Lipid nanoparticle coated MOTS-c peptide and derivative thereof

Abstract

The invention discloses a lipid nanoparticle-coated MOTS-c peptide and a derivative thereof, and is characterized by comprising a lipid nanoparticle preparation and drug loading process, which specifically comprises the steps of lipid nanoparticle preparation, targeting ligand grafting, liposome characterization verification and in-vivo evaluation of targeting efficiency. The preparation process of the lipid nanoparticles comprises the following steps: step 1, dissolving hydrogenated soybean phosphatidylcholine (HSPC), cholesterol (Chol) and polyethylene glycol modified phospholipid (DSPE-PEG-COOH) in a mixed solvent of chloroform and methanol according to a molar ratio of 10: (5-15): (2-10): (0.5-3); according to the invention, a specific type of lipid nanoparticles are designed to wrap the mitochondrial peptide and the derivative of the mitochondrial peptide. The nano-particles not only can effectively protect peptide drugs from being degraded by an in-vitro environment, but also can realize lung targeting delivery through optimized particle size and surface characteristics, and the targeting property of the drugs is enhanced by modifying specific lung targeting ligands on the surfaces of the lipid nano-particles, so that the lipid nano-particles are ensured to play a role preferentially in the lung.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technologies, and particularly to a lipid nanoparticle encapsulating MOTS-c peptide and its derivatives. Background Art

[0002] Acute lung injury (ALI) is an acute lung disease caused by various factors. Common inducing factors include infection, trauma, inhalation of harmful gases, surgical operations, etc. Its main characteristics are the damage of alveolar epithelial cells and pulmonary endothelial cells, which then trigger pulmonary inflammation, gas exchange disorders, and pulmonary edema.

[0003] MOTS-c is a small molecule peptide encoded by mitochondrial genes and has significant physiological effects. It can exhibit potential therapeutic effects on acute lung injury and chronic lung diseases by regulating metabolic pathways, having an anti-inflammatory effect, inhibiting ferroptosis, and promoting cell repair. MOTS-c activates the AMPK pathway, promotes glucose metabolism and fatty acid oxidation, enhances the energy metabolism of cells, and thus provides support in the face of cell metabolic disorders. In addition, MOTS-c can also inhibit the inflammatory response, reduce the release of inflammatory factors, and mitigate the damage to the lungs caused by excessive immune responses. Most importantly, MOTS-c has the ability to promote the repair of lung cells. It can promote the regeneration and repair of lung epithelial cells and pulmonary endothelial cells by improving mitochondrial function and cell metabolism. Multiple previous studies in our group have also preliminarily demonstrated the protective effect of MOTS-c in acute lung injury;

[0004] However, as a peptide drug, MOTS-c faces problems such as poor stability, low bioavailability, and insufficient targeting in actual clinical applications. The MOTS-c peptide is easily degraded in the body, and its short half-life makes it difficult to sustain the therapeutic effect. In addition, MOTS-c lacks a specific targeted delivery mechanism, resulting in wide distribution of the drug in the body, which may cause side effects, especially the accumulation in non-target organs.

[0005] Existing LNP systems can only improve the stability and targeting of drugs to a certain extent, but there is still a risk that some drugs are not fully released in the lungs or are prematurely metabolized. Especially for peptide drugs, they degrade rapidly in the body. Although LNP technology can provide certain protection, for some sensitive peptides, there are still problems with insufficient stability. At the same time, too large a particle size may cause the drug to deposit on the airways, and too small a particle size may be cleared or not effectively released into the alveoli. For special drugs such as the MOTS-c peptide, particle size and carrier design are crucial, and existing delivery systems usually lack sufficient targeting, resulting in the drug not being effectively localized to specific regions of the lungs. Summary of the Invention

[0006] The purpose of the present invention is to provide a lipid nanoparticle encapsulated MOTS-c peptide and its derivatives, which has the advantages of high stability and strong targeting, and solves the problems mentioned in the background technology.

