Preparation method of respiratory tract delivery preparation for severe respiratory system diseases

Abstract

The invention discloses a preparation method of a respiratory tract delivery preparation for severe respiratory system diseases, the respiratory tract delivery preparation comprises the following components by weight: 2 * 10 < 8 >-2 * 10 < 9 > pieces / ml of cell exosomes, normal saline with atomization concentration of 3%, and the balance of water. The cell exosome is composed of adipose-derived stem cell exosomes and bone marrow stem cell exosomes, the adipose-derived stem cell exosomes are 1 * 10 < 8 >-1 * 10 < 9 > / ml, the bone marrow stem cell exosomes are 1 * 10 < 8 >-1 * 10 < 9 > / ml, and the ratio of the adipose-derived stem cell exosomes to the bone marrow stem cell exosomes is 1: 1. The respiratory tract delivery preparation prepared by the invention can utilize nasal spray, aerosol inhalation or other suitable drug delivery technology, and the drug delivery method ensures that the preparation directly enters the respiratory system, nasal mucosa and bronchoalveolus, which generally relieves the curative effect of treating major respiratory system diseases at present. And a clinical basis is provided for research and development in a new direction of disease-targeted biological treatment.

Description

Technical Field

[0001] The present invention relates to the technical field of preparation of respiratory tract delivery preparations, and in particular to a method for preparing a respiratory tract delivery preparation for major respiratory diseases. Background Art

[0002] The information disclosed in this background technology section is only intended to enhance the understanding of the overall background of the invention, and should not be necessarily regarded as an admission or any form of suggestion that the information constitutes the prior art already known to those skilled in the art.

[0003] Respiratory diseases refer to various diseases that affect the normal function of the respiratory system, such as colds, pneumonia, and asthma. These diseases have a significant impact on people's health and life, so prevention and control of respiratory diseases is particularly important. Prevention and control of respiratory diseases requires us to pay attention to personal hygiene habits, maintain a good diet, avoid contact with pathogens, maintain a good indoor ventilation environment, strengthen physical exercise, and maintain a good mental state. The effective implementation of these measures can not only effectively prevent the occurrence of respiratory diseases, but also help improve overall health and quality of life. We should incorporate these preventive measures into our lives every day to protect the respiratory health of ourselves and others.

[0004] The present invention aims to address the major challenge of delivering drugs directly to the respiratory tract or nasal mucosa to prevent and control major respiratory diseases such as chronic bronchitis, cor pulmonale, idiopathic pulmonary fibrosis, bronchial asthma, bronchopulmonary dysplasia, chronic obstructive pulmonary disease and rhinitis. Current drug delivery methods are difficult to achieve the desired therapeutic effect. Summary of the invention

[0005] The purpose of the present invention is to provide a method for preparing a respiratory delivery preparation for major respiratory diseases, so as to solve the major challenge of delivering drugs directly to the respiratory tract or nasal mucosa to prevent and control major respiratory diseases, such as chronic bronchitis, cor pulmonale, idiopathic pulmonary fibrosis, bronchial asthma, bronchopulmonary dysplasia, chronic obstructive pulmonary disease and rhinitis proposed in the above background technology, and the problem that the current drug delivery method is difficult to achieve the ideal therapeutic effect. The present invention proposes a method for preparing a respiratory delivery preparation for major respiratory diseases, and the weight ratio of the respiratory delivery preparation is: 2×10 8 -2×10 9 / ml, physiological saline: Preferably, the atomization concentration of the physiological saline is 3%.

[0006] Preferably, the cell exosomes are composed of adipose stem cell exosomes and bone marrow stem cell exosomes, and the adipose stem cell exosomes are 1×10 8-1×10 9 The number of exosomes of bone marrow stem cells is 1×108-1×109 / ml, and the ratio of exosomes of adipose stem cells to exosomes of bone marrow stem cells is 1:1.

