Use of low molecular weight hyaluronic acid for the treatment of pulmonary mucosal inflammation

Abstract

In chronic inflammatory respiratory diseases such as cystic fibrosis or chronic obstructive pulmonary disease (COPD), the balance of cells and functions may be disrupted, resulting in re-epithelialization or remodeling areas with the presence of squamous metaplasia and / or hyperplasia of basal or secretory cells. Thus, there is a need to identify drugs capable of treating pulmonary mucosal inflammation. The present inventors have surprisingly shown that hyaluronic acid (HA) with a low molecular weight of 15,000-50,000 daltons has anti-inflammatory properties against pulmonary mucosal inflammation. Thus, the present invention relates to a method for treating pulmonary mucosal inflammation in a subject suffering from cystic fibrosis or chronic obstructive pulmonary disease (COPD), comprising administering to the subject a therapeutically effective amount of hyaluronic acid with a low molecular weight of 15,000-50,000 daltons.

Description

[Technical field]

[0001] Field of the invention: The present invention is in the field of medicine, in particular in respiratory medicine. [Background technology]

[0002] Background of the invention: The respiratory epithelium is in permanent contact with the external environment and has a total exchange surface area of ​​approximately 100–130 m 2 In non-pathological cases, the respiratory epithelium is continuously exposed through inhalation to various pathogens or particles that may induce epithelial lesions. When faced with these lesions, the airway epithelium should be able to restore its integrity through repair and regeneration mechanisms to regain all its functions, especially its defense and barrier functions. Thus, in chronic inflammatory respiratory diseases such as cystic fibrosis, chronic obstructive pulmonary disease, asthma or allergy, the balance of cells and functions may be disrupted, resulting in re-epithelialization or remodeling areas with the presence of squamous metaplasia and / or hyperplasia of basal or secretory cells. Therefore, there is a need to identify drugs that can treat lung mucosal inflammation.

[0003] Hyaluronic acid (HA) is an anionic, non-sulfated glycosaminoglycan that is widely distributed throughout connective, epithelial and neural tissues. There are various sources of hyaluronic acid, typically bacterial (e.g., from Streptococcus zooepidemicus), but also avian (e.g., from cockscomb) and bovine (e.g., from bovine vitreous humor). HA is said to be involved in inflammatory responses, the role of which is dependent on its molecular weight. Indeed, high molecular weight (HMW) hyaluronan has anti-inflammatory and immunosuppressive roles, whereas smaller fragments called hyaluronan oligosaccharides (o-HA) are pro-inflammatory and immunostimulatory (HUANG, T., CHAN, Y., CHENG, P., YOUNG, Y., LOU, P.et YOUNG, T. Increased mucociliary differentiation of human respiratory epithelial cells on hyaluronan-derivative membranes. Acta biomaterialia. 2010a. Vol.6, n 3, p.1191-1199.;OCHOA, C., GARG, H., HALES, C. et QUINN, D. Low molecular weight hyaluronan, via AP-1 and NF-κB signalling, induces IL-8 in transformed bronchial epithelial cells. Swiss medical weekly. 2011. Vol.141, p.13255.;RUPPERT, S., HAWN, T., ARRIGONI, A., WIGHT, T. et BOLLYKY, P. Tissue integrity signals communicated by high-molecular weight hyaluronan and the resolution of inflammation. Immunologic research. 2014. Vol.58, n 2-3, p.186-192.).In chondrocytes, HA and resveratrol gel has been shown to inhibit pathogen LPS-induced inflammation and reduce IL-1β secretion (SHEU, S., CHEN, W., SUN, J., LIN, F. et WU, T. Biological characterization of oxidized hyaluronic acid / resveratrol hydrogel for cartilage tissue engineering. Journal of biomedical materials research. Part A. 2013. Vol.101, n 12, p.3457-3466.). Recent studies in homozygous F508del-CFTR mice expressing the β subunit of the ENaC channel demonstrated that inhalation of a high molecular weight HA solution exerted an anti-inflammatory effect embodied by a reduction in the protein expression of pro-inflammatory cytokines such as TNF-α and MIP-2 (macrophage inflammatory protein-2) (GAVINA, M., LUCIANI, A., VILLELLA, V., ESPOSITO, S., FERRARI, E., BRESSANI, I., CASALE, A., BRUSCIA, E., MAIURI, L. et RAIA, V. Nebulized hyaluronan ameliorates lung inflammation in cystic fibrosis mice. Pediatric pulmonology. 2013. Vol.48, n 8, p.761-771.).

[0004] WO2004050187 and WO2009024677 disclose the pharmaceutical use of low molecular weight HA of 30,000-45,000 daltons for the treatment of respiratory diseases of the upper respiratory tract. In particular, WO2004050187 teaches that said HA may be suitable for repairing epithelium and for altering the surface properties of respiratory mucus to facilitate its transport by ciliary activity. WO2009024677 discloses the use of said HA to restore the defensive function of junctional complexes after an attack on the epithelium. Summary of the Invention [Problem to be solved by the invention]

[0005] Summary of the invention: The invention is defined by the claims. In particular, the invention relates to a method for treating pulmonary mucosal inflammation. [Means for solving the problem]

[0006] Detailed description of the invention: The present inventors have surprisingly shown that low molecular weight HA, between 15,000 and 50,000 daltons, has anti-inflammatory properties against pulmonary mucosal inflammation in subjects suffering from an inflammatory disease (e.g. cystic fibrosis or COPD).

[0007] Accordingly, a first object of the present invention relates to a method for treating pulmonary mucosal inflammation in a subject suffering from cystic fibrosis or chronic obstructive pulmonary disease (COPD), comprising administering to the subject a therapeutically effective amount of hyaluronic acid having a low molecular weight of 15,000 to 50,000 daltons. [Brief description of the drawings]

