A composition based on plant essential oil or active ingredient and its application in preparing medicine for preventing and treating pulmonary edema

Abstract

This invention belongs to the field of biomedical technology and relates to a composition based on plant essential oils or active ingredients and its application in the preparation of drugs for the prevention and treatment of pulmonary edema. It comprises at least three plant essential oils or active ingredients; said plant essential oils or active ingredients have agonistic activity against aquaporins and agonistic activity against sodium-potassium ATPase. The composition and its special formulation provided by this invention not only have good preventive and therapeutic effects on pulmonary edema, but can also be applied to the prevention and treatment of related lung diseases such as hypersputum secretion, acute lung injury, acute respiratory distress syndrome, pulmonary fibrosis, and COPD.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology and relates to a composition based on plant essential oils or active ingredients and its application in the preparation of drugs for the prevention and treatment of pulmonary edema. Background Technology

[0002] Acute respiratory distress syndrome (ARDS) is one of the major complications of COVID-19 infection, and pulmonary edema is a central link in the pathological process of ARDS. Pulmonary edema is associated with increased alveolar capillary permeability and reduced alveolar fluid clearance. Inflammatory stress, oxidative stress, and immune imbalance are the main factors inducing pulmonary edema. The anti-inflammatory and antioxidant effects of early-stage pulmonary edema treatment with anti-inflammatory drugs (such as the glucocorticoid dexamethasone) and antioxidant drugs (such as the antioxidants N-acetyl-L-cysteine ​​(NAC) and lipoic acid) are undeniable. However, their effectiveness in treating late-stage pulmonary edema, which affects systemic oxygenation and mortality, is very limited. This may be related to the toxicity of glucocorticoids, the choice of treatment regimen, and the limitations and compliance of antioxidants in anti-inflammatory treatment. Therefore, there is an urgent need to develop highly effective and low-toxicity drugs to improve the prevention and treatment of pulmonary edema.

[0003] Previous studies have found that some plant essential oils, extracts or active ingredients have a variety of pharmacological effects such as antibacterial, anti-inflammatory, antioxidant, lipid-regulating, blood pressure-lowering, blood sugar-lowering, liver-protecting, neuroprotective, immunomodulatory, and anti-tumor effects. They have also shown some efficacy in treating acute lung injury (ALI) / acute respiratory distress syndrome (ARDS). However, studies have shown that when a single plant essential oil, extract or active ingredient is used as an active ingredient, its anti-pulmonary edema effect is poor and difficult to achieve the expected results. Summary of the Invention

[0004] To address the shortcomings of existing technologies, the present invention aims to provide a composition based on plant essential oils or active ingredients and its application in the preparation of drugs for the prevention and treatment of pulmonary edema. The multi-component composition provided by the present invention can not only be used for the prevention and treatment of pulmonary edema, but also for the prevention and treatment of respiratory diseases such as COPD, pulmonary fibrosis, and acute respiratory distress syndrome.

[0005] To achieve the above objectives, the technical solution of the present invention is as follows:

[0006] In a first aspect, a composition based on plant essential oils or active ingredients comprises at least three plant essential oils or active ingredients; wherein the plant essential oils or active ingredients have agonistic activity against aquaporins and agonistic activity against sodium-potassium ATPase.

[0007] Intraalveolar fluid clearance is a crucial aspect of pulmonary edema treatment. Alveolar epithelial cells are the primary site for alveolar fluid clearance, and sodium ions (Na+) are the main component. +Active transport is the primary force in clearing fluid from the alveoli. Under normal circumstances, Na+ ions from the alveolar space enter the alveolar epithelial cells via sodium ion channels (ENaC), and are then transported to sodium-potassium ATPase (Na+). + -K + Na+ under the action of ATPase) + Water is pumped into the lung interstitium, creating an osmotic gradient, and then transferred out through aquaporins (AQPs). Therefore, this invention uses aquaporins and sodium-potassium ATPases as important drug targets for screening. Experiments unexpectedly revealed that some plant essential oils, extracts, or active ingredients simultaneously promote the activity of both aquaporins and sodium-potassium ATPases. However, further research showed that these plant essential oils, extracts, or active ingredients, when used as single active ingredients, while promoting the activity of both aquaporins and sodium-potassium ATPases, had limited therapeutic effects on pulmonary edema in animal models. To address this issue, this invention continued experimentation and discovered that when three or more plant essential oils or active ingredients simultaneously promote the activity of both aquaporins and sodium-potassium ATPases are combined, they have a synergistic effect, showing good therapeutic effects on pulmonary edema in animal models. Furthermore, they can be used for the prevention and treatment of respiratory diseases such as COPD, pulmonary fibrosis, and acute respiratory distress syndrome.

[0008] In some implementations, the plants in the essential oils or active ingredients include, but are not limited to, garlic, moringa, peppermint, camphor leaves, camphor tree, cinnamon, cloves, eucalyptus, white pearl tree, thyme, vanilla, and tea tree. Among them, garlic essential oil or its active ingredients are garlic oil and / or diallyl disulfide, etc.; moringa essential oil or its active ingredients are moringa oil and / or moringa isothiocyanates; peppermint essential oil or its active ingredients are peppermint oil and / or menthol, etc.; camphor leaf essential oil or its active ingredients are 1,8-cineole, etc.; borneol essential oil or its active ingredients are borneol oil and / or borneol, etc.; cinnamon essential oil or its active ingredients are cinnamaldehyde and / or eugenol, etc.; clove essential oil or its active ingredients are clove oil and / or eugenol, etc.; eucalyptus essential oil or its active ingredients are eucalyptus oil and / or 1,8-cineole, etc.; white pearl tree essential oil or its active ingredients are methyl salicylate glycoside, etc.; thyme essential oil or its active ingredients are thymol and / or carvacrol, etc.; vanilla essential oil or its active ingredients are vanillin, etc.; tea tree essential oil or its active ingredients are tea tree oil and / or 1,8-cineole, etc.

[0009] Specifically, plant essential oils or plant active ingredients include, but are not limited to, garlic oil, moringa oil, peppermint oil, borneol oil, clove oil, eucalyptus oil, cinnamon essential oil, tea tree essential oil, diallyl disulfide, menthol, moringa isothiocyanate, borneol, cinnamaldehyde, eugenol, 1,8-cineole, methyl salicylate glycoside, thymol, carvacrol, vanillin, etc.

[0010] In some implementations, plant essential oils or active ingredients are ranked by upregulating the expression levels of aquaporin AQP1 and sodium-potassium ATPase-α mRNA genes in A549 cells with pulmonary edema, and the top three from the top five are combined into a composition. The composition of plant essential oils or active ingredients is derived from screening for pulmonary edema aquaporin and sodium-potassium ATPase agonists. The results show that the order in which plant essential oils upregulate the expression levels of aquaporin AQP1 and sodium-potassium ATPase-α mRNA genes in A549 cells with pulmonary edema is garlic oil, moringa oil, peppermint oil, eucalyptus oil, clove oil, borneol oil, and cinnamon oil, while the order in which active ingredients upregulate the expression levels of aquaporin AQP1 and sodium-potassium ATPase-α mRNA genes in A549 cells with pulmonary edema is diallyl disulfide, moringa isothiocyanate-1, menthol, 1,8-cineole, eugenol, borneol, and cinnamaldehyde, etc. Studies have shown that the three-component combination of the top five plant essential oils or active ingredients is more effective than the two-component combination in upregulating the expression of pulmonary edema aquaporin AQP1 and sodium-potassium ATPase-α mRNA genes, indicating the synergistic effect of the components in the plant essential oil or active ingredient combination.

