter testing Blood was collected from mice in EDTA-Na anticoagulant tubes, and then routine blood tests were performed using a SYSMEX XN-1000V instrument.

[0096] 3.4 Measurement of bacterial load in the lungs The bronchoalveolar lavage fluid was serially diluted and spread onto LB agar plates. After incubation at 37°C for 24 hours, colony counting was performed.

[0097] 3.5 Detection of inflammatory factors ELISA kits for interleukin (IL)-6, IL-1β, and tumor necrosis factor-α (TNF-α) were purchased from Shanghai Enzyme-Linked Biotechnology Co., Ltd. (ml002095, ml063159, ml037873, Shanghai, China), and a nitric oxide (NO) kit was purchased from Beijing Beyotime Biotechnology Co., Ltd. (S0021S). Plasma biochemical analysis of these factors was performed using ELISA according to the manufacturer's instructions. Serum levels of IL-6, IL-1β, TNF-α, and NO were measured.

[0098] The expression of COX-2, IL-6 and IL-1β proteins in lung tissue was detected by Western blot.

[0099] 4. Results and Analysis 4.1 Synthesis and Characterization of Huashi Baidu Decoction Nanoparticles and Self-Assembled Nanoparticles like Figure 1As shown in Figure A, this invention employs a method simulating the traditional Chinese medicine decoction preparation process, involving filtration and gradient centrifugation to collect the Huashi Baidu Decoction nanoparticles (named HSBDN) and the corresponding supernatant after removing the nanoparticles (named HSBDT). Figure 1 The hydrated particle size results shown in B indicate that the hydrodynamic diameter (HD) of HSBDN is 379.2 nm. Transmission electron microscopy (TEM) images show that HSBDN has a spherical morphology. Figure 1 C). The surface charge of HSBDN is -12 mV, and the polydispersity index (PDI) is 0.22, indicating that the nanoparticles are uniform and stable in size. Figure 1 DE).

[0100] Subsequently, this invention developed a single carrier-free nano-formulation based on the main active ingredient of HSBDN. This invention first used ultra-high performance liquid chromatography combined with Orbitrap high-resolution mass spectrometry (UPLC-Q-Orbitrap MS) to comprehensively analyze its main components. Figure 1 F). Among the detected components, nine major compounds of HSBDN were selected for anti-inflammatory and antibacterial efficacy evaluation based on precise mass spectrometry and fragment spectroscopy. Notably, POG and RHE exhibited the best anti-inflammatory and antibacterial effects. Figure 1 (FH). Small molecule compounds in traditional Chinese medicine can self-assemble into carrier-free nanoparticles through non-covalent interactions, such as hydrophobic effects, hydrogen bonds, van der Waals forces, π-π stacking, and σ-π interactions. This carrier-free nanomedicine effectively addresses common drawbacks of conventional small molecule drugs, including poor solubility, high toxicity, and low bioavailability.

[0101] Given that POG and RHE are highly hydrophobic, abundant, and exhibit excellent pharmacological activity, this invention subsequently selected POG and RHE in a 1:1 molar ratio to prepare POG-RHE carrier-free self-assembled nanoparticles via reverse osmosis, named NPs( Figure 2 A). TEM images show that the synthesized NPs have a uniform and stable spherical morphology. Figure 2 BC). Dynamic light scattering further confirmed that the HD of the nanoparticles was 273.8 nm (BC). Figure 2 D), the ζ potential is –9.8 mV ( Figure 2 E). The stability of the NPs was then evaluated at room temperature, and the results showed that neither the HD nor PDI of the NPs changed significantly over a 7-day period. Figure 2F). This result further confirms the successful synthesis of stable NPs using POG and RHE. Subsequent characterization of the NPs by Fourier transform infrared spectroscopy (FTIR) and ultraviolet-visible spectroscopy (UV-Vis) confirmed the successful incorporation of POG and RHE. The nanoparticles exhibit characteristic peaks of the two constituent small molecules, POG and RHE. Figure 2 GH). Subsequently, quantitative analysis by liquid chromatography determined that the NPs consisted of 74% RHE and 20% POG. Figure 2 I).

[0102] To evaluate the synergistic anti-inflammatory and antibacterial effects of rhein and patchouli, this invention evaluated the anti-inflammatory and antibacterial efficacy of RHE, POG, RHE+POG, and self-assembled nanoparticles (NPs) formed by RHE and POG. ELISA analysis showed that RHE, POG, RHE+POG, and NPs all significantly downregulated the expression of inflammatory mediators (nitric oxide) in the supernatant of LPS-induced RAW264.7 cells, and the combination of POG and RHE was more effective than either drug alone, while NPs were more effective than the combination of POG and RHE. Figure 2 J).

