Novel omega-3 carrier formulations for inhaled drug delivery to treat pulmonary inflammation

Abstract

Inflammatory disorders of the lungs are induced by COVID-19 disease, asthma, and numerous other disorders. Omega-3 fatty acids (O3FA), particularly docosahexaenoic acid (DHA), docosapentaenoic acid (DPA) or eicosapentaenoic acid (EPA), are known to have anti-inflammatory and pro-resolving properties. Reduction of lung pathology and inflammation is achieved by a mixture of O3FAs suitable for nebulization using formulations and nebulization devices known in the art. O3FAs may be delivered as ethyl esters (EE) or free fatty acid (FFA) oils or non-phosphate-containing glycerolipids or phospholipids (PL) or any other form known in the art. The mixture can also be delivered either inhaled droplets or inhaled solids, which are other inhalation forms known in the art. The patient inhales for a therapeutically effective period of time to reduce inflammation and improve breathing.

Description

[Technical field]

[0001] The present invention provides novel forms of pharmaceutical grade omega-3 fatty acids (O3FAs or OFAs), specifically docosahexaenoic acid (DHA), docosapentaenoic acid (DPA) and / or eicosapentaenoic acid (EPA), that can be medicinally delivered to the lungs by nebulization or other means to reduce inflammation associated with conditions including COVID-19, asthma and numerous other disorders via direct inhalation. [Background technology]

[0002] The highly unsaturated fatty acids (HUFAs) omega-3 (ω3 or n-3), specifically docosahexaenoic acid (DHA, 22:6n-3), docosapentaenoic acid (DPA, 22:5n-3) and eicosapentaenoic acid (EPA, 20:5n-3), are naturally occurring healthy fats found in marine products such as fish, fish oil, squid oil, krill oil, marine oil supplements and microalgae. Plant-based omega-3s are obtained in the form of alpha-linolenic acid (ALA), the only omega-3 essential fatty acid. ALA is naturally converted by the body to the longer-chain omega-3 fatty acids docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA). Plant sources include nuts and seeds such as chia seeds, Brussels sprouts, walnuts, flaxseeds and soybean oil, which are rich in ALA, as well as green leafy vegetables and beans, which contain lesser amounts. Omega-3 is ubiquitous in mammalian tissues and is a bioactive component of cell membrane phospholipids, anchoring proteins to cell membranes and serving as a precursor for signaling molecules such as eicosanoids and docosanoids. Alteration of omega-3 through biosynthesis inhibition or receptor-mediated action remains a promising strategy for developing beneficial drug targets used in OTC and prescription drugs such as aspirin, nonsteroidal anti-inflammatory drugs (ibuprofen, naproxen) and leukotriene receptor inhibitors (zafirlukast, montelukast, zileuton). Eicosanoids and docosanoids have a wide range of functions in the cardiovascular, pulmonary, nervous, immune and endocrine systems of the body.

[0003] The global COVID-19 pandemic caused by the SARS-CoV-2 virus is partly enigmatic due to its extreme range of symptoms, from completely asymptomatic, possibly including through-air transmissibility from person to person, to respiratory failure and death within days of diagnosis. Since the first COVID-19 patient appeared in Wuhan, China, approximately 220 countries and territories have been affected, and as of July 2, 2021, more than 183,849,133 people have tested positive worldwide, with 3,979,872 deaths. The term "cytokine storm", previously confined to specialist journals, has begun to appear in the general press due to the severity of the out-of-control inflammation. A further striking event is widespread thrombosis with pulmonary pathology similar to SARS and MERS. Platelet-fibrin clots in small arterial vessels are consistent with coagulopathy. In the end stage, multiple organ failure with severe liver damage is noted, consistent with thrombotic microangiopathy. Both inflammation and thrombosis are mediated by signaling molecules derived from HUFA, or more precisely by their mutual mixtures that are always present in membranes.

[0004] (2003) Beyond the general inflammatory / prothrombotic potential of the HUFA environment, the spike protein of the SARS-CoV virus induces cyclooxygenase-2 (COX-2), one of the key synthetic enzymes in eicosanoid synthesis. Moreover, induction of COX-2 may be required for efficient early replication of mouse hepatitis virus, also a coronavirus. To the extent that this is true for SARS-CoV-2, individuals that exhibit a robust COX-2 response to viral COX-2 induction, thereby supporting rapid viral replication, may be particularly susceptible to a prothrombotic / proinflammatory HUFA environment. Inhibition of COX-2 early in infection by known inhibitors (e.g. celecoxib (Celebrex®)) is expected to reduce viral replication. COX-2 selective inhibitors are as effective in arthritis as high-dose omega-3 HUFA, and evidence from randomized controlled trials (RCTs) shows that regular consumption of omega-3 rich salmon in the context of an “anti-inflammatory dietary portfolio” reduces rheumatoid arthritis symptoms. Most COX-2 selective inhibitors have been removed from the market due to enhanced thrombotic events due to rebalancing the eicosanoid milieu by COX-1, which increases the production of thromboxane and thromboxane A2. Since severe COVID-19 is considered to be a thrombophilic event in nature, COX-2 selective inhibitors are not effective against it and may worsen the symptoms.

[0005] However, a balanced HUFA environment may be particularly important in avoiding a COX-2-enhanced cytokine storm, or hypercoagulopathy with characteristics of a thrombotic storm. Inherited genetic risk factors may increase the risk of thrombotic events or have a synergistic effect on the increased risk during hypercoagulable periods, such as severe COVID-19.

[0006] Drug options for treating lung inflammation may include corticosteroids or glucocorticoids or leukotriene receptor antagonists, as well as many other drugs, such as budesonide, prednisone, methylprednisolone, hydrocortisone, or montelukast. However, many of these drugs are associated with undesirable side effects that may cause additional health risks or physical discomfort. Omega-3 fatty acids (O3FAs) are edible in their natural form and are also available as supplements. Omega-3 fatty acids have been shown to prevent several types of lung disease, such as COVID-19, asthma, cystic fibrosis, COPD, pneumonia, tuberculosis, emphysema, pulmonary edema, lung cancer, acute respiratory distress syndrome (ARDS), asbestosis, bronchiectasis, interstitial lung disease (ILD) including sarcoidosis, idiopathic pulmonary fibrosis, and autoimmune diseases.

[0007] Omega-3 fatty acids are always administered systemically, primarily orally, but less commonly intravenously. When administered orally, omega-3 fatty acids are primarily provided in four common forms: as ethyl esters (EEs), as non-phosphate-containing glycerolipids selected from the group including triacylglycerols (TGs), diacylglycerols and / or monoacylglycerols, as phospholipids (PLs), and as free fatty acids (FFAs), also known as non-esterified fatty acids (NEFAs). In foods, TAGs and PLs are by far the predominant forms, with FFAs present in smaller amounts. In humans, only small amounts of FAs, EEs, are present and are usually endogenously synthesized by the ingestion of ethanol (alcohol). FAs EEs are usually synthesized by industrial processes from the natural form (mainly TAGs) and further purified. When administered intravenously, omega-3s are provided in emulsions, primarily as TAGs, but also in small amounts as PLs, which may also be derived from emulsifiers. FAs EEs may be administered IV as emulsions.

[0008] Systemic administration of any of these forms results in rapid hydrolysis ("lipolysis") of all forms, and the resulting free FFAs enter the normal biochemical pathways that exist for transporting and distributing FAs to the bloodstream, which perfuses all organs. Small amounts of O3FAs are trapped and re-esterified to various lipid classes within cells. In the bloodstream, O3FAs are rapidly taken up in a non-targeted manner by all tissues, thereby distributing the oral or intravenous dose to all organs. Thus, only a small fraction of the administered dose is taken up by certain tissues, such as the lungs.

[0009] The bioactivity / effectiveness of O3FAs against pathologies depends on the target tissue, more specifically the O3FA concentration in the target lipids of the target tissue. For example, the effectiveness against lung pathologies depends on the specific concentration of O3FAs in lung tissue, more specifically the concentration in PL present in cell membranes and possibly surfactant lipids. Due to the non-targeted nature of systemic administration, any particular dose is less effective than an equivalent dose delivered directly to the target organ. More specifically, any particular dose is less effective for treating lung pathologies when administered systemically as one of the common forms, compared to an equivalent dose administered directly to the lungs.

