WO2024149719A1 - Use of pentosan polysulfate for the treatment of dry cough - Google Patents

Abstract

Disclosed is a composition comprising an effective amount of pentosan polysulfate for use in the treatment of dry cough.

Description

Description

Title of the invention: USE OF PENTOSAN POLYSULFATE FOR THE TREATMENT OF DRY COUGH

[0001] The present invention relates to the use of a composition comprising pentosan polysulfate for the manufacture of a medication intended for the treatment of dry cough.

[0002] Coughing is a normal reaction to irritation of the respiratory tract. Cough is caused by irritation of the throat, trachea and bronchi. Cough is not a disease but a symptom caused by irritation. Cough is therefore generally an endogenous protective and cleansing mechanism.

[0003] Coughs can be grouped into two families: wet coughs and dry coughs.

[0004] A so-called wet cough is a productive or congested cough, intended to evacuate the airways of bodies obstructing these airways, due to the abundant production of phlegm and mucus.

[0005] The defense reflex due to bronchial or nasal secretions, from a wet cough, is reduced as soon as the bodies causing the irritation are expelled.

[0006] A so-called dry cough is a non-productive cough which is characterized by bouts of spasmodic and harsh coughing in which no phlegm is expelled.

[0007] The defense reflex due to the irritating sensations of a dry cough is difficult to alleviate.

[0008] The duration of a cough is variable; it is considered an acute cough if a person has been coughing for less than three weeks, a subacute cough, between three and eight weeks, and a chronic cough, if the cough continues for at least three weeks. beyond eight weeks.

[0009] A dry cough that does not disappear spontaneously can be disabling or harm the quality of life.

[0010] In general, medications to relieve dry cough are so-called antitussive medications. They reduce the cough reflex by acting via the central nervous system. Cough medications are divided into several categories, opiate cough medications, antihistamine cough medications, and non-opiate and non-antihistamine cough medications.

[0011] Opioid antitussive medications have a sedative effect and should not be used in cases of respiratory failure or cough related to asthma. They can only be purchased with a prescription. [0012] Antihistamine cough suppressants are recommended in cases of dry and irritating cough occurring at night, due to their sedative effect. They also have so-called atropinic effects, namely dry mouth, constipation, blockage of urine and visual disturbances.

[0013] Non-opiate and non-antihistamine cough suppressants are indicated in the short-term treatment of dry coughs. They are generally homeopathy medications traditionally offered for benign respiratory conditions.

[0014] Other alternatives have also been proposed, such as herbal supplements or the use of non-steroidal anti-inflammatory drugs.

[0015] Document EP 0 274 845 discloses the combination of ibuprofen as a non-steroidal anti-inflammatory with an opiate cough suppressant.

[0016] Document EP 2 106 258 describes the use of ibuprofen as a non-steroidal anti-inflammatory, as an antitussive, for the treatment of dry cough of viral or bacterial origin.

[0017] Document WO 2014/161811 discloses the use of an astringent substance and a mucilaginous substance, such as Icelandic moss, for the treatment of irritations of the mucous membranes of the mouth and throat, as well as dry cough .

[0018] Document EP 3 086 799 describes the treatment of persistent coughs by a mechanism for physical protection of the mucous membranes of the upper airways, through the use of agents of plant origin.

[0019] The exact mechanism of a chronic cough is not clearly explained to date.

[0020] However, it is widely recognized that a chronic cough is associated with inflammation of the airways.

Cough is generated by the cough reflex, a stimulus induced by the activation of afferent, sensory nerves innervating the respiratory tract. These signals are transmitted centrally via the vagus nerve, which is connected to brainstem networks. Such networks coordinate activation of motor output (e.g., phrenic, intercostal, and recurrent laryngeal nerves (RLN) and the ultimate expression of cough) (Taylor-Clark TE. Peripheral neural circuitry in cough. Curr Opin Pharmacol 2015 Jun;22:9-17.).