[0011] In some embodiments, at least one plant essential oil or plant active ingredient contains sulfur. Plant essential oils or plant active ingredients containing sulfur may include garlic oil, moringa oil, diallyl disulfide, moringa isothiocyanate-1, etc.

[0012] Based on the above research findings, the following implementation plan is further proposed:

[0013] In some implementations, the oil is composed of garlic oil, peppermint oil, and eucalyptus oil in a mass ratio of 4-8:1-2:4-12, with a preferred mass ratio of 4:1:4.

[0014] In some implementations, the oil is composed of garlic oil, peppermint oil, and clove oil in a mass ratio of 4-8:1-2:12-36, preferably 4:1:12.

[0015] In some implementations, the oil is composed of moringa oil, peppermint oil, and eucalyptus oil in a mass ratio of 4-8:1-2:4-12, preferably 4:1:4.

[0016] In some embodiments, the mixture consists of diallyl disulfide, L-menthol, and 1,8-cineole in a mass ratio of 4-8:1-2:4-12, preferably 4:1:4.

[0017] In some embodiments, the mixture consists of diallyl disulfide, L-menthol, and eugenol in a mass ratio of 4-8:1-2:12-36, preferably 4:1:12.

[0018] In some embodiments, the mixture consists of Moringa isothiocyanate-1, L-menthol, and 1,8-cineole in a mass ratio of 4-8:1-2:4-12, preferably 4:1:4.

[0019] To further improve the volatility (odor), irritation, and stability of compositions of plant essential oils or active ingredients, the following implementation scheme is proposed:

[0020] In some embodiments, the dosage form of the composition is one of the following: injectable liquid formulation, oral liquid formulation, microemulsion, nanoemulsion, liposome, nanopolymer formulation, soft capsule formulation, aerosol, nebulizer, powder aerosol, nasal spray, inclusion complex formulation, etc., preferably a soft capsule formulation, nanoemulsion or inclusion complex formulation.

[0021] In some embodiments, the composition further includes pharmaceutical excipients. Specifically, the excipients include one or more of fillers, emulsifiers, co-emulsifiers, capsule shell materials, preservatives, inclusion agents, etc.

[0022] The soft capsule formulation is a microemulsion soft capsule. Specifically, the excipients in the capsule core include an oil phase filler, an emulsifier, and a co-emulsifier. More specifically, plant organosulfur compounds and a flavor-masking and cooling system are used as active ingredients, and the mass ratio of the active ingredient, oil phase filler, emulsifier, and co-emulsifier is 1-10:1-10:1-10:1-5. More specifically, the oil phase filler is a medium-chain triglyceride. More specifically, the emulsifier is Tween-80. More specifically, the co-emulsifier is polyethylene glycol-400.

[0023] Specifically, the excipients of the nanoemulsion include emulsifiers, co-emulsifiers, and water. More specifically, plant organosulfur compounds and a flavor-masking and cooling system are used as active ingredients, and the mass ratio of the active ingredient, emulsifier, co-emulsifier, and water is 2-6:3-9:1-3:10-20. More specifically, the emulsifier is polyoxyethylene hydrogenated castor oil RH-40 (Cremophor RH-40), Tween-80, Span-80, and mixtures thereof, and the co-emulsifier is preferably selected from one or more mixtures of ethanol, 1,2-propanediol, glycerol, diethylene glycol monoethyl ether, and polyethylene glycol-400. More specifically, the co-emulsifier is ethanol.

[0024] Specifically, the excipient in the inclusion complex formulation is a solubilizing inclusion material. More specifically, the mass ratio of the active ingredient to the solubilizing inclusion material is 1:10-30. More specifically, the solubilizing inclusion material is methyl-β-cyclodextrin.

[0025] On the other hand, the use of the above-mentioned composition based on plant essential oils or active ingredients in the preparation of drugs for the prevention and treatment of pulmonary edema.

[0026] Thirdly, the use of the above-mentioned composition based on plant essential oils or active ingredients in the preparation of a medicament for preventing and treating hypersputum secretion, acute lung injury, acute respiratory distress syndrome, pulmonary fibrosis, or COPD.

[0027] Pharmacodynamic studies have shown that the composition contains diallyl disulfide and is in the form of a soft capsule. When taken orally, this composition has good therapeutic effects against pulmonary fibrosis, alveolar neutrophils, inflammatory stress, and oxidative stress. Its therapeutic effect is significantly better than that of the two-component composition and also significantly better than that of the positive control N-acetyl-L-cysteine ​​(NAC), demonstrating a significant therapeutic effect against pulmonary fibrosis.

[0028] Pharmacodynamic studies have shown that the composition containing diallyl disulfide, in the form of a nanoemulsion, has good therapeutic effects such as anti-lung injury, inhibition of mucin hypersecretion, downregulation of mucin MUC5AC, and anti-inflammatory stress. Moreover, the therapeutic effect is significantly better than that of the two-component composition group, and also significantly better than that of the positive control Myrtol, demonstrating significant therapeutic effects in inhibiting hypersputum secretion and anti-lung injury. Attached Figure Description

[0029] The accompanying drawings, which form part of this invention, are used to provide a further understanding of the invention. The illustrative embodiments of the invention and their descriptions are used to explain the invention and do not constitute an improper limitation of the invention.

[0030] Figure 1 This invention relates to the effect of garlic oil-based soft capsule CPD0351A on the lung tissue destruction index in rats with chronic obstructive pulmonary disease (COPD).

[0031] Figure 2 This invention relates to the effect of garlic oil-based soft capsule CPD0351A on the inflammatory factor TNF-α in lung homogenate of COPD rats.

[0032] Figure 3 This invention illustrates the effect of garlic oil-based soft capsule CPD0351A on the antioxidant factor GSH in the lung tissue of COPD rats.

[0033] Figure 4 The effect of garlic oil-based soft capsule CPD0351A on the spleen index of COPD rats in this embodiment of the invention;

[0034] Figure 5 This invention illustrates the effect of garlic oil-based soft capsule CPD0351A on relative pAkt in the lung tissue of COPD rats.

[0035] Figure 6 This invention illustrates the effect of the soft capsule formulation CPD0353A based on moringa oil on the dry-wet weight ratio of pulmonary edema in ALI mice.

[0036] Figure 7 The effect of the soft capsule formulation CPD0353A based on moringa oil in this embodiment of the invention on the inflammatory factor IL-6 in the lung tissue of ALI mice with pulmonary edema;

[0037] Figure 8 This invention illustrates the effect of diallyl disulfide-based soft capsule CPD0354A on the pulmonary fibrosis area of ​​fibrotic mice.

[0038] Figure 9 The effect of diallyl disulfide-based soft capsule CPD0354A on the number of neutrophils in the lung lavage fluid (BALF) of fibrotic mice in this embodiment of the invention;

[0039] Figure 10 This invention illustrates the effect of diallyl disulfide-based soft capsule CPD0354A on the inflammatory factor IL-1β in lung homogenate of mice with pulmonary fibrosis.

[0040] Figure 11 This invention illustrates the effect of diallyl disulfide-based soft capsule CPD0354A on the antioxidant SOD1 in lung homogenate of pulmonary fibrosis mice.

[0041] Figure 12 The lung injury resistance score of the soft capsule formulation based on Moringa isothiocyanate-1 in this embodiment of the invention is CPD0356A.

[0042] Figure 13 The effect of the soft capsule CPD0356A based on Moringa isothiocyanate-1 composition in this embodiment of the invention on the inflammatory factor IL-1β in the lung homogenate of ALI rats;

[0043] Figure 14 The effect of diallyl disulfide-based composition nanoemulsion CPD0357A on lung injury in ARDS mice in this embodiment of the invention;

[0044] Figure 15This invention illustrates the effect of diallyl disulfide-based nanoemulsion CPD0357A on goblet cell formation and mucin secretion in the lung tissue of ARDS mice.