[0010] Pulmonary lipids are unique among tissues due to the high secretion of pulmonary surfactant lipids, which is required to reduce surface tension and allow a large surface area for gas exchange. Pulmonary surfactant is composed of highly saturated PLs. Summary of the Invention

[0011] The present invention provides a novel pharmaceutical composition for reducing risk factors associated with pulmonary inflammation, which can be administered in the form of inhalation or spray. The present invention also relates to a method for treating lung disorders associated with inflammation, such as bronchiolitis, asthma, cystic fibrosis, COPD, pneumonia, tuberculosis, emphysema, pulmonary edema, lung cancer, acute respiratory distress syndrome (ARDS), interstitial lung disease (ILD), including asbestosis, bronchiectasis, sarcoidosis, idiopathic pulmonary fibrosis, autoimmune diseases, acute pulmonary thrombosis, acute or chronic lung inflammation, inflammatory conditions of the heart and its blood vessels, or to inhibit bronchoconstriction, optionally in combination with a fast-acting bronchodilator, and to inhibit conditions associated with acute respiratory distress, such as COVID-19, and also to central nervous system disorders, including inflammatory CNS disorders. Patients may be of any age, from newborns to the elderly.

[0012] The present invention provides novel pharmaceutical compositions of omega-3 fatty acids comprising a therapeutically effective amount of docosahexaenoic acid (DHA), docosapentaenoic acid (DPA), eicosapentaenoic acid (EPA), or a combination of any two or all three of DHA, DPA, EPA, present as free fatty acids (FFA) or ethyl esters (EE) or phospholipids (PL) or non-phosphate-containing glycerolipids selected from the group including triacylglycerols (TG), diacylglycerols and / or monoacylglycerols.

[0013] The pharmaceutical composition may include a pharma- ceutical acceptable carrier and O3FA or a combination of O3FA and melatonin or budesonide. In other embodiments, the pharmaceutical composition includes O3FA alone, O3FA and CBD, O3FA and phospholipids, O3FA and glycerolipids, and O3FA and other drugs. The pharmaceutical composition containing the active ingredient may be administered by inhalation or nebulization or other routes compatible with inhalation. The composition of the present invention may be administered to humans or animals to treat lung inflammation. The composition may be formulated as a suspension or emulsion and is safe because it contains edible, natural O3FA.

[0014] The formulations include phospholipids and / or glycerolipids to deliver therapeutically effective amounts of docosahexaenoic acid (DHA), docosapentaenoic acid (DPA), eicosapentaenoic acid (EPA), or a combination of any two or all three of DHA, DPA, and EPA. [Brief description of the drawings]

[0015] [Figure 1] Relative lung weights observed on days 8 and 15 [Figure 2-1] Microscopic images of lungs from individual animals [Figure 2-2] Same as above [Figure 2-3] Same as above [Figure 2-4] Same as above [Figure 2-5] Same as above [Figure 2-6] Same as above [Diagram 3] IL-6 in BALF on days 8 and 15 [Figure 4] TNF-α in BALF observed on days 8 and 15 [Diagram 5] Plasma IL-10 observed on days 8 and 15 [Figure 6] Plasma TGF-β observed on days 8 and 15 DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0016] Large amounts of proinflammatory omega-6 arachidonic acid (AA) suppress the synthesis and accumulation of anti-inflammatory omega-3 DHA, DPA, and EPA in cell membranes. AA-derived eicosanoids control immunopathological processes ranging from inflammatory responses to tissue remodeling. AA-derived prostaglandin (PG) synthesis occurs upon the liberation of AA in membranes by PLA2, so PG synthesis is limited by the supply of AA. DHA, an alternative to AA, is known to be an inhibitor of PG synthesis by cyclooxygenase. Dietary EPA and AA compete for incorporation into membrane phospholipids, but also for biosynthesis from their respective FA precursors, since the biosynthetic enzymes are the same. AA-derived metabolites mediate inflammation in lung diseases, whereas DHA-, DPA-, and EPA-derived metabolites resolve inflammation and coagulation.

[0017] The present invention is a simple and highly effective method for treating pulmonary inflammation via inhalation route. The formulation was tested in a lipopolysaccharide (LPS)-induced acute pulmonary inflammation Wistar rat model. Male rats aged 6-8 weeks at the time of dosing were selected for the study. Rats were selected and divided into groups based on their body weight using a stratified randomization method. After randomization, rats were divided into 11 groups with 6 rats in each group (Table 16).

[0018] Multiple studies in animals and humans have shown that both inhaled and systemic corticosteroids cause immunosuppression and inhibit the induction of antiviral type I interferon responses against various respiratory viruses, including COVID-19. These are usually undesirable side effects. In India and other parts of the world, a significant increase in the incidence of fungal infections such as invasive aspergillosis or mucormycosis, a life-threatening angioinvasive maxillofacial fungal infection, due to corticosteroid administration has been reported in many individuals suffering from COVID-19, especially in diabetic patients. In patients with very strong viral diseases, extensive immunosuppression may not be advisable. This novel invention has significant advantages over corticosteroids. O3FA is edible, natural, anti-inflammatory and antithrombotic, and the safety and efficacy of oral and intravenous lipid emulsions have been established in young children and adults. A double-blind randomized clinical trial showed that O3FA improves the levels of several respiratory and renal function parameters in severe COVID-19 patients. The invention involves the administration of O3FA without substantial adverse or side effects.

[0019] The inventors have found a novel anti-inflammatory formulation using O3FA. In rat experiments, the relative content of DHA, DPA, and EPA caused a significant reduction in lung pathology and inflammation. The inventors have also used combinations of O3FA with melatonin or budesonide, O3FA with cannabinoids (CBD), and O3FA with phospholipids and glycerolipids, which also showed a reduction in inflammation. Other combinations include combinations of O3FA with pirfenidone, apremilast, roflumilast, tiotropium bromide, nintedanib, isoniazid, streptomycin, tetrahydrocannabinol (THC), montelukast, and other additional active ingredients.

[0020] The pharmaceutical composition of the present invention can be used to treat acute symptoms or as a "continuation therapy". In the continuation therapy, the pharmaceutical composition can be administered as needed to suppress symptoms, and the dose of each administration can be the same or different depending on the improvement of symptoms. The pharmaceutical composition can also be used to normalize local deficiency of O3FA, which can normalize local inflammation caused by proinflammatory omega-6 fatty acids such as arachidonic acid.

[0021] The present invention provides methods for the treatment of pulmonary inflammatory disorders such as asthma, chronic obstructive pulmonary disease including cystic fibrosis and chronic sinusitis, interstitial fibrosis, COVID-19, and other disorders of the lung such as bronchiolitis, pneumonia, tuberculosis, emphysema, pulmonary edema, lung cancer, acute respiratory distress syndrome (ARDS), asbestosis, bronchiectasis, interstitial lung disease (ILD) including sarcoidosis, idiopathic pulmonary fibrosis, autoimmune diseases, CNS disorders, etc. According to an embodiment, the method includes administration of an anti-inflammatory and anti-thrombotic natural O3FA agent by inhaling a nebulizer. The anti-inflammatory and anti-thrombotic agent can be administered alone or with one or more additives, such as melatonin, CBD, phospholipids. The O3FA can be an esterified component of a phospholipid.

[0022] We have now discovered the first formulations of O3FA delivered by nebulization as a suspension or emulsion to reduce LPS-induced acute pulmonary inflammation in patients with contraindications to NSAIDs.

[0023] O3FAs were delivered primarily in the form of FFAs to enhance uptake into lung tissue and minimize lipoid pneumonia. Monotherapy with ethyl ester (O3EE) was the most common form of oral O3 supplementation. No symptoms associated with excess lipid accumulation were observed, and O3EE treatment was the most effective in treating the effects of LPS.

[0024] COVID-19 long haulers are people who survive the acute illness but have long-term symptoms. They have been found to suffer from fibromyalgia, fatigue, and sleep disorders. O3FA is known to be beneficial in treating arthritis and neuropathic pain associated with fibromyalgia syndrome (FMS) due to its anti-inflammatory properties. Melatonin helps reduce anxiety, pulmonary fibrosis, and manage insomnia in COVID-19 patients. In this embodiment, a combination of O3FA and melatonin can be administered to resolve FMS in long haulers and help patients get better sleep.