[0007] To achieve the above-mentioned object, the present invention provides the following technical solutions: a lipid nanoparticle encapsulating MOTS-c peptide and its derivatives, characterized by: comprising a lipid nanoparticle preparation and drug loading process, specifically: preparation of lipid nanoparticles, grafting of targeting ligands, characterization and verification of liposomes, and in vivo evaluation of targeting efficiency;

[0008] The lipid nanoparticle preparation process is as follows:

[0009] Step 1: Hydrogenated soybean phosphatidylcholine (HSPC), cholesterol (Chol) and polyethylene glycol-modified phospholipids (DSPE-PEG-COOH) were dissolved in a mixed solvent of chloroform and methanol at a molar ratio of 10:5–15:2–10:0.5–3 to form a uniform raw material solution;

[0010] Step 2: Add DiR fluorescent dye at a molar ratio of 0.1%–1.0% of the total lipids, mix it thoroughly with the lipids, evaporate the organic solvent in a rotary evaporator, and after forming a lipid film, place it in a vacuum drying oven for 6–24 hours to completely remove the residual solvent;

[0011] Step 3: The dried lipid film is re-dissolved by a hydration method. During the hydration process, 6-15 mL of a MOTS-c solution containing 0.01-0.02 M phosphate buffer (PBS, pH 7.4) is added, and the MOTS-c concentration range is 1-5 mg / mL. The mixed solution is stirred at 30-40°C for 30-120 minutes to form a crude liposome suspension containing MOTS-c;

[0012] Step 4: Place the liposome suspension in an ice bath and use an ultrasonic device with a power of 150-300W for 5-15 minutes to reduce the particle size and improve uniformity. Finally, filter through a 0.45μm filter membrane to remove large particles to obtain a liposome suspension with uniform particle size and containing MOTS-c. The liposome product is finally stored at 4°C for subsequent targeted modification or in vitro and in vivo experiments.

[0013] The targeting ligand grafting process is as follows:

[0014] To endow liposomes with lung tissue targeting function, a targeting ligand (such as the LTHSPWG peptide) is grafted onto the polyethylene glycol-modified phospholipid on the surface of liposomes through chemical modification. The LTHSPWG peptide is a targeting peptide sequence that can specifically recognize pulmonary surfactant protein A (SP-A). It achieves the targeted delivery of liposomes to lung tissue by binding to SP-A with high affinity. This peptide has a high targeting efficiency, a small molecular weight, and is not easily prone to triggering an immune response, making it suitable as a functionalized modification molecule for the delivery system of lung diseases.

[0015] Step 1: Place 2 mL of liposome suspension in a reaction vessel. Add 5–10 μmol of EDC and NHS per 1 μmol of polyethylene glycol-modified phospholipid, and stir at 25 - 40 °C for 2 - 4 hours to activate the carboxyl groups on the surface of the liposomes.

[0016] Step 2: Add the targeting ligand solution with a molar ratio of targeting ligand: polyethylene glycol-modified phospholipid of (0.0126 - 0.063):1 to the activated liposome solution, and gently stir at 4 °C for 12 - 18 hours to graft the targeting ligand onto the surface of the liposomes by forming stable amide bonds.

[0017] Step 3: After the reaction is completed, place the liposome solution in a dialysis bag with a molecular weight cut-off (MWCO) of 3500–10000, and dialyze at 4 °C for 24 hours, changing the buffer 3–5 times during this period to remove unreacted EDC, NHS, and free targeting ligand.

[0018] Step 4: The targeted-modified liposomes are detected by UV-Vis or HPLC to measure the concentration of unbound ligands and calculate the grafting efficiency. At the same time, dynamic light scattering (DLS) is used to detect changes in particle size and Zeta potential to verify the success of grafting.

[0019] The characterization and verification process of the liposomes is as follows:

[0020] The particle size and uniformity of the liposomes are measured by dynamic light scattering (DLS), with the particle size range being 80–150 nm and the polydispersity index (PDI) being less than 0.2. The surface potential is analyzed by Zeta potential to verify the success of targeted grafting, with the range being -10 mV to -30 mV.

[0021] The drug loading and encapsulation efficiency are detected and calculated by high-performance liquid chromatography (HPLC), representing the percentage of drug mass in the total mass of liposomes (DL) and the percentage of drug mass in liposomes in the initial added drug mass (EE), respectively.

[0022] The evaluation process of the targeting efficiency is as follows:

[0023] The targeted liposomes were delivered to the lungs of experimental animals through an aerosol inhalation device, and a small animal in vivo imaging system (such as IVIS Spectrum) was used to detect the fluorescence distribution of DiR-labeled liposomes in lung tissue. The experimental group was liposomes grafted with a targeting ligand, and the control group was liposomes without modification of the targeting ligand. By normalizing the fluorescence signal intensity ratio, the improvement of the enrichment efficiency of the targeted liposomes in lung tissue was quantitatively analyzed. Combining the fluorescence intensity in the lungs with the fluorescence distribution ratio in non-lung tissues (such as the liver and kidneys), the lung targeting and specificity of the liposomes were further verified.