[0007] A method for preparing a respiratory delivery preparation for serious respiratory diseases, characterized in that the preparation method comprises the following steps: S1. Collect peripheral blood and separate mononuclear cells by Eicoll density gradient centrifugation; S2, inoculating the mononuclear cells into a culture bottle at a certain density and placing it in an incubator for culture; S3. Observe the cells in the incubator for attachment to the wall and then change the medium for the first time. Change the medium at intervals thereafter. When the cells are low-generation cells, adhere to the wall and expand the culture, and when the cell fusion rate reaches more than 90%, collect the cell exosomes. S4, washing the cell exosomes with an appropriate separation buffer, and then centrifuging the washed cell exosomes at low temperature; S5. Wash the exosomes obtained in S5 again using an appropriate eluent to remove residual cell debris, serum proteins, and washing buffer; S6, resuspending the exosome precipitate after washing again, and using a purification technique to separate and purify the polypeptide factors in the exosomes, wherein the purification technique includes density gradient centrifugation, ultracentrifugation combined with column chromatography; S7. The concentration of purified exosomes was determined by nanoparticle tracking analysis (NTA). S8. Mix the purified and tested exosomes with normal saline in proportion to obtain a respiratory delivery preparation.

[0008] Preferably, in S2, the culture temperature is 37°C, the carbon dioxide concentration is 5%, and the culture time is 12-24 hours.

[0009] Preferably, in S4, the separation buffer is a phosphate buffered saline solution containing 0.1% bovine serum albumin; preferably, the separation buffer is sterilized by filtering with a 0.22 micron filter membrane in advance.

[0010] Preferably, in S4, the centrifugation factor is 2500-3000g, the centrifugation temperature is 3-4°C, and the centrifugation time is 50-60min.

[0011] Preferably, in S5, the eluent is a glycine solution with a pH of 2.8 to 3.5 and a glycine concentration of 0.1 to 0.2 mol / L.

[0012] Preferably, in S8, the addition ratio of the exosomes to the physiological saline is 1:9.

[0013] Preferably, in S1-S8, each result obtained is tested, recorded and uploaded to the traceability system.

[0014] Compared with the prior art, the present invention has the following beneficial effects: 1. The respiratory delivery preparation prepared by the present invention can be delivered by nasal spray, atomization inhalation or other suitable administration techniques. This administration method ensures direct access to the respiratory system, the nasal mucosa and the bronchial alveoli, which generally alleviates the current efficacy of treating major respiratory diseases and provides a clinical basis for the research and development of new directions of disease-targeted biological treatments; 2. The present invention is distinguished from previous inventions by the use of a combination of exosomes from two different types of cells and an innovative respiratory or intranasal delivery method, which provides a new treatment method for the prevention and control of chronic bronchitis, cor pulmonale, idiopathic pulmonary fibrosis, bronchial asthma, bronchopulmonary dysplasia, chronic obstructive pulmonary disease and rhinitis, and addresses the limitations of existing treatment methods in the treatment and prevention of major respiratory diseases. DETAILED DESCRIPTION

[0015] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0016] The present invention provides a method for preparing a respiratory delivery preparation for major respiratory diseases, wherein the weight ratio of the respiratory delivery preparation is as follows: 2×10 8 -2×10 9 pcs / ml, normal saline; In this embodiment, the atomization concentration of the physiological saline is 3%.

[0017] In this embodiment, the cell exosomes are composed of adipose stem cell exosomes and bone marrow stem cell exosomes. The adipose stem cell exosomes are 1×10 8 -1×10 9 The number of exosomes of bone marrow stem cells is 1×108-1×109 / ml, and the ratio of exosomes of adipose stem cells to exosomes of bone marrow stem cells is 1:1.

[0018] The preparation method of this embodiment comprises the following steps: S1. Collect peripheral blood and separate mononuclear cells by Eicoll density gradient centrifugation; S2, inoculating the mononuclear cells into a culture bottle at a certain density and placing it in an incubator for culture; S3. Observe the cells in the incubator for attachment to the wall and then change the medium for the first time. Change the medium at intervals thereafter. When the cells are low-generation cells, adhere to the wall and expand the culture, and when the cell fusion rate reaches more than 90%, collect the cell exosomes. S4, washing the cell exosomes with an appropriate separation buffer, and then centrifuging the washed cell exosomes at low temperature; S5. Wash the exosomes obtained in S5 again using an appropriate eluent to remove residual cell debris, serum proteins, and washing buffer; S6, resuspending the exosome precipitate after washing again, and using a purification technique to separate and purify the polypeptide factors in the exosomes, wherein the purification technique includes density gradient centrifugation, ultracentrifugation combined with column chromatography; S7. The concentration of purified exosomes was determined by nanoparticle tracking analysis (NTA). S8. Mix the purified and tested exosomes with normal saline in proportion to obtain a respiratory delivery preparation.