[0025] Omega-3FA is a precursor for the synthesis of endocannabinoids. Endocannabinoid epoxides derived from omega-3FA have potent anti-inflammatory properties. Cannabidiol exerts a wide range of anti-inflammatory and immunomodulatory effects and can suppress uncontrollable cytokine storm during acute lung injury. In this embodiment, dual administration of O3FA and CBD is used to resolve lung inflammation in COVID-19 patients.

[0026] COPD caused 3.23 million deaths in 2019 and is the third leading cause of death worldwide. Elevated IL-6 levels in exhaled breath condensate samples are associated with airway inflammation in COPD patients. Overexpression of TNF-α in both human and animal models showed pathological changes consistent with both emphysema and pulmonary fibrosis. Histology and computed tomography imaging of mouse lungs showed changes including enlarged airspaces, reduced small airspaces, increased collagen, and thickened pleural septa. Increased expression of TGF-β is seen in lung samples taken from COPD patients. IL-10 levels are elevated in COPD patients. Elevated serum IL-1β levels are associated with airway inflammation in COPD patients. High intakes of omega-3 fatty acids are associated with a reduced risk of severe exacerbations and improved health-related quality of life and reduced respiratory symptoms in COPD patients.

[0027] Several studies have shown IL-6 as an important tumor-promoting cytokine in NSCLC. IL-6 levels are elevated in serum and exhaled breath condensate samples from NSCLC patients and are associated with tumor size. By inducing epithelial-mesenchymal transition in lung cancer cells, IL-6 and TNF-α can promote invasion and metastasis of NSCLC. Increased expression of TGF-β has been found to be associated with lymph node metastasis and tumor angiogenesis in NSCLC. In patients with late-stage NSCLC, increased expression of IL-10 has been found in tumor-associated macrophages. IL-1β is a key mediator of inflammatory response initiation in NSCLC and is also a potent inducer of the COX2-PGE2 pathway, resulting in immunosuppression. Cachexia is a common finding in lung cancer. In NSCLC patients undergoing chemoradiotherapy, weight was maintained with oral omega-3 supplementation. Lung cancer patients with higher plasma phospholipid EPA concentrations maintained their weight better. Most of the treatments in our O3FA study significantly reduced levels of IL-6, TNF-α, TGF-β, IL-10, and IL-1β.

[0028] The following examples are provided to illustrate the present invention and are not intended to limit the scope of the invention. EXAMPLES

[0029] The following embodiments were tested in animal models. Syria Number Group Treatment Diluent / Formulation Group 1 Normal control Saline Group 2 Disease control Saline + LPS Group 3 Test formulation 1 Placebo emulsion Group 4 Test formulation 2 OFA emulsion (50 mg / mL) 5 Group 5 Test formulation 3 OFA emulsion diluted to 25 mg / mL with saline 6 Group 6 Test formulation 4 FO emulsion Group 7 Control formulation 1 Budesonide (0.25 mg / mL) Group 8 Control formulation 2 Montelukast sodium (4 mg / mL) 9 Group 9 Control formulation 3 Melatonin in OFA (1 mg / mL) Group 10 - Montelukast sodium (4mg / mL) + OFA emulsion 11 Group 11 Test formulation 5 OFA + LPC emulsion

[0030] Example 1: (Test Formulation 1, Group 3) [Table 1]

[0031] Manufacturing method All ingredients were weighed according to the manufacturing recipe. 90% Milli-Q water was placed in a manufacturing container, glycerol and sodium bicarbonate were added, and the mixture was dissolved by stirring at 400 rpm for 3 minutes. Lipoid E 80 S was added to the above solution, and the mixture was heated and dissolved in a water bath maintained at a temperature of 55°C (50°C to 60°C).

[0032] Preparation of emulsions High shear homogenization The resulting mixture was homogenized at 8500 rpm for 15 minutes while maintaining the product temperature at 45° C. (40° C. to 50° C.) The volume was then adjusted to the required level with the remaining amount of Milli-Q water, and homogenized at 8500 rpm for 15 minutes to produce a crude emulsion.

[0033] High Pressure Homogenization The resulting coarse emulsion was subjected to high pressure homogenization in three passes at different pressures, namely pass 1 at 10,000 psi, pass 2 at 18,000 psi, and pass 3 at 18,000 psi, after which the product was cooled to room temperature to obtain the emulsion.

[0034] [Table 2]

[0035] Example 2: (Test Formulation 2, Group 4) [Table 3]

[0036] Manufacturing method All ingredients were weighed according to the manufacturing recipe.

[0037] Preparation of the oil phase An accurately measured amount of omega fatty acid oil was placed in a manufacturing vessel, Lipoid E 80 S was added thereto, and the mixture was heated and dissolved in a water bath at 55°C (50°C to 60°C) to obtain an oil phase.

[0038] Preparation of the aqueous phase 90% Milli-Q water was placed in a separate manufacturing vessel, and stirred at 400 rpm on a magnetic stirrer while maintaining the temperature at 55°C (50°C to 60°C), and glycerol and sodium bicarbonate were dissolved to prepare an aqueous phase.

[0039] Preparation of emulsions High shear homogenization The oil phase was added to the water phase and homogenized at 8500 rpm for 15 minutes while maintaining the product temperature at 45°C (40°C to 50°C).Then, the volume was adjusted to the required level with the remaining amount of Milli-Q water and homogenized at 8500 rpm for 15 minutes to produce a crude emulsion.

[0040] High Pressure Homogenization The resulting coarse emulsion was subjected to high pressure homogenization in three passes at different pressures, namely pass 1 at 10,000 psi, pass 2 at 18,000 psi, and pass 3 at 18,000 psi, after which the product was cooled to room temperature to obtain the emulsion.

[0041] The physical parameters of the emulsions, particle size distribution and zeta potential (ZP) were measured. [Table 4]

[0042] [Table 5]

[0043] Example 3: (Test Formulation 3, Group 5) Test Formulation 2 diluted 50% with saline The physical parameters of the emulsions, particle size distribution and zeta potential (ZP) were measured.

[0044] [Table 6]

[0045] Example 4: (Test Formulation 4, Group 6) [Table 7]

[0046] Manufacturing method All ingredients were weighed according to the manufacturing recipe.

[0047] Preparation of the oil phase An accurately weighed amount of fish oil was placed in a production vessel, Lipoid E 80 S was added thereto, and the mixture was heated and dissolved in a water bath at 55°C (50°C to 60°C) to obtain an oil phase.

[0048] Preparation of the aqueous phase 90% Milli-Q water was placed in a separate manufacturing vessel, and stirred at 400 rpm on a magnetic stirrer while maintaining the temperature at 55°C (50°C to 60°C), and glycerol and sodium bicarbonate were dissolved to prepare an aqueous phase.

[0049] Preparation of emulsions High shear homogenization The oil phase was added to the water phase and homogenized at 8500 rpm for 15 minutes while maintaining the product temperature at 45°C (40°C to 50°C).Then, the volume was adjusted to the required level with the remaining amount of Milli-Q water and homogenized at 8500 rpm for 15 minutes to produce a crude emulsion.

[0050] High Pressure Homogenization The resulting coarse emulsion was subjected to high pressure homogenization in three passes at different pressures, namely pass 1 at 10,000 psi, pass 2 at 18,000 psi, and pass 3 at 18,000 psi, after which the product was cooled to room temperature to obtain the emulsion.

[0051] [Table 8]

[0052] [Table 9]

[0053] Example 5: (Control Formulation 2; Group 8) [Table 10]

[0054] Manufacturing method 90% Milli-Q water was placed in a manufacturing container. A measured amount of montelukast sodium was added and dissolved by stirring at 400 rpm. The remaining volume was adjusted with Milli-Q water. The sample was stored at 2-8°C, protected from light.

[0055] [Table 11]

[0056] Example 6: (Control Formulation 3; Group 9) [Table 12]

[0057] Manufacturing method 90% omega fatty acid oil was placed in a manufacturing vessel. A measured amount of melatonin was added and dissolved by stirring at 400 rpm. The volume was adjusted with the remaining OFA. The samples were stored at 2-8°C, protected from light.

[0058] Example 7: (Test Formulation 5, Group 11) [Table 13]

[0059] Manufacturing method All ingredients were weighed according to the manufacturing recipe.