[0008] [Figure 1]Measurement of the proinflammatory chemokine IL-8 secreted by F508del / F508del CF human airway epithelial cells cultured at the air-liquid interface (ALI) by ELISA assay. Airway epithelial cells were seeded in bicompartment chambers and cultured in liquid-liquid conditions until confluence was reached. At confluence, the culture medium from the upper chamber was removed to create an air-liquid interface (ALI) that favors epithelial cell differentiation. From the ALI creation, the different treatments (sodium hyaluronate 15-45 kDa (HA-Na 15-45 kDa) (ARD-Pomacle-France-prepared according to WO 2004050187), sodium hyaluronate 15-30 kDa (HA-Na 15-30 kDa) (Sigma Aldrich) or sodium hyaluronate 30-50 kDa (HA-Na 30-50 kDa) (Sigma Aldrich)) were added to the culture medium in the basal chamber at 1 mg / mL. Culture medium was freshly fed 3 times a week. On day 15 at ALI, the culture medium was replaced with fresh medium without any treatment. After 4 hours, this culture medium was collected and the IL-8 content was determined by ELISA assay. n=5 different F508del / F508del CF patients. p<0.001(***); p<0.01(**); p<0.05(*). ns: not significant. [Diagram 2]Measurement of the proinflammatory chemokine IL-8 secreted by F508del / F508del CF human airway epithelial cells cultured as monolayers by ELISA assay. Airway epithelial cells were seeded in 48-well plates and cultured until confluence was reached. Cells were then treated with a proinflammatory chemokine cocktail called Cytomix (Cy) (TNFα, IL1β, and IFNγ; 10 ng / mL each) for 24 hours. Cells were then treated for the following 24 hours with a combination containing Cytomix and the anti-inflammatory dexamethasone (Dexa; 10-6M) or 1 mg / mL of sodium hyaluronate 15-45 kDa (HA-Na 15-45 kDa) (prepared according to ARD-Pomacle-France-WO 2004050187), sodium hyaluronate 15-30 kDa (HA-Na 15-30 kDa) (Sigma Aldrich) or sodium hyaluronate 30-50 kDa (HA-Na 30-50 kDa) (Sigma Aldrich). Finally, culture medium was collected and IL-8 content was determined by ELISA assay. n=7 different F508del / F508del CF patients. p<0.001 (***); p<0.01 (**); p<0.05 (*). ns: not significant. [Diagram 3]Measurement of the proinflammatory chemokine IL-8 secreted by F508del / F508del CF human airway epithelial cells cultured as monolayers by ELISA assay. Airway epithelial cells were seeded in 48-well plates and cultured until confluence was reached. Cells were then treated for 24 hours with a combination containing a proinflammatory chemokine cocktail called Cytomix (Cy) (TNFα, IL1β and IFNγ; 10 ng / mL each) and the anti-inflammatory dexamethasone (Dexa; 10-6M), 1 mg / mL of sodium hyaluronate 15-45 kDa (HA-Na 15-45 kDa) (ARD-Pomacle-France-prepared according to WO 2004050187), sodium hyaluronate 15-30 kDa (HA-Na 15-30 kDa) (Sigma Aldrich), or sodium hyaluronate 30-50 kDa (HA-Na 30-50 kDa) (Sigma Aldrich). Culture medium was then collected and IL-8 content determined. n=7 F508del / F508del CF patients. p<0.001(***); p<0.01(**); p<0.05(*). ns: not significant. [Figure 4]Measurement of the proinflammatory chemokine IL-8 secreted by F508del / F508del CF human airway epithelial cells cultured at the air-liquid interface (ALI) by ELISA assay. Airway epithelial cells were seeded in bicompartment chambers and cultured in liquid-liquid conditions until confluence was reached. At confluence, the culture medium from the upper chamber was removed to create an air-liquid interface (ALI) that favors epithelial cell differentiation. From the ALI creation, the different treatments (sodium hyaluronate 15-45 kDa (HA-Na 15-45 kDa) (ARD-Pomacle-France - prepared according to WO 2004050187), calcium hyaluronate 15-45 kDa (HA-Ca 15-45 kDa; ARD), or potassium hyaluronate 15-45 kDa (HA-K 15-45 kDa; ARD)) were added to the culture medium in the basal chamber at 1 mg / mL. Culture medium was freshly fed three times a week. On day 15 at ALI, the culture medium was replaced with fresh medium without any treatment. After 4 hours, this culture medium was collected and the IL-8 content was determined by ELISA assay. n=4 different F508del / F508del CF patients. p<0.001(***); p<0.01(**); p<0.05(*). ns: not significant. [Diagram 5]Measurement of the proinflammatory chemokine IL-8 secreted by F508del / F508del CF human airway epithelial cells cultured as monolayers by ELISA assay. Airway epithelial cells were seeded in 48-well plates and cultured until confluence was reached. Cells were then treated with a proinflammatory chemokine cocktail called Cytomix (Cy) (TNFα, IL1β, and IFNγ; 10 ng / mL each) for 24 hours. Cells were then treated for the following 24 hours with a combination containing Cytomix and the anti-inflammatory dexamethasone (Dexa; 10-6M) or 1 mg / mL of sodium hyaluronate 15-45 kDa (HA-Na 15-45 kDa) (ARD-Pomacle-France-prepared according to WO 2004050187), calcium hyaluronate 15-45 kDa (HA-Ca 15-45 kDa; ARD) or potassium hyaluronate 15-45 kDa (HA-K 15-45 kDa; ARD). Finally, culture medium was collected and IL-8 content was determined. n=7 different F508del / F508del CF patients. p<0.001 (***); p<0.01 (**); p<0.05 (*). ns: not significant. [Figure 6]Measurement of the proinflammatory chemokine IL-8 secreted by F508del / F508del CF human airway epithelial cells cultured as monolayers by ELISA assay. Airway epithelial cells were seeded in 48-well plates and cultured until confluence was reached. Cells were then treated for 24 hours with a combination containing a proinflammatory chemokine cocktail called Cytomix (Cy) (TNFα, IL1β and IFNγ; 10 ng / mL each) and the anti-inflammatory dexamethasone (Dexa; 10-6M), or 1 mg / mL of sodium hyaluronate 15-45 kDa (HA-Na 15-45 kDa) (ARD-Pomacle-France-prepared according to WO 2004050187), calcium hyaluronate 15-45 kDa (HA-Ca 15-45 kDa; ARD) or potassium hyaluronate 15-45 kDa (HA-K 15-45 kDa; ARD). Culture medium was then collected and IL-8 content determined. n=7 different F508del / F508del CF patients. p<0.001(***); p<0.01(**); p<0.05(*). ns: not significant. [Figure 7]Measurement by ELISA assay of the proinflammatory chemokine IL-8 secreted by F508del / F508del CF human airway epithelial cells cultured at an air-liquid interface (ALI). Airway epithelial cells were seeded in bicompartment chambers and cultured in liquid-liquid conditions until confluence was reached. At confluence, the culture medium from the upper chamber was removed to create an air-liquid interface (ALI) favoring epithelial cell differentiation. From the ALI creation, sodium hyaluronate 15-45 kDa (HA-Na 15-45 kDa) (prepared according to ARD-Pomacle-France-WO 2004050187) was added to the culture medium in the basal chamber at 0.1 mg / mL, 0.5 mg / mL, 1 mg / mL and 2 mg / mL. The culture medium was freshly supplied three times a week. On the 15th day at ALI, the culture medium was replaced with fresh medium without any treatment. After 4 hours, the culture medium was collected and IL-8 content was determined by ELISA assay. n=3 different F508del / F508del CF patients. p<0.001(***); p<0.01(**); p<0.05(*). ns: not significant. [Figure 8]Measurement of mucin MUC-5B secreted by F508del / F508del CF human airway epithelial cells cultured at the air-liquid interface (ALI) by ELISA assay. Airway epithelial cells were seeded in bicompartment chambers and cultured in liquid-liquid conditions until confluence was reached. At confluence, the culture medium from the upper chamber was removed to create an air-liquid interface (ALI) that favors epithelial cell differentiation. From the ALI creation, the different treatments (15-45 kDa sodium hyaluronate (HA-Na 15-45 kDa) (ARD-Pomacle-France-prepared according to WO 2004050187), 15-30 kDa sodium hyaluronate (HA-Na 15-30 kDa) (Sigma Aldrich), or 30-50 kDa sodium hyaluronate (HA-Na 30-50 kDa) (Sigma Aldrich)) were added to the culture medium in the basal chamber at 1 mg / mL. The culture medium was freshly fed three times a week. On day 15 at ALI and day 25 at ALI, the culture medium in the upper chamber, without any treatment, was rinsed with PBS and then with a small amount of fresh medium and put in contact with the apical part of the epithelium. After 4 hours, this apical culture medium was collected and the MUC-5B content was determined by ELISA assay. n=3 different F508del / F508del CF patients. p<0.001(***); p<0.01(**); p<0.05(*). ns: not significant. [Figure 9]Measurement of mucin MUC-5B secreted by F508del / F508del CF human airway epithelial cells cultured at the air-liquid interface (ALI) by ELISA assay. Airway epithelial cells were seeded in bicompartment chambers and cultured in liquid-liquid conditions until confluence was reached. At confluence, the culture medium from the upper chamber was removed to create an air-liquid interface (ALI) that favors epithelial cell differentiation. From the ALI creation, the different treatments (sodium hyaluronate 15-45 kDa (HA-Na 15-45 kDa) (ARD-Pomacle-France-prepared according to WO 2004050187), calcium hyaluronate 15-45 kDa (HA-Ca 15-45 kDa; ARD), or potassium hyaluronate 15-45 kDa (HA-K 15-45 kDa; ARD)) were added to the culture medium in the basal chamber at 1 mg / mL. Culture medium was freshly fed three times a week. On day 15 at ALI and day 25 at ALI, the culture medium in the upper chamber, without any treatment, was rinsed with PBS and then with a small amount of fresh medium and put in contact with the apical part of the epithelium. After 4 hours, this apical culture medium was collected and the MUC-5B content was determined by ELISA assay. n=3 different F508del / F508del CF patients. p<0.001(***); p<0.01(**); p<0.05(*). ns: not significant. [Figure 10]Measurement by ELISA assay of mucin MUC-5B secreted by F508del / F508del CF human airway epithelial cells cultured at the air-liquid interface (ALI). Airway epithelial cells were seeded in bicompartment chambers and cultured in liquid-liquid conditions until confluence was reached. At confluence, the culture medium from the upper chamber was removed to create an air-liquid interface (ALI) favoring epithelial cell differentiation. From the ALI creation, sodium hyaluronate 15-45 kDa (HA-Na 15-45 kDa) (prepared according to ARD-Pomacle-France-WO 2004050187) was added to the culture medium in the