[0024] Preferably, in step one of the lipid nanoparticle preparation process, the volume ratio of the mixed solvent of chloroform and methanol is 1:1.

[0025] Preferably, in step two of the lipid nanoparticle preparation process, the environmental temperature in the rotary evaporator is set at 25–40 °C and the rotation speed is controlled at 20–50 rpm.

[0026] Preferably, in step one of the targeting ligand grafting process, the liposome suspension is 10–50 mg / mL of the total lipid amount.

[0027] Preferably, in step two of the targeting ligand grafting process, the concentration range of the targeting ligand solution is 1–3 mg / mL.

[0028] Preferably, the characterization and verification of the liposomes also include: studying the release characteristics of MOTS-c through an in vitro dialysis experiment simulating the liquid environment of lung tissue, plotting its release kinetic curve, and detecting the excitation and emission spectra of DiR-labeled liposomes by a fluorescence spectrophotometer to confirm the success and uniformity of the fluorescence labeling;

[0029] Preferably, it also includes an atomizing device that can be effectively atomized into aerosol particles with a particle size of 1-5 μm for inhalation by the patient through the respiratory system.

[0030] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0031] The present invention wraps mitochondrial peptides and their derivatives by designing specific types of lipid nanoparticles. These nanoparticles can not only effectively protect peptide drugs from degradation in the external environment, but also achieve lung-targeted delivery through optimized particle size and surface properties. By modifying specific lung-targeting ligands (such as lung surfactant protein like the short peptide LTHSPWG or ligands of lung endothelial cell receptors) on the surface of the lipid nanoparticles, the targeting of the drug is enhanced, ensuring that it can preferentially act on the lungs. By modifying the surface of the lipid nanoparticles with lung-specific ligands (such as specific ligands of lung surfactant protein receptors or lung endothelial cell receptors), these ligands are used to bind to the receptors of lung cells, thereby enhancing the targeting and ensuring that the drug can preferentially act on lung cells, reducing the distribution of the drug in non-target tissues and reducing side effects. The lipid nanoparticles encapsulating MOTS-c peptides and their derivatives have the advantages of high stability and strong targeting, solving the problem that existing LNP systems can only improve the stability and targeting of drugs to a certain extent, but there is still a risk that some drugs are not fully released in the lungs or are prematurely metabolized. Especially for peptide drugs, their degradation rate in the body is relatively fast. Although LNP technology can provide certain protection, for some sensitive peptides, there is still a problem of insufficient stability. At the same time, too large a particle size may cause the drug to deposit on the airway, and too small a particle size may be cleared or not effectively released into the alveoli. For special drugs such as MOTS-c peptides, the particle size and carrier design are crucial, and existing delivery systems usually lack sufficient targeting, resulting in the drug not being effectively localized to specific regions of the lungs. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 It is a schematic diagram of the process of the present invention;

[0033] Figure 2 It is a histological section diagram of the lungs of mice with LIRI and LIRI+MOTS aerosol-induced lung injury diseases of the present invention;

[0034] Figure 3 It is a histological section diagram of the lungs of mice with LPS and LPS+MOTS aerosol-induced lung injury diseases of the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0035] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

[0036] Please refer to Figure 3The present invention provides a technical solution: a lipid nanoparticle encapsulating MOTS-c peptide and its derivatives, characterized by: including a lipid nanoparticle preparation and drug loading process, specifically: lipid nanoparticle preparation, targeting ligand grafting, liposome characterization and verification, and in vivo evaluation of targeting efficiency.