[0103] Further cell polarization experiments demonstrated that the four drugs significantly inhibited LPS-induced M1 polarization in RAW264.7 cells, with the effect being NPs > RHE+POG > POG and RHE alone. Figure 2 HI).

[0104] To further describe the direct bactericidal activity of RHE, POG, RHE+POG, and NPs against Staphylococcus aureus, this invention evaluated the in vitro antibacterial effects of the four drugs. The results showed that RHE, POG, RHE+POG, and NPs all exhibited significant bactericidal activity against Staphylococcus aureus, with the bactericidal effect ranking as follows: NPs > RHE+POG > RHE > POG. Figure 2 These findings indicate that the combined use of RHE and POG has a synergistic anti-inflammatory and bactericidal effect, and that the anti-inflammatory and bactericidal effect of NPs composed of the two is much greater than that of the combined use of the two drugs.

[0105] In summary, these findings indicate the existence of stable Huashi Baidu Decoction nanoparticles (HSBDN) within Huashi Baidu Decoction (HSBD). Notably, the most abundant and potent anti-inflammatory and antibacterial components within the Huashi Baidu Decoction nanoparticles (HSBDN), namely POG and RHE, can be used to synthesize stable self-assembled nanoparticles (NPs). This study not only reveals the self-assembly phenomenon of bioactive nanoparticles during the decoction process of Huashi Baidu Decoction (HSBD), but also provides a feasible strategy for the nanodesign and preparation of its key active ingredients. Therefore, it lays an experimental foundation for the development of well-defined and efficacy-optimized nano-preparations based on complex traditional Chinese medicine systems.

[0106] 4.2 Mechanism of self-assembly of patchouli ketone and rhein into stable carrier-free nanoparticles This invention uses molecular dynamics (MD) simulations to elucidate the mechanism by which POG and RHE assemble into carrier-free nanoparticles. Snapshots are captured every 5 nanoseconds. Figure 3 A) illustrates the gradual accumulation of POG and RHE molecules during the simulation, ultimately forming a stable nanoparticle structure. After 30 ns in the simulation chamber, co-aggregation of POG and RHE (in a 1:1 ratio) with water molecules was observed. Figure 3 B). Molecular dynamics simulations show that the main driving forces for self-assembly within the POG-RHE system are π-π stacking and σ-π conjugation, and RHE molecules mainly self-assemble through π-π interactions. Figure 3 C). Root mean square deviation (RMSD) reflects conformational changes in the overall structure and is typically used to assess the overall structural stability of a molecular system. In a 30 ns self-assembly simulation of a POG and RHE mixture, the RMSD value stabilized after 15 ns and fluctuated around 3.256 ± 0.057 nm. Figure 3 D).

[0107] These results demonstrate that the system reaches equilibrium and remains stable throughout the MD simulation, supporting the reliability of subsequent trajectory analysis. The molecular surface areas of the POG and RHE mixture, as reflected by the solvent-accessible surface area (SASA) and solvent-inaccessible surface area (SEASA), gradually decrease during the 30 ns simulation, eventually stabilizing at approximately 102.501 ± 3.977. Figure 3 E). During the simulation, the number of hydrogen bonds formed in the POG and RHE mixture gradually increased, eventually stabilizing in the range of 4-7. As the dynamic simulation progressed, the fluctuations in the number of hydrogen bonds gradually reached equilibrium. Figure 3 F). Analysis of the molecular dynamic trajectory showed that the radius of gyration of the POG and RHE mixture decreased significantly during the 30 ns self-assembly simulation, eventually reaching equilibrium in the range of 2.092 ± 0.092 nm. Figure 3(G). Molecular dynamics simulations show that POG and RHE primarily self-assemble through non-covalent interactions, including π-π stacking, σ-π conjugation, and hydrogen bonding. In later stages of the simulations, the stabilization of key parameters such as RMSD, SASA, and radius of gyration confirmed that POG and RHE formed structurally stable nanoparticles.