basal chamber at 2 mg / mL, 1 mg / mL or 0.5 mg / mL. Culture medium was freshly supplied three times a week. On day 15 at ALI and day 25 at ALI, the cultures in the upper chamber were rinsed with PBS and then a small amount of fresh medium without any treatment and placed in contact with the apical part of the epithelium. After 4 hours, the apical culture medium was collected and MUC-5B content was determined by ELISA assay. n=3 different F508del / F508del CF patients. p<0.001 (***); p<0.01 (**); p<0.05 (*). ns: not significant. [Figure 11]Measurement of the proinflammatory chemokine IL-8 secreted by non-CF human airway epithelial cells cultured at the air-liquid interface (ALI) by ELISA assay. Airway epithelial cells were seeded in bicompartment chambers and cultured in liquid-liquid conditions until confluence was reached. At confluence, the culture medium from the upper chamber was removed to create an air-liquid interface (ALI) that favors epithelial cell differentiation. From the ALI creation, the different treatments (sodium hyaluronate 15-45 kDa (HA-Na 15-45 kDa) (ARD-Pomacle-France-prepared according to WO 2004050187), sodium hyaluronate 15-30 kDa (HA-Na 15-30 kDa) (Sigma Aldrich), or sodium hyaluronate 30-50 kDa (HA-Na 30-50 kDa) (Sigma Aldrich)) were added to the culture medium in the basal chamber at 1 mg / mL. Culture medium was freshly fed three times a week. On the 15th day at ALI, the culture medium was replaced with fresh medium without any treatment. After 4 hours, this culture medium was collected and the IL-8 content was determined by ELISA assay. n=1 non-CF patient. p<0.001 (***); p<0.01 (**); p<0.05 (*). ns: not significant. [Figure 12]Measurement of the proinflammatory chemokine IL-8 secreted by non-CF human airway epithelial cells cultured as monolayers by ELISA assay. Airway epithelial cells were seeded in 48-well plates and cultured until confluence was reached. Cells were then treated with a proinflammatory chemokine cocktail called Cytomix (Cy) (TNFα, IL1β, and IFNγ; 10 ng / mL each) for 24 hours. Cells were then treated for the following 24 hours with a combination containing Cytomix and the anti-inflammatory dexamethasone (Dexa; 10-6M), 1 mg / mL of sodium hyaluronate 15-45 kDa (HA-Na 15-45 kDa) (prepared according to ARD-Pomacle-France-WO 2004050187), sodium hyaluronate 15-30 kDa (HA-Na 15-30 kDa) (Sigma Aldrich), or sodium hyaluronate 30-50 kDa (HA-Na 30-50 kDa) (Sigma Aldrich). Finally, the culture medium was collected and the IL-8 content was determined by ELISA assay. n=8 different non-CF patients. p<0.001 (***); p<0.01 (**); p<0.05 (*). ns: not significant. [Figure 13]Measurement of the proinflammatory chemokine IL-8 secreted by non-CF human airway epithelial cells cultured as monolayers by ELISA assay. Airway epithelial cells were seeded in 48-well plates and cultured until confluence was reached. Cells were then treated for 24 hours with a combination containing a proinflammatory chemokine cocktail called Cytomix (Cy) (TNFα, IL1β and IFNγ; 10 ng / mL each) and the anti-inflammatory dexamethasone (Dexa; 10-6M), 1 mg / mL of sodium hyaluronate 15-45 kDa (HA-Na 15-45 kDa) (ARD-Pomacle-France-prepared according to WO 2004050187), sodium hyaluronate 15-30 kDa (HA-Na 15-30 kDa) (Sigma Aldrich), or sodium hyaluronate 30-50 kDa (HA-Na 30-50 kDa) (Sigma Aldrich). Culture medium was then collected and IL-8 content determined. n=8 different non-CF patients. p<0.001(***); p<0.01(**); p<0.05(*). ns: not significant. [Figure 14]Measurement by ELISA assay of the proinflammatory chemokine IL-8 secreted by non-CF human airway epithelial cells cultured at an air-liquid interface (ALI). Airway epithelial cells were seeded in bicompartment chambers and cultured in liquid-liquid conditions until confluence was reached. At confluence, the culture medium from the upper chamber was removed to create an air-liquid interface (ALI) favoring epithelial cell differentiation. From the ALI creation, different treatments (sodium hyaluronate 15-45 kDa (HA-Na 15-45 kDa) (ARD-Pomacle-France-prepared according to WO 2004050187), calcium hyaluronate 15-45 kDa (HA-Ca 15-45 kDa; ARD) or potassium hyaluronate 15-45 kDa (HA-K 15-45 kDa; ARD)) were added at 1 mg / mL to the culture medium in the basal chamber. Culture medium was refreshed three times a week. On day 15 at ALI, culture medium was replaced with fresh medium without any treatment. After 4 hours, the culture medium was collected and IL-8 content was determined by ELISA assay. n=1 non-CF patient. p<0.001(***); p<0.01(**); p<0.05(*). ns: not significant. [Figure 15]Measurement of the proinflammatory chemokine IL-8 secreted by non-CF human airway epithelial cells cultured as monolayers by ELISA assay. Airway epithelial cells were seeded in 48-well plates and cultured until confluence was reached. Cells were then treated with a proinflammatory chemokine cocktail called Cytomix (Cy) (TNFα, IL1β, and IFNγ; 10 ng / mL each) for 24 hours. The cells were then treated for the following 24 hours with a combination containing Cytomix and the anti-inflammatory dexamethasone (Dexa; 10-6M), 1mg / mL of sodium hyaluronate 15-45kDa (HA-Na 15-45kDa) (ARD-Pomacle-France-prepared according to WO 2004050187), calcium hyaluronate 15-45kDa (HA-Ca 15-45kDa; ARD), or potassium hyaluronate 15-45kDa (HA-K 15-45kDa; ARD). Finally, the culture medium was collected and the IL-8 content was determined. n=8 different non-CF patients. p<0.001 (***); p<0.01 (**); p<0.05 (*). ns: not significant. [Figure 16] Measurement of the proinflammatory chemokine IL-8 secreted by non-CF human airway epithelial cells cultured as monolayers by ELISA assay. Airway epithelial cells were seeded in 48-well plates and cultured until confluence was reached. The cells were then treated for 24 hours with a combination containing a proinflammatory chemokine cocktail called Cytomix (Cy) (TNFα, IL1β and IFNγ; 10 ng / mL each) and the anti-inflammatory dexamethasone (Dexa; 10-6M), 1 mg / mL of sodium hyaluronate 15-45 kDa (HA-Na 15-45 kDa) (ARD-Pomacle-France-prepared according to WO 2004050187), calcium hyaluronate 15-45 kDa (HA-Ca 15-45 kDa; ARD), or potassium hyaluronate 15-45 kDa (HA-K 15-45 kDa; ARD). Culture medium was then collected and IL-8 content determined. n=8 different non-CF patients. p<0.001(***); p<0.01(**); p<0.05(*). ns: not significant. [Figure 17] Measurement of the proinflammatory chemokine IL-8 secreted by human COPD airway epithelial cells cultured as monolayers by ELISA assay. Airway epithelial cells were seeded in 48-well plates and cultured until confluence was reached. Cells were then treated with a proinflammatory chemokine cocktail called Cytomix (Cy) (TNFα, IL1β, and IFNγ; 10 ng / mL each) for 24 hours. The cells were then treated for the following 24 hours with a combination containing Cytomix and the anti-inflammatory dexamethasone (Dexa; 10-6M), 1 mg / mL of sodium hyaluronate 15-45 kDa (HA-Na 15-45 kDa) (prepared according to ARD-Pomacle-France-WO 2004050187), sodium hyaluronate 15-30 kDa (HA-Na 15-30 kDa) (Sigma Aldrich), or sodium hyaluronate 30-50 kDa (HA-Na 30-50 kDa) (Sigma Aldrich). Finally, the culture medium was collected and the IL-8 content was determined by ELISA assay. n=5 different COPD patients. p<0.001 (***); p<0.01 (**); p<0.05 (*). ns: not significant. [Figure 18]Measurement of the proinflammatory chemokine IL-8 secreted by human COPD airway epithelial cells cultured as monolayers by ELISA assay. Airway epithelial cells were seeded in 48-well plates and cultured until confluence was reached. Cells were then treated for 24 hours with a combination containing a proinflammatory chemokine cocktail called Cytomix (Cy) (TNFα, IL1β and IFNγ; 10 ng / mL each) and the anti-inflammatory dexamethasone (Dexa; 10-6M), 1 mg / mL of sodium hyaluronate 15-45 kDa (HA-Na 15-45 kDa) (ARD-Pomacle-France-prepared according to WO 2004050187), sodium hyaluronate 15-30 kDa (HA-Na 15-30 kDa) (Sigma Aldrich), or sodium hyaluronate 30-50 kDa (HA-Na 30-50 kDa) (Sigma Aldrich). Culture medium was then collected and IL-8 content determined. n=5 COPD patients. p<0.001(***); p<0.01(**); p<0.05(*). ns: not significant. [Figure 19]Measurement of the proinflammatory chemokine IL-8 secreted by human COPD airway epithelial cells cultured as monolayers by ELISA assay. Airway epithelial cells were seeded in 48-well plates and cultured until confluence was reached. Cells were then treated with a proinflammatory chemokine cocktail called Cytomix (Cy) (TNFα, IL1β, and IFNγ; 10 ng / mL each) for 24 hours. The cells were then treated for the following 24 hours with a combination containing Cytomix and the anti-inflammatory dexamethasone (Dexa; 10-6M) or 1 mg / mL of sodium hyaluronate 15-45 kDa (HA-Na 15-45 kDa) (ARD-Pomacle-France-prepared according to WO 2004050187), calcium hyaluronate 15-45 kDa (HA-Ca 15-45 kDa; ARD) or potassium hyaluronate 15-45 kDa (HA-K 15-45 kDa; ARD). Finally, the culture medium was collected and the IL-8 content was determined. n=5 different COPD patients. p<0.001 (***); p<0.01 (**); p<0.05 (*). ns: not significant. [Figure 20] Measurement of the proinflammatory chemokine IL-8 secreted by human COPD airway epithelial cells cultured as monolayers by ELISA assay. Airway epithelial cells were seeded in 48-well plates and cultured until confluence was reached. The cells were then treated for 24 hours with a combination containing a proinflammatory chemokine cocktail called Cytomix (Cy) (TNFα, IL1β and IFNγ; 10 ng / mL each) and the anti-inflammatory dexamethasone (Dexa; 10-6M), 1 mg / mL of sodium hyaluronate 15-45 kDa (HA-Na 15-45 kDa) (ARD-Pomacle-France-prepared according to WO 2004050187), calcium hyaluronate 15-45 kDa (HA-Ca 15-45 kDa; ARD), or potassium hyaluronate 15-45 kDa (HA-K 15-45 kDa; ARD). Culture medium was then collected and IL-8 content determined. n=5 different COPD patients. p<0.001(***); p<0.01(**); p<0.05(*). ns: not significant. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0009] As used herein, the term "subject" refers to mammals such as rodents, cats, dogs, and primates. In particular, subjects according to the present invention are humans. The term "subject" encompasses the term "patient."