[0060] Preparation of the oil phase An accurately measured amount of omega fatty acid oil was placed in a manufacturing vessel, egg lecithin was added to it, and it was heated and dissolved in a water bath at 55°C (50°C to 60°C). The mixture was cooled to 40°C, and cannabidiol (LPC) was added and dissolved to obtain an oil phase.

[0061] Preparation of the aqueous phase 90% Milli-Q water was placed in a separate manufacturing vessel, and stirred at 400 rpm on a magnetic stirrer while maintaining the temperature at 55°C (50°C to 60°C), and glycerol and sodium bicarbonate were dissolved to prepare an aqueous phase.

[0062] Preparation of emulsions High shear homogenization The oil phase was added to the water phase and homogenized at 8500 rpm for 15 minutes while maintaining the product temperature at 45°C (40°C to 50°C).Then, the volume was adjusted to the required level with the remaining amount of Milli-Q water and homogenized at 8500 rpm for 15 minutes to produce a crude emulsion.

[0063] High Pressure Homogenization The resulting coarse emulsion was subjected to high pressure homogenization in three passes at different pressures, namely pass 1 at 10,000 psi, pass 2 at 18,000 psi, and pass 3 at 18,000 psi, after which the product was cooled to room temperature to obtain the emulsion.

[0064] The physical parameters of the emulsions, particle size distribution and zeta potential (ZP) were measured. [Table 14]

[0065] Table 15: Physical parameter data for omega fatty acids and LPC emulsion [Table 15]

[0066] Acute Pulmonary Inflammation Study Evaluation of the efficacy of the test formulation against LPS-induced acute pulmonary inflammation in a Wistar rat model.

[0067] A total of 66 male Wistar rats were used and assigned to 11 groups (Groups 1 (G1) to G11) with 6 animals (06) per group (Group (G) description and treatments are shown in Table 16). The experts performing the experiment were blinded to the treatment / control drugs and only knew the group number (G1, G2, etc.).

[0068] On each dosing day, a fresh LPS solution of 2 mg / mL was prepared using sterile saline. Animals in groups G2 to G11 were intraperitoneally administered LPS at a dose of 2 mg / kg body weight once daily. Animals in group G1 did not receive LPS. LPS was administered once daily, and test or control products were administered twice daily.

[0069] LPS injections were given once daily in the morning. One hour later, each group received either the test or control treatment. They were given the test or control treatment again in the afternoon. This continued daily for 7 or 14 days.

[0070] All animals were restrained and exposed to the test or control articles by inhalation using a standard nebulizer. Animals were observed for overt clinical signs throughout the treatment period and until the end of the study, as well as for local effects during and after administration of the test articles.

[0071] All animals gained weight during the study. No mortality was observed during the study.

[0072] After arriving at the laboratory, the rats were allowed to acclimate in cages for 7 days without treatment. During the acclimatization period, none of the animals showed any clinical signs. During the treatment period, some clinical signs were observed in animals from groups G6 (treatment 4), G9 (treatment 7) and G11 (treatment 9). Immediate eye irritation and lacrimation were observed after nebulization treatment. These clinical effects subsided within 30 minutes after nebulization. In the treatment apparatus, the rats' eyes are exposed to the test / control drug vapors by a nebulizer.

[0073] One day after the seven-day treatment was completed, i.e., the eighth day after the start of treatment, three rats from each group were anesthetized with isoflurane and blood was collected. Plasma was separated for further analysis. The animals were then euthanized and bronchoalveolar lavage fluid (BALF) and lung tissue samples were collected. The lung tissue samples were promptly collected and stored in 10% neutral buffered formalin (NBF) for further histopathological analysis process.

[0074] One day after the 14-day treatment was completed, i.e., 15 days after the start of treatment, the remaining three rats from each group were anesthetized with isoflurane and blood was collected. Plasma was separated for further analysis. The animals were then euthanized and bronchoalveolar lavage fluid (BALF) and lung tissue samples were collected. Lung tissue samples were promptly collected and stored in 10% neutral buffered formalin (NBF) for further histopathological analysis process.

[0075] lung weight Results are shown in Figure 1 as the mean ± SD / time point (days 8 and 15) of 3 rats. Relative lung weights in the control group (G1) were compared with all treatment groups (G2–G11) using one-way ANOVA and Dunnett's multiple comparisons. *p<0.05; **p<0.01; ***p<0.001.

[0076] Lung histology Histopathological observations were performed on lung tissue samples fixed in 10% NBF. One slide was prepared for each animal and stained with H&E stain. All gross lesions were examined.

[0077] Histopathological evaluation External findings - Gross pathology External examination of male animals from the normal control group (G1), disease control group (G2) and treatment groups (G3, G4, G5, G6, G7, G8, G9, G10, G11) revealed no lesions of pathological significance.

[0078] Internal findings - gross pathology Internal examination of male animals belonging to normal controls (G1), disease controls (G2) and treatment groups (G3, G4, G5, G6, G7, G8, G9, G10, G11) revealed no pathological abnormalities.

[0079] Microscopic findings Various microscopic changes in normal controls (G1), disease controls (G2), and treatment groups (G3–G11) are shown in Figure 2.

[0080] The lungs showed mild alveolar histiocytosis [2 / 6 (G2), 3 / 6 (G3), 3 / 6 (G4), 3 / 6 (G5), 1 / 6 (G6), 1 / 6 (G7), 2 / 6 (G9), 4 / 6 (G10), 2 / 6 (G11)], mild alveolar histiocytosis [3 / 6 (G2), 1 / 6 (G4), 1 / 6 (G5), 1 / 6 (G6), 3 / 6 (G8)], and moderate alveolar histiocytosis [1 / 6 (G2)].

[0081] In the lungs, alveolar septa with slight vacuolation [2 / 6 (G3), 1 / 6 (G4), 2 / 6 (G5)] and alveolar septa with mild vacuolation [1 / 6 (G6)] were observed.

[0082] The mean severity scores of alveolar histiocytosis in different groups were G1-0.0, G2-1.83, G3-0.50, G4-0.83, G5-0.83, G6-0.50, G7-0.17, G8-1.0, G9-0.33, G10-0.67, and G11-0.33.

[0083] The mean severity scores of alveolar septa with vacuolation in the different groups were G1-0.0, G2-0.0, G3-0.33, G4-0.17, G5-0.33, G6-0.33, G7-0.0, G8-0.0, G9-0.0, G10-0.0, and G11-0.0.

[0084] For all results, statistical significance is expressed as follows: *p<0.05; **p<0.01; ***p<0.001.

[0085] Animals on budesonide (B-ref) had the lowest scores, based on one animal having a mild score and the remaining scores being normal. Treatments including O3 had less than half the severity scores compared to Cd.

[0086] Minor vacuolization was observed in 1–2 animals in the EPL, O3, O3-0.5, and O3EE groups.

[0087] Immune parameters Collected BALF samples were analyzed for IL-6, IL-1β and TNF-α levels in all animals, while IL-10 and TGF-β were estimated in plasma samples.

[0088] IL-6 The results of IL-6 levels are shown in Figure 3. Mean ± SD / time point (days 8 and 15) of 3 rats are shown in Figure 3. IL-6 values ​​in the LPS control group (G2) were compared with all treatment groups (G3-G11) using one-way ANOVA and Dunnett's multiple comparisons. *p<0.05; **p<0.01; ***p<0.001.

[0089] TNF-α The results of TNF-α levels are shown in Figure 4. Mean ± SD / time point (days 8 and 15) of 3 rats are shown in Figure 4. TNF-α values ​​in the LPS control group (G2) were compared with all treatment groups (G3-G11) using one-way ANOVA and Dunnett's multiple comparisons. *p<0.05; **p<0.01; ***p<0.001.

[0090] IL-10 The results of IL-10 levels are shown in Figure 5. Mean ± SD / time point (days 8 and 15) of 3 rats are shown in Figure 5. IL-10 values ​​of the LPS control group (G2) were compared with all treatment groups (G3-G11) using one-way ANOVA and Dunnett's multiple comparisons. *p<0.05; **p<0.01; ***p<0.001.

[0091] TGF-β The results of TGF-β levels are shown in Figure 6. Mean ± SD / time point (days 8 and 15) of 3 rats are shown in Figure 6. TGF-β values ​​in the LPS control group (G2) were compared with all treatment groups (G3-G11) using one-way ANOVA and Dunnett's multiple comparisons. *p<0.05; ***p<0.001.