[0108] 4.3 HSBDT, HSBDN, and NPs alleviate Staphylococcus aureus-induced pneumonia in mice To evaluate whether the Huashi Baidu Decoction nanoparticles (HSBDN) represent the main active ingredient of Huashi Baidu Decoction (HSBD), this invention established a mouse pneumonia model by intranasal infection with Staphylococcus aureus and implemented a therapeutic intervention via nebulized inhalation. The nebulized delivery of nanomedicine helps enhance deposition within lung lesions and prolongs retention time at the infection site. Animal experiments are as follows... Figure 4 As shown in Figure A, the mice were first administered medication via nebulization for three consecutive days. On day 4, Staphylococcus aureus was administered via nasal drops, with medication continuing at 1 hour and 24 hours post-infection. Lung function was then assessed at 48 hours, and lung tissue and blood samples were collected for subsequent histopathological and biochemical analysis. Lung function results showed a significant decrease in key lung function parameters, including maximum inspiratory flow, inspiratory volume, tidal volume, and respiratory rate, following Staphylococcus aureus infection. Simultaneously, a significant increase in relaxation time indicated severe lung function impairment in the mice. Treatment with HSBDT, HSBDN, and NPs all showed varying degrees of improvement in lung function parameters. Figure 4 BF). Histopathological evaluation by hematoxylin and eosin (H&E) staining confirmed that intranasal inoculation with Staphylococcus aureus induced severe pulmonary lesions, including alveolar hemorrhage, edema, significant thickening of the alveolar septa, and extensive inflammatory cell infiltration. Treatment with HSBDT, HSBDN, or NPs significantly improved these infection-induced pathological changes. Figure 4 G). Complete blood count (CBC) results, a key clinical indicator for diagnosing bacterial pneumonia, showed varying degrees of improvement in Staphylococcus aureus-induced lesions after treatment with HSBDT, HSBDN, or NPs. This included a reduction in the elevated total white blood cell count, neutrophil percentage, and monocyte percentage, while also mitigating the decrease in the percentage of lymphocytes caused by the infection. Figure 4 HK).

[0109] In summary, these results indicate that all three agents—HSBDT, HSBDN, and NPs—significantly alleviated Staphylococcus aureus-induced pneumonia, with NPs showing superior therapeutic efficacy compared to HSBDN, which in turn was superior to HBDT.

[0110] This invention further investigated whether HSBDT, HSBDN, and NPs exert their therapeutic effects on bacterial pneumonia through a dual anti-inflammatory and antibacterial mechanism. In vivo antibacterial efficacy was determined by plate count to identify the number of Staphylococcus aureus colonies in mouse bronchoalveolar lavage fluid (BALF). Compared with the model group, treatment with HSBDT, HSBDN, or NPs significantly reduced the number of surviving Staphylococcus aureus colonies in BALF. Figure 5 A). These results indicate that all three drugs can inhibit the colonization and survival of Staphylococcus aureus in lung tissue in vivo, demonstrating effective antibacterial activity. A hallmark of Staphylococcus aureus infection is the infiltration of inflammatory cells such as neutrophils into the bloodstream, accompanied by the release of inflammatory mediators. This invention subsequently used enzyme-linked immunosorbent assay (ELISA) and Western blot (WB) analysis to comprehensively assess the levels of inflammatory factors in blood and lung tissue to evaluate the anti-inflammatory effects of HSBDT, HSBDN, and NPs in the treatment of bacterial pneumonia. The results of this invention show that all three significantly attenuated the increase in key inflammatory mediators in the blood induced by Staphylococcus aureus infection, including IL-6 (…). Figure 5 B), IL-1β Figure 5 C), TNF-α Figure 5 D) and NO( Figure 5 E). Western blot analysis further confirmed these findings, showing that treatment with HSBDT, HSBDN, or NPs effectively suppressed the upregulation of COX-2, IL-6, and IL-1β expression in lung tissue following Staphylococcus aureus infection. Figure 5 FG).

[0111] In summary, these results indicate that HSBDT, HSBDN, and NPs all exert therapeutic effects on bacterial pneumonia through anti-inflammatory and antibacterial activities, with the efficacy ranking as NPs > HSBDN > HSBDT. This confirms that HSBDN is the main active pharmacologically active substance of HSBD. Furthermore, self-assembled nanoparticles constructed from its main active ingredients POG and RHE exhibit superior efficacy in treating Staphylococcus aureus pneumonia.