[0045] Figure 16 This invention illustrates the effect of diallyl disulfide-based nanoemulsion CPD0357A on mucin MUC5AC in the lung tissue of ARDS mice.

[0046] Figure 17 This invention illustrates the effect of diallyl disulfide-based nanoemulsion CPD0357A on the inflammatory factor IL-1β in the lung homogenate of ARDS mice.

[0047] Figure 18 The effect of diallyl disulfide-based composition inclusion complex formulation CPD0358A on lung injury in ALI mice in this embodiment of the invention;

[0048] Figure 19 This invention illustrates the effect of diallyl disulfide-based composition inclusion complex formulation CPD0358A on the dry-wet weight ratio of pulmonary edema in ALI mice.

[0049] Figure 20 This invention relates to the effect of diallyl disulfide-based composition inclusion complex formulation CPD0358A on the expression of AQP3 mRNA in pulmonary edema of ALI mice.

[0050] Figure 21 This invention illustrates the effect of diallyl disulfide-based composition inclusion complex formulation CPD0358A on the expression of sodium-potassium channel ATPase-α2 mRNA in ALI mice with pulmonary edema.

[0051] Figure 22 This invention relates to the effect of diallyl disulfide-based composition inclusion complex formulation CPD0358A on the inflammatory factor TNF-α in the serum of ALI mice. Detailed Implementation

[0052] To enable those skilled in the art to better understand the technical solution of the present invention, the technical solution of the present invention will be described in detail below with reference to specific embodiments.

[0053] Example 1: Activity determination of single plant essential oils and their active ingredient, aquaporin agonists.

[0054] Based on literature review and preliminary experiments, the selected single plant essential oils include garlic oil, moringa oil, peppermint oil, eucalyptus oil, clove oil, borneol oil, and cinnamon oil; the selected single plant active ingredients include diallyl disulfide, moringa isothiocyanate-1, menthol, 1,8-cineole, eugenol, borneol, and cinnamaldehyde.

[0055] Aquaporin agonist activity test: (1) Pulmonary edema modeling: 20 μg / mL lipopolysaccharide (LPS) solution was used to stimulate each well 4×10 5 (2) Drug administration and sample preparation: Plant essential oil or its active ingredients were added to a 96-well plate containing LPS-stimulated lung edema A549 cell culture medium at a concentration of 30 µg / mL and cultured for 24 hours. The relevant test samples were prepared according to the experimental steps of RNA extraction, RNA reverse transcription and RT-PCR reaction in the conventional RT-PCR detection technology. (3) Determination of the expression level of aquaporin APQ1 mRNA gene: The expression level of aquaporin AQP1 mRNA gene in lung edema A549 cells was detected by RT-PCR technology according to the experimental steps of RT-PCR detection.

[0056] The order in which plant essential oils upregulated the expression level of aquaporin AQP1 mRNA in A549 cells with pulmonary edema was garlic oil, moringa oil, peppermint oil, eucalyptus oil, clove oil, borneol oil, and cinnamon oil, while the order in which the active ingredients of plant essential oils upregulated the expression level of aquaporin AQP1 mRNA in A549 cells with pulmonary edema was diallyl disulfide, moringa isothiocyanate-1, menthol, 1,8-cineole, eugenol, borneol, and cinnamaldehyde, among others.

[0057] Example 2: Determination of the activity of a single plant essential oil and its active ingredient, sodium-potassium ATPase agonist.

[0058] The preferred single plant essential oil and its active ingredients in this embodiment are described in Example 1.

[0059] Activity test of sodium-potassium ATPase agonist: The relevant experimental operations were completed according to the experimental steps of pulmonary edema modeling, drug administration and sample preparation in Example 1; Determination of sodium-potassium ATPase-α mRNA level: The gene expression level of sodium-potassium ATPase-α mRNA in pulmonary edema A549 cells was detected by RT-PCR technology according to the experimental steps of RT-PCR detection.

[0060] The order in which plant essential oils upregulated the expression level of sodium-potassium ATPase-α mRNA gene in A549 cells with pulmonary edema was garlic oil, moringa oil, peppermint oil, eucalyptus oil, clove oil, borneol oil, and cinnamon oil, while the order in which the active ingredients of plant essential oils upregulated the expression level of sodium-potassium ATPase-α mRNA gene in A549 cells with pulmonary edema was diallyl disulfide, moringa isothiocyanate-1, menthol, 1,8-cineole, eugenol, borneol, and cinnamaldehyde, among others.

[0061] Example 3: Determination of the synergistic effect of plant essential oils and their multi-component active ingredients as aquaporin agonists.

[0062] Based on the activity assay results of single plant essential oils and their active ingredient, aquaporin agonists, in Example 1, garlic oil, moringa oil, peppermint oil, eucalyptus oil, and clove oil were identified as the top five optimal aquaporin APQ1 agonists. Using essential oils containing organosulfur compounds as the primary components, the following three-component (1:1:1 ratio) and two-component essential oil compositions (1:1 ratio) were optimized: garlic oil + peppermint oil + eucalyptus oil, garlic oil + peppermint oil + clove oil, moringa oil + peppermint oil + eucalyptus oil, moringa oil + peppermint oil + clove oil, garlic oil + peppermint oil, garlic oil + eucalyptus oil, garlic oil + clove oil, moringa oil + peppermint oil, moringa oil + eucalyptus oil, and moringa oil + clove oil. Following the aquaporin agonist activity assay procedures in Example 1, RT-PCR was used to detect the effects of the above three-component and two-component essential oil compositions on aquaporin AQP1 in A549 cells with pulmonary edema. The effect of mRNA gene expression level was measured, and the results showed that the activity of the above three-component composition was better than that of the corresponding two-component composition, indicating that the preferred multi-component essential oil composition has a synergistic effect in upregulating aquaporin AQP1.

[0063] Based on the activity determination results of single plant essential oils and their active ingredient aquaporin agonists in Example 1, the top 5 active ingredients of the plant essential oils include diallyl disulfide, moringa isothiocyanate-1, L-menthol, 1,8-cineole, and eugenol; using the active ingredients containing organosulfur compounds as the primary components, the following three-component (ratio 1:1:1) and two-component active ingredient compositions (ratio 1:1) were optimized: diallyl disulfide + L-menthol + 1,8-cineole, diallyl disulfide + L-menthol + eugenol, moringa isothiocyanate-1 + The following compounds were selected: L-menthol + 1,8-cineole, Moringa isothiocyanate-1 + L-menthol + eugenol, diallyl disulfide + L-menthol, diallyl disulfide + 1,8-cineole, diallyl disulfide + eugenol, Moringa isothiocyanate-1 + L-menthol, Moringa isothiocyanate-1 + 1,8-cineole, Moringa isothiocyanate-1 + eugenol. Following the aquaporin agonist activity assay procedure in Example 1, RT-PCR was used to detect the effects of the above three-component and two-component active ingredient compositions on the expression level of aquaporin AQP1 mRNA in pulmonary edema A549 cells. The results showed that the activity of the three-component compositions was superior to that of the corresponding two-component compositions, indicating that the preferred multi-component active ingredient compositions have an additive effect in upregulating aquaporin AQP1.

[0064] Example 4: Determination of the synergistic effect of plant essential oils and their multi-component active ingredients as aquaporin agonists.

