Formulation of an inhalable quercetin solution for the management of respiratory diseases

Abstract

The present invention describes the formulation of a phytotherapy product, namely an inhalable solution developed from a natural extract of quercetin, either alone or in combination with resveratrol, rigorously selected for their synergistic effects on the respiratory system and their potential in alleviating symptoms associated with respiratory diseases such as asthma, COPD, lung cancer, COVID-19, and long COVID. The solution is intended to be inhaled using a nebuliser or other inhalation device intended to transform the solution into an aerosol in order to have a direct effect on the bronchi and minimise side effects. Quercetin has anti-inflammatory, antioxidant, immunomodulatory, and antiviral properties. This document describes, through various examples, the manufacturing process developed to obtain a solution for inhalation.

Description

[0001] Title of the invention

[0002] Formulation of a Quercetin solution for inhalation for the management of respiratory diseases.

[0003] Technical field to which the invention relates

[0004] The present invention relates to an inhalation solution based on quercetin alone or combined with other polyphenols such as resveratrol, intended for pulmonary administration for the relief of breathing difficulties linked to asthma, chronic obstructive pulmonary disease "COPD", lung cancer, Covid 19 and the prevention of acute respiratory distress syndrome responsible for the majority of deaths in patients with SARS-COV 2 as well as long covid.

[0005] Purpose of the invention

[0006] Chronic respiratory diseases are illnesses that affect the airways and other parts of the lungs. Among the most common are asthma, COPD, and lung cancer. In addition, with the COVID-19 epidemic, a number of patients are suffering from persistent symptoms called long COVID.

[0007] In recent years, considerable attention has been paid to natural compounds due to their numerous biological effects. Polyphenols are a class of plant derivatives that have been extensively studied to prevent and treat numerous pathologies related to oxidative stress, such as neurodegenerative, respiratory, cardiovascular, inflammatory diseases, and certain cancers.

[0008] Among the most interesting polyphenols, quercetin is the most active, many medicinal plants owe their effectiveness to their high quercetin content. It has demonstrated anti-inflammatory, immunomodulatory, antioxidant and antiviral effects, particularly in rhinitis and numerous respiratory diseases, but quercetin has a low and variable bioavailability by oral route, an intense metabolism and a rapid and complete elimination which means that the oral route is not the most appropriate route for the administration of quercetin; hence the interest of the present invention which aims to administer quercetin by inhalation, therefore directly into the lungs in order to benefit from the anti-inflammatory, immunomodulatory, antioxidant and antiviral properties of this substance for the relief of pulmonary disorders.

[0009] Natural products have become more popular in recent decades due to their natural origin and proven safety. Indeed, several studies have indicated that plant-based products are able to downregulate: the expression, production of inflammatory mediators and their receptors and to inhibit the expression of transcription factors that promote the secretion of mediators. Quercetin and resveratrol have distinguished themselves by their numerous beneficial effects and are the subject of several advanced researches on asthma and COPD, lung cancer and covid 19 as well as long covid in order to further determine their action on the different inflammatory pathologies.Studies have shown that quercetin reduces increased levels of IL-4 and Th2 cytokine production in an OVA-sensitized induced asthma mouse model, while increasing the production of IFN-γ, a Th1 cytokine in quercetin-treated mice. In this model, quercetin administration resulted in significant inhibition of all asthmatic reactions (Park et al 2009).

[0010] Additionally, a pilot clinical trial evaluated the effects of quercetin formulated with the Phytosome® delivery system in subjects with mild to moderate asthma attacks. In this study, quercetin demonstrated excellent results in controlling, preventing, and reducing daily and nighttime symptoms, maintaining a higher peak expiratory flow rate (PEF), and decreasing PEF variability. The additional use of quercetin improved additional asthma management measures, decreasing the use of inhalers, nasal drops, rescue medications, and improving the rhinitis score. Quercetin demonstrated a significant reduction in oxidative stress and a very good safety profile.

