Mesenchymal stem cell preparation in-airway drug delivery system and application thereof

Abstract

The invention provides a mesenchymal stem cell preparation in-airway drug delivery system and application thereof. In particular to a mesenchymal stem cell preparation in-airway drug delivery system for treating idiopathic pulmonary fibrosis and application of the mesenchymal stem cell preparation in-airway drug delivery system. The drug delivery system can be used for clinical safe and accurate mesenchymal stem cell preparation drug delivery, and can enable FVC of an idiopathic pulmonary fibrosis patient to keep a growth trend for a long time; and the lung function of a patient can be continuously improved by single administration.

Description

Technical Field

[0001] The present invention relates to the field of medicine, and in particular to an intra-airway drug delivery system for a mesenchymal stem cell preparation and applications thereof. Background Art

[0002] Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive, fibrosing interstitial pneumonia of unknown etiology. It is primarily manifested by progressively worsening dyspnea, accompanied by restrictive ventilation dysfunction and gas exchange disorders, ultimately leading to hypoxemia and even respiratory failure. IPF has a poor prognosis, with a median survival of only 2 to 3 years after diagnosis. High-resolution chest CT (HRCT) of IPF demonstrates usual interstitial pneumonia (UIP), and the main pathological features of lung histology include diffuse alveolitis and pulmonary interstitial fibrosis. However, the lung damage caused by IPF is persistent and irreversible, and the lack of alveolar count ultimately leads to decreased or even loss of pulmonary ventilation function, which in turn causes hypoxia, decreased mobility, and even death in patients.

[0003] The treatment of IPF can be divided into two categories: drug therapy and non-drug therapy. The IPF treatment drugs pirfenidone and nintedanib have a certain effect on delaying the decline of lung function in mild to moderate patients, but it is difficult to improve lung function or reverse the course of the disease. This is because there is a lack of effective methods to regenerate and repair damaged alveolar structures, and it is impossible to truly prevent the occurrence of fibrosis. For IPF patients with severe lung dysfunction, whether taking the above two drugs is beneficial is still under study. Glucocorticoids combined with azathioprine and N-acetylcysteine ​​were once the "standard treatment" for IPF. Later, due to the lack of theoretical basis for improving pulmonary fibrosis, it could not delay the progression of the disease, but had many side effects or worsened existing comorbidities such as diabetes, cardiovascular and cerebrovascular diseases, and osteoporosis. In recent years, it is no longer recommended for stable IPF patients.

[0004] Non-drug treatments mainly include smoking cessation, oxygen therapy, mechanical ventilation, pulmonary rehabilitation, and lung transplantation. Lung transplantation is the only option for patients with end-stage IPF, which can improve the quality of life of IPF patients and increase the 5-year survival rate to 50% to 56%. Lung transplantation, like other organ transplants, has many limitations:

[0005] 1. The source of lung organs is limited, and the age and physiological conditions of the donor and recipient are very demanding, and the matching success rate is low;

[0006] 2. Compared with other solid organ transplants, the incidence of acute rejection confirmed by biopsy after lung transplantation is still as high as 80% one year after transplantation. Rejection is associated with chronic allograft lung dysfunction and bronchiolar obstructive syndrome, which is the main cause of death in adult lung transplant recipients.

[0007] 3. Postoperative complications such as infection, pain, wound bleeding and dehiscence also limit the effectiveness of lung transplantation;

[0008] 4. The operation time must be short, with warm ischemia of the ex vivo lung not exceeding 35 minutes and cold ischemia not exceeding 10 hours. The utilization rate of the donor lung must be only 20% of that of the donor kidney.

[0009] Although the number of lung transplants is increasing every year, no more than 1,000 lung transplants can be performed each year, which is seriously insufficient compared to the number of IPF patients who need lung transplants.

[0010] Therefore, limited by the shortcomings of existing drug and non-drug treatments, IPF patients urgently need treatments that can improve pulmonary fibrosis.

[0011] Mesenchymal stem cells (MSCs) are found in tissues such as bone marrow, fat, umbilical cord, placenta, and dental pulp. MSCs isolated from various sources possess self-renewal capacity, multipotential differentiation potential, and specific surface biomarkers. Under certain induction conditions, MSCs can differentiate into various cell types, such as chondrocytes, osteoblasts, adipocytes, and neuron-like cells.

[0012] Human umbilical cord mesenchymal stem cells (hUCMSCs) are a type of adult mesenchymal stem cell found in the umbilical cord's Wharton's jelly and perivascular tissue. They have physiological characteristics such as low immunogenicity, rapid self-replication and renewal, and strong multidirectional proliferation and differentiation capabilities. They participate in regulating cell proliferation and apoptosis through their unique cytokine secretion function, exerting immunomodulatory and anti-inflammatory effects. They are also easy to obtain and have no ethical controversy.

[0013] In nonclinical animal studies, existing MSCs have been shown to aid tissue regeneration in lung injury, repairing damaged lung sites, alleviating chronic airway inflammation, and restoring alveolar fluid balance. Several clinical trials using hUCMSCs have been conducted in lung diseases such as chronic obstructive pulmonary disease, asthma, and pulmonary fibrosis. Existing research has demonstrated that allogeneic mesenchymal stem cells (hMSCs) have good safety and tolerability in the treatment of IPF. Summary of the Invention

[0014] The purpose of the present invention is to provide an intra-airway drug delivery system for a mesenchymal stem cell preparation and its application in treating idiopathic pulmonary fibrosis.

[0015] In a first aspect, the present invention provides an intra-airway drug delivery system for a mesenchymal stem cell preparation for treating idiopathic pulmonary fibrosis, comprising:

[0016] (a) a first container containing a first preparation comprising mesenchymal stem cells and a pharmaceutically acceptable carrier;

[0017] (b) a bronchoscope injection module, comprising: an injection catheter, an injection pump, and a bronchoscope, wherein one end of the catheter contacts the first preparation in the first container, and the other end is used to deliver the preparation to the lungs; the injection pump is used to push the preparation in a quantitative manner; the bronchoscope lens is located near the end of the catheter that extends into the lungs and is used to capture images and / or identify the location;

[0018] (c) a high-resolution computed tomography (HRCT) imager, comprising a tomography module and a data processing module, wherein the tomography module is configured to scan the subject's lungs to obtain lung images capable of displaying pulmonary fibrosis, and the data processing module is configured to calculate, based on the obtained lung images, the total volume of each lung lobe of the subject and the volume and / or proportion of fibrotic tissue;

[0019] (d) a control module, configured to calculate the injection volume at each injection site based on the total volume of each lung lobe and the volume and / or proportion of fibrotic tissue of the subject calculated by the data processing module of the high-resolution computed tomography scanner (input), and control the bronchoscope injection module to inject a corresponding volume of the first agent into each injection site (output);

[0020] Wherein, the total volume of the first preparation is 10 mL to 50 mL.

[0021] In another preferred embodiment, in the data processing module, when the high-density attenuation value of lung tissue is set to be ≥-700HU (preferably -600 to -250HU), it is included in the calculation of the fibrotic tissue volume.

[0022] In another preferred embodiment, the total volume of the first preparation is 35 mL to 45 mL, preferably 40±2 mL.

[0023] In another preferred example, the data processing module of the high-resolution computed tomography scanner is configured to calculate the volume of the subject's left lung fibrosis tissue and the total volume of the left lung based on the obtained lung image, and calculate the ratio P1 of the volume of the left lung fibrosis tissue to the total volume of the left lung; accordingly, the volume of the subject's right lung fibrosis tissue and the total volume of the right lung are calculated based on the obtained lung image, and calculate the ratio P2 of the volume of the right lung fibrosis tissue to the total volume of the right lung; then, a proportional coefficient is obtained based on the ratio of the volume of the left lung fibrosis tissue to the total volume of the left lung / the ratio of the volume of the right lung fibrosis tissue to the total volume of the right lung (P1 / P2), and the proportional coefficient is input into the control module.