[0065] Based on the experimental procedures for determining the activity of a single plant essential oil and its active ingredient, sodium-potassium ATPase agonist, in Example 2, the additive effect of the plant essential oil or its multi-component composition of active ingredients in Example 3 on the activity of sodium-potassium ATPase-α agonist was evaluated. The test results showed that the activity of the above three-component plant essential oil or its active ingredient composition was better than that of the corresponding two-component plant essential oil or its active ingredient composition, indicating that the preferred multi-component essential oil composition has additive effect in upregulating sodium-potassium ATPase-α.

[0066] Example 5: Screening of active ingredients and excipients for soft capsules CPD0351 based on garlic oil composition.

[0067] Based on the screening results of aquaporins and sodium-potassium ATPase agonists in Examples 1-4 above, and the pharmacological effects of related components, and taking into account the target indications for the prevention and treatment of pulmonary edema of the developed soft capsules, such as oxidative stress and inflammatory stress, the effective components of the soft capsule CPD0351 based on garlic oil composition were determined to be garlic oil, peppermint oil, and eucalyptus oil. According to the Chinese Pharmacopoeia, the recommended dosage of relevant traditional Chinese medicines (or extracts, etc.) in domestic and foreign studies, the content of effective components, and their preliminary experiments, the mass ratio of medicinal plant essential oils in the formulation of soft capsule CPD0351 was determined to be approximately 4-8:1-2:4-12.

[0068] Microemulsion soft capsules have stringent formulation requirements. The oil phase, emulsifiers, and co-emulsifiers must be non-toxic, non-irritating, and biocompatible. The type and ratio of these components can affect the final product's performance, such as droplet size, zeta potential, and polydispersity index. Therefore, the optimal composition and ratio are crucial for soft capsule formulation. The oil phase used in microemulsion soft capsules should allow for appropriate penetration and interaction with emulsifier molecules on the emulsion interface film, and readily form an interfacial film with the emulsifier. Thus, the molecular size of the oil phase is significant for the formation of the soft capsule microemulsion. Suitable oil phases for selection include soybean oil, corn oil, olive oil, rapeseed oil, peanut oil, isopropyl myristate, castor oil, isopropyl laurate, ethyl oleate, and medium-chain triglycerides. Emulsifiers can be classified according to the type of ion, including anionic, cationic, amphoteric, and nonionic types. Considering safety and irritation in application, nonionic emulsifiers are more widely used because they have the lowest toxicity and are less affected by changes in pH and ionic strength. The hemolytic effect of emulsifiers must also be considered. Nonionic emulsifiers available for screening include fatty acid sorbitan, polysorbates, polyoxyethylene fatty acid esters (trade name Myrj), polyoxyethylene fatty alcohol ethers (trade name Brij), polyoxyethylene polyoxypropylene copolymers (polyether type), sucrose fatty acid esters, and glyceryl monostearate. Preferred emulsifiers include polysorbates (such as Tween-20, Tween-40, Tween-60, and Tween-80), fatty acid sorbates (such as Span-20, Span-40, Span-60, and Span-80), and castor oil polyoxyethylene ethers (such as Kolliphor EL, Marlowet 40, and Emulgin RO). 40) Polyoxyethylene hydrogenated castor oil (such as Kolliphor RH-40, Kolliphor RH-60), etc. Co-emulsifiers can improve the strength and flexibility of the film, reduce emulsifier consumption, and facilitate microemulsion formation. Co-emulsifiers should be pharmaceutical-grade short-chain alcohols or nonionic surfactants with suitable hydrophilic-lipophilic balance (HLB). Suitable co-emulsifiers include ethanol, ethylene glycol, propylene glycol, glycerol, n-butanol, diethylene glycol monoethyl ether, glycerol, polyethylene glycol (such as PEG 400, PEG 600), polyglycerol esters, etc.

[0069] The screening of the oil phase used in the soft capsule CPD0351 based on garlic oil composition was divided into two parts: (1) The miscibility of the mixture of CPD0351 essential oil composition and different oil phases was observed after 48 hours. Soybean oil, corn oil, olive oil, rapeseed oil, peanut oil, castor oil, isopropyl myristate, isopropyl laurate, ethyl oleate, medium chain triglycerides and other oil phases did not show layering or discoloration; (2) The mixture of CPD0351 essential oil composition, different oil phases, emulsifier Tween-80 or Span-80 was observed after 48 hours. The Span-80 group did not show discoloration or layering, while the mixture of (1) medium oil phase and Tween-80 showed different degrees of discoloration or layering except for ethyl oleate and medium chain triglycerides.

[0070] The garlic oil-based soft capsule CPD0351 was mixed with the oil phase, emulsifier, and co-emulsifier in a pre-set 1:1:1 ratio and shaken to form a clear, transparent solution. 0.1 g of the above mixture was taken and added to 50 mL of preheated 37°C pure water. The mixture was shaken, and the combinations that formed a transparent solution were recorded. The types of oil phase, emulsifier, and co-emulsifier were determined using a pseudo-three-phase diagram. The preferred emulsifiers were Tween-80, Span-80, Kolliphor RH-40, and mixtures thereof, while ethanol, diethylene glycol monoethyl ether, polyethylene glycol-400, and mixtures thereof were preferred co-emulsifiers.

[0071] The ideal formulation of essential oils, filler oil phase, emulsifier and co-emulsifier was studied using pseudo-three-phase diagrams. The optimal ratio was determined to be 1-10:1-10:1-10:1-5. The preferred filler oil phase, emulsifier and co-emulsifier were medium-chain triglycerides, Tween-80 and polyethylene glycol-400, respectively.

[0072] Example 6: Screening and preparation of capsule material for garlic oil-based soft capsules CPD0351 series.

[0073] Based on the screening in Example 5, the mass ratio of the active ingredients—garlic oil, peppermint oil, and eucalyptus oil—in the garlic oil-based soft capsule composition CPD0351 was determined to be 4:1:4. The initial ratio of the drug essential oil, medium-chain triglycerides in the oil phase, and emulsifier Tween-80 in the contents was set at 9:4:12. Commonly used soft capsule materials include gelatin, plasticizer (glycerin), and water. The capsule shell was prepared according to the conventional ratio of 3:1:3. By conducting compatibility studies between the capsule shell and the contents, gelatin from different sources and with different gel strengths was screened. The specific ingredients for CPD0351A are shown in Table 1. The capsule shell was cut into 0.5 × 1 cm pieces and placed in a sample vial. 1 mL of CPD0351 contents was added. According to the influencing factor test conditions, the capsules were placed under high temperature, high humidity, and light conditions. A separate gelatin block was placed as a blank control under each condition. Samples were taken on days 5 and 10 to observe and photograph changes. Three samples were taken in parallel for each group. The selected gelatin sources and gel strengths included Fengyuan 150 gel strength, Fengyuan 180 gel strength, Pingda 150 gel strength, and Pingda 180 gel strength. Observation of the appearance changes of the capsule shell showed that none of the capsule shells melted or deformed under various conditions. The color of the capsule shells deepened under high temperature conditions, while the color change was minimal under high humidity and light conditions. Fengyuan 180 gel strength showed almost no change.

[0074] Table 1. Feeding table for the preparation of garlic oil-based soft capsules CPD0351A

[0075]

[0076] The preparation of soft capsules includes the following steps:

[0077] (1) Preparation of sol: Methylparaben, propylparaben and glycerin are dissolved in 95% pharmaceutical alcohol, and then dissolved in boiling hot water under vacuum stirring. The mixed sol is stirred for about 45 minutes at a temperature of 75-78℃ and a pressure of 0.2 MPa. When the temperature is reduced to 58-62℃, the material is discharged and weighed.