[0010] As used herein, the term "inflammation" has its general meaning in the art and is used to describe an underlying pathological process consisting of a dynamic set of cytological and histological reactions that occur in tissues in response to injury or abnormal stimuli caused by physical, chemical or biological agents (e.g. bacteria, viruses...), including local reactions and resulting morphological changes, destruction or removal of injurious materials, and responses that lead to repair and healing. The so-called cardinal signs of inflammation are swelling, pain, and in certain cases, inhibition or loss of function of the target organ. Swelling usually results from congestion and exudation; compression (or stretching) of nerve endings and changes in osmolarity and pH can lead to significant pain; inhibition of function can result in impaired movement or actual destruction of an anatomical site or organ. In some embodiments, inflammation is localized to the pulmonary tract, particularly the lungs, and is well treated by the methods of the present invention. In particular, inflammation localized to the oral mucosa, e.g. buccal and sublingual; nasal mucosa; pulmonary mucosa; bronchial mucosa is well treated by the methods of the present invention.

[0011] As used herein, the phrase "pulmonary mucosa inflammation" has its ordinary meaning in the art and refers to swelling or irritation of the pulmonary mucosa. As used, the term "mucosa" has its ordinary meaning in the art and refers to the moist, mucus-secreting, epithelial-covered tissue that lines body cavities.