[0019] In this embodiment, in S2, the culture temperature is 37°C, the carbon dioxide concentration is 5%, and the culture time is 12-24 hours.

[0020] In this embodiment, in S4, the separation buffer is a phosphate buffered saline solution containing 0.1% bovine serum albumin; preferably, the separation buffer is sterilized by filtering with a 0.22 micron filter membrane in advance.

[0021] In this embodiment, in S4, the centrifugation factor is 2500-3000g, the centrifugation temperature is 3-4°C, and the centrifugation time is 50-60min.

[0022] In this embodiment, in S5, the eluent is a glycine solution with a pH of 2.8 to 3.5 and a glycine concentration of 0.1 to 0.2 mol / L.

[0023] In this embodiment, in S8, the addition ratio of the exosomes to the physiological saline is 1:9.

[0024] In this embodiment, in S1-S8, each result obtained is tested, recorded and uploaded to the traceability system.

[0025] Embodiment 2: The difference from the first embodiment is that: The weight ratio of the respiratory delivery preparation is as follows: 2×10 8 -2×10 9 pcs / ml, normal saline; In this embodiment, the atomization concentration of the physiological saline is 5%.

[0026] In this embodiment, the cell exosomes are composed of adipose stem cell exosomes and bone marrow stem cell exosomes. The adipose stem cell exosomes are 1×10 8 -1×10 9 The number of exosomes of bone marrow stem cells is 1×108-1×109 / ml, and the ratio of exosomes of adipose stem cells to exosomes of bone marrow stem cells is 1:1.

[0027] The preparation method of this embodiment comprises the following steps: S1. Collect peripheral blood and separate mononuclear cells by Eicoll density gradient centrifugation; S2, inoculating the mononuclear cells into a culture bottle at a certain density and placing it in an incubator for culture; S3. Observe the cells in the incubator for attachment to the wall and then change the medium for the first time. Change the medium at intervals thereafter. When the cells are low-generation cells, adhere to the wall and expand the culture, and when the cell fusion rate reaches more than 90%, collect the cell exosomes. S4, washing the cell exosomes with an appropriate separation buffer, and then centrifuging the washed cell exosomes at low temperature; S5. Wash the exosomes obtained in S5 again using an appropriate eluent to remove residual cell debris, serum proteins, and washing buffer; S6, resuspending the exosome precipitate after washing again, and using a purification technique to separate and purify the polypeptide factors in the exosomes, wherein the purification technique includes density gradient centrifugation, ultracentrifugation combined with column chromatography; S7. The concentration of purified exosomes was determined by nanoparticle tracking analysis (NTA). S8. Mix the purified and tested exosomes with normal saline in proportion to obtain a respiratory delivery preparation.

[0028] In this embodiment, in S2, the culture temperature is 37°C, the carbon dioxide concentration is 5%, and the culture time is 12-24 hours.

[0029] In this embodiment, in S4, the separation buffer is a phosphate buffered saline solution containing 0.1% bovine serum albumin; preferably, the separation buffer is sterilized by filtering with a 0.22 micron filter membrane in advance.

[0030] In this embodiment, in S4, the centrifugation factor is 2500-3000g, the centrifugation temperature is 3-4°C, and the centrifugation time is 50-60min.

[0031] In this embodiment, in S5, the eluent is a glycine solution with a pH of 2.8 to 3.5 and a glycine concentration of 0.1 to 0.2 mol / L.

[0032] In this embodiment, in S8, the addition ratio of the exosomes to the physiological saline is 1:9.

[0033] In this embodiment, in S1-S8, each result obtained is tested, recorded and uploaded to the traceability system.

[0034] Embodiment three: The difference between the first embodiment and the second embodiment is that: The weight ratio of the respiratory delivery preparation is as follows: 2×10 8 -2×10 9 pcs / ml, normal saline; In this embodiment, the atomization concentration of the physiological saline is 1%.