[0022] The majority of sensory nerve fibers innervating the airways are capsaicin-sensitive pulmonary vagal afferents (CSLV) (Agostoni, E., Chinnock, J. E., De Daly, M. B., and Murray, J. G. (1957). Functional and histological studies of the vagus nerve and its branches to the heart, lungs and abdominal viscera in the cat. J. Physiol.), nociceptive sensory nerve endings.

[0023] These fibers are very chemosensitive and can be activated by several mechanisms. endogenous diators whose releases are elevated during pulmonary inflammation (Carr, MJ, and Undem, BJ (2003). Bronchopulmonary afferent nerves. Respirology 8, 291-301.; Hsu, CC, Lin, RL, Lee, L. Y, and Lin, Y.

S. (2013). Hydrogen sulfide induces hypersensitivity of rat capsaicin-sensitive lung vagal neurons: role of TRPA1 receptors. Am. J. Physiol. Reg. Integrator Comp. Physiol. 305, R769-R779.).

[0024] Anatomically speaking, there are two types of neurons: sensory neurons with myelinated Aô fibers and sensory neurons with C fibers. Myelinated Aô fibers play a role in generating cough. The Aô fibers are responsible for the violent and sudden cough which occurs during aspiration (Location

T, Undem B J: Neuroplasticity in vagal afferent neurons involved in cough. Pulm Pharmacol Ther 2011, 24(3): 276-279. ; Canning BJ, Mori N, Mazzone SB: Vagal afferent nerves regulating the cough reflex. Respir Physiol Neurobiol 2006, 152(3):223—242.).

[0025] In recent years, enormous progress has been made in our understanding of the neuronal mechanism of itching. As an endogenous neurotransmitter, serotonin has emerged as a key player in both acute and chronic itch (X. Dong (2018): “Peripheral and central mechanisms of itch”. Neuron 98, 482-494.).

[0026] In our body, it seems that 95% of serotonin is synthesized and released by enterochromaffin cells in the intestine, in response to mechanical or chemical stimulation (Linan-Rico, A., et al. (2016) , Front.

10:564.)

[0027] Recently, serotonin and its receptor 5HT7 were identified as a key factor in mediating the sensation of acute and chronic itching of the skin (Morita T, McClain SP, Batia LM, et al. 2015: HTR7 Mediates Serotonergic Acute and Chronic Itch.

[0028] It has been demonstrated that serotonin produces strong and robust itching in mice and rats, as well as in humans (Akiyama et al., 2009: T. Akiyama, A.W. Merrill, M.I. Carstens, et al. Activation of superficial dorsal horn neurons in the mouse by a PAR-2 agonist and 5-HT: potential role in itch J Neurosci, 29, pp. 6691-6699; Wilson Sarah R, Kienzler Michael A, Lyman K, Olsen Anne Sofie B, Wong Justin F, Stucky Cheryl L, et al. 138.)

[0029] C fibers are also involved in the generation of cough. C fiber is believed to be important for dry type cough, which is used to get rid of an itchy feeling in the throat. This feeling is more typical of chronic cough.

[0030] Serotonin and its receptors can also play a similar role in the lungs (Pecova T, Kocan I, Vysehradsky R, Pecova R. Itch and Cough - Similar Role of Sensory Nerves in Their Pathogenesis. Physiol Res. 2020 Mar 27;69(Suppl 1):S43-S54.).

[0031] Indeed, serotonin exerts an intense stimulating effect on the terminals of the pulmonary C fiber via activation of the serotonin 5-HT3 receptor, which can contribute to the hypersensitivity of the respiratory tract under pulmonary inflammation (Potenzieri C, Meeker S, Undem BJ. Activation of mouse bronchopulmonary C-fibers by serotonin and allergen-oval-bumin challenge. J Physiol. 2012 Nov l;590(21):5449-59. ; Hsu CC, Ruan T, Lee LY, Lin YS.

[0032] During infections and tissue inflammation, the number of cells of myeloid origin with a suppressive nature increases very significantly (Ost, M., Singh, A., Peschel, A., Mehling, R., Rieber , N., and Hartl, D. (2016).