[0037] The lipid nanoparticle preparation process is as follows:

[0038] Step 1: Hydrogenated soybean phosphatidylcholine (HSPC), cholesterol (Chol) and polyethylene glycol-modified phospholipids (DSPE-PEG-COOH) were dissolved in a mixed solvent of chloroform and methanol at a molar ratio of 10:5–15:2–10:0.5–3 to form a uniform raw material solution;

[0039] Step 2: Add DiR fluorescent dye at a molar ratio of 0.1%–1.0% of the total lipids, mix it thoroughly with the lipids, evaporate the organic solvent in a rotary evaporator, and after forming a lipid film, place it in a vacuum drying oven for 6–24 hours to completely remove the residual solvent;

[0040] Step 3: The dried lipid film is re-dissolved by a hydration method. During the hydration process, 6-15 mL of a MOTS-c solution containing 0.01-0.02 M phosphate buffer (PBS, pH 7.4) is added, and the MOTS-c concentration range is 1-5 mg / mL. The mixed solution is stirred at 30-40°C for 30-120 minutes to form a crude liposome suspension containing MOTS-c;

[0041] Step 4: Place the liposome suspension in an ice bath and use an ultrasonic device with a power of 150-300W for 5-15 minutes to reduce the particle size and improve uniformity. Finally, filter through a 0.45μm filter membrane to remove large particles to obtain a liposome suspension with uniform particle size and containing MOTS-c. The liposome product is finally stored at 4°C for subsequent targeted modification or in vitro and in vivo experiments.

[0042] The targeting ligand grafting process is as follows:

[0043] Step 1: Place 2 mL of liposome suspension in a reaction vessel, add 5–10 μmol of EDC and NHS per 1 μmol of polyethylene glycol-modified phospholipid, and stir at 25–40 °C for 2–4 hours to activate the carboxyl groups on the liposome surface;

[0044] Step 2: Add the targeting ligand solution: polyethylene glycol-modified phospholipids at a molar ratio of (0.0126-0.063):1 to the activated liposome solution, gently stir at 4°C for 12-18 hours, and graft the targeting ligand to the liposome surface by forming a stable amide bond;

[0045] Step 3: After the reaction is completed, place the liposome solution in a dialysis bag with a molecular weight cut-off (MWCO) of 3500–10000, and dialyze it at 4 °C for 24 hours, changing the buffer 3–5 times during this period to remove unreacted EDC, NHS, and free targeting ligands;

[0046] Step 4: The concentration of unbound ligands in the target-modified liposomes is detected by UV-Vis or HPLC to calculate the grafting efficiency. At the same time, dynamic light scattering (DLS) is combined to detect the particle size change and Zeta potential to verify the success of grafting.

[0047] The characterization and verification process of liposomes is as follows:

[0048] The particle size and uniformity of liposomes are determined by dynamic light scattering (DLS), with the particle size range of 80–150 nm and the polydispersity index (PDI) less than 0.2; the surface potential is analyzed by Zeta potential to verify the success of target grafting, with the range of -10 mV to -30 mV;

[0049] The drug loading and encapsulation efficiency are detected and calculated by high-performance liquid chromatography (HPLC), which represent the percentage of drug mass in the total mass of liposomes (DL) and the percentage of drug mass in liposomes in the initial added drug mass (EE), respectively.

[0050] The evaluation process of the targeting efficiency is as follows:

[0051] The target liposomes are delivered to the lungs of experimental animals through an aerosol inhalation device, and the fluorescence distribution of DiR-labeled liposomes in lung tissue is detected using a small animal in vivo imaging system (such as IVIS Spectrum). The experimental group is the liposomes grafted with target ligands, and the control group is the liposomes without modified target ligands. By normalizing the fluorescence signal intensity ratio, the enrichment efficiency improvement of target liposomes in lung tissue is quantitatively analyzed. Combining the fluorescence intensity ratio in the lungs with the fluorescence distribution ratio in non-lung tissues (such as the liver and kidneys) further verifies the lung targeting and specificity of liposomes.

[0052] In the present invention: In step 1 of the lipid nanoparticle preparation process, the volume ratio of the mixed solvent of chloroform and methanol is 1:1.

[0053] In the present invention: In step 2 of the lipid nanoparticle preparation process, the environmental temperature in the rotary evaporator is set at 25–40 °C, and the rotation speed is controlled at 20–50 rpm.

[0054] In the present invention: In step 1 of the target ligand grafting process, the liposome suspension is 10–50 mg / mL of the total lipid amount.

[0055] In the present invention: In step 2 of the target ligand grafting process, the concentration range of the target ligand solution is 1–3 mg / mL.

[0056] In the present invention, the characterization and verification of liposomes further include: studying the release characteristics of MOTS-c through an in vitro dialysis experiment simulating the liquid environment of lung tissue, plotting its release kinetic curve, and detecting the excitation and emission spectra of DiR-labeled liposomes by a fluorescence spectrophotometer to confirm the success and uniformity of fluorescence labeling;

[0057] In the present invention, it further includes an atomization device that can be effectively atomized into aerosol particles with a particle size of 1 - 5 μm for inhalation by patients through the respiratory system.