[0035] In this embodiment, the cell exosomes are composed of adipose stem cell exosomes and bone marrow stem cell exosomes. The adipose stem cell exosomes are 1×10 8 -1×10 9 The number of exosomes of bone marrow stem cells is 1×108-1×109 / ml, and the ratio of exosomes of adipose stem cells to exosomes of bone marrow stem cells is 1:1.

[0036] The preparation method of this embodiment comprises the following steps: S1. Collect peripheral blood and separate mononuclear cells by Eicoll density gradient centrifugation; S2, inoculating the mononuclear cells into a culture bottle at a certain density and placing it in an incubator for culture; S3. Observe the cells in the incubator for attachment to the wall and then change the medium for the first time. Change the medium at intervals thereafter. When the cells are low-generation cells, adhere to the wall and expand the culture, and when the cell fusion rate reaches more than 90%, collect the cell exosomes. S4, washing the cell exosomes with an appropriate separation buffer, and then centrifuging the washed cell exosomes at low temperature; S5. Wash the exosomes obtained in S5 again using an appropriate eluent to remove residual cell debris, serum proteins, and washing buffer; S6, resuspending the exosome precipitate after washing again, and using a purification technique to separate and purify the polypeptide factors in the exosomes, wherein the purification technique includes density gradient centrifugation, ultracentrifugation combined with column chromatography; S7. The concentration of purified exosomes was determined by nanoparticle tracking analysis (NTA). S8. Mix the purified and tested exosomes with normal saline in proportion to obtain a respiratory delivery preparation.

[0037] In this embodiment, in S2, the culture temperature is 37°C, the carbon dioxide concentration is 5%, and the culture time is 12-24 hours.

[0038] In this embodiment, in S4, the separation buffer is a phosphate buffered saline solution containing 0.1% bovine serum albumin; preferably, the separation buffer is sterilized by filtering with a 0.22 micron filter membrane in advance.

[0039] In this embodiment, in S4, the centrifugation factor is 2500-3000g, the centrifugation temperature is 3-4°C, and the centrifugation time is 50-60min.

[0040] In this embodiment, in S5, the eluent is a glycine solution with a pH of 2.8 to 3.5 and a glycine concentration of 0.1 to 0.2 mol / L.

[0041] In this embodiment, in S8, the addition ratio of the exosomes to the physiological saline is 1:9.

[0042] In this embodiment, in S1-S8, each result obtained is tested, recorded and uploaded to the traceability system.

[0043] Embodiment 4: The difference between the first embodiment, the second embodiment and the third embodiment is that: The weight ratio of the respiratory delivery preparation is as follows: 2×10 8 -2×10 9 pcs / ml, normal saline; In this embodiment, the atomization concentration of the physiological saline is 3%.

[0044] In this embodiment, the cell exosomes are composed of adipose stem cell exosomes and bone marrow stem cell exosomes. The adipose stem cell exosomes are 1×10 8 -1×10 9 The number of exosomes of bone marrow stem cells is 1×108-1×109 / ml, and the ratio of exosomes of adipose stem cells to exosomes of bone marrow stem cells is 1:1.

[0045] The preparation method of this embodiment comprises the following steps: S1. Collect peripheral blood and separate mononuclear cells by Eicoll density gradient centrifugation; S2, inoculating the mononuclear cells into a culture bottle at a certain density and placing it in an incubator for culture; S3. Observe the cells in the incubator for attachment to the wall and then change the medium for the first time. Change the medium at intervals thereafter. When the cells are low-generation cells, adhere to the wall and expand the culture, and when the cell fusion rate reaches more than 90%, collect the cell exosomes. S4, washing the cell exosomes with an appropriate separation buffer, and then centrifuging the washed cell exosomes at low temperature; S5. Wash the exosomes obtained in S5 again using an appropriate eluent to remove residual cell debris, serum proteins, and washing buffer; S6, resuspending the exosome precipitate after washing again, and using a purification technique to separate and purify the polypeptide factors in the exosomes, wherein the purification technique includes density gradient centrifugation, ultracentrifugation combined with column chromatography; S7. The concentration of purified exosomes was determined by nanoparticle tracking analysis (NTA). S8. Mix the purified and tested exosomes with normal saline in proportion to obtain a respiratory delivery preparation.

[0046] In this embodiment, in S2, the culture temperature is 37°C, the carbon dioxide concentration is 5%, and the culture time is 12-24 hours.