[0033] In response to an inflammatory signal, the myeloid cells produced by the bone marrow migrate into the blood and become functionally active, with the aim of restoring or replacing damaged peripheral populations (Panopoulos, A.D., and Watowich, S. S. ( 2008). Granulocyte coloniestimulating factor: molecular mechanisms of action during steady state and emergency hematopoiesis.

[0034] In addition to these conventional immune effector cells, it has been demonstrated that hematopoietic stem cells participate directly in the inflammatory process in respiratory diseases, such as asthma, allergies, etc. (Massberg S, Schaerli P, Knezevic-Maramica I, Kollnberger M, Tubo N, Moseman EA, Huff IV, Junt T, Wagers AJ, Mazo IB, von Andrian UH (2007) “Immunosurveillance by hematopoietic progenitor cells trafficking through blood, lymph, and peripheral tissues. .” Cell. 131(5):994-1008; Allakhverdi, Z., Comeau, M. R., Smith, D. E., Mfuna Endam, L., Desrosiers, M., Liu, ., Howie, K. J., Denburg, J. A., Gauvreau, G. M., Delespesse, G. (2009): “CD34+ hemopoietic progenitor cells are potent effectors of allergic inflammation”. -478; Blanchet, M. R., McNagny, K. M. (2009): “Stem cells, inflammation and allergy”. Hematopoietic stem and progenitor cells in inflammation and allergy”. Frontiers in immunology, 4, 428.).

[0035] These results are confirmed by the work of the inventor: a large quantity of hematopoietic stem cells are visible on biopsies taken from patients suffering from inflammatory diseases. Additionally, serotonin was detected in these hematopoietic stem cells. As mentioned previously, during inflammation, the hematopoietic cells aggregate and produce serotonin, partly responsible for itching, the stimulus for dry cough.

The applicant discovered that pentosan poly sulfate modified the morphology of hematopoietic cells. The modification of the morphology of these cells limits or even prevents their aggregations during an inflammation process, limiting the release of serotonin.

[0038] Pentosan poly sulfate, also called pentosan poly sulfate or pentosan sulfuric polyester, has been used in various therapeutic applications.

Pentosan poly sulfate is an analogue of glucosaminoglycans, derived from heparin, with anticoagulant, fibrinolytic and anti-inflammatory properties.

[0040] Document WO 93/23059 describes the use of composition comprising pentosan polysulfate in the treatment of thrombocytopenia.

[0041] Document WO 2019/023761 discloses the use of pentosan polysulfate in the treatment of diseases or disorders associated with bone marrow pathologies.

[0042] Document EP 1 038 528 describes the use of pentosan polysulfate in the treatment of fibrosis.

[0043] Document EP 1 488 788 describes the use of composition comprising pentosan polysulfate in the treatment of gastrointestinal disorders.

[0044] Document EP 2 164 501 discloses the use of pentosan polysulfate in the treatment of diseases such as asthma, allergic rhinitis and chronic obstructive pulmonary disease.

[0045] Pentosan polysulfate is also used as an active ingredient in creams aimed at combating inflammation. They are used in the local treatment of bruises and hematomas, even muscle and joint pain.

[0046] Pentosan polysulfate is also indicated in the treatment of painful bladder syndrome characterized by glomerulations or Hunner's ulcers in adults suffering from moderate to severe pain, urinary urgency and urinary frequency. Its mechanism of action in the treatment of painful bladder syndrome is poorly understood; pentosan polysulfate seems to act through a local effect in the bladder by binding of glycosaminoglycans to the bladder mucosa and by an anti-inflammatory effect.

[0047] Excessive or chronic cough is a major problem which is poorly managed by currently available therapies.

[0048] Thus, a need for the development of new drugs is felt.

[0049] The present invention proposes to treat excessive dry or chronic coughs. nics by the use of compositions comprising pentosan polysulfate.