[0078] (2) Preparation of capsule core: Weigh the amount of garlic oil, peppermint oil and eucalyptus oil, and mix them with a magnetic stirrer to obtain a compound raw material mixture. Add the amount of medium chain triglycerides, Tween 80 and PEG400 to the compound raw material mixture and stir with a magnetic stirrer for 12 h to obtain the soft capsule core.

[0079] (3) Soft Capsule Machine Debugging: The prepared gelatin solution is piped into the gelatin box of the soft capsule machine and spread on the rubber wheel to form a rubber sheet. The medicine solution is injected into the soft capsules through the feeding system and the filling pump is adjusted to ensure that the soft capsule filling amount meets the requirements. Wuxi Zhongyi RJWJ-100G soft capsule filling machine is used. The cleanliness of the mixing room is 100,000 class, the indoor temperature is controlled at 22-24℃, and the relative humidity is ≤45-60%. Main control settings: gelatin box temperature 46.5℃, left and right gelatin box temperatures 55℃, hopper temperature 40℃, main control speed 2.6, rubber wheel left and right speed ratio 0.5, fan 30 Hz, soft capsule filling amount should be between 582-583mg.

[0080] (4) Soft capsule compression: The soft capsules are compressed using a rotary compression method with a high degree of automation and low material loss.

[0081] (5) Capsule shaping: In order to remove the moisture in the capsule shell, a first-stage rolling dry is usually used. The humidity of the hot air should be low, and the drying time is 1.5 to 3 hours.

[0082] (6) Washing soft capsules: Wash soft capsules with petroleum ether and blow dry.

[0083] (7) Secondary drying: Dry the soft capsules at 35±2℃ and relative humidity of 35-45% for 5 hours.

[0084] (8) The prepared soft capsules are inspected, sorted and screened.

[0085] Example 7: Formulation and preparation of garlic oil-based soft capsules CPD0352 series.

[0086] Based on the screening results of masking agents / analgesics / antiirritants and the pharmacological effects of related components in Examples 1-4 above, and taking into account the target indications for the prevention and treatment of pulmonary edema in the developed soft capsules, including oxidative stress and inflammatory stress, the effective components of the CPD0352 series of soft capsules based on garlic oil were determined to be garlic oil, peppermint oil, and clove oil. According to the Chinese Pharmacopoeia, the recommended dosages of relevant traditional Chinese medicines (or extracts, etc.) in domestic and international research, the content of effective components, and their screening experiments, the mass ratio of plant essential oils in the CPD0352 series of soft capsules was determined to be approximately 4-8:1-2:12-36. The specific ingredient dosages for CPD0352A are shown in Table 2. The preparation process of the soft capsules is described in Example 6.

[0087] Table 2. Feeding table for the preparation of garlic oil-based soft capsules CPD0352A

[0088]

[0089] Example 8: Formulation and preparation of soft capsules CPD0353 series based on moringa oil.

[0090] Based on the screening results of aquaporins and sodium-potassium ATPase agonists in Examples 1-4 above, and considering the pharmacological effects of related components, and taking into account the target indications for the prevention and treatment of pulmonary edema in the developed soft capsules, including oxidative stress and inflammatory stress, the effective components of the CPD0353 series of soft capsules based on moringa oil were determined to be moringa oil, peppermint oil, and eucalyptus oil. According to the Chinese Pharmacopoeia, the recommended dosages of relevant traditional Chinese medicines (or extracts, etc.) in domestic and international studies, the content of effective components, and their screening experiments, the mass ratio of plant essential oils in the CPD0353 series of soft capsules was determined to be approximately 4-8:1-2:4-12. The specific ingredient dosages for CPD0353A are shown in Table 3. The preparation process of the soft capsules is described in Example 6.

[0091] Table 3. Material feeding table for the preparation of Moringa oil-based soft capsules CPD0353A

[0092]

[0093] Example 9: Formulation and preparation of soft capsules CPD0354 series based on diallyl disulfide compositions.

[0094] Based on the screening results of aquaporins and sodium-potassium ATPase agonists in Examples 1-4 above, and considering the pharmacological efficacy of related components, and taking into account the target indications for the prevention and treatment of pulmonary edema in the developed soft capsules, including oxidative stress and inflammatory stress, the active ingredients of the soft capsule CPD0354 based on diallyl disulfide were determined to be diallyl disulfide, L-menthol, and 1,8-cineole. According to the Chinese Pharmacopoeia, the recommended dosages of relevant traditional Chinese medicines (or extracts, etc.) in domestic and international studies, the content of active ingredients, and their screening experiments, the mass ratio of the three active ingredients in the CPD0354 soft capsule formulation was determined to be 4-8:1-2:4-12. The specific ingredient dosages for CPD0354A are shown in Table 4. The preparation process of the soft capsules is described in Example 6.

[0095] Table 4. Material feeding table for the preparation of soft capsules CPD0354A based on diallyl disulfide.

[0096]

[0097] Example 10: Formulation and preparation of soft capsules CPD0355 series based on diallyl disulfide compositions.

[0098] Based on the screening results of aquaporins and sodium-potassium ATPase agonists in Examples 1-4 above, and considering the pharmacological efficacy of related components, and taking into account the target indications for the prevention and treatment of pulmonary edema in the developed soft capsules, including oxidative stress and inflammatory stress, the active ingredients of the soft capsule CPD0355 based on diallyl disulfide were determined to be diallyl disulfide, L-menthol, and eugenol. According to the Chinese Pharmacopoeia, recommended dosages of relevant traditional Chinese medicines (or extracts, etc.) in domestic and international research, the content of active ingredients, and screening experiments, the mass ratio of the three active ingredients in the CPD0355 soft capsule formulation was determined to be 4-8:1-2:12-36. The specific ingredient dosages for CPD0355A are shown in Table 5. The preparation process of the soft capsules is described in Example 6.

[0099] Table 5. Material feeding table for the preparation of soft capsules CPD0355A based on diallyl disulfide.

[0100]

[0101] Example 11: Formulation and preparation of soft capsules CPD0356 series based on Moringa isothiocyanate-1.

[0102] Based on the screening results of aquaporins and sodium-potassium ATPase agonists in Examples 1-4 above, and considering the pharmacological efficacy of related components, and taking into account the target indications for the prevention and treatment of pulmonary edema in the developed soft capsules, including oxidative stress and inflammatory stress, the active ingredients of the soft capsule CPD0356 based on Moringa isothiocyanate-1 were determined to be Moringa isothiocyanate-1, L-menthol, and 1,8-cineole. According to the Chinese Pharmacopoeia, the recommended dosages of relevant traditional Chinese medicines (or extracts, etc.) in domestic and international studies, the content of active ingredients, and their screening experiments, the mass ratio of the three essential oils in the CPD0356 soft capsule formulation was determined to be 4-8:1-2:4-12. The specific ingredient dosages for CPD0356A are shown in Table 6. The preparation process of the soft capsules is described in Example 6.

[0103] Table 6. Material feeding table for the preparation of soft capsules CPD0356A based on Moringa isothiocyanate-1.

[0104]

[0105] Example 12: Formulation and preparation of CPD0357 series of nanoemulsions based on diallyl disulfide.

[0106] Based on the screening results of aquaporins and sodium-potassium ATPase agonists in Examples 1-4 above, and considering the pharmacological efficacy of related components, and taking into account the target indications for the prevention and treatment of pulmonary edema in the developed soft capsules, including oxidative stress and inflammatory stress, the active ingredients of the diallyl disulfide-based nanoemulsion CPD0357 were determined to be diallyl disulfide, L-menthol, and 1,8-cineole. According to the Chinese Pharmacopoeia, recommended dosages of relevant traditional Chinese medicines (or extracts, etc.) in domestic and international studies, the content of active ingredients, and screening experiments, the mass ratio of the three active ingredients in the CPD0357 soft capsule formulation was determined to be 4-8:1-2:4-12. The specific dosage of CPD0357A is shown in Table 7.