[0024] In another preferred example, the control module is configured to control the bronchoscope injection module to inject 50% volume of the first preparation into the bronchi of the left lung and the right lung, respectively, when the proportional coefficient calculated by the high-resolution computed tomography imager is less than a preset value k; and to control the bronchoscope injection module to inject 40±2% volume of the first preparation into the left bronchus and 60±2% volume of the first preparation into the right bronchus (the sum of the injection volumes of the left and right lungs is 100% of the first preparation), when the proportional coefficient calculated by the high-resolution computed tomography imager is greater than or equal to the preset value k.

[0025] In another preferred embodiment, the injection into the left lung and the right lung is administered into the left lower lobe and the right lower lobe of the subject, respectively.

[0026] In another preferred embodiment, the injection site is the root of the secondary bronchus or tertiary bronchus of the left lower lobe and the right lower lobe, and in particular, the drug is evenly administered to the secondary bronchus or tertiary bronchus of each lung lobe.

[0027] In another preferred example, when injecting the first agent into the lungs, the control module is configured to control the bronchoscope injection module to inject the first preparation into the bronchi of the dorsal segment, anterior medial basal segment, lateral basal segment and posterior basal segment of the left lower lobe, and the dorsal segment, medial basal segment, anterior basal segment, lateral basal segment and posterior basal segment of the right lower lobe, wherein the administration volume of the anterior medial basal segment of the left lower lobe is 2 / 5 of the administration volume of the left lung, and the administration volume of other lung segments is 1 / 5 of the administration volume of the corresponding lung lobes.

[0028] In another preferred embodiment, the bronchoscope injection module further includes a display module, and the display module is used to display images captured by the bronchoscope, such as a display screen.

[0029] In another preferred embodiment, one end of the conduit may have a dripper.

[0030] In another preferred embodiment, the injection pump is a peristaltic pump or a dosage pump, and the peristaltic pump or the dosage pump is configured to pump the first preparation.

[0031] In another preferred embodiment, the bronchoscope includes a flexible bronchoscope, preferably an electronic fiber bronchoscope.

[0032] In another preferred embodiment, the mesenchymal stem cells are derived from umbilical cord, bone marrow, fat, placental tissue, or a combination thereof, and are preferably human umbilical cord mesenchymal stem cells.

[0033] In another preferred embodiment, a single dose of the first preparation contains 3×10 7 ~7×10 7 Mesenchymal stem cells, preferably 6±0.5×10 7 cells.

[0034] In another preferred example, the compound electrolyte injection comprises: sodium chloride 5.26±0.2 g / L; sodium gluconate 5.02±0.2 g / L; sodium acetate 3.68±0.2 g / L; potassium chloride 0.37±0.02 g / L; magnesium chloride 0.30±0.02 g / L, the solvent is water, and the pH is 7±0.5.

[0035] In another preferred embodiment, the concentration of mesenchymal stem cells in the first preparation is 6×10 5 ~6×10 6 cells / mL, preferably 1×10 6 ~3×10 6 cells / mL, more preferably 1.5×10 6 ~2×10 6 cells / mL.

[0036] In another preferred embodiment, the first preparation comprises: mesenchymal stem cells, human serum albumin and compound electrolyte injection.

[0037] In another preferred embodiment, the concentration of human serum albumin in the first preparation is 0.5-2 wt %, preferably 0.8-2 wt %, and more preferably 1-1.8 wt %. In another preferred embodiment, the subject is a human or non-human mammal, such as a rat, mouse, New Zealand rabbit, or cynomolgus monkey.

[0038] In another preferred embodiment, the subject suffers from idiopathic pulmonary fibrosis.

[0039] In another preferred embodiment, the subject may also suffer from a disease selected from the group consisting of: hyperlipidemia, thyroid nodules, pulmonary bullae, emphysema, hypertension, coronary heart disease, arrhythmia, cerebral infarction, type 2 diabetes, or a combination thereof.

[0040] In another preferred embodiment, the subject receives the intra-airway administration of the present invention only once every 24 to 48 weeks.

[0041] In another preferred embodiment, the subject receives intratracheal administration of the present invention once every 24 weeks, with a dosage of 3×10 7 ~4×10 7 Mesenchymal stem cells.

[0042] In another preferred embodiment, the subject receives intratracheal administration of the present invention once every 48 weeks, with a dosage of 6×10 7 ~7×10 7 Mesenchymal stem cells.

[0043] In another preferred embodiment, the administration of the present invention is non-invasive administration.

[0044] In another preferred embodiment, the "injection" is administration by injection into the bronchus.

[0045] In another preferred embodiment, the first preparation is a liquid preparation, usually a solution. Preferably, the preparation is non-irritating to the airways and lungs.

[0046] In another preferred embodiment, the catheter is passed through the patient's mouth, and one end for injecting the drug is delivered to the bronchus.

[0047] In another preferred embodiment, the pH value of the first preparation is 7-7.4, preferably 7.1-7.2.

[0048] In another preferred embodiment, the osmotic pressure of the first preparation is 310-340 mOsmol / Kg, preferably 330±5 mOsmol / Kg.

[0049] In a second aspect, the present invention provides use of the drug delivery system according to the first aspect of the present invention in preparing a medical device for treating idiopathic pulmonary fibrosis.

[0050] In a third aspect, the present invention provides a method for treating idiopathic pulmonary fibrosis in a subject, comprising the steps of: (i) scanning the subject's lungs using a high-resolution computed tomography (HRCT) imager to obtain a lung image capable of demonstrating pulmonary fibrosis;

[0051] (ii) calculating the volume of the fibrotic tissue in the left lung and the total volume of the left lung of the subject based on the obtained lung image, and calculating the ratio P1 of the volume of the fibrotic tissue in the left lung to the total volume of the left lung; correspondingly, calculating the volume of the fibrotic tissue in the right lung and the total volume of the right lung of the subject based on the obtained lung image, and calculating the ratio P2 of the volume of the fibrotic tissue in the right lung to the total volume of the right lung; and then obtaining a proportional coefficient based on the ratio of the volume of the fibrotic tissue in the left lung to the total volume of the left lung / the ratio of the volume of the fibrotic tissue in the right lung to the total volume of the right lung (P1 / P2);

[0052] (iii) when the preset value of the proportional coefficient is less than k, 50% of the volume of the first preparation is injected into the bronchi of the left lung and the right lung, respectively; when the proportional coefficient is greater than or equal to the preset value k, 40±2% of the volume of the first preparation is injected into the left lung bronchi, and 60±2% of the volume of the first preparation is injected into the right lung bronchi (the sum of the injection volumes of the left and right lungs is 100%); wherein the preset value k is 1.0-1.25;

[0053] Wherein, the first preparation contains mesenchymal stem cells and a pharmaceutically acceptable carrier, with a total volume of 10 mL to 50 mL;

[0054] A single dose of the first formulation contains 3×10 7 ~7×10 7 Mesenchymal stem cells.

[0055] In another preferred embodiment, the injection is administered to the left lower lobe bronchus and the right lower lobe bronchus of the subject respectively.

[0056] In another preferred embodiment, the injection site is the secondary bronchus or tertiary bronchus of each lung lobe, especially the root of the bronchus.

[0057] In another preferred embodiment, the injection is administered to the dorsal segment, inner anterior basal segment, outer basal segment and posterior basal segment of the left lower lobe, and the dorsal segment, inner basal segment, anterior basal segment, outer basal segment and posterior basal segment of the right lower lobe through the corresponding secondary or tertiary bronchial roots, wherein the administration volume of the inner anterior basal segment of the left lower lobe is 2 / 5 of the administration volume of the left lung, and the administration volume of other lung segments is 1 / 5 of the administration volume of the corresponding lung lobe.