[0058] In summary: The lipid nanoparticles encapsulate MOTS-c peptide and its derivatives by designing specific types of lipid nanoparticles to encapsulate mitochondrial peptides and their derivatives. These nanoparticles can not only effectively protect peptide drugs from degradation in the external environment but also achieve lung-targeted delivery through optimized particle size and surface properties. The targeting of the drug is enhanced by modifying specific lung-targeting ligands (such as lung surfactant protein like the LTHSPWG short peptide or ligands of lung endothelial cell receptors) on the surface of the lipid nanoparticles, ensuring that it can preferentially act in the lungs. By modifying the surface of the lipid nanoparticles with lung-specific ligands (such as specific ligands of lung surfactant receptors or lung endothelial cell receptors), these ligands bind to the receptors of lung cells, thereby enhancing the targeting and ensuring that the drug can preferentially act on lung cells, reducing the distribution of the drug in non-target tissues and reducing side effects. The lipid nanoparticles encapsulating MOTS-c peptide and its derivatives have the advantages of high stability and strong targeting, solving the problem that existing LNP systems can only improve the stability and targeting of drugs to a certain extent, but there is still a risk that some drugs are not fully released or are prematurely metabolized in the lungs, especially for peptide drugs, which have a relatively fast degradation rate in the body. Although LNP technology can provide certain protection, for some sensitive peptides, there is still a problem of insufficient stability. At the same time, too large a particle size may cause the drug to deposit on the airway, and too small a particle size may be cleared or not effectively released into the alveoli. For special drugs such as MOTS-c peptide, particle size and carrier design are crucial, and existing delivery systems usually lack sufficient targeting, resulting in the drug not being effectively localized to specific regions of the lungs.

[0059] It should be noted that, in this text, relational terms such as first and second are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any actual relationship or order between these entities or operations. Moreover, the terms "comprising", "including" or any other variant thereof are intended to cover non-exclusive inclusion, such that a process, method, article or device comprising a series of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such process, method, article or device.

[0060] Although the embodiments of the present invention have been shown and described, those of ordinary skill in the art can understand that various changes, modifications, substitutions and variations can be made to these embodiments without departing from the principles and spirit of the present invention. The scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A lipid nanoparticle encapsulating MOTS-c peptide and its derivatives, characterized in that: It includes a lipid nanoparticle preparation and drug loading process, specifically: lipid nanoparticle preparation, targeting ligand grafting, liposome characterization and verification, and in vivo evaluation of targeting efficiency The lipid nanoparticle preparation process is as follows: Step 1: Dissolve hydrogenated soybean phosphatidylcholine (HSPC), cholesterol (Chol) and polyethylene glycol-modified phospholipid (DSPE-PEG-COOH) in a mixed solvent of chloroform and methanol at a molar ratio of 10:5–15:2–10:0.5–3 to form a uniform raw material solution; GC-MS is used to detect the residual solvent concentration; Step 2: Add DiR fluorescent dye at a molar ratio of 0.1%–1.0% of the total lipids, mix it thoroughly with the lipids, evaporate the organic solvent in a rotary evaporator, and after forming a lipid film, place it in a vacuum drying oven for 6–24 hours to completely remove the residual solvent; Step 3: The dried lipid film is re-dissolved by a hydration method. During the hydration process, 6-15 mL of a MOTS-c solution containing 0.01-0.02 M phosphate buffer (PBS, pH 7.4) is added, and the MOTS-c concentration range is 1-5 mg / mL. The mixed solution is stirred at 30-40°C for 30-120 minutes to form a crude liposome suspension containing MOTS-c; Step 4: Place the liposome suspension in an ice bath and use an ultrasonic device with a power of 150-300W for 5-15 minutes to reduce the particle size and improve uniformity. Finally, filter through a 0.45μm filter membrane to remove large particles to obtain a liposome suspension with uniform particle size and containing MOTS-c. The liposome product is finally stored at 4°C for subsequent targeted modification or in vitro and in vivo experiments. The targeting ligand grafting process is as follows: Step 1: Place 2 mL of liposome suspension in a reaction vessel, add 5–10 μmol of EDC and NHS per 1 μmol of polyethylene glycol-modified phospholipid, and stir at 25–40 °C for 2–4 hours to activate the carboxyl groups on the liposome surface; Step 2: Add the targeting ligand solution: polyethylene glycol-modified phospholipids at a molar ratio of (0.0126-0.063):1 to the activated liposome solution, gently stir at 4°C for 12-18 hours, and graft the targeting ligand to the liposome surface by forming a stable amide bond; Step 3: After the reaction is completed, the liposome solution is placed in a dialysis bag with a molecular weight cutoff (MWCO) of 3500-10000 and dialyzed at 4°C for 24 hours, during which the buffer is replaced 3-5 times to remove unreacted EDC, NHS and free targeting ligand; Step 4: The concentration of unbound ligand in the targeted modified liposomes is detected by UV-Vis or HPLC to calculate the grafting efficiency. At the same time, the particle size change and Zeta potential are detected by dynamic light scattering (DLS) to verify the success of the grafting. The characterization and verification process of the liposomes is as follows: The particle size and uniformity of the liposomes were determined by dynamic light scattering (DLS), with a particle size range of 80–150 nm and a polydispersity index (PDI) of less than 0.

