SI26055A - Nano-micellar pharmaceutical synergetic composition with antioxidant, anti-inflammatory, immunomodulating, antiviral properties for multiple therapeutic applications - Google Patents

Abstract

The invention relates to a nanomicellar synergistic pharmaceutical composition consisting of curcumin (0.1-5%), boswellia (0.1-5%), artemisinin (0.1-3%), vitamin C (0.1-6%) ) and optionally also cannabinoids (0.1-5%) and / or nitroxides (0.1-2%). The composition can be produced in liquid form (oral spray for mucous membranes, drops) or pharmaceutically acceptable solid form, with antioxidant, anti-inflammatory, immunomodulatory, antiviral, anti-cancer properties and is intended for several therapeutic applications.

Description

Nanomicella pharmaceutical synergistic composition with antioxidant, anti-inflammatory, immunomodulatory, antiviral properties intended for several therapeutic applications

DESCRIPTION

TECHNICAL AREA

The invention relates to a nanomicellar pharmaceutical synergistic composition consisting of curcumin (0.1-5%), boswellia (0.1-5%), artemisinin (0.1-3%), vitamin C (0.1-6% ) and optionally also cannabinoids (0.1-5%) and/or nitroxides (0.1-2%).

This invention relates to a nanomicellar pharmaceutical synergistic composition with antioxidant, anti-inflammatory, immunomodulating, antiviral, anti-cancer properties, and is intended for several therapeutic applications.

This invention relates to a micellar pharmaceutical synergistic composition produced in liquid form (oral mucosal spray, drops) or in a pharmaceutically acceptable solid form (powder).

BACKGROUND OF THE INVENTION

Plants have long been recognized for their therapeutic properties. For centuries, indigenous cultures around the world have used traditional herbal remedies to treat many ailments. In contrast, the rise of the modern pharmaceutical industry in the past century was based on the exploitation of individual active compounds with precise modes of action. This direction in pharmacy has produced highly effective drugs widely used in health care, including many plant-based natural products and analogues derived from these products, but has failed to provide effective drugs for complex human diseases with complex causes, such as cancer, diabetes , autoimmune disorders and degenerative diseases. While plants continue to be an important source of chemical compounds that support drug discovery, the rich traditions of herbal medicine, developed over thousands of years through trial and error on humans themselves, contain invaluable biomedical information waiting to be discovered by modern scientific approaches.

In many traditional herbal healing systems, a recipe often contains several ingredients mixed in a specific ratio in a single formula, with a single ingredient in isolation sometimes lacking the therapeutic activities seen in the entire formulation, a phenomenon known as the combination effect. It is hypothesized that pharmacological efficacy may be enhanced by simultaneous action of multiple chemicals targeting multiple sites and/or synergistic action at a single site. Given the limited success of modern single-compound pharmaceuticals in treating complex diseases such as influenza viruses, cancer, type II diabetes, autoimmune disorders, and degenerative diseases, elucidating the mechanistic basis for the combination effect of well-established holistic traditional herbal recipes will shed light on the complex biology diseases and helped in the design of new drugs.

Updating comprehensive traditional herbal recipes

Since each plant species contains many metabolites in its metabolome, consuming the whole plant as medicine is a form of combinatorial medicine. In addition, in many traditional herbal medicine systems, especially those in Asian countries, well-established recipes often contain several ingredients mixed in a certain ratio, suggesting that the synergistic effect of several active ingredients contained in different plant parts is based on the effectiveness of these methods of treatment. Although the initial development of these comprehensive traditional herbal recipes was known before modern science, the process was based on millennia of human-adapted clinical trials based on phenotype. Meanwhile, generations of herbalists also recorded precise descriptions of disease symptoms and systematic medical theories that linked the therapeutic properties of various medicinal plants to their utility in treating specific symptoms. But most fundamental concepts in traditional medical systems—for example, the concepts of yin versus yang and cold versus heat in traditional Chinese medicine—are divorced from modern descriptions of normal and disease states in the language of physiology and molecular biology. The lack of modern scientific and clinical evidence of safety, efficacy and mode of action prevents these holistic herbal recipes from being accepted outside of their culture of origin. Thus, detailed phytochemical, pharmacological and clinical studies of traditional holistic herbal recipes are imperative to determine modern guidelines for their use. Moreover, understanding the molecular basis of the synergistic effects of these comprehensive herbal medicines is likely to provide new insights into the complex mechanisms of disease and new clues for future pharmaceutical development.