[0011] In a preclinical model of chronic obstructive pulmonary disease (COPD), quercetin reduced markers of oxidative stress and lung inflammation, contributing to the decline of this respiratory disease. It is also important to note that quercetin was safely tolerated orally up to 2000 mg / day.

[0012] Molecular docking studies have highlighted that quercetin, a natural polyphenol belonging to the flavanol class, inhibits the 3CLpro, PLpro and S proteins of SARS-CoV-2. Biophysical techniques then confirmed very recently that quercetin is reasonably a potent inhibitor of the 3CLpro protease (Abian O, et al 2020). Previously, Khaerunnisa et al., performed a molecular docking study to examine the affinity between several natural compounds and SARS-CoV-2 proteases, in particular the 6LU7 protein. Their results showed a potential for binding affinity between quercetin and the 6LU7 protein of the SARS-CoV-2 3CLpro protease, which may lead to its inhibition and thus disrupt the virus life cycle.

[0013] Quercetin can be used alone or in combination with other polyphenols. Among the polyphenols that can be combined with quercetin, resveratrol has attracted considerable interest due to its antioxidant, free radical scavenger, and anti-inflammatory activity.

[0014] In the experimental model of ovalbumin-induced allergic asthma, resveratrol significantly reduced IL-5, IL-13, and TGF-[3 in serum and BALF and decreased CD3+CD4+, CD3+CD8+, and CD4+IL-4+ cells with an increase in CD4+CD25+FOXP3+ cells in the lung inflammatory cell infiltrate in the OVA-res group. MiRNA expression arrays using lung infiltrating cells showed that resveratrol caused significant alterations in miRNA expression, specifically downregulating miR-34a expression. In addition, miR-34a was found to target FOXP3, as evidenced by enhanced FOXP3 expression in lung tissue.

[0015] Collectively, these data indicated that resveratrol is a promising polyphenol in the relief of symptoms related to respiratory diseases. However, its low water solubility and increased metabolism lead to low bioavailability, thus limiting its clinical efficacy by oral route, therefore, the route of administration of quercetin solution is ideally suited for this active ingredient. An invention (W02002032410) cited the use of resveratrol for the treatment of inflammatory respiratory pathologies, such as asthma and chronic obstructive pulmonary disease by administering it, among other things, by pulmonary route for these diseases; hence the interest of the present invention, to combine quercetin and resveratrol which will act in synergy in order to maximize the anti-inflammatory, immunomodulatory and antiviral effect for a more effective relief of symptoms associated with respiratory conditions.Taken together, these data indicate significant potential efficacy of using phytochemicals as inhalation therapy for respiratory pathologies.

[0016] In an effort to improve the bioavailability of these substances in order to potentiate their action and offer a product that can help in the management of patients with respiratory diseases, this invention offers a new route of administration of the quercetin solution alone or in combination with resveratrol. This new formula is administered by inhalation via a nebulizer in order to improve the bioavailability of these two very promising molecules.

[0017] State of the prior art

[0018] Quercetin and resveratrol are natural compounds found in certain foods and are being studied for their anti-inflammatory, antioxidant, immunomodulatory, antiviral properties and their potential health benefits. Although they are available as supplements in different formulations (capsules, tablets, etc.), a specific formulation for inhalation was not widely available according to the literature. To our knowledge, an invention (W02002032410) cited the use of resveratrol for the treatment of inflammatory respiratory pathologies, such as asthma and chronic obstructive pulmonary disease by administering it, among other things, via the pulmonary route for these diseases.Also, clinical studies on the use of resveratrol in a nasal spray formula for the treatment of allergic rhinitis and the common cold have been published demonstrating the improvement of respiratory symptoms with this molecule (Miraglia Del Giudice et al. 2014), (ME Baldassarre et al. 2020). Quercetin, alone or combined with resveratrol, will act synergistically to maximize the anti-inflammatory, antioxidant, immunomodulatory and antiviral effect for more effective relief of symptoms associated with respiratory conditions. However, the low bioavailability of these two compounds makes their use difficult. The lack of an inhaled formula of these compounds prompted the development of the present invention.In an effort to improve the bioavailability of these compounds in order to potentiate their action and offer a product that can help in the management of patients with respiratory diseases, this invention offers a new route of administration of the solution of quercetin alone or combined with resveratrol. This new formula is administered by inhalation via a nebulizer in order to improve the bioavailability of these two very promising molecules.