[0092] Interpretation of immune data COVID-19 is an enigma. Dhar et al. reviewed that multiple studies have shown that both pro- and anti-inflammatory cytokines are elevated in COVID-19 patients. IL-6 and IL-10 were found to predict COVID-19 disease severity. Dramatic elevation of IL-6 and IL-10 levels is a hallmark of cytokine storm in COVID-19 patients. Persistent viral stimulation and IL-6, IL-10 and TNF-α levels are indicators of T cell exhaustion in COVID-19 patients. Elevated IL-6 and TNF-α levels are significant predictors of COVID-19 severity and death.

[0093] In COVID-19 patients, elevated levels of proinflammatory IL-6 are associated with elevated body temperature, elevated CRP and ferritin inflammatory markers, pulmonary inflammation, and widespread lung damage. The test therapeutic agent significantly reduced IL-6 levels (Figure 3).

[0094] Tissue necrosis factor alpha (TNF-α) is a well-known pro-inflammatory molecule. TNF-α is upregulated in most inflammatory conditions and contributes to alterations in blood clotting. Elevated TNF-α along with IL-6 and IL-10 are indicators of a hyperinflammatory response, contributing to the cytokine storm in COVID-19 patients. Excessive amounts of ferritin in COVID-19 patients also reflect excess TNF-α levels. The test therapeutic agent significantly reduced TNF-α levels (Figure 4).

[0095] IL-10 is a pleiotropic cytokine whose primary function in most tissues is to suppress inflammatory responses. However, in COVID-19, IL-10 is dramatically elevated. This phenomenon in COVID-19 is thought to be a negative feedback mechanism to suppress inflammation. IL-10 is known to provide energy to T cells during viral infection. The test therapeutic agent significantly reduced IL-10 levels compared to the positive control LPS group without therapeutic agent (Figure 5).

[0096] Transforming growth factor beta (TGF-β) is a multifunctional cytokine that plays a major role in inflammatory conditions. TGF-β, together with IL-6, promotes the differentiation of T helper 17 (Th17) cells that promote inflammation and reinforce autoimmune conditions. In addition, TGF-β promotes the differentiation of IL-10-producing T cells that lack suppressive function, thus promoting tissue inflammation. TGF-β promotes pulmonary fibrosis in COVID-19 patients. The tested therapeutic agent reduced TGF-β levels (Figure 6).

[0097] IL-1β is a proinflammatory cytokine crucial for host defense responses against infections, antimicrobial immune inflammation and autoimmune inflammation. IL-1β levels are associated with cytokine storm in a subset of COVID-19 patients. IL-1β expression levels have been found to be significantly increased in the bronchial walls of asthmatic patients. Meanwhile, in healthy human volunteers, fish oil treatments reduced IL-1β production. Our O3FA test treatment O3EE significantly reduced IL-1β levels at both time points, and the reduction was better than that in the B-Ref group.

[0098] Table 16: Test and control groups and treatments After randomization, the rats were divided into 11 groups with 6 rats in each group. Animals in each group were administered their respective treatments by nebulization twice daily for 7 days (3 rats / group) or 14 days (3 rats / group). The concentration and dose of nebulization for each group are shown in the table below. [Table 16]

[0099] Example 8: [Table 17]

[0100] Manufacturing method All ingredients were weighed according to the manufacturing recipe.

[0101] Preparation of the oil phase An accurately measured amount of omega-3 acid ethyl ester was placed in a manufacturing vessel, egg lecithin was added thereto, and the mixture was heated and dissolved in a water bath at 55°C (50°C to 60°C) to obtain an oil phase.

[0102] Preparation of the aqueous phase 90% Milli-Q water was placed in a separate manufacturing container, and stirred at 400 rpm on a magnetic stirrer while maintaining the temperature at 55°C (50°C to 60°C), and glycerol was dissolved to prepare an aqueous phase.

[0103] Preparation of emulsions High shear homogenization The oil phase was added to the water phase and homogenized at 8500 rpm for 15 minutes while maintaining the product temperature at 45°C (40°C to 50°C).Then, the volume was adjusted to the required level with the remaining amount of Milli-Q water and homogenized at 8500 rpm for 15 minutes to produce a crude emulsion.

[0104] High Pressure Homogenization The resulting coarse emulsion was subjected to high pressure homogenization in three passes at different pressures, namely pass 1 at 10,000 psi, pass 2 at 18,000 psi, and pass 3 at 18,000 psi, after which the product was cooled to room temperature to obtain the emulsion.

[0105] The physical parameters of the emulsions, particle size distribution and zeta potential (ZP) were measured. [Table 18]

[0106] [Table 19]

[0107] Example 9: [Table 20]

[0108] Manufacturing method All ingredients were weighed according to the manufacturing recipe.

[0109] Preparation of the oil phase An accurately measured amount of omega-3 acid ethyl ester was placed in a manufacturing vessel, egg lecithin was added thereto, and the mixture was heated and dissolved in a water bath at 55°C (50°C to 60°C). A measured amount of cannabidiol was added to the above mixture, and the mixture was heated and dissolved in a water bath at 45°C (40°C to 50°C) to obtain an oil phase.

[0110] Preparation of the aqueous phase 90% Milli-Q water was placed in a separate manufacturing container, and stirred at 400 rpm on a magnetic stirrer while maintaining the temperature at 55°C (50°C to 60°C). Glycerol and sodium bicarbonate were dissolved in that order to prepare an aqueous phase.

[0111] Preparation of emulsions High shear homogenization The oil phase was added to the water phase and homogenized at 8500 rpm for 15 minutes while maintaining the product temperature at 45°C (40°C to 50°C).Then, the volume was adjusted to the required level with the remaining amount of Milli-Q water and homogenized at 8500 rpm for 15 minutes to produce a crude emulsion.

[0112] High Pressure Homogenization The resulting coarse emulsion was subjected to high pressure homogenization in three passes at different pressures, namely pass 1 at 10,000 psi, pass 2 at 18,000 psi, and pass 3 at 18,000 psi, after which the product was cooled to room temperature to obtain the emulsion.

[0113] The physical parameters of the emulsions, particle size distribution and zeta potential (ZP) were measured. [Table 21]

[0114] [Table 22]

[0115] Example 10: [Table 23]

[0116] Manufacturing method All ingredients were weighed according to the manufacturing recipe.

[0117] Preparation of the oil phase An accurately measured amount of omega-3 acid ethyl ester was placed in a manufacturing vessel, egg lecithin was added thereto, and the mixture was heated and dissolved in a water bath at 55°C (50°C to 60°C). A measured amount of cannabidiol was added to the above mixture, and the mixture was heated and dissolved in a water bath at 45°C (40°C to 50°C) to obtain an oil phase.

[0118] Preparation of the aqueous phase 90% Milli-Q water was placed in a separate manufacturing container, and stirred at 400 rpm on a magnetic stirrer while maintaining the temperature at 55°C (50°C to 60°C). Glycerol and sodium bicarbonate were dissolved in that order to prepare an aqueous phase.

[0119] Preparation of emulsions High shear homogenization The oil phase was added to the water phase and homogenized at 8500 rpm for 15 minutes while maintaining the product temperature at 45°C (40°C to 50°C).Then, the volume was adjusted to the required level with the remaining amount of Milli-Q water and homogenized at 8500 rpm for 15 minutes to produce a crude emulsion.

[0120] High Pressure Homogenization The resulting coarse emulsion was subjected to high pressure homogenization in three passes at different pressures, namely pass 1 at 10,000 psi, pass 2 at 18,000 psi, and pass 3 at 18,000 psi, after which the product was cooled to room temperature to obtain the emulsion.

[0121] The physical parameters of the emulsions, particle size distribution and zeta potential (ZP) were measured. [Table 24]

[0122] [Table 25]

[0123] Example 11: [Table 26]

[0124] Manufacturing method All ingredients were weighed according to the manufacturing recipe.

[0125] Preparation of the oil phase An accurately measured amount of omega-3 acid ethyl ester was placed in a manufacturing vessel, egg lecithin was added thereto, and the mixture was heated and dissolved in a water bath at 55°C (50°C to 60°C). A measured amount of cannabidiol was added to the above mixture, and the mixture was heated and dissolved in a water bath at 45°C (40°C to 50°C) to obtain an oil phase.