[0107] Preliminary experiments established reverse emulsification as the preparation technique for nanoemulsions. A pseudo-ternary phase diagram was constructed using a dropwise method, and the area and stability of the nanoemulsion regions were used as evaluation indicators to screen for the optimal formulation. The optimal preparation process involves weighing an appropriate amount of the active ingredient (medicinal essential oil) of CPD0357 and thoroughly mixing it with emulsifiers and co-emulsifiers, while simultaneously adding distilled water dropwise until a homogeneous, clear, and transparent nanoemulsion is formed. Screening results showed that the optimal excipients for CPD0357 nanoemulsions include polyoxyethylene hydrogenated castor oil RH-40 (Cremophor RH-40), Tween-80, Span-80, and mixtures thereof as emulsifiers, and preferably ethanol, 1,2-propanediol, glycerol, diethylene glycol monoethyl ether, polyethylene glycol-400, and mixtures thereof as co-emulsifiers.

[0108] The ideal formulation of essential oil, emulsifier, co-emulsifier and aqueous phase was studied using pseudo-three-phase diagram. The optimal ratio was determined to be 2-6:3-9:1-3:10-20. The optimal emulsifier and co-emulsifier were selected as polyoxyethylene hydrogenated castor oil RH-40 (Cremophor RH-40) and ethanol, respectively.

[0109] Table 7. Feeding Table for the Preparation of Diallyl Disulfide-Based Nanoemulsion CPD0357A

[0110]

[0111] Preparation process of CPD0357 series nanoemulsion products: First, diallyl disulfide, L-menthol, 1,8-cineole, polyoxyethylene hydrogenated castor oil RH-40, and ethanol are added to a suitable mixing tank and stirred evenly with a stirrer. Purified water is continuously added dropwise while stirring. CPD0357 nanoemulsion is prepared and transferred to a high-speed centrifuge and centrifuged at 4000 rpm for 20-40 minutes.

[0112] The prepared nanoemulsions were characterized in terms of appearance, particle size, zeta potential, drug loading, content, and stability. The particle size, zeta potential, and drug loading of nanoemulsion CPD0357A were 32.77±0.38 nm, -2.65±0.25 mV, and 10.80%, respectively; while the particle size, zeta potential, and drug loading of nanoemulsion CPD0357B were 35.50±0.58 nm, -2.70±0.30 mV, and 11.10%, respectively. After being placed at 4℃ and room temperature for 15 days, the nanoemulsions of CPD0357A and CPD0357B remained clear and transparent with minimal particle size fluctuation, indicating good stability.

[0113] Example 13: Formulation and preparation of the CPD0358 series of composition inclusion complex formulations based on diallyl disulfide.

[0114] Based on the screening results of aquaporins and sodium-potassium ATPase agonists in Examples 1-4 above, and considering the pharmacological efficacy of related components, and taking into account the target indications for the prevention and treatment of pulmonary edema in the developed soft capsules, including oxidative stress and inflammatory stress, the active ingredients of the compositional encapsulation formulation CPD0358 based on diallyl disulfide are determined to be diallyl disulfide, L-menthol, and 1,8-cineole. According to the Chinese Pharmacopoeia, the recommended dosages of relevant traditional Chinese medicines (or extracts, etc.) in domestic and international studies, the content of active ingredients, and their screening experiments, the mass ratio of the three active ingredients in the CPD0358 soft capsule formulation is determined to be 4-8:1-2:4-12. The specific ingredient dosages for CPD0358A are shown in Table 8.

[0115] Using the phase solubility method, the solubilizing inclusion materials for the essential oil components of the drugs were determined to be hydroxypropyl-β-cyclodextrin, sulfobutyl ether-β-cyclodextrin, methyl-β-cyclodextrin, etc. The optimal inclusion ratio between the inclusion material and the essential oil was determined to be 10-30:1 (mass ratio) using single-factor experiments, and methyl-β-cyclodextrin was selected as the preferred inclusion material.

[0116] Table 8. Feeding table for the preparation of diallyl disulfide-based inclusion formulation CPD0358A.

[0117]

[0118] Preparation process of inclusion complex formulation CPD03578 series products: Through single-factor investigation experiments, ultrasonic method was selected as the best preparation method from ultrasonic method, stirring method, single-phase method and grinding method; according to the above-mentioned relevant feeding table, methyl-β-cyclodextrin was first dissolved in pure water, and active ingredients such as diallyl disulfide, L-menthol, and 1,8-cineole were added. The inclusion complex was prepared by ultrasonication for 40-60 minutes under key process conditions such as temperature 20°C and power 300 W.

[0119] The quality evaluation study of the prepared inclusion agent included appearance, inclusion rate, content, and stability. The quality evaluation study showed that the quality and stability of the prepared inclusion agent CPD0358A met the requirements.

[0120] Example 14: Evaluation of the anti-COPD efficacy of garlic oil-based soft capsule CPD0351A.

[0121] A rat model of COPD was established by intraperitoneal injection of cigarette smoke extract solution (CSE). Male Sprague-Dawley rats (weighing 130-150 g) were selected as the research subjects, and budesonide was used as the positive control drug. Male Sprague-Dawley rats (28 in total) were randomly divided into 4 groups of 7 rats each: (1) Control group: While rats in other groups received CSE injection to establish the model, rats in the control group received the same volume of phosphate injection (PBS); (2) COPD model group: 1 mL of CSE solution was injected intraperitoneally on days 1, 8, 15 and 22 of the experiment; (3) CPD0351A treatment group: 150 mg / kg (calculated based on garlic oil content) of CPD0351A soft capsules, which were dispersed in an aqueous solution to prepare a suspension, and were administered orally once a day starting from day 2; (4) Budesonide positive control group: 10 mg / kg of budesonide suspension, administered at the same time as CPD0351A. All animals were euthanized painlessly, and bronchoalveolar lavage fluid (BALF) and lung tissue were collected as samples for subsequent experimental studies.

[0122] The main studies evaluating the efficacy of CPD0351A in treating COPD included: (1) assessing alveolar damage in lung tissue using hematoxylin-eosin (HE) staining; (2) measuring inflammatory cytokines in the supernatant of lung tissue homogenate using ELISA according to the supplier's kit instructions; (3) measuring oxidative stress factors in lung (or liver) homogenate according to the supplier's BCA kit instructions; (4) dissecting the animals after the experiment, removing the spleen, removing adipose tissue, washing away blood in ice-cold saline, gently absorbing surface moisture with filter paper, weighing and recording the spleen weight, and calculating the spleen index to evaluate the immunomodulatory effect; (5) using Western blotting to detect the relative expression of p-Akt, total-Akt, and mTOR in rats, and exploring the feasibility of reversing budesonide resistance. The efficacy of CPD0351A in treating alveolar damage, anti-inflammatory stress, anti-oxidative stress, immunomodulation, and reversing budesonide resistance are as follows: Figure 1 , Figure 2 , Figure 3 , Figure 4 , Figure 5 As shown, these results indicate that CPD0351A is significantly more effective than budesonide in terms of antioxidant and immunomodulatory effects; furthermore, CPD0351A reverses budesonide resistance by regulating the PI3K signaling pathway (it is ineffective against p-Akt overexpression in COPD). Figure 5 This indicates that CPD0351A can be used in combination with hormone drugs such as budesonide to achieve a special effect of synergistic effect and reduced toxicity.