[0050] The present invention therefore relates to a composition comprising an effective amount of pentosan polysulfate (PPS) for use in the treatment of dry cough.

[0051] The implementation of the invention will be better understood with the aid of the detailed description which is set out below with reference to the appended figures in which: [0052] Figures 1 to 3 illustrate images of lungs, piece of pulmonary parenchyma and pieces of bronchial rings.

[0053] [Fig 1] is an image representing normal lungs, uniform in color and smooth on the surface.

[0054] [Fig 2] is an image representing a piece of pulmonary parenchyma from a patient suffering from pulmonary fibrosis.

[0055] [Fig 3] is an image representing two pieces of bronchial rings, on the left, of an organ donor without respiratory disease; on the right, of a patient who received a lung transplant.

[0056] [Fig 4] is a close-up image of blood congestion in the lumen of the bronchi of a patient with chronic pulmonary disease.

[0057] Figures 5 and 6 illustrate images obtained by optical microscope of blood congestion cells from an allergic donor, with a x20 magnification.

[0058] [Fig 5] is a contrast phase image.

[0059] [Fig 6] is a fluorescent image showing a subset of blood congestion cells expressing a large quantity of serotonin represented by a red color in real color, and illustrated by light areas.

[0060] Figures 7 and 8 illustrate images obtained by optical microscope of blood congestion cells stained positive for the serotonin 5HT2C receptor by a specific monoclonal antibody, with a x20 magnification.

[0061] [Fig 7] is an image of the contrast phase.

[0062] [Fig 8] is a fluorescent image of the 5-HT2C coloring, green in real color, and illustrated by light areas.

[0063] Figures 9 to 11 illustrate images obtained by optical microscope of blood congestion cells.

[0064] [Fig 9] illustrates a control image of blood congestion cells without treatment, magnification x20.

[0065] [Fig 10] illustrates an image of blood congestion cells after being exposed to an aqueous solution of 0.5% pentosan polysulfate, for 24 hours, the blood congestion cells took the shape of needles, x20 magnification.

[0066] [Fig 11] is an enlargement of Figure 10. [0067] Thus the invention relates to the treatment of dry cough comprising the administration of a composition comprising an effective quantity of pentosan polysulfate, for a duration and under conditions sufficient for the treatment of dry cough.

[0068] More precisely, the invention relates to the use of pentosan polysulfate in the manufacture of a medicine for the treatment of dry cough.

[0069] The term "effective amount" of pentosan polysulfate as used means a sufficient amount of pentosan polysulfate to provide the desired therapeutic or physiological effect.

[0070] The exact quantity required varies from one subject to another, the age and general condition of the subject, the method of administration, etc.

[0071] An appropriate "effective amount" can be determined using routine experimentation. The effective amount in this context includes an amount required to treat dry cough.

[0072] These compositions are proposed for use as pharmaceutical compositions or formulations.

[0073] The compositions include advantageously aqueous and sterile solutions.

[0074] Sterile solutions are prepared by incorporating a required amount of pentosan polysulfate in a suitable solvent, optionally with other active ingredients, followed by sterilization or at least a process to reduce viruses, bacteria or the like. biological entities contaminants at acceptable levels.

[0075] Pentosan polysulfate (PPS) is preferably chosen from the group consisting of: the sodium salt of pentosan polysulfate (NaPPS), the magnesium salt of pentosan polysulfate (MgPPS), the calcium salt of polysulfate pentosan (CaPPS), and the zinc salt of pentosan polysulfate (ZnPPS).

Preferably, the pentosan polysulfate (PPS) is sodium pentosan polysulfate (NaPPS).

[0077] According to embodiments, the medication is administered by inhalation or in the form of a nasal spray, delivered by an inhaler.

According to one characteristic, the mass concentration of pentosan polysulfate, expressed as a percentage, is between 0.5 and 3%.

[0079] According to one embodiment, the mass concentration of pentosan polysulfate, expressed as a percentage, is equal to 1%.