[0019] Statement of figures

[0020] The invention will be better understood with the aid of the following description, with reference to the attached drawings, by way of examples:

[0021] - Figure 1: Chemical formula of Quercetin

[0022] - Figure 2: Chemical formula of resveratrol

[0023] - Figure 3: Conversion of trans-resveratrol to cis-resveratrol

[0024] Presentation of the invention and method of implementation

[0025] The present invention consists of the development of a natural product based on quercetin alone or associated with other polyphenols capable of interfering in the replication of the Sars-cov 2 virus, and of acting as an anti-inflammatory and modulator of the immune response.

[0026] The present invention helps to relieve respiratory symptoms linked to covid 19 and helps prevent the development of acute respiratory distress syndrome (ARDS), the main fatal complication of Covid 19.

[0027] The present invention is in the form of a solution for inhalation by nebulization based on quercetin alone or associated with another polyphenol in order to improve its bioavailability and potentiate its local effect on the lungs.

[0028] Nebulized inhalation involves transforming the solution into a cloud of very fine particles. This route of administration promotes the local action of the inhaled product while reducing systemic side effects. It has the advantage of avoiding the intense hepatic metabolism that some polyphenols undergo.

[0029] In addition to improving the bioavailability of the substances in question, the specificity of the inhalation form for respiratory conditions is to have a direct pulmonary deposition at the level of the target organ and thus to act directly on the lungs for a rapid action on the bronchi. The doses administered are low compared to other forms, particularly the oral form, which greatly reduces the possible occurrence of side effects.

[0030] Due to the low solubility of quercetin in water, several cosolvents can be used for the formulation of inhalation solutions, preferably monopropylene glycol.

[0031]

[0001] The present invention relates to the field of formulation of phytotherapy products.

[0032]

[0002] More specifically, the invention relates to the formulation of an inhalation solution based on quercetin alone or combined with other polyphenols.

[0033]

[0003] The solution of the present invention is indicated in the relief of symptoms related to respiratory diseases

[0034]

[0004] Since inflammation is highly present and plays an important role in the development of respiratory diseases, we have been looking for natural compounds that can have an anti-inflammatory and immunomodulatory effect. Several compounds, such as polyphenols, from medicinal plants have been reported to have anti-inflammatory and immunomodulatory bioactivities. The present invention focused on two polyphenols that have great potential in the management of respiratory diseases due to their anti-inflammatory and immunomodulatory effect.

[0035]

[0005] Lung infections caused by viruses affect lung health and promote exacerbation of respiratory diseases, quercetin has demonstrated beneficial effects against respiratory viruses, including influenza virus, parainfluenza virus, respiratory syncytial virus, adenovirus, rhinovirus and coronaviruses.

[0036]

[0006] Quercetin (Figure 1) is a flavonoid from the polyphenol group with anti-inflammatory, antioxidant, antihistamine and antiviral properties,

[0037]

[0007] Discovered in 1937 by Albert Szent-Gyorgyi, quercetin belongs to the family of flavonol-type flavonoids, pigments naturally present in plants. It is particularly concentrated in certain fruits and vegetables

[0038]

[0008] Numerous in vitro studies have shown that quercetin is an excellent antioxidant. Of all flavonoids, it is even the most powerful scavenger of reactive oxygen species ERO (or free oxygen radicals). Previous studies have suggested that consuming a diet rich in quercetin reduces the risk of developing asthma, bronchial hyperreactivity and chronic obstructive pulmonary disease. In addition to its antiviral action against rhinoviruses, coronaviruses, influenza, and inflammatory syncytial virus.