[0058] In another preferred embodiment, the mesenchymal stem cells are derived from umbilical cord, bone marrow, fat, placental tissue, or a combination thereof, and are preferably human umbilical cord mesenchymal stem cells.

[0059] In another preferred embodiment, the mesenchymal stem cells are derived from the subject itself or are allogeneic.

[0060] In another preferred embodiment, a single dose of the first preparation contains 3×10 7 ~7×107 Mesenchymal stem cells, preferably 6±0.5×10 7 cells.

[0061] In another preferred embodiment, the pharmaceutically acceptable carrier is selected from the following group: physiological saline and compound electrolyte injection.

[0062] In another preferred embodiment, the administration is by injection through a bronchoscope catheter.

[0063] In another preferred embodiment, the administration is performed through the above-mentioned bronchoscope injection module.

[0064] In another preferred embodiment, the administration is via the above-mentioned administration system.

[0065] In another preferred example, the compound electrolyte injection comprises: sodium chloride 5.26±0.2 g / L; sodium gluconate 5.02±0.2 g / L; sodium acetate 3.68±0.2 g / L; potassium chloride 0.37±0.02 g / L; magnesium chloride 0.30±0.02 g / L, the solvent is water, and the pH is 7±0.5.

[0066] In another preferred embodiment, the concentration of mesenchymal stem cells in the first preparation is 6×10 5 ~6×10 6 cells / mL, preferably 1×10 6 ~3×10 6 cells / mL, more preferably 1.5×10 6 ~2×10 6 cells / mL.

[0067] In another preferred embodiment, the subject is a human or non-human mammal, such as a rat, a mouse, a New Zealand rabbit, or a cynomolgus monkey.

[0068] In another preferred embodiment, the subject suffers from idiopathic pulmonary fibrosis.

[0069] In another preferred embodiment, the subject may also suffer from a disease selected from the group consisting of: hyperlipidemia, thyroid nodules, pulmonary bullae, emphysema, hypertension, coronary heart disease, arrhythmia, cerebral infarction, type 2 diabetes, or a combination thereof.

[0070] In another preferred embodiment, the subject receives the intra-airway administration of the present invention only once every 24 weeks to every 48 weeks.

[0071] In another preferred embodiment, the subject receives intratracheal administration of the present invention once every 24 weeks, with a dosage of 3×10 7 ~4×10 7 Mesenchymal stem cells.

[0072] In another preferred embodiment, the subject receives intratracheal administration of the present invention once every 48 weeks, with a dosage of 6×10 7 ~7×10 7 Mesenchymal stem cells.

[0073] In another preferred embodiment, the "injection" is infusion / drip into the bronchus (non-invasive) administration.

[0074] In another preferred embodiment, the administration of the present invention is non-invasive administration.

[0075] In another preferred embodiment, each step or condition of the present invention can be independently the step or condition described in the embodiments.

[0076] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features described in detail below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be listed here one by one. BRIEF DESCRIPTION OF THE DRAWINGS

[0077] Figure 1 Middle a is a morphological diagram of mesenchymal stem cells; b is the front view of the lung segment.

[0078] Figure 2 It shows that umbilical cord mesenchymal stem cells can be induced to differentiate into a variety of cell types.

[0079] Figure 3 Flow cytometry of umbilical cord mesenchymal stem cells.

[0080] Figure 4 The figure shows the changes in FVC after 24 and 48 weeks of treatment in the low-dose and high-dose groups. * indicates a significant difference between the two groups, and △ indicates a significant difference between the baseline and the baseline in either group between the observation periods after dosing, indicating statistically significant differences within the groups.

[0081] Figure 5 Disease progression in subjects in the low-dose and high-dose groups after 24 and 48 weeks of treatment is shown.

[0082] Figure 6 CT images of subjects in the low-dose group.

[0083] Figure 7 CT images of subjects in the high-dose group. DETAILED DESCRIPTION

[0084] After extensive and in-depth research, screening, and testing, the present inventors have developed a mesenchymal stem cell preparation intra-airway delivery system and its use in treating idiopathic pulmonary fibrosis. Surprisingly, the delivery system and method of the present invention can significantly improve clinical efficacy indicators in patients. This is the basis for the completion of the present invention.

[0085] the term

[0086] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0087] As used herein, the terms "comprising" or "including" may be open, semi-closed, or closed. In other words, the terms also include "consisting essentially of" or "consisting of."

[0088] Administration of cell preparations

[0089] Existing research using MSCs for IPF treatment has mostly used intravenous infusion. Because MSCs are slightly larger than pulmonary capillaries, intravenously administered MSCs first return to the right side of the heart with venous blood, then enter the pulmonary circulation. They briefly linger in the lungs, then gradually return to the left side of the heart after about 6-8 hours, passing through the lungs, and then enter the systemic circulation. MSCs that enter the systemic circulation gradually home to the liver and spleen, rarely homing to the lungs.

[0090] When MSCs remain in the pulmonary capillaries, they are trapped by their large diameter and are unrelated to the chemokine-induced MSC homing. IPF lesions occur on the alveolar wall near the alveolar cavity, separated from the pulmonary capillaries by thickened interstitial tissue. MSCs are trapped within these vessels, unable to penetrate the capillary wall and interstitial tissue layer, and unable to reach the affected area to exert their biological effects.

[0091] Clinical studies have attempted to treat radiation-induced pulmonary fibrosis by intrapulmonary instillation of MSCs through bronchoscopy. Imaging changes were observed, but there was no difference in lung function. This is because radiation-induced pulmonary fibrosis is usually a local lung tissue change caused by radiotherapy, which has little impact on overall lung function. In addition, radiation-induced pulmonary fibrosis may heal on its own, which is fundamentally different from the situation in IPF patients.

[0092] MSC preparations utilize autologous cells, which are then harvested, cultured, and then reinfused to avoid rejection. However, the patient's health must be considered when procuring these cells. The average age of IPF patients is over 60, making autologous cell collection difficult, as autologous cells function declines with age. Failure in autologous cell preparation is also a potential risk, and secondary cell collection after such failures increases the burden on patients. Furthermore, the preparation process can be lengthy, potentially leaving patients at a late stage of treatment.

[0093] Currently, existing technologies lack clinical-grade MSC preparation standards and standard operating procedures for the application process, and the main clinical efficacy indicators have failed to achieve significant results.

[0094] Mesenchymal stem cell preparations

[0095] The mesenchymal stem cell preparation or first preparation of the present invention refers to a liquid preparation containing mesenchymal stem cells as therapeutic cells.

[0096] Preferably, the mesenchymal stem cells are derived from umbilical cord, bone marrow, adipose tissue, placental tissue, or a combination thereof, preferably human umbilical cord mesenchymal stem cells. Preferably, the mesenchymal stem cells are derived from the subject itself or allogeneic.

[0097] Typically, in the preparation, mesenchymal stem cells are dispersed in a medium suitable for pulmonary administration, as long as the medium is non-irritating to the lungs, does not destroy cell activity, and is easy to disperse and administer, such as physiological saline, compound electrolyte injection, etc.

[0098] Since mesenchymal stem cells are difficult to obtain, it is necessary to pay attention to the amount of mesenchymal stem cells used to save cells. According to the research of the present invention, for a single dose, the preferred number of cells is 3×10 7 ~7×10 7 mesenchymal stem cells, more preferably 6±0.5×10 7 cells. For example, 3×10 7 ~4×10 7 The therapeutic effect was able to delay the progression of pulmonary fibrosis to 24 weeks at a low dose of 6±0.5×10 7 High and low doses of individual cells can significantly change the development trend of pulmonary fibrosis, and the improvement effect can last up to 48 weeks. High dose administration is preferred.

[0099] In addition, since accumulation of liquid in the lungs during intrapulmonary administration may cause breathing difficulties, the volume of pulmonary liquid preparation administration should be taken seriously to ensure safety. Therefore, in the present invention, the single administration volume is limited to a safe range of 10 mL to 50 mL.