[0080] The solvent is preferably an aqueous saline solution, for example comprising a mass concentration, expressed as a percentage, of sodium chloride of 0.9%, buffered with 10mM HEPES (or 4-(2-hydroxyethyl)-1 acid). -piperazine ethane sulfonic acid).

[0081] Other active ingredients mean an antibiotic or antiviral molecule. [0082] Healthy lungs are more or less uniform in color and shape, as illustrated in Figure 1.

[0083] The pulmonary parenchymas as well as the bronchi of patients suffering from chronic respiratory diseases, such as cystic fibrosis, idiopathic pulmonary fibrosis, sarcoidosis, present areas of aggregation of hematopoietic cells of red colors in real colors and illustrated by dark areas, as shown in Figures 2 to 4.

[0084] These areas of cellular aggregation are commonly called blood congestion of the lungs.

[0085] These blood congestions thus participate directly in inflammatory responses and tissue repair.

[0086] By immunocytochemistry, the applicant discovered that the aggregation of hematopoietic cells in the lungs express a large quantity of serotonin, as illustrated in Figure 8.

[0087] These observations of the presence of serotonin were confirmed by two different antibodies in several independent experiments, with the same results; an example of such confirmation is illustrated in Figure 6.

[0088] Thus, not only the aggregation of hematopoietic cells in the lungs express serotonin, but also a serotonin receptor, 5-HT2C (figure 8).

[0089] On the basis of these data, the applicant proposes the following scheme for chronic cough: during viral or bacterial infections, or tissue damage, pro-inflammatory mediators, including serotonin, are released in the affected area. .

[0090] These mediators will recruit hematopoietic cells from the surrounding lung tissues as well as from the blood circulation.

[0091] Serotonin acts on hematopoietic cells mainly via 5HT2C receptors in a paracrine and autocrine manner, amplifying the inflammatory response.

[0092] A large number of hematopoietic cells will quickly accumulate in the affected area, causing the affected area to appear red in color.

[0093] The hematopoietic cells release an increasing quantity of serotonin which activates the C fibers causing an itching sensation via the 5HT3 or 5HT7 receptor, activating the cough reflex.

[0094] As long as significant areas of aggregation of hematopoietic cells are present, for example as illustrated in Figures 2 and 3, the dry cough should not stop, due to the continued release of serotonin from the aggregates. of hematopoietic cells.

[0095] In addition, dry cough can further increase serotonin secretion, creating a positive feedback loop.

[0096] The applicant has discovered that pentosan polysulfate has a direct effect on hematopoietic cells located in cell aggregates: as an anticoagulant.

[0097] Indeed, pentosan polysulfate considerably modifies the morphology of hematopoietic cells in vitro, as illustrated in Figure 10.

[0098] During treatment with pentosan polysulfate, the hematopoietic cells extracted from the aggregates changed in morphology, towards a needle or nail shape, as illustrated in Figures 10 and 11.

[0099] To aggregate, the cells must have the shape of a disk, as illustrated in Figure 9.

[0100] This morphological change induced by pentosan polysulfate would prevent hematopoietic cells from forming cellular aggregates.

[0101] This hypothesis provides not only an explanation of the functioning of pentosan polysulfate, but also a justification for its use as a drug to break the vicious circle of chronic inflammation in various inflammatory diseases, in particular respiratory diseases, frequently associated to chronic cough.

[0102] Local and direct application of pentosan polysulfate by inhalation or in the form of a nasal spray, to the respiratory tract, will have a double benefit, namely increased drug effectiveness and a reduction in the toxicity of the drug.

[0103] Furthermore, by acting directly on the aggregates of hematopoietic cells, pentosan polysulfate does not alter the cough reflex, thus preserving the protective effects of cough against harmful agents of the respiratory tract.

[0104] Materials and methods:

[0105] The lungs and lung biopsies:

[0106] The pieces of organs illustrated in Figures 2 to 4 were obtained from patients who had undergone a lung transplant or a polypectomy.