[0012] In some embodiments, the subject suffers from an inflammatory disease of the mucous membranes that affects the respiratory system and typically includes cystic fibrosis and chronic obstructive pulmonary disease.

[0013] Thus, in some embodiments, the subject has cystic fibrosis.

[0014] As used herein, the term "cystic fibrosis" has its general meaning in the art and refers to an inherited autosomal disease with mutations in the gene encoding cystic fibrosis transmembrane conductor regulator (CFTR).The method of the present invention can be carried out for any type of cystic fibrosis, such as those revised in the World Health Organization classification of cystic fibrosis and selected from group E84: mucoviscidosis, cystic fibrosis with pulmonary symptoms, cystic fibrosis with intestinal symptoms and cystic fibrosis with other symptoms.In some embodiments, the subject carries at least one mutation in the CFTR gene, including but not limited to F508del-CFTR, R117H CFTR and G551D CFTR (for CFTR mutations, see, for example, http: / / www.genet.sickkids.on.ca / cftr).

[0015] In some embodiments, the subject has chronic obstructive pulmonary disease (COPD).

[0016] As used herein, the term "COPD" refers to chronic obstructive pulmonary disease. The term "COPD" generally refers to a chronic respiratory disease process characterized by persistent obstruction of bronchial airflow. COPD patients may suffer from conditions such as bronchitis or emphysema.

[0017] In some aspects, the pulmonary mucosal inflammation can result from a pulmonary infection.

[0018] As used herein, the term "pulmonary infection" has its general meaning in the art and refers to the invasion of a patient's lung tissue by pathogenic microorganisms, their proliferation, and the response of the lung tissue to those microorganisms and the toxins they produce. In some aspects, the patient suffers from a chronic lung infection.

[0019] As used herein, the term "chronic infection" refers to a long-term infection that may be an overt, subclinical or latent infection. In some aspects, the patient suffers from an acute pulmonary infection.

[0020] In some aspects, the pulmonary infection is a bacterial infection, such as bacterial pneumonia.

[0021] In some embodiments, the bacterial infection is caused by Streptococcus pneumoniae (also known as pneumococcus), Staphylococcus aureus, Streptococcus agalactiae, Streptococcus pyogenes, Haemophilus influenzae, Haemophilus parainfluenzae, Klebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa, Moraxella catarrhalis, Chlamydophila pneumoniae, or any of a number of other pathogenic bacteria. pneumoniae, Mycoplasma pneumoniae, Legionella pneumophila, Serratia marcescens, Burkholderia cepacia, Burkholderia pseudomallei, Bacillus anthracis, Bacillus cereus, Bordetella pertussis, Stenotrophomonas maltophilia, bacteria from the Citrobacter family, bacteria from the Acinetobacter family, and Mycobacterium tuberculosis or Mycobacterium abscessus. In some embodiments, the pulmonary infection is caused by a bacterium selected from the group consisting of: Bacillus subtilis, Bacillus abscessus, Bacillus subtilis ...In some embodiments, the fungal infection is caused by a fungus selected from the group consisting of Histoplasma capsulatum, Cryptococcus neoformans, Pneumocystis jiroveci, Coccidioides immitis, Candida albicans, and Pneumocystis jirovecii (causing Pneumocystis pneumonia (PCP), also known as pneumocystiosis) or Aspergillus fumigatus. In some embodiments, the pulmonary infection is a viral infection, e.g., viral pneumonia. In some embodiments, the viral infection is caused by a virus selected from the group consisting of influenza virus (e.g., influenza virus A, influenza virus B), respiratory syncytial virus, adenovirus, metapneumovirus, cytomegalovirus, parainfluenza virus (e.g., hPIV-1, hPIV-2, hPIV-3, hPIV-4), rhinovirus, coxsackievirus, echovirus, herpes simplex virus, coronavirus (SARS-coronavirus, such as SARS-Cov1 or SARS-Cov2), and smallpox. In some embodiments, the viral lung infection may be due to a member of the Pneumoviridae, Paramyxoviridae and / or Coronaviridae families, in particular selected from the group consisting of upper and lower respiratory tract infections due to human respiratory syncytial virus (hRSV) types A and B, human metapneumovirus (hMPV) types A and B; parainfluenza virus type 3 (PIV-3), measles virus, endemic human coronaviruses (HCoV-229E, -NL63, -OC43, and -HKU1), severe acute respiratory syndrome (SARS) and Middle East respiratory syndrome (MERS) coronaviruses. In particular, the method of the present invention is suitable for the treatment of severe acute respiratory syndrome (SARS). More particularly, the method of the present invention is suitable for the treatment of lung mucosal inflammation in patients suffering from COVID-19.

[0022] As used herein, the term "treatment" or "treating" refers to both preventative or prophylactic treatments and curative or disease-curative treatments, including treatments of patients at risk of or suspected of having a disease, as well as patients who are ill or have been diagnosed with a disease or medical condition, including suppression of clinical recurrence. Treatments may be administered to subjects with a medical disorder or at risk of eventually acquiring a disorder, to prevent, cure, delay the onset of, reduce the severity of, or ameliorate one or more symptoms of the disorder or recurrent disorder, or to prolong the survival of the subject beyond that expected in the absence of such treatment. "Therapeutic regimen" refers to a pattern of treatment of a disease, e.g., a pattern of dosing used during treatment. Therapeutic regimens may include induction regimens and maintenance regimens. The phrase "induction regimen" or "induction period" refers to a therapeutic regimen (or a portion of a therapeutic regimen) used for the initial treatment of a disease. The general goal of an induction regimen is to provide high levels of drug to the patient during the initial period of the treatment regimen. The induction regimen may employ (partially or entirely) a "loading regimen", which may involve administering a higher dose of the drug than the physician would employ during a maintenance regimen, administering the drug more frequently than the physician would administer during a maintenance regimen, or both. The phrase "maintenance regimen" or "maintenance period" refers to a therapeutic regimen (or a portion of a therapeutic regimen) used to maintain a patient during disease treatment, for example, to keep the patient in remission for an extended period of time (months or years). A maintenance regimen may employ continuous therapy (e.g., administering the drug at regular intervals, e.g., weekly, monthly, yearly, etc.) or intermittent therapy (e.g., discontinued treatment, intermittent treatment, treatment upon relapse, or treatment upon reaching a certain predetermined criterion (e.g., symptoms of disease, etc.).