[0047] In this embodiment, in S4, the separation buffer is a phosphate buffered saline solution containing 0.1% bovine serum albumin; preferably, the separation buffer is sterilized by filtering with a 0.22 micron filter membrane in advance.

[0048] In this embodiment, in S4, the centrifugation factor is 2500-3000g, the centrifugation temperature is 3-4°C, and the centrifugation time is 50-60min.

[0049] In this embodiment, in S5, the eluent is a glycine solution with a pH of 2.8 to 3.5 and a glycine concentration of 0.1 to 0.2 mol / L.

[0050] In this embodiment, in S8, the addition ratio of the exosomes to the physiological saline is 1:15.

[0051] In this embodiment, in S1-S8, each result obtained is tested, recorded and uploaded to the traceability system.

[0052] Embodiment five: The difference between the first embodiment, the second embodiment, the third embodiment and the fourth embodiment is that: The weight ratio of the respiratory delivery preparation is as follows: 2×10 8 -2×10 9 pcs / ml, normal saline; In this embodiment, the atomization concentration of the physiological saline is 3%.

[0053] In this embodiment, the cell exosomes are composed of adipose stem cell exosomes and bone marrow stem cell exosomes. The adipose stem cell exosomes are 1×10 8 -1×10 9The number of exosomes of bone marrow stem cells is 1×108-1×109 / ml, and the ratio of exosomes of adipose stem cells to exosomes of bone marrow stem cells is 1:1.

[0054] The preparation method of this embodiment comprises the following steps: S1. Collect peripheral blood and separate mononuclear cells by Eicoll density gradient centrifugation; S2, inoculating the mononuclear cells into a culture bottle at a certain density and placing it in an incubator for culture; S3. Observe the cells in the incubator for attachment to the wall and then change the medium for the first time. Change the medium at intervals thereafter. When the cells are low-generation cells, adhere to the wall and expand the culture, and when the cell fusion rate reaches more than 90%, collect the cell exosomes. S4, washing the cell exosomes with an appropriate separation buffer, and then centrifuging the washed cell exosomes at low temperature; S5. Wash the exosomes obtained in S5 again using an appropriate eluent to remove residual cell debris, serum proteins, and washing buffer; S6, resuspending the exosome precipitate after washing again, and using a purification technique to separate and purify the polypeptide factors in the exosomes, wherein the purification technique includes density gradient centrifugation, ultracentrifugation combined with column chromatography; S7. The concentration of purified exosomes was determined by nanoparticle tracking analysis (NTA). S8. Mix the purified and tested exosomes with normal saline in proportion to obtain a respiratory delivery preparation.

[0055] In this embodiment, in S2, the culture temperature is 37°C, the carbon dioxide concentration is 5%, and the culture time is 12-24 hours.

[0056] In this embodiment, in S4, the separation buffer is a phosphate buffered saline solution containing 0.1% bovine serum albumin; preferably, the separation buffer is sterilized by filtering with a 0.22 micron filter membrane in advance.

[0057] In this embodiment, in S4, the centrifugation factor is 2500-3000g, the centrifugation temperature is 3-4°C, and the centrifugation time is 50-60min.

[0058] In this embodiment, in S5, the eluent is a glycine solution with a pH of 2.8 to 3.5 and a glycine concentration of 0.1 to 0.2 mol / L.

[0059] In this embodiment, in S8, the addition ratio of the exosomes to the physiological saline is 1:5.

[0060] In this embodiment, in S1-S8, each result obtained is tested, recorded and uploaded to the traceability system.

[0061] In summary: the preparation method of a respiratory delivery preparation for major respiratory diseases of the present invention is that the respiratory delivery preparation obtained by the proportioning method of Example 1 can not only inhibit inflammation when acting on the respiratory system, but also improve the immune regulation ability, the preparation time is short, and it helps the establishment of capillaries, thereby potentially alleviating the symptoms and progression of asthma, acute lung injury and anti-pulmonary fibrosis. Increasing the atomization concentration of physiological saline to 5% for the respiratory delivery preparation obtained by the proportioning method of Example 2 will reduce the ability to inhibit inflammation, while the respiratory delivery preparation obtained by the proportioning method of Example 3, The atomization concentration of normal saline is reduced to 1%, and the ability to inhibit inflammation is improved, but it is not good for the health of the patient. The respiratory tract delivery preparation obtained by the ratio method of Example 4, the addition ratio of exosomes to normal saline is 1:15, which will lead to a decrease in the concentration of exosomes in the respiratory tract delivery preparation, thereby reducing the therapeutic effect of the respiratory tract delivery preparation. The respiratory tract delivery preparation obtained by the ratio method of Example 5, the addition ratio of exosomes to normal saline is 1:5, which will lead to an excessively high concentration of exosomes in the respiratory tract delivery preparation, which is not good for the establishment of the patient's own capillaries and will reduce the patient's immune regulation ability.