[0100] Drug delivery system

[0101] The present invention provides an intra-airway drug delivery system for a mesenchymal stem cell preparation for treating idiopathic pulmonary fibrosis, comprising:

[0102] (a) a first container containing a first preparation comprising mesenchymal stem cells and a pharmaceutically acceptable carrier;

[0103] (b) a bronchoscope injection module, comprising: an injection catheter, an injection pump, and a bronchoscope, wherein one end of the catheter contacts the first preparation in the first container, and the other end is used to deliver the preparation to the lungs; the injection pump is used to push the preparation in a quantitative manner; the bronchoscope lens is located near the end of the catheter that extends into the lungs and is used to capture images and / or identify the location;

[0104] (c) a high-resolution computed tomography (HRCT) imager, comprising a tomography module and a data processing module, wherein the tomography module is configured to scan the subject's lungs to obtain lung images capable of displaying pulmonary fibrosis, and the data processing module is configured to calculate, based on the obtained lung images, the total volume of each lung lobe of the subject and the volume and / or proportion of fibrotic tissue;

[0105] (d) a control module, configured to calculate the injection volume at each injection site based on the total volume of each lung lobe and the volume and / or proportion of fibrotic tissue of the subject calculated by the data processing module of the high-resolution computed tomography scanner (input), and control the bronchoscope injection module to inject a corresponding volume of the first agent into each injection site (output);

[0106] Wherein, the total volume of the first preparation is 10 mL to 50 mL.

[0107] In another preferred embodiment, in the data processing module, when the high-density attenuation value of lung tissue is set to be ≥-700HU (preferably -650 to -200HU or -600 to -250HU), it is included in the calculation of the fibrotic tissue volume.

[0108] In another preferred embodiment, the total volume of the first preparation is 35 mL to 45 mL, preferably 40±2 mL.

[0109] In another preferred example, the data processing module of the high-resolution computed tomography scanner is configured to calculate the volume of the subject's left lung fibrosis tissue and the total volume of the left lung based on the obtained lung image, and calculate the ratio P1 of the volume of the left lung fibrosis tissue to the total volume of the left lung; accordingly, the volume of the subject's right lung fibrosis tissue and the total volume of the right lung are calculated based on the obtained lung image, and calculate the ratio P2 of the volume of the right lung fibrosis tissue to the total volume of the right lung; then, a proportional coefficient is obtained based on the ratio of the volume of the left lung fibrosis tissue to the total volume of the left lung / the ratio of the volume of the right lung fibrosis tissue to the total volume of the right lung (P1 / P2), and the proportional coefficient is input into the control module.

[0110] In another preferred example, the control module is configured to control the bronchoscope injection module to inject 50% volume of the first preparation into the bronchi of the left lung and the right lung, respectively, when the proportional coefficient calculated by the high-resolution computed tomography imager is less than a preset value k; and to control the bronchoscope injection module to inject 40±2% volume of the first preparation into the left bronchus and 60±2% volume of the first preparation into the right bronchus (the sum of the injection volumes of the left and right lungs is 100% of the first preparation, excluding normal losses), when the proportional coefficient calculated by the high-resolution computed tomography imager is greater than or equal to the preset value k.

[0111] In another preferred embodiment, the injection into the left lung and the right lung is administered into the left lower lobe and the right lower lobe of the subject, respectively.

[0112] In another preferred embodiment, the injection site is the root of the secondary bronchus or tertiary bronchus of the left lower lobe and the right lower lobe, and in particular, the drug is evenly administered to the secondary bronchus or tertiary bronchus of each lung lobe.

[0113] In another preferred example, when injecting the first agent into the lungs, the control module is configured to control the bronchoscope injection module to inject the first preparation into the dorsal segment, anterior medial basal segment, lateral basal segment and posterior basal segment of the left lower lobe, and the dorsal segment, medial basal segment, anterior basal segment, lateral basal segment and posterior basal segment of the right lower lobe, wherein the administration volume of the anterior medial basal segment of the left lower lobe is 2 / 5 of the administration volume of the left lung (because the anterior medial basal segment shares a common trunk, different lung segments are actually controlled and administered as two lung segments), and the administration volume of other lung segments is 1 / 5 of the administration volume of the corresponding lung lobes.

[0114] In another preferred embodiment, the bronchoscope injection module further includes a display module, and the display module is used to display images captured by the bronchoscope, such as a display screen.

[0115] In another preferred embodiment, one end of the conduit may have a dripper.

[0116] In another preferred embodiment, the injection pump is a peristaltic pump or a dosage pump, and the peristaltic pump or the dosage pump is configured to pump the first preparation.

[0117] In another preferred embodiment, the bronchoscope includes a flexible bronchoscope, preferably an electronic fiber bronchoscope.

[0118] The modules of the system of the present invention are electrically connected, mechanically connected or pipe-connected as needed to achieve corresponding functions.

[0119] use

[0120] The present invention also provides use of the drug delivery system of the present invention in preparing a medical device for treating idiopathic pulmonary fibrosis.

[0121] and a method for treating idiopathic pulmonary fibrosis in a subject, comprising the steps of: (i) scanning the subject's lungs using a high-resolution computed tomography (HRCT) imager to obtain a lung image capable of demonstrating pulmonary fibrosis;

[0122] (ii) calculating the volume of the fibrotic tissue in the left lung and the total volume of the left lung of the subject based on the obtained lung image, and calculating the ratio P1 of the volume of the fibrotic tissue in the left lung to the total volume of the left lung; correspondingly, calculating the volume of the fibrotic tissue in the right lung and the total volume of the right lung of the subject based on the obtained lung image, and calculating the ratio P2 of the volume of the fibrotic tissue in the right lung to the total volume of the right lung; and then obtaining a proportional coefficient based on the ratio of the volume of the fibrotic tissue in the left lung to the total volume of the left lung / the ratio of the volume of the fibrotic tissue in the right lung to the total volume of the right lung (P1 / P2);

[0123] (iii) when the preset value of the proportional coefficient is less than k, 50% of the volume of the first preparation is injected into the bronchi of the left lung and the right lung, respectively; when the proportional coefficient is greater than or equal to the preset value k, 40±2% of the volume of the first preparation is injected into the left lung bronchi, and 60±2% of the volume of the first preparation is injected into the right lung bronchi (the sum of the injection volumes of the left and right lungs is 100%); wherein the preset value k is 1.0-1.25;

[0124] Wherein, the first preparation contains mesenchymal stem cells and a pharmaceutically acceptable carrier, with a total volume of 10 mL to 50 mL;

[0125] A single dose of the first formulation contains 3×10 7 ~7×10 7 Mesenchymal stem cells.

[0126] The methods of the present invention may be administered with or without the administration system of the present invention.

[0127] In another preferred embodiment, the subject is a human or non-human mammal, such as a rat, a mouse, a New Zealand rabbit, or a cynomolgus monkey.

[0128] In another preferred embodiment, the subject suffers from idiopathic pulmonary fibrosis.

[0129] In another preferred embodiment, the subject may also suffer from a disease selected from the group consisting of: hyperlipidemia, thyroid nodules, pulmonary bullae, emphysema, hypertension, coronary heart disease, arrhythmia, cerebral infarction, type 2 diabetes, or a combination thereof. Experiments have shown that common diseases do not significantly affect the administration and treatment of the present invention.

[0130] It should be emphasized that, in the drug delivery system or method of the present invention, the "injection" should be understood as non-invasive administration by infusion / drip into the bronchus.

[0131] The main advantages of the present invention include:

[0132] The present invention provides a safe, precise, and universal mesenchymal stem cell preparation delivery system and method that can be used clinically, and can be used to treat idiopathic pulmonary fibrosis through bronchoscopic intrapulmonary injection.

[0133] The drug delivery system and drug delivery method of the present invention can maintain a long-term growth trend in the FVC of patients with idiopathic pulmonary fibrosis, and a single administration can help patients continuously improve their lung function.