All experimental procedures were explained in full, and all subjects received informed consent. The study was conducted in accordance with the Declaration of Helsinki on Biomedical Research (Hong Kong Amendment, 1989), and received approval from the local ethics committee.

[0107] Isolation of hematopoietic stem cells from the lung parenchyma: [0108] The lung tissue samples were dissected into pieces of approximately 5 mm ³ . Approximately 10 g of minced tissue was placed in 50 ml conical tubes, filled with washing buffer (Dulbecco's Phosphate Buffered Saline (DPBS) each containing 100 ml of penicillin and streptomycin). The tubes were inverted to wash the tissue and the supernatant removed, repeating these operations three times. As much liquid as possible was removed, chopped and washed tissues were filtered through a layer of sterile gauze. The tissues on the gauze were distributed into 15 ml Falcon tubes, comprising approximately 1.5 g of tissue per tube. The filtrates were passed through a 10 μm cell strainer. THE Cells were collected by centrifugation.

[0109] Immuno-col oration, Figures 5, 6 and 8:

[0110] The cells are cultured on a coverslip, then fixed with a solution of formaldehyde titrated to 2%, and glutaraldehyde titrated to 2.5% in 0.1 M sodium phosphate buffer, for 30 minutes at room temperature. . The fixed cells are then rinsed several times (3X) with PBS (phosphate-buffered saline). To facilitate antibody access, the fixed cells are then permeabilized with 0.25% Triton-X-100. The fixed cells are rinsed several times (3X) with PBS. A solution of 100 μl of PBS supplemented with 10% serum (bovine, goat, or human, depending on the primary antibody used), was applied for 60 minutes at room temperature. 100 µl, or a volume sufficient to cover the cells, of primary antibody was applied and incubated for 30-60 minutes. Cells were rinsed with PBS 3 times for 2 min. 100 μl, or a volume sufficient to cover the cells was applied, of a solution of secondary antibody conjugated with a fluorophore (Alexa 488, Alexa 560). The coverslip is then mounted on a glass slide for observation under a fluorescence microscope.

[0111] For the sample illustrated in Figure 6, the primary antibody is a specific antibody directed against serotonin (Sigma, S5545). This antibody was produced in rabbits (dilution 1 in 50 in PBS). The secondary antibody used is therefore an anti-rabbit IgG antibody, produced in goats, conjugated with a red fluorochrome [Alexa Fluor 568, Goat anti-rabbit IgG (H+L), dilution 1 in 500 in PBS], ( Invitrogen, Al 1011). The photo was taken on a fluorescence microscope.

[0112] For the sample illustrated in Figure 8, the primary antibody is a specific antibody directed against the Serotonin receptor, 5HT2C (R&D Systems, Minneapolis, MAB5764). This antibody was produced in mice (dilution 1 in 50 in PBS). The secondary antibody used is therefore an anti-mouse IgG antibody, produced in goats, conjugated with a green fluorochrome [Alexa Fluor 488, Goat anti-rabbit IgG (H+L), dilution 1 in 500 in PBS], (Invitrogen, Al 1011). The photo was taken on a fluorescence microscope.

Claims

Claims [Claim 1] Composition comprising an effective amount of poly pentosan sulfate for use in the treatment of dry cough. [Claim 2] Composition for use according to claim 1, characterized in that the mass concentration, expressed as a percentage, of pentosan polysulfate is between 0.5 and O zZo. [Claim 3] Composition for use according to claim 1 or 2, characterized in that it comprises an antibiotic or antiviral molecule. [Claim 4] Composition for use according to any one of claims 1 to 3, characterized in that it is capable of being packaged in an inhaler to be capable of being administered by inhalation or in the form of a nasal spray. [Claim 5] Composition for use according to any one of claims 1 to 4, characterized in that the treatment of dry cough comprises the administration of pentosan polysulfate by inhalation or nasal spray. [Claim 6] Pentosan polysulfate for use in the treatment of dry cough.