[0126] Preparation of the aqueous phase 90% Milli-Q water was placed in a separate manufacturing container, and stirred at 400 rpm on a magnetic stirrer while maintaining the temperature at 55°C (50°C to 60°C). Glycerol and sodium bicarbonate were dissolved in that order to prepare an aqueous phase.

[0127] Preparation of emulsions High shear homogenization The oil phase was added to the water phase and homogenized at 8500 rpm for 15 minutes while maintaining the product temperature at 45°C (40°C to 50°C).Then, the volume was adjusted to the required level with the remaining amount of Milli-Q water and homogenized at 8500 rpm for 15 minutes to produce a crude emulsion.

[0128] High Pressure Homogenization The resulting coarse emulsion was subjected to high pressure homogenization in three passes at different pressures, namely pass 1 at 10,000 psi, pass 2 at 18,000 psi, and pass 3 at 18,000 psi, after which the product was cooled to room temperature to obtain the emulsion.

[0129] The physical parameters of the emulsions, particle size distribution and zeta potential (ZP) were measured. [Table 27]

[0130] [Table 28]

[0131] Example 12: [Table 29]

[0132] Manufacturing method All ingredients were weighed according to the manufacturing recipe.

[0133] Preparation of the oil phase An accurately measured amount of omega-3 acid ethyl ester was placed in a manufacturing vessel, egg lecithin was added thereto, and the mixture was heated and dissolved in a water bath at 55°C (50°C to 60°C). A measured amount of cannabidiol was added to the above mixture, and the mixture was heated and dissolved in a water bath at 45°C (40°C to 50°C) to obtain an oil phase.

[0134] Preparation of the aqueous phase 90% Milli-Q water was placed in a separate manufacturing container, and stirred at 400 rpm on a magnetic stirrer while maintaining the temperature at 55°C (50°C to 60°C). Glycerol and sodium bicarbonate were dissolved in that order to prepare an aqueous phase.

[0135] Preparation of emulsions High shear homogenization The oil phase was added to the water phase and homogenized at 8500 rpm for 15 minutes while maintaining the product temperature at 45°C (40°C to 50°C).Then, the volume was adjusted to the required level with the remaining amount of Milli-Q water and homogenized at 8500 rpm for 15 minutes to produce a crude emulsion.

[0136] High Pressure Homogenization The resulting coarse emulsion was subjected to high pressure homogenization in three passes at different pressures, namely pass 1 at 10,000 psi, pass 2 at 18,000 psi, and pass 3 at 18,000 psi, after which the product was cooled to room temperature to obtain the emulsion.

[0137] The physical parameters of the emulsions, particle size distribution and zeta potential (ZP) were measured. [Table 30]

[0138] [Table 31]

[0139] Example 13: [Table 32]

[0140] Manufacturing method All ingredients were weighed according to the manufacturing recipe.

[0141] Preparation of the oil phase An accurately measured amount of omega-3 acid ethyl ester was placed in a manufacturing vessel, egg lecithin was added thereto, and the mixture was heated and dissolved in a water bath at 55°C (50°C to 60°C). A measured amount of cannabidiol was added to the above mixture, and the mixture was heated and dissolved in a water bath at 45°C (40°C to 50°C) to obtain an oil phase.

[0142] Preparation of the aqueous phase 90% Milli-Q water was placed in a separate manufacturing container, and stirred at 400 rpm on a magnetic stirrer while maintaining the temperature at 55°C (50°C to 60°C). Glycerol and sodium bicarbonate were dissolved in that order to prepare an aqueous phase.

[0143] Preparation of emulsions High shear homogenization The oil phase was added to the water phase and homogenized at 8500 rpm for 15 minutes while maintaining the product temperature at 45°C (40°C to 50°C).Then, the volume was adjusted to the required level with the remaining amount of Milli-Q water and homogenized at 8500 rpm for 15 minutes to produce a crude emulsion.

[0144] High Pressure Homogenization The resulting coarse emulsion was subjected to high pressure homogenization in three passes at different pressures, namely pass 1 at 10,000 psi, pass 2 at 18,000 psi, and pass 3 at 18,000 psi, after which the product was cooled to room temperature to obtain the emulsion.

[0145] The physical parameters of the emulsions, particle size distribution and zeta potential (ZP) were measured. [Table 33]

[0146] [Table 34]

[0147] Example 14: [Table 35]

[0148] Manufacturing method All ingredients were weighed according to the manufacturing recipe.

[0149] Preparation of the oil phase An accurately measured amount of omega-3 acid ethyl ester was placed in a manufacturing vessel, egg lecithin was added thereto, and the mixture was heated and dissolved in a water bath at 55°C (50°C to 60°C). A measured amount of cannabidiol was added to the above mixture, and the mixture was heated and dissolved in a water bath at 45°C (40°C to 50°C) to obtain an oil phase.

[0150] Preparation of the aqueous phase 90% Milli-Q water was placed in a separate manufacturing container, and stirred at 400 rpm on a magnetic stirrer while maintaining the temperature at 55°C (50°C to 60°C) to dissolve glycerol and sodium bicarbonate, and then pirfenidone was dispersed therein to prepare an aqueous phase.

[0151] Preparation of emulsions High shear homogenization The oil phase was added to the water phase and homogenized at 8500 rpm for 15 minutes while maintaining the product temperature at 45°C (40°C to 50°C).Then, the volume was adjusted to the required level with the remaining amount of Milli-Q water and homogenized at 8500 rpm for 15 minutes to produce a crude emulsion.

[0152] High Pressure Homogenization The resulting coarse emulsion was subjected to high pressure homogenization in three passes at different pressures, namely pass 1 at 10,000 psi, pass 2 at 18,000 psi, and pass 3 at 18,000 psi, after which the product was cooled to room temperature to obtain the emulsion.

[0153] Example 15: [Table 36]

[0154] Manufacturing method All ingredients were weighed according to the manufacturing recipe.

[0155] Preparation of the oil phase An accurately measured amount of omega-3 acid ethyl ester was placed in a manufacturing vessel, to which egg lecithin was added, and the mixture was heated and dissolved in a water bath at 55°C (50°C to 60°C). A measured amount of cannabidiol was added to the above mixture, and the mixture was heated and dissolved in a water bath at 45°C (40°C to 50°C), and then apremilast was dissolved / dispersed to obtain an oil phase.

[0156] Preparation of the aqueous phase 90% Milli-Q water was placed in a separate manufacturing vessel, and stirred at 400 rpm on a magnetic stirrer while maintaining the temperature at 55°C (50°C to 60°C), and glycerol and sodium bicarbonate were dissolved to prepare an aqueous phase.

[0157] Preparation of emulsions High shear homogenization The oil phase was added to the water phase and homogenized at 8500 rpm for 15 minutes while maintaining the product temperature at 45°C (40°C to 50°C).Then, the volume was adjusted to the required level with the remaining amount of Milli-Q water and homogenized at 8500 rpm for 15 minutes to produce a crude emulsion.

[0158] High Pressure Homogenization The resulting coarse emulsion was subjected to high pressure homogenization in three passes at different pressures, namely pass 1 at 10,000 psi, pass 2 at 18,000 psi, and pass 3 at 18,000 psi, after which the product was cooled to room temperature to obtain the emulsion.

[0159] Example 16: [Table 37]

[0160] Manufacturing method All ingredients were weighed according to the manufacturing recipe.

[0161] Preparation of the oil phase An accurately measured amount of omega-3 acid ethyl ester was placed in a manufacturing vessel, to which egg lecithin was added and dissolved by heating on a water bath at 55°C (50°C to 60°C). A measured amount of cannabidiol was added to the above mixture and dissolved by heating on a water bath at 45°C (40°C to 50°C), and then roflumilast was dissolved / dispersed to obtain an oil phase.

[0162] Preparation of the aqueous phase 90% Milli-Q water was placed in a separate manufacturing vessel, and stirred at 400 rpm on a magnetic stirrer while maintaining the temperature at 55°C (50°C to 60°C), and glycerol and sodium bicarbonate were dissolved to prepare an aqueous phase.

[0163] Preparation of emulsions High shear homogenization The oil phase was added to the water phase and homogenized at 8500 rpm for 15 minutes while maintaining the product temperature at 45°C (40°C to 50°C).Then, the volume was adjusted to the required level with the remaining amount of Milli-Q water and homogenized at 8500 rpm for 15 minutes to produce a crude emulsion.