[0112] 4.4 HSBDT, HSBDN, and NPs exhibited good biocompatibility and safety in vivo. Safety evaluation is crucial for drug development and clinical translation. This invention subsequently further evaluated the in vivo safety of three formulations: HSBDT, HSBDN, and NPs. Figure 6As shown in Figure A, H&E staining revealed no histopathological changes or tissue damage in the heart, liver, spleen, or kidneys of the treatment groups receiving HSBDT, HSBDN, or NPs compared to the control group. These findings indicate that the three formulations do not produce toxicity to the major organs of mice. The present invention then conducted a comprehensive in vivo safety assessment of HSBDT, HSBDN, and NPs from multiple perspectives, including liver function, kidney function, myocardial injury, and lipid metabolism. The results showed that after administration, serum markers of liver function (alanine aminotransferase, ALT aspartate aminotransferase; total bilirubin, TBIL), kidney function (creatinine, renal blood urea nitrogen (BUN), myocardial injury (creatine kinase, CK), and lipid metabolism (triglycerides, TG; total cholesterol (TCHO)) remained within the normal physiological range, with no statistically significant differences compared to the normal control group. Figure 6 (BI). Systemic in vivo safety assessments confirmed that effective therapeutic doses of HSBDT, HSBDN, and NPs did not cause toxic damage to major organs. All serum biochemical parameters remained within normal physiological ranges, collectively demonstrating their good biocompatibility, thus laying a crucial foundation for subsequent clinical translation.

[0113] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A self-assembled nanoparticle derived from the Huashi Baidu Decoction nanoparticles, characterized in that, The active ingredients of the self-assembled nanoparticles include patchouli ketone and rhein.

2. The self-assembled nanoparticles derived from the Huashi Baidu Decoction nanoparticles according to claim 1, characterized in that, The molar ratio of patchouli ketone to rhein is 1:1, the particle size of the self-assembled nanoparticles is 200-300 nm, and the zeta potential of the self-assembled nanoparticles is -8 to -12 mV.

3. The method for preparing self-assembled nanoparticles derived from the Huashi Baidu Decoction nanoparticles according to claim 1, characterized in that, Includes the following steps: (1) Dissolve patchouli and rhein in an organic solvent in proportion, mix well to obtain a mixture; (2) Add the mixture dropwise to preheated water and stir; (3) Dialyze the obtained solution and freeze-dry it to obtain the self-assembled nanoparticles derived from Huashi Baidu Decoction nanoparticles.

4. A type of nanoparticle for dispelling dampness and detoxifying, characterized in that, The self-assembled nanoparticles are derived from the decoction of Huashi Baidu Tang.

5. The nanoparticles for the Detoxifying and Dampness-Clearing Decoction according to claim 4, characterized in that, The self-assembled nanoparticles have a particle size of 300-400 nm, a Zeta potential of -10 to -15 mV, and a spherical shape.

6. The method for preparing the Huashi Baidu Decoction nanoparticles according to claim 4, characterized in that, Includes the following steps: (1) Boil the raw materials of Huashi Baidu Decoction with water and collect the decoction; (2) The decoction is subjected to gradient centrifugation and the precipitate is collected to obtain the Huashi Baidu Decoction nanoparticles.

7. The method for preparing the Huashi Baidu Decoction nanoparticles according to claim 6, characterized in that, Step (2) involves centrifuging at 2000×g, 4000×g, 6000×g and 8000×g in sequence, collecting the supernatant; centrifuging the supernatant at 20000×g, and collecting the precipitate.

8. The use of the self-assembled nanoparticles derived from Huashi Baidu Decoction nanoparticles according to any one of claims 1-2, or the self-assembled nanoparticles derived from Huashi Baidu Decoction nanoparticles prepared by the method according to claim 3, or the Huashi Baidu Decoction nanoparticles according to claims 4-5, or the Huashi Baidu Decoction nanoparticles prepared by the method according to any one of claims 6-7 in the preparation of medicaments for the treatment and / or prevention of bacterial pneumonia.

9. The application according to claim 8, characterized in that, The bacterial pneumonia mentioned refers to pneumonia caused by Staphylococcus aureus infection.

10. The application according to claim 8, characterized in that, The drug has the following properties: it improves lung function indicators in mice with bacterial pneumonia, reduces lung tissue pathological damage, reduces the total number of white blood cells, the percentage of neutrophils and monocytes in the blood, increases the percentage of lymphocytes in the blood, reduces the level of inflammatory factors in serum and / or lung tissue, and inhibits the expression of COX-2, IL-6 and IL-1β proteins in lung tissue.

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