[0134] The present invention provides a drug use operation process for treating idiopathic pulmonary fibrosis, dosage range selection, and significant long-term clinical effect.

[0135] The present invention will be further described below in conjunction with specific implementation. It should be understood that these examples are intended to illustrate the present invention only and are not intended to limit the scope of the present invention. The experimental methods in the following examples, for which specific conditions are not specified, are generally based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight.

[0136] Example 1

[0137] 1. Preparation of umbilical cord mesenchymal stem cells:

[0138] The umbilical cord after delivery was collected aseptically and stored in DMEM containing 1% double-antibody at 4°C. Mesenchymal stem cells were isolated and prepared within 48 hours. The umbilical cord was removed in a biosafety cabinet and rinsed several times with 1% double-antibody + PBS. After removing the blood vessels, the umbilical cord or fat was cut into 1-3 mm using sterile scissors. 3 Add a small amount of culture medium to the tissue fragments and mix well. Use a sterile plastic dropper to draw an appropriate amount of the mixture with tissue fragments into a 10cm sterile culture dish. Place the inoculated culture dish in a saturated humidity, 37℃, 5% CO2 incubator and culture. Change half of the medium every two days until the cell confluence reaches 60%. Discard the tissue fragments and change the medium with the full amount until the cell confluence reaches 80%. Add digestive enzyme to digest the cells. After digestion, collect the cell pellet by centrifugation and centrifuge at 8000 cells / cm 2 The cells were inoculated at a high density and cultured for 3 to 4 days. When the confluence reached 80% to 90%, the cells were harvested and continued to be passaged and expanded to the required cell number.

[0139] 2. Preparation of Mesenchymal Stem Cell Preparations:

[0140] The harvested cells were suspended in DMEM and filtered through a filter. The filtered cell suspension was centrifuged to obtain a cell pellet. The cell pellet was suspended in a cell preservation solution containing 5% dimethyl sulfoxide (DMSO), 35% human serum albumin, and 60% compound electrolyte solution. The cell preservation concentration was 1.0×10 6 ~1.0×10 7 The cell suspension is then filled into cell freezing bags, with each bag containing 10ml to 30ml of storage volume. The filled bags are heat-sealed. Finally, the cell preparation is placed in a programmed cooling device and frozen according to the freezing procedure. After freezing, the frozen cell preparation is transferred to liquid nitrogen (-196°C to -160°C) for storage.

[0141] 3. Mesenchymal stem cell quality standards

[0142] Mesenchymal stem cells adhere to the surface of the cell, growing in long, spindle-shaped spirals. They can differentiate into osteoblasts, adipocytes, and chondrocytes under specific conditions. Cellular immunophenotype: CD14, CD19, CD34, CD45, and HLA-DR are all ≤ 2.0%; CD73, CD90, CD105, and CD166 are all ≥ 95.0%. Free of mycoplasma, fungal, and bacterial microbial impurities.

[0143] 3.1 Cell morphology:

[0144] Mesenchymal stem cells grow adherently and in a long spindle-shaped spiral. Figure 1 As shown in a.

[0145] 3.2 Induced differentiation:

[0146] Mesenchymal stem cells can differentiate into adipogenic, osteogenic, and chondrogenic cells, such as Figure 2 shown.

[0147] 3.3 Mesenchymal stem cell surface markers:

[0148] Mesenchymal stem cell surface markers CD73, CD90, CD105 ≥ 95%, CD14, CD19, CD34, CD45, HLA-DR ≤ 2%, such as Figure 3 shown.

[0149] 3.4 Impurity detection:

[0150] The cells and preparations were tested for sterility using the pharmacopoeia method (membrane filtration method), and no bacteria or fungi were detected; the cells and preparations were tested for mycoplasma using the pharmacopoeia method (culture method and indicator cell culture method), and the results were all negative; the cells and preparations were tested for bacterial endotoxins using the pharmacopoeia method (gel method), and the results were all <0.5EU / mL, which met the requirements.

[0151] Example 2 Human umbilical cord mesenchymal stem cell preparation for the treatment of idiopathic pulmonary fibrosis

[0152] 2.1 Subject Screening

[0153] Subjects were selected if they were diagnosed with IPF according to the 2018 diagnostic guidelines for idiopathic pulmonary fibrosis published by the American Thoracic Society (ATS) in collaboration with the European Respiratory Society (ERS), the Japanese Respiratory Society (JRS), and the Latin American Thoracic Association (ALAT). The preferred age range was 50 to 70 years. Patients had a diffusion capacity of the lung for carbon monoxide (DLCO) of 30% to 79% of the predicted value (corrected for Hb), a forced vital capacity (FVC) ratio of ≥50% of the predicted value, and were classified as mild (measured / predicted FEV1 ≥70%) or moderate (measured / predicted FEV1 ≥60% and <70%) IPF based on the measured / predicted forced expiratory volume in first second (FEV1).

[0154] Exclusion criteria for recruiting subjects;

[0155] 1) Subjects with a history of mechanical ventilation or concurrent infectious pneumonia or asthma within one month before screening;

[0156] 2) Patients with malignant tumors within 5 years before screening;

[0157] 3) Those with a known history of immune system disease (such as thymic disease, systemic lupus erythematosus);

[0158] 4) Those who are allergic to human albumin, anesthetic drugs or their components;

[0159] 5) Subjects who participated in any other clinical trials within 3 months before screening;

[0160] 6) Subjects who cannot tolerate bronchoscopy;

[0161] 7) Patients judged by the researchers to have a higher risk of general anesthesia / local anesthesia;

[0162] Subjects with idiopathic pulmonary fibrosis may also suffer from hyperlipidemia, thyroid nodules, pulmonary bullae, emphysema, hypertension, coronary heart disease, arrhythmia, cerebral infarction, and type 2 diabetes.

[0163] The subjects were divided into two groups, three in each group, and received different doses of human umbilical cord mesenchymal stem cells, with low and high doses. The low dose was 3×10 7 cells per person, and the high dose is 6×10 7 Each subject received only one dose.

[0164] 1) Preparation transportation

[0165] Mesenchymal stem cell medications should be delivered to the clinical research center on the day of dosing. If administration is premature, the medications may be delivered to the research center for temporary storage 24 hours prior to resuscitation and incorporation. During this temporary storage period, the mesenchymal stem cell medications must be stored in a liquid nitrogen tank at a temperature not exceeding -160°C.

[0166] 2) Resuscitation and compatibility

[0167] Prior to administration, mesenchymal stem cell drugs stored in liquid nitrogen need to be resuscitated and formulated. The packaging bag that comes into direct contact with the mesenchymal stem cell drug can be placed directly in a water bath set at 37°C. Once the contents are completely thawed and liquidized, the entire resuscitation process takes no more than one minute.

[0168] The mesenchymal stem cell preparation is prepared and diluted with compound electrolyte injection (1000ml contains: 5.26g sodium chloride; 5.02g sodium gluconate; 3.68g sodium acetate; 0.37g potassium chloride; 0.30g magnesium chloride, in water, pH 7.0±1.0). Use a dispensing syringe to withdraw the required volume of medication from the drug package, then withdraw compound electrolyte injection to a fixed volume of 40mL. Specifically, the low-dose group requires first withdrawing 5mL of mesenchymal stem cell preparation, followed by 35mL of compound electrolyte injection, while the high-dose group requires first withdrawing 10mL of mesenchymal stem cell preparation, followed by 30mL of compound electrolyte injection. The final volume after preparation is 40mL for both the low-dose and high-dose groups. After withdrawing the compound electrolyte injection, gently shake the syringe horizontally to thoroughly mix the mesenchymal stem cell preparation and compound electrolyte injection.

[0169] The ambient temperature during the preparation process was maintained below 26°C, and the time from the start of preparation to the end of administration did not exceed 6 hours.