[0062] It should be noted that, in this article, relational terms such as first and second, etc. are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, the terms "include", "comprise" or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device.

[0063] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A method for preparing a respiratory delivery preparation for serious respiratory diseases, characterized in that: The weight ratio of the respiratory tract delivery preparation is as follows: 2×10 8 -2×10 9 / ml, normal saline.

2. The method for preparing a respiratory delivery preparation for serious respiratory diseases according to claim 1, characterized in that: The atomization concentration of the physiological saline is 3%.

3. The method for preparing a respiratory delivery preparation for serious respiratory diseases according to claim 1, characterized in that: The cell exosomes are composed of adipose stem cell exosomes and bone marrow stem cell exosomes. The adipose stem cell exosomes are 1×10 8 -1×10 9 The number of exosomes of bone marrow stem cells is 1×108-1×109 / ml, and the ratio of exosomes of adipose stem cells to exosomes of bone marrow stem cells is 1:

1.

4. The method for preparing a respiratory delivery preparation for serious respiratory diseases according to claim 1, characterized in that: The preparation method comprises the following steps: S1. Collect peripheral blood and separate mononuclear cells by Eicoll density gradient centrifugation; S2, inoculating the mononuclear cells into a culture bottle at a certain density and placing it in an incubator for culture; S3. Observe the cells in the incubator for attachment to the wall and then change the medium for the first time. Change the medium at intervals thereafter. When the cells are low-generation cells, adhere to the wall and expand the culture, and when the cell fusion rate reaches more than 90%, collect the cell exosomes. S4, washing the cell exosomes with an appropriate separation buffer, and then centrifuging the washed cell exosomes at low temperature; S5. Wash the exosomes obtained in S5 again using an appropriate eluent to remove residual cell debris, serum proteins, and washing buffer; S6, resuspending the exosome precipitate after washing again, and using a purification technique to separate and purify the polypeptide factors in the exosomes, wherein the purification technique includes density gradient centrifugation, ultracentrifugation combined with column chromatography; S7. The concentration of purified exosomes was determined by nanoparticle tracking analysis (NTA). S8. Mix the purified and tested exosomes with normal saline in proportion to obtain a respiratory delivery preparation.

5. The method for preparing a respiratory delivery preparation for serious respiratory diseases according to claim 1, characterized in that: In S2, the culture temperature is 37°C, the carbon dioxide concentration is 5%, and the culture time is 12-24 hours.

6. The method for preparing a respiratory delivery preparation for serious respiratory diseases according to claim 1, characterized in that: In S4, the separation buffer is a phosphate buffered saline solution containing 0.1% bovine serum albumin; preferably, the separation buffer is sterilized by filtering with a 0.22 micron filter membrane in advance.

7. The method for preparing a respiratory delivery preparation for serious respiratory diseases according to claim 1, characterized in that: In the S4, the centrifugation factor is 2500-3000g, the centrifugation temperature is 3-4°C, and the centrifugation time is 50-60min.

8. The method for preparing a respiratory delivery preparation for serious respiratory diseases according to claim 1, characterized in that: In the S5, the eluent is a glycine solution with a pH of 2.8 to 3.5 and a glycine concentration of 0.1 to 0.2 mol / L.

9. The method for preparing a respiratory delivery preparation for serious respiratory diseases according to claim 1, characterized in that: In the S8, the addition ratio of the exosomes to the physiological saline is 1:

9.

10. The method for preparing a respiratory delivery preparation for serious respiratory diseases according to claim 4, characterized in that: In S1-S8, each result obtained is tested, recorded and uploaded to the traceability system.