[0023] As used herein, the term "hyaluronic acid" or "HA" refers to a compound having the formula: [ka] (wherein n is the number of repeat units). It refers to a polymer having the following structure. All sources of hyaluronic acid, including bacterial and avian sources, are useful in the present invention. However, hyaluronic acid of bacterial origin is preferred. The hyaluronic acid useful in the present invention has a molecular weight of 15,000 to 50,000 daltons ("low molecular weight"). Preferably, the hyaluronic acid of the present invention has a molecular weight of 25,000 daltons. In some embodiments, the hyaluronic acid of the present invention is administered to a subject in the form of a salt. In some embodiments, sodium, potassium, lithium, calcium, barium, strontium, magnesium, aluminum, or ammonium salts are used. In particular, the hyaluronic acid of the present invention is used in the form of a sodium salt. Commercial sources of hyaluronic acid typically include those from Sigma-Aldrich (e.g., CAS Number: 9067-32-7 or CAS Number: 9067-32-7).

[0024] By "therapeutically effective amount" is meant an amount of HA of the present invention sufficient for the treatment of pulmonary mucosal inflammation with a reasonable benefit / risk ratio applicable to any medical treatment. It will be understood that the total daily usage of the compound will be determined by the attending physician within the scope of sound medical judgment. The specific therapeutically effective dose level for any particular subject will depend on a wide variety of factors, including the subject's age, weight, general health, sex, and diet; the time of administration, the route of administration, and the rate of excretion of the specific compound employed; the duration of treatment; drugs used in combination with or simultaneously with the specific polypeptide employed; and similar factors well known in the medical arts. For example, it is well within the skill of the art to start the dose of the compound at a level lower than that required to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved. However, the daily dosage of the drug may vary over a wide range, from 0.01 to 1,000 mg per adult per day. Preferably, the composition contains 0.01, 0.05, 0.1, 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, 50.0, 100, 250 and 500 mg of active ingredient for symptomatic adjustment of dosage to the subject to be treated. The medicament typically contains about 0.01 mg to about 500 mg of active ingredient, preferably 1 mg to about 100 mg of active ingredient. An effective amount of the drug is usually supplied at a dosage level of 0.0002 mg / kg to about 20 mg / kg (body weight) per day, in particular about 0.001 mg / kg to 7 mg / kg (body weight) per day.

[0025] Moreover, the use of the hyaluronic acid of the present invention for reducing lung mucosal inflammation can combine additional effects.In particular, the hyaluronic acid of the present invention can regulate the differentiation of secretory cells and therefore participate in the regulation of mucin secretion, and can modulate the secretion of chloride ions via CFTR, in particular via F508del / F508del mutant CFTR.

[0026] In some embodiments, hyaluronic acid is administered in combination with a corticosteroid. As used, the term "corticosteroid" has its usual meaning in the art and refers to a class of active ingredients that have a hydrogenated cyclopentoperhydrophenanthrene ring system with anti-inflammatory activity. Corticosteroids typically include cortisone, cortisol, hydrocortisone (11β,17-dihydroxy,21-(phosphonooxy)-pregn-4-ene,3,20-dione disodium), dihydroxycortisone, dexamethasone (21-(acetyloxy)-9-fluoro-1β,17-dihydroxy-16α-m-ethylpregna-1,4-diene-3,20-dione), and highly derivatized steroids such as Beconase (beclomethasone dipropionate, which is 9-chloro-11-β,17,21,trihydroxy-16β-methylpregna-1,4-diene-3,20-dione 17,21-dipropionate). Other examples of corticosteroids include flunisolide, prednisone, prednisolone, methylprednisolone, triamcinolone, deflazacort and betamethasone, and corticosteroids such as cortisone, hydrocortisone, methylprednisolone, prednisone, prednisolone, betamethasone, beclomethasone dipropionate, budesonide, dexamethasone sodium phosphate, flunisolide, fluticasone propionate, triamcinolone acetonide, betamethasone, fluocinolone, fluocinonide, betamethasone dipropionate, betamethasone valerate, desonide, desoximetasone, fluocinolone, triamcinolone, triamcinolone acetonide, clobetasol propionate, and dexamethasone.

[0027] Typically, the active ingredient of the present invention (i.e., the HA of the present invention) is combined with a pharma- ceutically acceptable excipient and, optionally, a sustained release matrix, such as a biodegradable polymer, to form a pharmaceutical composition. The term "pharmaceutical" or "pharmaceutically acceptable" refers to molecular entities and compositions that do not produce adverse, allergic or other untoward reactions when administered to a mammal, particularly a human, as appropriate. A pharma- ceutically acceptable carrier or excipient refers to any kind of non-toxic solid, semi-solid or liquid filler, diluent, encapsulating material or formulation aid. The carrier can also be a solvent or dispersion medium, including, for example, water, ethanol, polyol (e.g., glycerol, propylene glycol, and liquid polyethylene glycol, etc.), suitable mixtures thereof, and vegetable oils. Proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants. The inhibition of microbial action can be achieved by various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include an isotonic agent, such as sugar or sodium chloride. Prolonged absorption of an injectable composition can be achieved by using an agent that delays absorption, such as aluminum monostearate and gelatin, in the composition. In the pharmaceutical composition of the present invention, the active ingredient of the present invention can be administered in unit dosage form as a mixture with a conventional pharmaceutical support. Suitable unit dosage forms include oral route forms, such as tablets, gel capsules, powders, granules, and oral suspensions or solutions, sublingual and buccal dosage forms, aerosols, implants, subcutaneous, transdermal, topical, intraperitoneal, intramuscular, intravenous, subdermal, transdermal, intrathecal, and intranasal dosage forms, and rectal dosage forms. In some embodiments, the pharmaceutical composition of the present invention is administered locally (i.e., to the airways of a subject). Thus, the composition can be formulated in the form of a spray, aerosol, solution, emulsion, or other form known to those skilled in the art. When the method of the present invention involves intranasal administration of the composition, the composition can be formulated in the form of an aerosol, spray, mist, or droplets.In particular, the active ingredient for use according to the present invention can be conveniently delivered in the form of an aerosol spray presentation from a pressurized pack or nebulizer using a suitable propellant (e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas).In the case of a pressurized aerosol, the dosage unit can be determined by providing a valve to deliver a metered amount.Capsules and cartridges (e.g., composed of gelatin) for use in an inhaler or insufflator can be formulated containing a powder mix of the compound and a suitable powder base, such as lactose or starch.