[0170] The rationale for diluting to a fixed 40 mL volume is as follows: ① The human albumin in the preparation has a viscous consistency. If injected directly into the airway without dilution, it will mix with the air in the airway under respiratory action to form a highly viscous foam, which hinders drug dispersion. ② Undiluted human albumin can affect the viability of stem cells in the lungs. A preparation containing 20% ​​human albumin has a filling volume of 35%. When a 10 mL preparation is diluted to 40 mL, the human albumin content is 1.75% (20% × 35% × 25%). When a 5 mL preparation is diluted to 40 mL, the human albumin content is 0.875% (20% × 35% × 12.5%). Human albumin concentrations of approximately 1% and 2% both have a protective effect on stem cell survival and biological activity, and the protective effect is similar within 3 hours. ③ There is still a lack of accurate clinical experience regarding the maximum tolerated volume for intrapulmonary administration. Excessive fluid retention in the bronchi and alveolar cavities may cause a certain degree of dyspnea. Preclinical animal studies used healthy cynomolgus macaques, with a safe dosing volume of 1 mL / kg. Considering the lung atrophy and reduced peak expiratory volume in patients with idiopathic pulmonary fibrosis, and considering the inclusion criterion of a measured / predicted forced vital capacity ≥ 50%, the clinical dosing volume was set at 50% of the preclinical study volume. Based on the results of a subgroup analysis (Japanese population) in the nintedanib study of idiopathic pulmonary fibrosis, with a mean body weight of 60.2 kg and a standard deviation of 10.0 kg, the upper limit of the body weight covering 95% of the population was approximately 80 kg. Taking all these factors into consideration, the dosing volume was 80 kg x 1 mL / kg x 50%, or 40 mL. The dosing volume range is 10 mL to 50 mL.

[0171] 3) Medication process

[0172] After evenly mixing and matching, the preparation is connected to the bronchoscope injection catheter port, and the mesenchymal stem cell preparation is pushed to the designated lung lobe at the root of the secondary bronchus of each lung lobe through the bronchoscope. The lungs are divided into left and right sides. The left lung has two lobes, the upper and lower lobes, and the right lung has three lobes, the upper, middle, and lower lobes. The left lung is smaller than the right lung. Idiopathic pulmonary fibrosis lesions are more common in the lower lobes of both lungs, and occasionally in the middle and upper lobes. The left lower lobe is divided into the dorsal segment, the anterior medial basal segment, the lateral basal segment, and the posterior basal segment. The right lower lobe is divided into the dorsal segment, the medial basal segment, the anterior basal segment, the lateral basal segment, and the posterior basal segment (the front view of the lung segments is as follows Figure 1 (As shown in b). The anteromedial basal segment of the left lower lobe shared a common trunk, effectively controlling different lung segments and administering the drug as two separate segments. Therefore, a total of 10 segments were used as dosing areas across the left and right lobes. Lesion severity in each dosing area was scored using high-resolution computed tomography (HRCT) imaging, and the dosing volume for each lobe was calculated based on the scores.

[0173] The volume distribution of the drug is determined by considering two factors: ① the degree of atrophy of the lung lobes; ② the degree of fibrosis affecting the lung lobes.

[0174] Lung lobes with more severe fibrosis have a reduced volume, a more severe loss of regenerative function, and a poorer response to treatment, so a smaller volume of drug is administered. In the absence of severe atrophy, the administered volume is split 50% each.

[0175] HRCT manifestations were analyzed using image post-processing analysis software (3D Slicer image computing platform). First, 3D Slicer analysis software was used to perform three-dimensional reconstruction of the subjects' baseline HRCT image data, and the total volume of the lungs and each lobe was calculated. The Parenchyma Analysis plug-in was then used to set the lower limit of the high-density attenuation value (HAA) of pulmonary fibrosis tissue in Hounsfield units (HU). The lower limit of HAA was -700 HU, and a more accurate high-density attenuation value was -600 to -250 HU. Therefore, -600 to -250 HU was defined as the value range for fibrotic tissue. Through 3D Slicer analysis software, the proportion of fibrotic tissue in each lung lobe can be calculated based on the calculated lung lobe (HAA% -600 to -250), thereby calculating the total volume of fibrotic tissue in the entire lung, the volume of fibrotic tissue in the left and right lungs, and the total volume of fibrotic tissue in the left and right lower lobes, thereby obtaining the ratio of fibrotic tissue in the left and right lower lobes to the left and right lungs and the entire lung.

[0176] The proportional coefficient of the left and right lung fibrosis tissues is used as the basis for allocating the volume of mesenchymal stem cell preparations. The proportional coefficient is equal to the ratio of the high-density attenuation value area of ​​the left lung (HAA% -600 to -250) to the high-density attenuation value area of ​​the right lung (HAA% -600 to -250). The higher the proportional coefficient, the more severe the fibrosis of the left lung. The critical value of the proportional coefficient is 1.20: when it is less than 1.20, the proportion of human umbilical cord mesenchymal stem cell preparations allocated to the left and right lungs is 50% each. When the proportional coefficient is greater than or equal to 1.20, the proportion of human umbilical cord mesenchymal stem cell preparations allocated to the left lung is 40%, and the proportion of the volume allocated to the right lung is 60%. Since the lesions are mainly concentrated in the left and right lower lobes, the allocated human umbilical cord mesenchymal stem cell preparations are only administered to the left and right lower lobes, and each lung segment of the left and right lower lobes is evenly distributed. The actual administration situation is shown in Table 1 below:

[0177] Table 1 Dosage regimen

[0178]

[0179] After the infusion of the mesenchymal stem cell preparation is completed, each treated lung lobe is flushed with 5 mL of compound electrolyte injection or normal saline to help the mesenchymal stem cell preparation be evenly dispersed and pushed to the bronchi and alveoli at all levels deep in the lungs.

[0180] To prevent bronchial reflux of the infused mesenchymal stem cell preparation, subjects should sit with their upper body upright or lie flat with their head elevated and feet down within 2 hours of the infusion. Severe or persistent coughing may cause the infused mesenchymal stem cell preparation to be excreted as sputum; codeine can be taken orally.

[0181] 4) Clinical observation

[0182] Subjects received only one treatment. After treatment, they were required to return to the hospital for follow-up observations at 24 and 48 weeks. Follow-up observations were divided into safety and efficacy indicators.

[0183] Safety indicators include physical examination, blood routine, blood biochemistry, urine routine, electrocardiogram, abdominal B-ultrasound, and myocardial enzyme spectrum.

[0184] The efficacy indicators include the actual measured value of forced vital capacity (FVC), the percentage of the actual measured value of forced vital capacity (FVC) to the predicted value, the actual measured value of forced expiratory volume (FEV1) in one second, the percentage of the actual measured value of forced expiratory volume (FEV1) in one second to the predicted value, the actual measured value of total lung capacity (TLC), the percentage of the actual measured value of total lung capacity (TLC) to the predicted value, and the actual measured value of alveolar ventilation (VA) per minute.

[0185] 5) Treatment effect

[0186] The efficacy indicators of the subjects who received the low-dose and high-dose mesenchymal stem cell preparations were analyzed. The analysis content and methods are as follows:

[0187] First, the baseline indicators of the two groups of subjects were statistically analyzed;

[0188] Secondly, a statistical analysis was performed on the changes in the efficacy indicators of measured vital capacity (FVC) and the percentage of measured forced vital capacity (FVC) to predicted values ​​at week 24 and week 48 after treatment compared with the baseline period.

[0189] Secondly, 3D Slicer was used to compare and analyze the changes in the total lung volume, left and right lung volume, left and right lower lobe volume, lung fibrosis tissue ratio, left and right lung fibrosis tissue ratio, and left and right lower lobe fibrosis tissue ratio compared with the baseline period.

[0190] Numerical data were analyzed using the Student's t test for significant differences, while categorical data were analyzed using the chi-square test for significant differences. A P value less than 0.05 was considered significant between the two groups. The results are shown in Table 2.