[0164] High Pressure Homogenization The resulting coarse emulsion was subjected to high pressure homogenization in three passes at different pressures, namely pass 1 at 10,000 psi, pass 2 at 18,000 psi, and pass 3 at 18,000 psi, after which the product was cooled to room temperature to obtain the emulsion.

[0165] Example 17: [Table 38]

[0166] Manufacturing method All ingredients were weighed according to the manufacturing recipe.

[0167] Preparation of the oil phase An accurately measured amount of omega-3 acid ethyl ester was placed in a manufacturing vessel, egg lecithin was added thereto, and the mixture was heated and dissolved in a water bath at 55°C (50°C to 60°C). A measured amount of cannabidiol was added to the above mixture, and the mixture was heated and dissolved in a water bath at 45°C (40°C to 50°C) to obtain an oil phase.

[0168] Preparation of the aqueous phase 90% Milli-Q water was placed in a separate manufacturing vessel, and stirred at 400 rpm on a magnetic stirrer while maintaining the temperature at 55°C (50°C to 60°C) to dissolve glycerol and sodium bicarbonate, and then tiotropium bromide was dissolved to prepare an aqueous phase.

[0169] Preparation of emulsions High shear homogenization The oil phase was added to the water phase and homogenized at 8500 rpm for 15 minutes while maintaining the product temperature at 45°C (40°C to 50°C).Then, the volume was adjusted to the required level with the remaining amount of Milli-Q water and homogenized at 8500 rpm for 15 minutes to produce a crude emulsion.

[0170] High Pressure Homogenization The resulting coarse emulsion was subjected to high pressure homogenization in three passes at different pressures, namely pass 1 at 10,000 psi, pass 2 at 18,000 psi, and pass 3 at 18,000 psi, after which the product was cooled to room temperature to obtain the emulsion.

[0171] Example 18: [Table 39]

[0172] Manufacturing method All ingredients were weighed according to the manufacturing recipe.

[0173] Preparation of the oil phase An accurately measured amount of omega-3 acid ethyl ester was taken into a manufacturing vessel, to which egg lecithin was added and dissolved by heating on a water bath at 55°C (50°C to 60°C). A measured amount of cannabidiol was added to the above mixture and dissolved by heating on a water bath at 45°C (40°C to 50°C), and then nintedanib was dissolved / dispersed to obtain an oil phase.

[0174] Preparation of the aqueous phase 90% Milli-Q water was placed in a separate manufacturing vessel, and stirred at 400 rpm on a magnetic stirrer while maintaining the temperature at 55°C (50°C to 60°C), and glycerol and sodium bicarbonate were dissolved to prepare an aqueous phase.

[0175] Preparation of emulsions High shear homogenization The oil phase was added to the water phase and homogenized at 8500 rpm for 15 minutes while maintaining the product temperature at 45°C (40°C to 50°C).Then, the volume was adjusted to the required level with the remaining amount of Milli-Q water and homogenized at 8500 rpm for 15 minutes to produce a crude emulsion.

[0176] High Pressure Homogenization The resulting coarse emulsion was subjected to high pressure homogenization in three passes at different pressures, namely pass 1 at 10,000 psi, pass 2 at 18,000 psi, and pass 3 at 18,000 psi, after which the product was cooled to room temperature to obtain the emulsion.

[0177] Example 19: [Table 40]

[0178] Manufacturing method All ingredients were weighed according to the manufacturing recipe.

[0179] Preparation of the oil phase An accurately measured amount of omega-3 acid ethyl ester was placed in a manufacturing vessel, to which egg lecithin was added and dissolved by heating on a water bath at 55°C (50°C to 60°C). A measured amount of cannabidiol was added to the above mixture and dissolved by heating on a water bath at 45°C (40°C to 50°C), and then streptomycin was dissolved / dispersed to obtain an oil phase.

[0180] Preparation of the aqueous phase 90% Milli-Q water was placed in a separate manufacturing container, and stirred at 400 rpm on a magnetic stirrer while maintaining the temperature at 55°C (50°C to 60°C). Glycerol, drug, and sodium bicarbonate were dissolved in that order to prepare the aqueous phase.

[0181] Preparation of emulsions High shear homogenization The oil phase containing the drug was added to the aqueous phase and homogenized at 8500 rpm for 15 minutes while maintaining the product temperature at 45°C (40°C to 50°C).Then, the volume was adjusted to the required level with the remaining amount of Milli-Q water and homogenized at 8500 rpm for 15 minutes to produce a crude emulsion.

[0182] High Pressure Homogenization The resulting coarse emulsion was subjected to high pressure homogenization in the first to third passes at different pressures ranging from 10,000 psi to 18,000 psi, and then the product was cooled to room temperature.

[0183] Example 20: [Table 41]

[0184] Manufacturing method All ingredients were weighed according to the manufacturing recipe.

[0185] Preparation of the oil phase An accurately measured amount of omega-3 acid ethyl ester was placed in a manufacturing vessel, to which egg lecithin was added and dissolved by heating on a water bath at 55°C (50°C to 60°C). A measured amount of cannabidiol was added to the above mixture and dissolved by heating on a water bath at 45°C (40°C to 50°C) to obtain an oil phase. THC was dissolved in the oil mixture.

[0186] Preparation of the aqueous phase 90% Milli-Q water was placed in a separate manufacturing container, and stirred at 400 rpm on a magnetic stirrer while maintaining the temperature at 55°C (50°C to 60°C). Glycerol and sodium bicarbonate were dissolved in that order to prepare an aqueous phase.

[0187] Preparation of emulsions High shear homogenization The oil phase containing the drug was added to the aqueous phase and homogenized at 8500 rpm for 15 minutes while maintaining the product temperature at 45°C (40°C to 50°C).Then, the volume was adjusted to the required level with the remaining amount of Milli-Q water and homogenized at 8500 rpm for 15 minutes to produce a crude emulsion.

[0188] High Pressure Homogenization The resulting coarse emulsion was subjected to high pressure homogenization in the first to third passes at different pressures ranging from 10,000 psi to 18,000 psi, and then the product was cooled to room temperature.

[0189] Example 21: [Table 42]

[0190] Manufacturing method All ingredients were weighed according to the manufacturing recipe.

[0191] Preparation of the oil phase An accurately measured amount of omega-3 acid ethyl ester was placed in a manufacturing vessel, to which egg lecithin was added and dissolved by heating on a water bath at 55°C (50°C to 60°C). A measured amount of cannabidiol was added to the above mixture and dissolved by heating on a water bath at 45°C (40°C to 50°C), and then pirfenidone was dispersed / dissolved to obtain an oil phase.

[0192] Preparation of the aqueous phase 90% Milli-Q water was placed in a separate manufacturing container, and stirred at 400 rpm on a magnetic stirrer while maintaining the temperature at 55°C (50°C to 60°C). Glycerol and sodium bicarbonate were dissolved in that order to prepare an aqueous phase.

[0193] Preparation of Suspensions High shear homogenization The oil phase containing the drug was added to the water phase and homogenized at 8500 rpm for 15 minutes while maintaining the product temperature at 45°C (40°C to 50°C).Then, the volume was adjusted to the required level with the remaining amount of Milli-Q water and homogenized at 8500 rpm for 15 minutes to produce a crude dispersion.

[0194] High Pressure Homogenization The resulting crude dispersion was subjected to high pressure homogenization in 1st to 3rd passes at different pressures ranging from 10,000 psi to 18,000 psi, and then the product was cooled to room temperature.

[0195] Example 22: [Table 43]

[0196] Manufacturing method All ingredients were weighed according to the manufacturing recipe.

[0197] Preparation of the oil phase An accurately measured amount of omega-3 acid ethyl ester was taken into a manufacturing vessel, to which egg lecithin was added and dissolved by heating on a water bath at 55°C (50°C to 60°C). A measured amount of cannabidiol was added to the above mixture and dissolved by heating on a water bath at 45°C (40°C to 50°C), and then nintedanib was dispersed / dissolved to obtain an oil phase.

[0198] Preparation of the aqueous phase 90% Milli-Q water was placed in a separate manufacturing container, and stirred at 400 rpm on a magnetic stirrer while maintaining the temperature at 55°C (50°C to 60°C). Glycerol and sodium bicarbonate were dissolved in that order to prepare an aqueous phase.