[0123] Example 15: Evaluation of the anti-pulmonary edema effect of the moringa oil-based composition inclusion complex formulation CPD0353A.

[0124] An acute lung injury (ALI) model was established in male SD rats (weighing 180-220 g) by intratracheal instillation of lipopolysaccharide (LPS) using a high-pressure spray needle. CPD0353A and the positive control Myrtol were administered by gavage. The therapeutic effects of CPD0353A on lung tissue damage, pulmonary edema, oxidative stress, and inflammatory stress were investigated by recording changes in body weight. Rats were randomly divided into a blank control group, an ALI model group (5 mg / kg LPS), a CPD0353A group (100 mg / kg, calculated as Moringa oil), and a Myrtol positive control group (400 mg / kg). After anesthetizing the rats, a 5 mg / kg LPS sulfate solution was instilled into the lungs via a high-pressure spray needle. The rats were kept in a sitting position and gently rocked from side to side to ensure even distribution of LPS and saline in the lungs, thus establishing a rat pulmonary fibrosis model. First, administer the drug by gavage according to the above protocol, once a day for 7 consecutive days. On the eighth day, use a micro-needle to drip LPS or saline into the trachea of ​​the rats. After each group of rats is exposed to saline or LPS for 24 hours, they are euthanized. 24 hours after drug withdrawal, all animals are sacrificed painlessly. Bronchoalveolar lavage fluid (BALF), serum and lung tissue are collected as samples for subsequent experimental studies.

[0125] The main studies on the efficacy of anti-lung injury and anti-pulmonary edema included: (1) washing away residual blood on the surface of a small piece of rat lung tissue with physiological saline, wiping away excess water on the surface of the lung tissue with filter paper, weighing to obtain wet weight, then baking in an 80°C oven for 48 h to constant weight, weighing to obtain dry weight, and calculating the dry-wet weight ratio; (2) using RT-PCR to detect inflammatory cytokines in lung tissue.

[0126] CPD0353A's therapeutic effects against pulmonary edema, inflammatory stress, and other lung injuries and pulmonary edema are listed in [the table / list of data]. Figure 6 and Figure 7 .like Figure 6 and Figure 7As shown, the oral dose of CPD0353A at 100 mg / kg showed significantly better therapeutic effects against pulmonary edema and inflammatory stress than Myrtol at 400 mg / kg, demonstrating a significant therapeutic effect against pulmonary edema injury.

[0127] Example 16: Evaluation of the anti-pulmonary fibrosis activity of soft capsule CPD0354A based on diallyl disulfide composition.

[0128] A mouse model of pulmonary fibrosis was established by intratracheal instillation of bleomycin sulfate (BLM) using a high-pressure spray needle. CPD0354A and the positive control allylcysteine ​​(NAC) were administered by gavage. The therapeutic effects of CPD0354A on pulmonary fibrosis, lung tissue damage, pulmonary edema, and cascade inflammation were assessed by recording changes in body weight. Mice were randomly divided into five groups: a blank control group (Ctrl), a BLM model group (BLM), a BLM + two-component combination group (100 mg / kg, DADS+MENT, diallyl disulfide + L-menthol), a BLM + CPD0354A group (100 mg / kg, CPD0354A), and a BLM + NAC positive control group (600 mg / kg, NAC). After anesthetizing mice, a 5 mg / kg BLM sulfuric acid solution was instilled into the lungs of the mice via intratracheal drip using a high-pressure spray needle. The mice were kept in a sitting position and gently rocked from side to side to ensure even distribution of BLM and saline in the lungs, thus establishing a mouse model of pulmonary fibrosis. Twenty-four hours later, the mice were administered saline, DADS+MENT, CPD0354A, and NAC by gavage, respectively. After several days of continuous administration, all animals were sacrificed painlessly, and bronchoalveolar lavage fluid (BALF) and lung tissue were collected as samples for subsequent experimental studies.

[0129] The main studies evaluating the efficacy against pulmonary fibrosis included: (1) assessing the area of ​​pulmonary fibrosis in lung tissue using Masson staining; (2) after collecting BLAF, centrifuging and washing with phosphate-buffered saline (PBS), and then counting neutrophils and total cells using a hemocytometer using the Swiss-Gymsa staining method; (3) measuring inflammatory cytokines in the supernatant of lung tissue homogenate using ELISA according to the supplier's kit instructions; and (4) measuring oxidative stress factors in lung (or liver) homogenate according to the supplier's BCA kit instructions.

[0130] The anti-fibrotic effects of CPD0354A on pulmonary fibrosis area, changes in alveolar neutrophil count, and inflammation and oxidative stress in lung homogenate were listed in [the table / reference]. Figure 8 , Figure 9 , Figure 10 ,and Figure 11 .like Figures 8-11As shown, the oral dose (100 mg / kg) of CPD0354A showed significantly better therapeutic effects against pulmonary fibrosis, alveolar neutrophils, inflammatory stress, and oxidative stress than the two-component combination group (100 mg / kg, DADS+MENT) and the positive control allylcysteine ​​(NAC) at 600 mg / kg, demonstrating significant anti-pulmonary fibrosis efficacy.

[0131] Example 17: Evaluation of the anti-lung injury effect of soft capsule CPD0356A based on Moringa isothiocyanate-1 composition.

[0132] An acute lung injury (ALI) model was established in male SD rats (weighing 180-220 g) by intratracheal instillation of lipopolysaccharide (LPS) using a high-pressure spray needle. CPD0356A and the positive control Myrtol were administered by gavage. The therapeutic effects of CPD0356A on lung tissue damage, pulmonary edema, oxidative stress, and inflammatory stress were investigated by recording changes in body weight. Rats were randomly divided into a blank control group, an ALI model group (ALI, 5 mg / kg LPS), a CPD0356A group (100 mg / kg, calculated as Moringa isothiocyanate-1), and a Myrtol positive control group (400 mg / kg). After anesthetizing the rats, a 5 mg / kg LPS sulfate solution was instilled into the lungs via a high-pressure spray needle. The rats were kept in a sitting position and gently rocked from side to side to ensure even distribution of LPS and saline in the lungs, thus establishing a rat pulmonary fibrosis model. First, administer the drug by gavage according to the above protocol, once a day for 7 consecutive days. On the eighth day, use a micro-needle to drip LPS or saline into the trachea of ​​the rats. After each group of rats is exposed to saline or LPS for 24 hours, they are euthanized. 24 hours after drug withdrawal, all animals are sacrificed painlessly. Bronchoalveolar lavage fluid (BALF), serum and lung tissue are collected as samples for subsequent experimental studies.

[0133] The main studies evaluating the efficacy of anti-lung injury included: (1) assessing alveolar damage in lung tissue using hematoxylin-eosin (HE) staining; and (2) detecting inflammatory cytokines in lung tissue using RT-PCR.

[0134] CPD0356A's therapeutic effects on lung injury, including anti-lung injury and anti-inflammatory stress, are listed in the following... Figure 12 and Figure 13 .like Figures 12-13 As shown, the oral dose of CPD0356A at 100 mg / kg showed significantly better therapeutic effects against lung injury and inflammatory stress than Myrtol at 400 mg / kg, demonstrating a significant therapeutic effect against injury.

[0135] Example 18: Evaluation of the expectorant and anti-lung injury efficacy of diallyl disulfide-based composition nanoemulsion CPD0357A.