[0028] The present invention will be further illustrated by the following figures and examples, which should not, however, be construed as limiting the scope of the present invention in any way. EXAMPLES

[0029] Anti-inflammatory effects of 15-50 kDa hyaluronic acid on airway epithelium in patients with cystic fibrosis: Sodium forms of 15-50 kDa hyaluronan exhibit anti-inflammatory effects during regeneration and differentiation of human cystic fibrosis (CF) airway epithelia harboring the F508del / F508del class II mutation. Figure 1 Notably, treatment with HA-Na 15-45kDa led to a significant decrease (52%) in IL-8 secretion by human CF airway epithelial cells compared to control conditions, as did treatment with Ha-Na 15-30kDa (57% decrease) and HA-Na 30-50kDa (67.2% decrease).

[0030] Sodium forms of 15-50 kDa hyaluronan exhibit therapeutic anti-inflammatory effects on human cystic fibrosis (CF) airway epithelia harboring the F508del / F508del class II mutation. Figure 2Notably, treatment with HA-Na 15-45kDa led to a significant decrease (32.1%) in IL-8 secretion by human CF airway epithelial cells, as did HA-Na 15-30kDa (44.2% decrease), HA-Na 30-50kDa (42.2% decrease) and the control anti-inflammatory dexamethasone (30.9% decrease).

[0031] Sodium forms of 15-50 kDa hyaluronan show anti-inflammatory effects in combination treatments on human cystic fibrosis (CF) airway epithelia harboring the F508del / F508del class II mutation ( Figure 3 In particular, treatment with HA-Na 15-45kDa led to a significant decrease (33.9%) in IL-8 secretion by human CF airway epithelial cells, as did HA-Na 15-30kDa (40.2% decrease), HA-Na 30-50kDa (30.1% decrease) and the control anti-inflammatory dexamethasone (21.4% decrease).

[0032] Hyaluronan of 15-50 kDa, in its sodium and calcium forms, exhibits anti-inflammatory effects during regeneration and differentiation of human cystic fibrosis (CF) airway epithelia harboring the F508del / F508del class II mutation. Figure 4 Notably, treatment with HA-Na 15-45kDa led to a significant decrease (55.5%) in IL-8 secretion by human CF airway epithelial cells compared to control conditions, as did treatment with Ha-Ca 15-45kDa (59.25% decrease).

[0033] Hyaluronan of 15-50 kDa, in its sodium and potassium forms, exerts healing anti-inflammatory effects on human cystic fibrosis (CF) airway epithelium harboring the F508del / F508del class II mutation ( Figure 5In particular, treatment with 15-45 kDa sodium hyaluronate (HA-Na) led to a significant decrease (32.1%) in IL-8 secretion by human CF airway epithelial cells, as did dexamethasone (30.9% decrease) and 15-45 kDa potassium hyaluronate (47.5% decrease). 15-45 kDa calcium hyaluronate led to a non-significant decrease in IL-8 secretion of 33.6%.

[0034] Hyaluronan of 15-50 kDa, in its sodium and potassium forms, exhibits anti-inflammatory effects in combination treatments on human cystic fibrosis (CF) airway epithelia harboring the F508del / F508del class II mutation ( Figure 6 In particular, treatment with sodium hyaluronate 15-45 kDa (HA-Na 15-45 kDa) led to a significant decrease (33.9%) in IL-8 secretion by human CF airway epithelial cells, as did dexamethasone (21.4% decrease) and potassium hyaluronate 15-45 kDa (30% decrease). Calcium hyaluronate 15-45 kDa led to a non-significant decrease in IL-8 secretion of 11.6%.

[0035] Sodium forms of 15-50 kDa hyaluronan exhibit dose-responsive anti-inflammatory effects during regeneration and differentiation of human cystic fibrosis (CF) airway epithelia harboring the F508del / F508del class II mutation. Figure 7 In particular, treatment with 15-45 kDa sodium hyaluronate (HA-Na) leads to a significant decrease in IL-8 secretion by human CF airway epithelial cells at 2 mg / mL (65% decrease), 1 mg / mL (62% decrease), 0.5 mg / mL (47.3%), but has no further effect at 0.1 mg / mL.

[0036] The sodium form of 15-50 kDa hyaluronan leads to a decrease in MUC-5B mucin secretion during regeneration of human cystic fibrosis (CF) airway epithelium harboring the F508del / F508del class II mutation. Figure 8In particular, treatment with HA-Na 30-50 kDa led to a significant decrease in MUC-5B secretion by human CF airway epithelial cells compared to control conditions (day 15 at ALI: 68.7%; day 25 at ALI: 67.1%), whereas treatment with HA-Na 15-45 kDa and HA-Na 15-30 kDa led to a non-significant decrease in MUC-5B secretion (day 15 at ALI: 37.3% and 14.1%, respectively; day 25 at ALI: 33.2% and 17.3%, respectively).

[0037] Hyaluronan of 15-50 kDa, in its sodium, calcium and potassium forms, leads to a decrease in MUC-5B mucin secretion during regeneration of human cystic fibrosis (CF) airway epithelium harboring the F508del / F508del class II mutation. Figure 9 In particular, treatment with HA-Ca 15-45kDa leads to a significant decrease in MUC-5B secretion by human CF airway epithelial cells at day 15 at ALI (85.5% decrease) and a non-significant decrease at day 25 at ALI (58.8%) compared to control conditions. Treatment with HA-Na 15-45kDa and HA-K 15-45kDa leads to a non-significant decrease in MUC-5B secretion at day 15 at ALI (37.3% and 34.7% decrease, respectively) compared to control conditions. Treatment with HA-Na 15-45kDa leads to a non-significant decrease in MUC-5B secretion by human CF airway epithelial cells at day 25 at ALI (33.2% decrease) compared to control conditions.

[0038] The sodium form of 15-50 kDa hyaluronan leads to a dose-dependent decrease in MUC-5B mucin secretion during regeneration of human cystic fibrosis (CF) airway epithelia harboring the F508del / F508del class II mutation. Figure 10In particular, treatment with HA-Na 15-45kDa leads to a significant decrease in MUC-5B secretion by human CF airway epithelial cells at day 15 at ALI and day 25 at ALI, respectively, at 2 mg / mL, a non-significant decrease in MUC-5B secretion at day 15 at ALI: 37.3% decrease; day 25 at ALI: 33.2% decrease, and no effect at 0.5 mg / mL, compared to control conditions.

[0039] Anti-inflammatory effects of 15-50 kDa hyaluronan on airway epithelium in non-CF patients: Sodium forms of hyaluronan of 15-50 kDa exhibit anti-inflammatory effects during regeneration and differentiation of human airway epithelium from non-CF patients ( Figure 11 Notably, treatment with HA-Na 15-45kDa led to a decrease in IL-8 secretion by human non-CF airway epithelial cells (69.7%) compared to control conditions, as did treatments with HA-Na 15-30 (60.6% decrease) and HA-Na 30-50kDa (66.6% decrease).

[0040] Sodium forms of hyaluronan of 15–50 kDa exhibit therapeutic anti-inflammatory effects on human airway epithelium in non-CF patients suffering from pulmonary inflammation ( Figure 12 Notably, treatment with HA-Na 15-45kDa led to a significant decrease (39.6%) in IL-8 secretion by human non-CF airway epithelial cells, as did HA-Na 15-30kDa (46.1% decrease), HA-Na 30-50kDa (34.8% decrease) and the control anti-inflammatory dexamethasone (21.1% decrease).