[0039] Quercetin has been reported as a long-lasting anti-inflammatory substance that possesses strong anti-inflammatory capabilities. Its anti-inflammatory potential can be expressed on different cell types, both in animal and human models. It can also play a modulatory, biphasic and regulatory action on inflammation and immunity by interfering with the expression and activation of transcription factors and protein kinases. Indeed, several studies have demonstrated an inhibitory action of quercetin on the transcription factor NFkB, STAT, as well as on several protein kinases along the molecular signaling pathways of immune mechanisms.

[0040] Furthermore, several clinical trials are underway to evaluate the potential of quercetin as a therapeutic agent in the management of respiratory diseases.

[0041] These data support the use of quercetin as an antiviral and immunomodulatory agent.

[0042] The effect of quercetin can be enhanced by combining it with other polyphenols such as resveratrol:

[0043] Combined with resveratrol, these two polyphenols will act synergistically to relieve respiratory symptoms and modulate the immune response linked to respiratory diseases and infections.

[0044]

[0009] Quercetin has a low and variable bioavailability by oral route, moreover, it is rapidly eliminated by the body, with an elimination half-life of 1 to 2 hours, it undergoes a rapid and extensive metabolism which makes the presumed biological effects of in vitro studies unlikely to apply in vivo, hence the interest of the present invention, which aims to administer quercetin by inhalation, therefore directly into the lungs in order to benefit from the anti-inflammatory and antiviral properties of this substance for the relief of symptoms linked to pulmonary damage.

[0045]

[0010] The effect of quercetin can be enhanced by other polyphenols, such as resveratrol

[0046]

[0011] Resveratrol (Figure 2) is a polyphenol of the stilbene family.

[0047] The chemical formula of resveratrol is: C14H12O3. This molecule contains double bonds and exists in two isoforms, trans-resveratrol which is the bioactive form and cis-resveratrol, with the transition from the trans form to the Cis form by UV irradiation,

[0048]

[0012] The form used in the present invention is the trans-resveratrol form (figure 3), it is also possible to use the cis form or a combination of cis and trans.

[0013] Resveratrol has extensive biological properties, it is a powerful antioxidant with multiple benefits. It has an anti-inflammatory, immunomodulatory effect, as well as an effect on cardiovascular protection, not to mention an antiviral effect against rhinoviruses, coronaviruses, influenzae, and other respiratory viruses. Numerous publications based on in vivo and in vitro tests attest to the benefits of resveratrol on respiratory diseases, including rhinitis and broncho-obstructive diseases, asthma, allergies, lung cancers, viral infections, hence the indication of the present invention.

[0049] Several in vitro and in vivo studies have demonstrated the role that resveratrol plays in inhibiting viral infections due to, among others, Respiratory Syncytial Virus, Chickenpox virus, Influenza and many others.

[0050] Resveratrol has also been reported to inhibit the production of proinflammatory cytokines, indicating that it is both an antiviral and anti-inflammatory therapeutic agent for the treatment of infections.

[0051] The mechanism by which resveratrol expresses these actions is located at the level of cellular signaling pathways in response to pathogen infections. Indeed, resveratrol has an inhibitory action on the nuclear translocation of the nuclear transcription factor NFkB. NFkB represents a family of inducible transcription factors that regulate a wide range of genes involved in the main cellular mechanisms; among others in the various processes of immune and inflammatory responses. The overactivation of NF-KB is closely linked to the deregulation of inflammatory responses and their exacerbation, hence the interest in developing therapy capable of modulating the expression of this factor.

[0052] Numerous studies have demonstrated the inhibitory effects of resveratrol on NF-kB activation and NF-kB-induced target gene expression (primarily genes that encode proinflammatory cytokine synthesis, lymphocyte recruitment, and various other immune responses). Resveratrol's action appears to target components along the NFkB signaling pathway, rather than the factor itself.

[0053] Furthermore, several clinical trials are underway to evaluate the potential of resveratrol in the management of respiratory diseases.

[0054] Based on these studies, it can be concluded that resveratrol has great potential in the management of respiratory diseases, making it the ideal candidate to enhance the effects of quercetin.