[0136] Twenty male BALB / c mice were randomly divided into four groups of six each: a blank control group (Ctrl), an acute respiratory distress syndrome (ARDS) model group (ARDS), a two-component combination group (100 mg / kg, DADS+MENT, diallyl disulfide + L-menthol), a CPD0357A group (100 mg / kg, calculated as diallyl disulfide), and a Myrtol positive control group (300 mg / kg, Myrtol). Mice were anesthetized, and except for the blank control group, an ARDS mouse model was established by intratracheal instillation of lipopolysaccharide (LPS, 5 mg / kg) solution into the lungs. The blank control group received the same volume of saline solution. Six hours later, mice were administered saline, DADS+MENT, CPD0357A, and Myrtol via gavage, respectively. All mice were euthanized 24 hours later, and lung tissue and bronchoalveolar lavage fluid were collected for further studies.

[0137] The main studies on the efficacy of anti-lung injury and expectoration included: (1) assessing alveolar injury in lung tissue using hematoxylin-eosin (HE) staining; (2) examining the inhibition of goblet cell production and mucin secretion in lung tissue using AB-PAS staining; (3) measuring the expression of mucin MUC5AC in lung tissue using Western blotting; and (4) measuring inflammatory cytokines in lung tissue homogenate supernatant using ELISA according to the supplier's kit instructions.

[0138] The therapeutic effects of CPD0357A on lung injury, goblet cell formation and mucin secretion in lung tissue, expression of mucin MUC5AC in lung tissue, and inflammatory stress in lung tissue homogenate supernatant are listed in the following categories: Figure 14 , Figure 15 , Figure 16 ,and Figure 17 .like Figures 14-17 As shown, the oral dose (100 mg / kg) of CPD0357A showed significantly better therapeutic effects than the two-component combination group (100 mg / kg, DADS+MENT) and the positive control Myrtol (300 mg / kg) in terms of inhibiting lung injury, suppressing mucin hypersecretion, downregulating mucin MUC5AC, and anti-inflammatory stress, demonstrating significant efficacy in inhibiting hypersputum secretion and preventing lung injury.

[0139] Example 19: Evaluation of the anti-pulmonary edema and anti-pulmonary injury effects of the diallyl disulfide-based composition inclusion complex formulation CPD0358A.

[0140] An acute lung injury (ALI) model was established in male SD rats (weighing 180-220 g) by intratracheal instillation of lipopolysaccharide (LPS) using a high-pressure spray needle. CPD0358A and the positive control Myrtol were administered by gavage. The therapeutic effects of CPD0358A on lung tissue damage, pulmonary edema, oxidative stress, and inflammatory stress were investigated by recording changes in body weight. Rats were randomly divided into a blank control group, an ALI model group (5 mg / kg LPS), a CPD0358A group (80 mg / kg, calculated as diallyl disulfide), and a Myrtol positive control group (400 mg / kg). After anesthetizing the rats, LPS sulfate (5 mg / kg) solution was instilled into the lungs via a high-pressure spray needle. The rats were kept in a sitting position and gently rocked from side to side to ensure even distribution of LPS and saline in the lungs, thus establishing a rat pulmonary fibrosis model. First, administer the drug by gavage according to the above protocol, once a day for 7 consecutive days. On the eighth day, use a micro-needle to drip LPS or saline into the trachea of ​​the rats. After each group of rats is exposed to saline or LPS for 24 hours, they are euthanized. 24 hours after drug withdrawal, all animals are sacrificed painlessly. Bronchoalveolar lavage fluid (BALF), serum and lung tissue are collected as samples for subsequent experimental studies.

[0141] The main studies evaluating the efficacy of anti-lung injury and anti-pulmonary edema included: (1) assessing alveolar damage in lung tissue using hematoxylin-eosin (HE) staining; (2) washing away residual blood on the surface of a small piece of rat lung tissue with physiological saline, wiping away excess moisture on the surface of the lung tissue with filter paper, weighing to obtain wet weight, then baking in an 80°C oven for 48 h to constant weight, weighing to obtain dry weight, and calculating the wet-dry weight ratio; (3) using RT-PCR to detect the relevant aquaporins AQP3 and Na+ in lung tissue. + -K + (3) ATPase-α2 mRNA expression level; (4) ELISA method was used to measure inflammatory cytokines in serum according to the supplier's kit instructions.

[0142] CPD0358A has effects on lung injury, pulmonary edema, and upregulation of aquaporin AQP3 and Na+. + -K + The therapeutic effects of ATPase-α2 level, anti-inflammatory stress, and other anti-lung injury and anti-pulmonary edema treatments are listed in the following categories. Figure 18 , Figure 19 , Figure 20 , Figure 21 and Figure 22 .like Figures 18-22 As shown, an oral dose of 80 mg / kg of CPD0358A exhibits anti-lung injury, anti-pulmonary edema, and upregulation of aquaporin AQP3 and Na+.+ -K + The therapeutic effects of ATPase-α2 level and anti-inflammatory stress were significantly better than those of Myrtol at 400 mg / kg, demonstrating significant efficacy in treating lung injury and pulmonary edema.

[0143] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A composition based on plant essential oils or active ingredients, characterized in that, It includes at least three plant essential oils or plant active ingredients; the plant essential oils or plant active ingredients have agonistic activity against aquaporins and agonistic activity against sodium-potassium ATPase; The composition consists of garlic oil, peppermint oil, and eucalyptus oil in a mass ratio of 4-8:1-2:4-12. Alternatively, it can be composed of garlic oil, peppermint oil, and clove oil in a mass ratio of 4-8:1-2:12-36. Alternatively, it can be composed of moringa oil, peppermint oil, and eucalyptus oil in a mass ratio of 4-8:1-2:4-12; Alternatively, it is composed of diallyl disulfide, L-menthol, and 1,8-cineole in a mass ratio of 4-8:1-2:4-12; Alternatively, it is composed of diallyl disulfide, L-menthol, and eugenol in a mass ratio of 4-8:1-2:12-36; Alternatively, it is composed of Moringa isothiocyanate-1, L-menthol, and 1,8-cineole in a mass ratio of 4-8:1-2:4-12.

2. The composition according to claim 1, characterized in that, Plant essential oils or active ingredients were ranked by upregulating the expression levels of aquaporin AQP1 and sodium-potassium ATPase-α mRNA genes in A549 cells with pulmonary edema, and the top three were selected to form a combination.

3. The composition according to claim 1, characterized in that, At least one plant essential oil or plant active ingredient contains sulfur.

4. The composition according to claim 1, characterized in that, The composition further includes pharmaceutical excipients, which include one or more of fillers, emulsifiers, co-emulsifiers, capsule shell materials, preservatives, and inclusion agents.

5. The composition according to claim 1, characterized in that, The dosage form of the composition is an injectable liquid formulation, an oral liquid formulation, a microemulsion, a nanoemulsion, a liposome, a nanopolymer formulation, a soft capsule formulation, an aerosol, a nebulizer, a powder aerosol, a nasal spray, or an inclusion complex formulation.

6. The composition of claim 5, characterized in that, The excipients for the core material in soft capsule formulations include oil phase fillers, emulsifiers, and co-emulsifiers; Alternatively, the excipients for nanoemulsions include emulsifiers, co-emulsifiers, and water; Alternatively, the excipients in inclusion complex formulations may be solubilizing inclusion materials.

7. The use of a composition based on plant essential oils or active ingredients as described in any one of claims 1 to 6 in the preparation of a drug for preventing and treating pulmonary edema.

8. The use of a composition based on plant essential oils or active ingredients as described in any one of claims 1 to 6 in the preparation of a medicament for preventing and treating hypersputum secretion, acute lung injury, acute respiratory distress syndrome, pulmonary fibrosis, or COPD.

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