[0041] Sodium forms of hyaluronan of 15-50 kDa show anti-inflammatory effects in combination treatments on human airway epithelium from non-CF patients suffering from pulmonary inflammation ( Figure 13Notably, treatment with HA-Na 15-45kDa led to a significant decrease (41.6%) in IL-8 secretion by human non-CF airway epithelial cells, as did HA-Na 15-30kDa (38.1% decrease), HA-Na 30-50kDa (32.1% decrease) and the control anti-inflammatory dexamethasone (33.9% decrease).

[0042] Hyaluronan of 15-50 kDa, in its sodium, calcium and potassium forms, exhibits anti-inflammatory effects during regeneration and differentiation of human airway epithelium from non-CF patients ( Figure 14 In particular, treatment with HA-Na 15-45kDa led to a decrease in IL-8 secretion by human non-CF airway epithelial cells (69.7%) compared to control conditions, as did treatments with Ha-Ca 15-45kDa (70.5% decrease) and HA-K 15-45kDa (57.9% decrease).

[0043] Hyaluronan of 15-50 kDa, in its sodium form, exhibits healing anti-inflammatory effects on human airway epithelium in non-CF patients suffering from pulmonary inflammation ( Figure 15 In particular, treatment with 15-45 kDa sodium hyaluronate (HA-Na) led to a significant decrease (39.6%) in IL-8 secretion by human non-CF airway epithelial cells, as did dexamethasone (a 21.1% decrease). Calcium hyaluronate 15-45 kDa and potassium hyaluronate 15-45 kDa led to a non-significant decrease (6.5% and 17.8%, respectively) in IL-8 secretion.

[0044] Hyaluronan of 15-50 kDa, in its sodium form, exhibits anti-inflammatory effects in combination treatments on human airway epithelium from non-CF patients suffering from pulmonary inflammation ( Figure 16In particular, treatment with sodium hyaluronate 15-45 kDa (HA-Na 15-45 kDa) led to a significant decrease (41.6%) in IL-8 secretion by human non-CF airway epithelial cells, as did dexamethasone (33.9% decrease). Calcium hyaluronate 15-45 kDa and potassium hyaluronate 15-45 kDa led to a non-significant decrease in IL-8 secretion (8.5% and 22.2%, respectively).

[0045] Anti-inflammatory effects of 15-50 kDa hyaluronic acid on airway epithelium in patients with COPD: The sodium form of hyaluronic acid of 15-50 kDa exhibits therapeutic anti-inflammatory effects on human airway epithelium in COPD patients ( Figure 17 In particular, treatment with HA-Na 15-45kDa led to a significant decrease (60.2%) in IL-8 secretion by human COPD airway epithelial cells, as did HA-Na 15-30kDa (65% decrease), HA-Na 30-50kDa (69.5% decrease) and the control anti-inflammatory dexamethasone (68.8% decrease).

[0046] Sodium forms of hyaluronic acid of 15-50 kDa exhibit anti-inflammatory effects in combination treatments on human airway epithelium in COPD patients ( Figure 18 In particular, treatment with HA-Na 15-45kDa led to a significant decrease (33.4%) in IL-8 secretion by human COPD airway epithelial cells, as did HA-Na 15-30kDa (43.1% decrease) and the control anti-inflammatory dexamethasone (32.1% decrease). HA-Na 30-50kDa led to a non-significant (p=0.0511) decrease (30.8% decrease) in IL-8 secretion.

[0047] Hyaluronic acid of 15-50 kDa, in its sodium, calcium and potassium forms, exhibits healing anti-inflammatory effects on human airway epithelium in COPD patients ( Figure 19In particular, treatment with sodium hyaluronate 15-45 kDa (HA-Na) led to a significant decrease (60.2%) in IL-8 secretion by human COPD airway epithelial cells, as did dexamethasone (68.8% decrease) and calcium or potassium hyaluronate 15-45 kDa (HA-Ca 15-45 kDa: 61.5% decrease; HA-K 15-45 kDa: 61.1% decrease).

[0048] Hyaluronic acid of 15-50 kDa, in its sodium form, exhibits anti-inflammatory effects in combination treatments on human airway epithelium in COPD patients ( Figure 20 In particular, treatment with sodium hyaluronate 15-45 kDa (HA-Na 15-45 kDa) led to a significant decrease (33.4%) in IL-8 secretion by human COPD airway epithelial cells, as did dexamethasone (32.1% decrease). Calcium or potassium hyaluronate 15-45 kDa led to a non-significant decrease in IL-8 secretion (12.9% and 27.5% decrease, respectively).

[0049] References: Throughout this application, various references describe the state of the art to which this invention pertains. The disclosures of these references are incorporated by reference into this disclosure.

Claims

1. A pharmaceutical composition for treating pulmonary mucosal inflammation in a subject suffering from cystic fibrosis or chronic obstructive pulmonary disease, comprising hyaluronic acid having a low molecular weight of 15,000 to 50,000 daltons.

2. 10. The pharmaceutical composition of claim 1, wherein the subject suffering from cystic fibrosis carries at least one mutation in the CFTR gene, including, but not limited to, F508del-CFTR, R117H CFTR, and G551D CFTR.

3. The pharmaceutical composition according to claim 1, wherein the pulmonary mucosal inflammation results from a pulmonary infection, particularly a viral infection.

4. The pharmaceutical composition of claim 3, wherein the subject is suffering from COVID-19.

5. The pharmaceutical composition of claim 1 , wherein the hyaluronic acid is administered to the subject in the form of a salt.

6. 6. The pharmaceutical composition of claim 5, wherein the salt is a sodium, potassium, lithium, calcium, barium, strontium, magnesium, aluminum or ammonium salt.

7. 6. The pharmaceutical composition according to claim 5, wherein hyaluronic acid is used in the form of its sodium salt.

8. 2. The pharmaceutical composition of claim 1, wherein hyaluronic acid regulates the differentiation of secretory cells; is involved in the regulation of mucin secretion; and / or modulates the secretion of chloride ions via CFTR, in particular via F508del / F508del mutant CFTR.

9. 10. The pharmaceutical composition of claim 1, wherein the hyaluronic acid is administered in combination with a corticosteroid.

10. 10. The pharmaceutical composition of claim 9, wherein the corticosteroid is selected from the group consisting of flunisolide, prednisone, prednisolone, methylprednisolone, triamcinolone, deflazacort, and betamethasone, and the corticosteroid is, for example, cortisone, hydrocortisone, methylprednisolone, prednisone, prednisolone, betamethasone, beclomethasone dipropionate, budesonide, dexamethasone sodium phosphate, flunisolide, fluticasone propionate, triamcinolone acetonide, betamethasone, fluocinolone, fluocinonide, betamethasone dipropionate, betamethasone valerate, desonide, desoximetasone, fluocinolone, triamcinolone, triamcinolone acetonide, clobetasol propionate, and dexamethasone.

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