[0055]

[0014] The present invention consists of a solution based on quercetin alone or combined with resveratrol, carefully chosen for their biological effects and their safety. Given the low bioavailability of quercetin, the solution is intended to be nebulized or inhaled in the form of vapor.

[0056] Furthermore, an invention (WO2002032410) has claimed the use of resveratrol for the treatment of inflammatory respiratory pathologies, such as asthma and chronic obstructive pulmonary disease by administering, preferably orally or pulmonarily, an active agent selected from the group consisting of resveratrol, pharmacologically acceptable salts, esters, amides, prodrugs and analogues thereof, or any combinations thereof. The invention also relates to pharmaceutical formulations, designed to be used in conjunction with the above method. The invention claims synergy with corticosteroids, NSAIDs, antibiotics, bronchodilators and not with quercetin or other natural products.

[0057]

[0015] In a second aspect, the present invention relates to the formulation of the inhalation solution, in particular the choice of solvent and the manufacturing process.

[0058]

[0016] The subject of the present invention is a solution for inhalation, preferably by nebulization, where the active substances are solubilized in a Co solvent.

[0059]

[0017] In one embodiment, the solvent used to solubilize the active ingredients is propylene glycol.

[0060]

[0018] Propylene glycol is a well-known and widely used excipient in the food and pharmaceutical fields. It is recognized as harmless and safe (Generally recognized as safe: GRAS) by the FDA (21 CFR184, 1666) as a food additive. It is used in the pharmaceutical field by several routes of administration such as: topical, intramuscular, intravenous, nasal as well as by inhalation, which proves the safe nature of this excipient.

[0061]

[0019] In other embodiments, other excipients may also be introduced into the solution for the purposes of the formulations such as an osmolality adjuster such as glucose or sodium chloride, a pH adjuster such as citric acid or sodium hydroxide, or a humectant such as glycerin, or a surfactant such as polysorbate 20 or 80.

[0062]

[0020] Flavorings can also be added in very small quantities, in order to make nebulization more pleasant and promote good compliance with the treatment. A natural mint-type flavor is preferable because in addition to the safe nature of the substance, it also has a nasal decongestant effect, promoting inhalation of the product.

[0063]

[0021] In an additional embodiment, other vehicles may be used to dissolve the active substances, including polyethylene glycol 400 and Di ethylene glycol ethyl ether, caprylocaproyl Polyoxyl-8 glycerides, glycerin

[0022] Within the scope of the present invention, the quercetin and resveratrol solution may be nebulized directly, although it may be diluted in a volume of physiological serum before administration.

[0064]

[0023] Since the solubility of the active ingredients is low in water, it is important to respect a ratio: active solution / physiological serum to prevent the active ingredients from becoming resuspended.

[0065]

[0024] The ratio of active solution / physiological serum can be 3 / 2, 5 / 4 but preferably 5 / 3 for the reconstitution of an aqueous solution of quercetin alone or associated with resveratrol. It can also be used as is, more particularly in the case where the solution already contains physiological saline or water for injectable preparation.

[0066]

[0025] , The concentration of polyphenols in the solution can range from 0.1 to 3% for each active ingredient, preferably 0.3 - 1% and more preferably 0.35% for each substance.

[0067]

[0026] In view of the low solubility of the active ingredients, a micronized grade is highly desirable to promote the dissolution of the substances in the vehicles.

[0068]

[0027] Examples:

[0069] Table 01: Examples of formulation of quercetin-based inhalation solution: The above formulation examples relate to particular embodiments which make it possible to illustrate the general description of the present invention; it is clear that the invention is in no way limited to said examples:

[0070] The manufacturing steps are summarized below:

[0071] Example F1:

[0072] (i) Dissolve the large mint crystals in propylene glycol by heating to a temperature of 40 - 45 °C and stir until completely dissolved.

[0073] (ii) Add quercetin and stir until completely dissolved

[0074] (iii) Take 3 ml of the solution and add 2 ml of physiological serum or EPPI and ensure the formation of a clear solution and the absence of suspended particles.