[0191] There were no statistical differences between the low-dose group and the high-dose group in age, gender, disease severity, whether or not concurrent fibrosis treatment was used, the measured value of forced vital capacity (FVC), the percentage of the measured value of forced vital capacity (FVC) to the predicted value, the measured value of forced expiratory volume (FEV1) in the first second, the percentage of the measured value of forced expiratory volume (FEV1) in the first second, the measured value of total lung capacity (TLC), the percentage of the measured value of total lung capacity (TLC), and the measured value of alveolar ventilation (VA) per minute, indicating that the baseline levels of the two groups were consistent and comparable.

[0192] Table 2 Clinical indicators of subjects before treatment

[0193]

[0194]

[0195] As shown in Table 3 and Figure 4 As shown, 24 weeks after treatment, the subjects in the high-dose group showed better responses than those in the low-dose group. The FVC measured / predicted %, FEV1 measured, and FEV1 measured / predicted % in the high-dose group were improved compared with the baseline period, while those in the low-dose group were reduced.

[0196] Table 3 Clinical indicators of subjects after 24 weeks of treatment

[0197] 24w Low-dose group (n=3) High-dose group (n=3) p-value between groups FVC change ± mL -86.7 86.7 0.066 FVC change ±% -3.08 3.59 0.068 FVC_pre% change ± -3.66 3.50 0.002 FEV1 change ± mL -100 67 0.029 FEV1 change ±% -4.18 3.29 0.010 FEV1_pre% change value ± -4.91 4.08 0.001 TLC_pre% change value ± 8.29 0.09 0.104 VA change ±mL 410 -30 0.079

[0198] As shown in Table 4 and Figure 4 As shown, after 48 weeks of treatment, the FVC and FEV1 of the subjects in the high-dose group continued to rise compared to baseline, remaining above baseline levels. In contrast, these continued to decline in the low-dose group, with a significant decrease in FVC compared to baseline. The data between the two groups were statistically significant.

[0199] The total lung capacity (TLC) and minute alveolar ventilation (VA) values ​​of both groups were improved.

[0200] Table 4 Clinical indicators of subjects after 48 weeks of treatment

[0201]

[0202]

[0203] The above results show that the low-dose group is helpful to subjects with idiopathic pulmonary fibrosis in terms of total lung capacity TLC and minute alveolar ventilation VA. After 48 weeks of observation, FVC and FEV1 still decreased, and the downward trend and degree were similar to those of the approved pirfenidone or nintedanib.

[0204] The high-dose group showed a therapeutic effect superior to that of marketed drugs, with FVC and FEV1 improved compared to baseline and lasting for up to 48 weeks.

[0205] A decrease of more than 5% in FVC compared with the baseline level was defined as disease progression, and an increase of more than 5% in FVC compared with the baseline level was defined as disease improvement. Figure 5 It can be seen that two-thirds of the subjects in the low-dose group experienced disease progression at 48 weeks, while no one in the high-dose group experienced disease progression. In addition, one-third of the subjects in the high-dose group had their disease under control, and the disease improved in another two-thirds of the subjects.

[0206] HRCT images are used to assist in the evaluation of treatment efficacy in three planes: the aortic arch, the bronchial bifurcation, and the 1-cm plane of the diaphragmatic apex. Efficacy is assessed by observing changes in the area of ​​ground-glass opacity, lattice opacity, and honeycombing at each plane.

[0207] like Figure 6 As shown in the figure, S01007 (low-dose group) had large ground-glass opacities on all three levels at baseline. After 48 weeks of treatment, the ground-glass opacities disappeared, while the lattice and honeycomb shadows increased and worsened slightly.

[0208] like Figure 7 As shown in the figure, the ground-glass opacities at the bronchial bifurcation level and 1 cm level of the diaphragmatic dome at baseline in the S01008 (high-dose group) were reduced at 48 weeks after administration. Other lesions were basically stable without increasing or worsening.

[0209] Neither the aortic arch nor the bronchial bifurcation are target areas for drug administration. Only the 1 cm layer above the diaphragm is exposed to the stem cell drug, and it can be seen that areas not treated are also affected. Human umbilical cord stem cell therapy is highly effective against ground-glass opacities. Ground-glass opacities are inflammatory lesions, and stem cell therapy can modulate the immune response and inhibit excessive infiltration of inflammatory cells. It can also maintain honeycomb and lattice opacities, preventing them from worsening or expanding.

[0210] The changes in total lung volume (TLV), right lung volume (RV), left lung volume (LV), left lower lobe volume (LLLV) and right lower lobe volume (RLLV) (unit: L) are shown in Table 5:

[0211] Table 5

[0212]

[0213]

[0214] Observation up to 24 weeks after administration:

[0215] In the low-dose group, one patient (low-dose-1) showed increases in total lung volume, right lung volume, left lung volume, left lower lobe volume, and right lower lobe volume compared to baseline. In the other two patients (low-dose-2 and low-dose-3), total lung volume, right lung volume, left lung volume, left lower lobe volume, and right lower lobe volume remained stable compared to baseline.

[0216] In the high-dose group, the total lung volume, right lung volume, left lung volume, left lower lobe volume and right lower lobe volume of 3 cases increased compared with the baseline period.

[0217] This indicates that administering low or high doses of human umbilical cord mesenchymal stem cell injection only to the lower lobe of the lung can improve the recovery of left and right lung volume and total lung volume in the short term.

[0218] Observation up to 48 weeks after administration:

[0219] In the low-dose group, one patient (low-dose-1) had an increase in total lung volume, right lung volume, left lung volume, left lower lobe volume, and right lower lobe volume compared to baseline. Another patient (low-dose-3) had a stable total lung volume, right lung volume, left lung volume, left lower lobe volume, and right lower lobe volume compared to baseline.

[0220] In the high-dose group, the total lung volume, right lung volume, left lung volume, left lower lobe volume and right lower lobe volume of two patients (high-dose-1 and high-dose-3) increased compared with the baseline period.

[0221] This indicates that administering a high dose of human umbilical cord mesenchymal stem cell injection only in the lower lobe of the lung can improve the recovery of the left and right lungs and the total lung volume, and the improvement can last for a long time.

[0222] The changes in the total lung fibrosis ratio (T-HAA%), right lung fibrosis ratio (R-HAA%), left lung fibrosis ratio (L-HAA%), left lower lobe fibrosis ratio (LL-HAA%), and right lower lobe fibrosis ratio (RL-HAA%) (unit: %) are shown in Table 6:

[0223] Table 6

[0224]

[0225]

[0226] Observation up to 24 weeks after administration:

[0227] The overall lung fibrosis ratio, right lung fibrosis ratio, left lung fibrosis ratio, left lower lobe fibrosis ratio and right lower lobe fibrosis ratio of the three cases in the low-dose group were all reduced compared with the baseline period.

[0228] In the high-dose group, two cases (high-dose-1 and high-dose-3) had lower overall lung fibrosis ratio, right lung fibrosis ratio, left lung fibrosis ratio, left lower lobe fibrosis ratio, and right lower lobe fibrosis ratio compared with the baseline period.

[0229] This indicates that administering low or high doses of human umbilical cord mesenchymal stem cell injection only to the lower lobe of the lung can help reduce the proportion of fibrosis in the left and right lungs and the total lung in the short term.

[0230] Observation up to 48 weeks after administration:

[0231] In the low-dose group, one patient (low-dose-3) had stable overall lung fibrosis ratio, right lung fibrosis ratio, left lung fibrosis ratio, left lower lobe fibrosis ratio, and right lower lobe fibrosis ratio compared with the baseline period.

[0232] In the high-dose group, two cases (high-dose-1 and high-dose-3) had lower overall lung fibrosis ratio, right lung fibrosis ratio, left lung fibrosis ratio, left lower lobe fibrosis ratio, and right lower lobe fibrosis ratio compared with the baseline period.