[0199] Preparation of Suspensions High shear homogenization The oil phase containing the drug was added to the water phase and homogenized at 8500 rpm for 15 minutes while maintaining the product temperature at 45°C (40°C to 50°C).Then, the volume was adjusted to the required level with the remaining amount of Milli-Q water and homogenized at 8500 rpm for 15 minutes to produce a crude dispersion.

[0200] High Pressure Homogenization The resulting crude dispersion was subjected to high pressure homogenization in 1st to 3rd passes at different pressures ranging from 10,000 psi to 18,000 psi, and then the product was cooled to room temperature.

[0201] Example 23: [Table 44]

[0202] Manufacturing method All ingredients were weighed according to the manufacturing recipe.

[0203] Preparation of the oil phase An accurately measured amount of omega-3 acid ethyl ester was placed in a manufacturing vessel, to which egg lecithin was added and dissolved by heating on a water bath at 55°C (50°C to 60°C). A measured amount of cannabidiol was added to the above mixture and dissolved by heating on a water bath at 45°C (40°C to 50°C), and then apremilast was dispersed / dissolved to obtain an oil phase.

[0204] Preparation of the aqueous phase 90% Milli-Q water was placed in a separate manufacturing container, and stirred at 400 rpm on a magnetic stirrer while maintaining the temperature at 55°C (50°C to 60°C). Glycerol and sodium bicarbonate were dissolved in that order to prepare an aqueous phase.

[0205] Preparation of Suspensions High shear homogenization The oil phase containing the drug was added to the water phase and homogenized at 8500 rpm for 15 minutes while maintaining the product temperature at 45°C (40°C to 50°C).Then, the volume was adjusted to the required level with the remaining amount of Milli-Q water and homogenized at 8500 rpm for 15 minutes to produce a crude dispersion.

[0206] High Pressure Homogenization The resulting crude dispersion was subjected to high pressure homogenization in 1st to 3rd passes at different pressures ranging from 10,000 psi to 18,000 psi, and then the product was cooled to room temperature.

[0207] Example 24: [Table 45]

[0208] Manufacturing method All ingredients were weighed according to the manufacturing recipe.

[0209] Preparation of the oil phase An accurately measured amount of omega-3 acid ethyl ester was placed in a manufacturing vessel, to which egg lecithin was added and dissolved by heating on a water bath at 55°C (50°C to 60°C). A measured amount of cannabidiol was added to the above mixture and dissolved by heating on a water bath at 45°C (40°C to 50°C), and then roflumilast was dispersed / dissolved to obtain an oil phase.

[0210] Preparation of the aqueous phase 90% Milli-Q water was placed in a separate manufacturing container, and stirred at 400 rpm on a magnetic stirrer while maintaining the temperature at 55°C (50°C to 60°C). Glycerol and sodium bicarbonate were dissolved in that order to prepare an aqueous phase.

[0211] Preparation of Suspensions High shear homogenization The oil phase containing the drug was added to the water phase and homogenized at 8500 rpm for 15 minutes while maintaining the product temperature at 45°C (40°C to 50°C).Then, the volume was adjusted to the required level with the remaining amount of Milli-Q water and homogenized at 8500 rpm for 15 minutes to produce a crude dispersion.

[0212] High Pressure Homogenization The resulting crude dispersion was subjected to high pressure homogenization in 1st to 3rd passes at different pressures ranging from 10,000 psi to 18,000 psi, and then the product was cooled to room temperature.

[0213] Example 25: [Table 46]

[0214] Manufacturing method All ingredients were weighed according to the manufacturing recipe.

[0215] Preparation of the oil phase An accurately measured amount of omega-3 acid ethyl ester was placed in a manufacturing vessel, to which egg lecithin was added and dissolved by heating on a water bath at 55°C (50°C to 60°C). A measured amount of cannabidiol was added to the above mixture and dissolved by heating on a water bath at 45°C (40°C to 50°C), and then isoniazid was dispersed / dissolved to obtain an oil phase.

[0216] Preparation of the aqueous phase 90% Milli-Q water was placed in a separate manufacturing container, and stirred at 400 rpm on a magnetic stirrer while maintaining the temperature at 55°C (50°C to 60°C). Glycerol and sodium bicarbonate were dissolved in that order to prepare an aqueous phase.

[0217] Preparation of Suspensions High shear homogenization The oil phase containing the drug was added to the water phase and homogenized at 8500 rpm for 15 minutes while maintaining the product temperature at 45°C (40°C to 50°C).Then, the volume was adjusted to the required level with the remaining amount of Milli-Q water and homogenized at 8500 rpm for 15 minutes to produce a crude dispersion.

[0218] High Pressure Homogenization The resulting crude dispersion was subjected to high pressure homogenization in 1st to 3rd passes at different pressures ranging from 10,000 psi to 18,000 psi, and then the product was cooled to room temperature.

Claims

1. A novel inhaled formulation of omega-3 fatty acids comprising a therapeutically effective amount of docosahexaenoic acid (DHA), docosapentaenoic acid (DPA), eicosapentaenoic acid (EPA), or a combination of any two or all three of DHA, DPA, and EPA.

2. 2. The formulation of claim 1, wherein the fatty acids are present as free fatty acids (FFA), or ethyl esters (EE), or phospholipids (PL), or non-phosphate-containing glycerolipids selected from the group comprising triacylglycerols, diacylglycerols and / or monoacylglycerols.

3. 10. The formulation of claim 1, wherein the formulation is a suspension or emulsion that can be administered by inhalation.

4. 10. The formulation of claim 1, wherein the formulation further comprises a therapeutically active ingredient selected from the group consisting of pirfenidone, apremilast, roflumilast, tiotropium bromide, nintedanib, isoniazid, streptomycin, montelukast, tetrahydrocannabinol, and the like.

5. The formulation of claim 1, used for (i) the treatment of COVID-19 and other lung disorders, or (ii) to reduce pathological levels of IL-6 or IL-10 in the lung, or (iii) to reduce pathological levels of TNF-α / TGF-β in the lung.

6. A formulation comprising a therapeutically effective amount of a phospholipid, wherein the phospholipid delivers docosahexaenoic acid (DHA), docosapentaenoic acid (DPA), eicosapentaenoic acid (EPA), or a combination of any two or all three of DHA, DPA, and EPA.

7. A formulation comprising a therapeutically effective amount of a glycerolipid, wherein the glycerolipid delivers docosahexaenoic acid (DHA), docosapentaenoic acid (DPA), eicosapentaenoic acid (EPA), or a combination of any two or all three of DHA, DPA, and EPA.

8. 8. The formulation of any one of claims 1, 6 and 7, further comprising a therapeutically effective amount of melatonin.

9. 8. The formulation of any one of claims 1, 6 and 7, further comprising a therapeutically effective amount of budesonide.

10. 8. The formulation of any one of claims 1, 6 and 7, further comprising a therapeutically effective amount of cannabidiol.

11. The formulation of any one of claims 1, 6 and 7 for use in the treatment of a disorder selected from the group consisting of bronchiolitis, asthma, cystic fibrosis, COPD, pneumonia, tuberculosis, emphysema, pulmonary edema, lung cancer, acute respiratory distress syndrome (ARDS), asbestosis, bronchiectasis, interstitial lung diseases (ILDs) including sarcoidosis, idiopathic pulmonary fibrosis and CNS disorders.

12. The formulation described in claim 8 for administration to patients for whom NSAIDs are contraindicated.

13. The formulation of claim 9 for administration to patients for whom NSAIDs are contraindicated.

14. The formulation of claim 10 for administration to patients for whom NSAIDs are contraindicated.

15. A formulation according to any one of claims 1, 6 and 7 for administration to a patient for the prevention or treatment of a condition selected from the group consisting of acute pulmonary thrombosis, acute or chronic pulmonary inflammation, inflammatory conditions of the heart and its blood vessels, or optionally in combination with a fast-acting bronchodilator to inhibit bronchoconstriction.

16. A novel inhalation formulation of omega-3 fatty acids, comprising a therapeutically effective amount of docosahexaenoic acid (DHA), docosapentaenoic acid (DPA), eicosapentaenoic acid (EPA), or a combination of any two or all three of DHA, DPA, and EPA, and which is a suspension or emulsion.

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