[0075] Example F2:

[0076] (i) Mix propylene glycol and glycerin

[0077] (ii) Dissolve the large mint crystals in the mixture from step (i) by heating to a temperature of 40 - 45 °C and stir until dissolved.

[0078] (iii) Add quercetin and stir until completely dissolved

[0079] (iv) Take 5 ml of the solution and add 3 ml of physiological serum to it and ensure the formation of a clear solution and the absence of suspended particles.

[0080] Example F3:

[0081] (i) Dissolve the large mint crystals in PEG 400 by heating to a temperature of 60-65°C and stir until completely dissolved.

[0082] (ii) Allow to cool to 40°C then add the quercetin and stir until completely dissolved.

[0083] (iii) Take 5 ml of the solution and add 3 ml of physiological serum to it and ensure that a clear solution is formed and that there are no suspended particles.

[0084] Example F4:

[0085] (i) Dissolve the large mint crystals in monopropylene glycol by heating to a temperature of 40-45°C and stir until completely dissolved.

[0086] (ii) Add quercetin and stir until completely dissolved

[0087] (iii) Once the quercetin is dissolved, add the resveratrol and stir until completely dissolved. (iv) Take 5 ml of the solution and add 3 ml of physiological saline to it and ensure that a clear solution is formed and that there are no suspended particles.

[0088] (v)

[0028] In another formulation mode, it is also possible to add water as a solvent in the formula, in this case, the ratio, cosolvent / Water must be studied in order to ensure total dissolution of the active ingredients and avoid their resuspension.

[0089] (vi)

[0029] In this embodiment the water used can be purified water, physiological water or even water for injection.

[0090] (vii)

[0030] Given the route of administration, the solution obtained must undergo sterilization

[0091] (viii)

[0031] Different sterilization methods can be used, in particular, sterilizing filtration as well as sterilization by heat (dry or wet).

[0092] (ix)

[0032] In the present invention, the solution obtained is inhaled by nebulization using a nebulizer which will transform the preparation into an aerosol, different nebulizers can be used: pneumatic or ultrasonic

[0093] (x)

[0094] (xi)

[0033] A nebulizer consists of different parts: compressor, pumping tube, nebulization chamber, mouthpiece and mask. The compressor's role is to push the air into the nebulization chamber, the latter converts the solution into an aerosol which will be inhaled through the mask or mouthpiece.

[0095] (xü)

[0096]

[0034] It is possible to use any other type of inhalation device generating a cold nebulizer but also inhalation devices generating a vapor of the solution by heating the solution such as electric thermal vaporization devices.

Claims

Claims 1- The formulation of a solution for inhalation by nebulization based on quercetin alone or associated with other polyphenols in order to benefit from the advantages of this (its) substance(s) known to undergo hepatic metabolism which strongly limits their effectiveness by oral route. 2- The formulation, according to claim 1, is characterized by the use of a Co-solvent to solubilize quercetin and the associated active ingredients. 3- The composition of the formulation according to claim 1 is characterized by the solubilization of the active ingredient(s) in monopropylene glycol. 4- The composition of the formulation according to claim 1, is characterized by the solubilization of the active ingredient(s) in diethylene glycol ethyl ether, PEG 400, glycerin, or caprylocaproyl polyoxyl 8 glyceride. 5- The composition of the formulation according to claim 1, 3 and 4 is characterized by the association of these cosolvents. 6- The composition of the formulation according to claim 1 is characterized by the association with a second polyphenol which would be resveratrol. 7- The composition of the formulation according to claim 1 is characterized by a concentration of polyphenols in the solution ranging from 0.1 to 3% for each polyphenol. 8- The use of the solution of this present invention according to claim 1, is characterized for the relief of respiratory conditions related to respiratory diseases such as asthma, COPD and lung cancer. 9- The use of the solution of this present invention according to claim 1, is characterized by the relief of symptoms related to pulmonary infections caused by viruses such as Corona virus. 10 - The inhalation of the solution according to claim 1, is characterized by the use of any type of inhalation device generating a nebulizer or a vapor of the solution such as electric thermal vaporization devices.

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