2. The surface potential was verified by Zeta potential analysis to verify the success of targeted grafting, ranging from -10 mV to -30 mV. The drug loading and encapsulation efficiency were calculated by high performance liquid chromatography (HPLC) detection, which respectively represent the percentage of drug mass to the total mass of liposomes (DL) and the percentage of drug mass within liposomes to the mass of initially added drug (EE); In vitro release characteristics: The release characteristics of MOTS-c were studied by in vitro dialysis experiments simulating the liquid environment of lung tissue, and its release kinetics curve was drawn; The evaluation process of the targeting efficiency is as follows: In vivo experiments: Targeted liposomes were delivered to the lungs of experimental animals through an atomizer inhalation device, and the fluorescence distribution of DiR-labeled liposomes in lung tissue was detected using a small animal in vivo imaging system (such as IVISSpectrum). The experimental group was liposomes grafted with targeting ligands, and the control group was liposomes without modified targeting ligands. The enrichment efficiency of targeted liposomes in lung tissue was quantitatively analyzed by normalizing the fluorescence signal intensity ratio. The lung fluorescence intensity was combined with the fluorescence distribution ratio of non-lung tissues (such as liver and kidney) to further verify the lung targeting and specificity of the liposomes. In vitro experiment: The binding rate of targeted liposomes to lung epithelial cells was detected by flow cytometry.

2. The lipid nanoparticles encapsulating MOTS-c peptide and its derivatives according to claim 1, characterized in that: In step 1 of the lipid nanoparticle preparation process, the volume ratio of the mixed solvent of chloroform and methanol is 1:

1.

3. The lipid nanoparticles encapsulating MOTS-c peptide and its derivatives according to claim 1, characterized in that: In step 2 of the lipid nanoparticle preparation process, the ambient temperature in the rotary evaporator is set to 25-40°C and the rotation speed is controlled to 20-50 rpm.

4. The lipid nanoparticles encapsulating MOTS-c peptide and its derivatives according to claim 1, characterized in that: In step 1 of the targeting ligand grafting process, the liposome suspension has a total lipid content of 10-50 mg / mL.

5. The lipid nanoparticles encapsulating MOTS-c peptide and its derivatives according to claim 1, characterized in that: The concentration range of the targeting ligand solution in step 2 of the targeting ligand grafting process is 1-3 mg / mL.

6. The lipid nanoparticles encapsulating MOTS-c peptide and its derivatives according to claim 1, characterized in that: The characterization and verification of the liposomes also includes: studying the release characteristics of MOTS-c through an in vitro dialysis experiment simulating the liquid environment of lung tissue, drawing its release kinetics curve, and detecting the excitation and emission spectra of DiR-labeled liposomes through a fluorescence spectrophotometer to confirm the success and uniformity of the fluorescence labeling.

7. The lipid nanoparticles encapsulating MOTS-c peptide and its derivatives according to claim 1, characterized in that: It also includes a nebulizer that can be effectively nebulized into aerosol particles with a particle size of 1-5 μm, and the alveolar deposition rate after nebulization is >30% and the encapsulation rate decreases by <5%, so that the particles can be inhaled by the patient through the respiratory system.

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