[0233] This indicates that administering high doses of human umbilical cord mesenchymal stem cell injection only in the lower lobe of the lung can help reduce the proportion of fibrosis in the left and right lungs and the total lung, and the improvement effect can last for a longer period of time.

[0234] The changes in the total lung fibrosis volume (T-HAA-V), right lung fibrosis volume (R-HAA-V), left lung fibrosis volume (L-HAA-V), left lower lobe fibrosis volume (LL-HAA-V), and right lower lobe fibrosis volume (RL-HAA-V) (unit: mL) are shown in Table 7:

[0235] Table 7

[0236]

[0237] Observation up to 24 weeks after administration:

[0238] In the low-dose group, the lung fibrosis volume, right lung fibrosis volume, left lung fibrosis volume, left lower lobe fibrosis volume, and right lower lobe fibrosis volume of the three cases were reduced compared with the baseline period.

[0239] In the high-dose group, the lung fibrosis volume, right lung fibrosis volume, left lung fibrosis volume, left lower lobe fibrosis volume and right lower lobe fibrosis volume of two cases (high-dose-1 and high-dose-3) were reduced compared with the baseline period.

[0240] This indicates that administering low or high doses of human umbilical cord mesenchymal stem cell injection only to the lower lobe of the lung can improve the reduction of fibrosis volume in the left and right lungs and the total lung in the short term.

[0241] Observation up to 48 weeks after administration:

[0242] In the low-dose group, one patient (low-dose-2) showed reductions in lung fibrosis volume, right lung fibrosis volume, left lung fibrosis volume, left lower lobe fibrosis volume, and right lower lobe fibrosis volume compared to baseline. Another patient (low-dose-3) showed stable lung fibrosis volume, right lung fibrosis volume, left lung fibrosis volume, left lower lobe fibrosis volume, and right lower lobe fibrosis volume compared to baseline.

[0243] In the high-dose group, two cases (high-dose-1 and high-dose-2) had increased lung fibrosis volume, right lung fibrosis volume, left lung fibrosis volume, left lower lobe fibrosis volume, and right lower lobe fibrosis volume compared with the baseline period.

[0244] This indicates that administering a high dose of human umbilical cord mesenchymal stem cell injection only in the lower lobe of the lung can improve the reduction of fibrosis volume in the left and right lungs and the total lung, reverse the trend of fibrosis, and the improvement effect can be sustained for a long time.

[0245] In summary, the present invention provides a method for administering a mesenchymal stem cell preparation through bronchoscopy, which allows the drug to pass through the airway into the alveolar cavity, effectively contact the lesion, and remain in the lesion for a long time. MSCs have a long survival period and can continuously exert a therapeutic effect.

[0246] Clinical trial results of the present invention demonstrate that the method can effectively improve the lung function and quality of life of IPF patients, with significant clinical efficacy. Administration of low-dose mesenchymal stem cells can delay the progression of pulmonary fibrosis for up to 24 weeks. While other treatments can only slow the decline in lung function, administration of high-dose mesenchymal stem cells according to the present invention can significantly alter the progression of pulmonary fibrosis, achieving significantly better lung function improvements than other treatments, with sustained effects lasting up to 48 weeks.

[0247] All documents mentioned in this application are incorporated herein by reference, just as if each document were incorporated herein by reference individually. It should also be understood that after reading the above teachings of the present invention, those skilled in the art may make various changes or modifications to the present invention, and that such equivalents also fall within the scope of the claims appended hereto.

Claims

1. An intra-airway delivery system for a mesenchymal stem cell preparation for treating idiopathic pulmonary fibrosis, characterized in that: include: (a) a first container containing a first preparation comprising mesenchymal stem cells and a pharmaceutically acceptable carrier; (b) a bronchoscope injection module, comprising: an injection catheter, an injection pump, and a bronchoscope, wherein one end of the catheter contacts the first preparation in the first container, and the other end is used to deliver the preparation to the lungs; the injection pump is used to push the preparation in a quantitative manner; the bronchoscope lens is located near the end of the catheter that extends into the lungs and is used to capture images and / or identify the location; (c) a high-resolution computed tomography (HRCT) imager, comprising a tomography module and a data processing module, wherein the tomography module is configured to scan the subject's lungs to obtain lung images capable of displaying pulmonary fibrosis, and the data processing module is configured to calculate, based on the obtained lung images, the total volume of each lung lobe of the subject and the volume and / or proportion of fibrotic tissue; (d) a control module, configured to calculate the injection volume at each injection site based on the total volume of each lung lobe and the volume and / or proportion of fibrotic tissue of the subject calculated by the data processing module of the high-resolution computed tomography scanner (input), and control the bronchoscope injection module to inject a corresponding volume of the first agent into each injection site (output); Wherein, the total volume of the first preparation is 10 mL to 50 mL.

2. The drug delivery system according to claim 1, wherein In the data processing module, when the high-density attenuation value of lung tissue is set to be ≥-700HU (preferably -600 to -250HU), it is included in the calculation of the fibrotic tissue volume.

3. The drug delivery system according to claim 1, wherein The data processing module of the high-resolution computed tomography scanner is configured to calculate the volume of the subject's left lung fibrosis tissue and the total volume of the left lung based on the obtained lung image, and calculate the ratio P1 of the volume of the left lung fibrosis tissue to the total volume of the left lung; correspondingly, calculate the volume of the subject's right lung fibrosis tissue and the total volume of the right lung based on the obtained lung image, and calculate the ratio P2 of the volume of the right lung fibrosis tissue to the total volume of the right lung; then obtain a proportional coefficient based on the ratio of the volume of the left lung fibrosis tissue to the total volume of the left lung / the ratio of the volume of the right lung fibrosis tissue to the total volume of the right lung (P1 / P2), and input the proportional coefficient into the control module.

4. The drug delivery system according to claim 1, wherein The control module is configured to control the bronchoscope injection module to inject 50% of the volume of the first preparation into the bronchi of the left lung and the right lung, respectively, when the proportional coefficient calculated by the high-resolution computed tomography imager is less than a preset value k; and to control the bronchoscope injection module to inject 40±2% of the volume of the first preparation into the left bronchus and 60±2% of the volume of the first preparation into the right bronchus (the sum of the injection volumes of the left and right lungs is 100%) when the proportional coefficient calculated by the high-resolution computed tomography imager is greater than or equal to the preset value k. The preset value k is 1.0 to 1.25, preferably 1.1 to 1.

2.

5. The drug delivery system according to claim 1, wherein When injecting the first agent into the lungs, the control module is configured to control the bronchoscope injection module to inject the first preparation into the bronchi of the dorsal segment, anterior medial basal segment, lateral basal segment and posterior basal segment of the left lower lobe, and the dorsal segment, medial basal segment, anterior basal segment, lateral basal segment and posterior basal segment of the right lower lobe, wherein the administration volume of the anterior medial basal segment of the left lower lobe is 2 / 5 of the administration volume of the left lung, and the administration volume of other lung segments is 1 / 5 of the administration volume of the corresponding lung lobes.

6. The drug delivery system according to claim 1, wherein The mesenchymal stem cells are derived from umbilical cord, bone marrow, fat, placental tissue, or a combination thereof, and are preferably human umbilical cord mesenchymal stem cells.

7. The drug delivery system according to claim 1, wherein A single dose of the first formulation contains 3×10 7 ~7×10 7 Mesenchymal stem cells, preferably 6±0.5×10 7 cells.

8. The method according to claim 1, wherein The subject suffers from idiopathic pulmonary fibrosis.

9. The drug delivery system according to claim 1, wherein The subject receives intra-airway administration of the present invention only once every 24 to 48 weeks.

10. Use of the drug delivery system according to any one of claims 1 to 9 in preparing a medical device for treating idiopathic pulmonary fibrosis.

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