Most natural plant compounds such as curcuminoids, boswellic acids, artemisinin, cannabinoids, resveratrol, hypericin, bacosides, cmo seed, ginseng extract and many others have been proven to have multiple therapeutic benefits, such as anti-inflammatory, antioxidant, anti-obesity, they improve memory, have an anti-allergic, antimicrobial effect, help in the fight against cancer and have many other medicinal properties. However, due to their poor bioavailability and lack of stability in the body, little has been achieved with these molecules for disease prevention and treatment.

However, many plant molecules mentioned above are hydrophobic in nature and therefore not soluble in water. Despite their traditionally known benefits, the poor bioavailability of these molecules is the reason that these effective natural remedies are in short supply on the market. On the other hand, biopharmaceuticals are not as effective because they are unstable and biodegradable before reaching the target site. In the last two decades, therefore, several formulation strategies have been designed to improve drug solubility in order to increase the rate and extent of drug absorption from the gastro-intestinal tract - GIT [109; 76; 49; 93],

In addition, there is a great need for a pharmaceutical composition with synergistic properties suitable for human patients, namely with stronger effects than the simple sum of the effects of the individual components, and at the same time synergistic with the standard clinical treatment of various diseases.

Inflammation

Inflammation plays a fundamental role in host defense and the progression of immune-mediated disease. The inflammatory response is initiated in response to injury (e.g., wounds, ischemia, and foreign bodies) and infection (e.g., bacterial or viral infection) by multiple events, including chemical mediators (e.g., cytokines and prostaglandins) and inflammatory cells (e.g., leukocytes). . It is characterized by increased blood flow to the tissue, causing pyrexia, erythema, induration and pain.

Cytokines

Cytokines, especially IL-Ιβ, IL-6, IL-8 and TNF-α, play an important role in the inflammatory process. TNF-α, a pleiotropic cytokine, is produced primarily by macrophages, but can also be produced by other cell types. TNF-α exhibits beneficial as well as pathological activities. In addition, it has growth-promoting and growth-inhibiting properties and self-regulatory effects. Beneficial functions of TNFα include maintaining homeostasis by regulating the body's circadian rhythm, enhancing the immune response to bacterial, viral, fungal, and parasitic infections, replacing or remodeling damaged tissue by stimulating fibroblast growth, and, as the name suggests, destroying certain tumors.

Although TNF-α plays an important role in innate and acquired immune responses, inappropriate production of TNF-α can lead to pathological changes that result in chronic inflammation and tissue damage. TNF-α has been shown to play a key role in the pathogenesis of many chronic inflammatory diseases such as inflammatory bowel disease, rheumatoid arthritis, juvenile rheumatoid arthritis, psoriatic arthritis, osteoarthritis, refractory rheumatoid arthritis, chronic non-rheumatoid arthritis, osteoporosis/bone resorption, coronary heart disease disease, vasculitis, ulcerative colitis, psoriasis, adult respiratory distress syndrome, hypersensitivity type diabetes and Alzheimer's disease.

Interleukin-1 (IL-1) is an important part of the innate immune system that regulates the functions of the adaptive immune system. The balance between IL-1 and IL-1 receptor antagonist (IL-1 ra) in local tissues influences the possible development of inflammatory disease and consequent structural damage. In the presence of excessive amounts of IL-1, inflammatory and autoimmune disorders can develop in the joints, lungs, gastrointestinal tract, central nervous system (CNS), or blood vessels. Two clinically important cytokines released in the synovium are IL-Ιβ and TNF-α. TNF-α can increase its effect and facilitate the expression of other genes involved in RA, including ILΙβ, IL-6, IL-8, cyclooxygenase-2 (C0X-2), inducible nitric oxide synthetase (iNOS), intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VC AM-1) and E-Selectin. This type of positive regulatory loop can amplify and maintain local inflammatory responses. Therefore, inappropriate or overexpression of TNF-α leads to a coordinated increase in the expression of many genes whose products mediate inflammatory and immune responses and thus cause clinical disease manifestations.

Leukocyte recruitment and retention is a key event in the pathogenesis of all chronic inflammatory disorders, including RA. In addition, the adhesion of circulating leukocytes, especially monocytes, to endothelial vasculature is also crucial in the development of atherosclerosis. This process depends on the interaction between adhesion molecules expressed on the surface of endothelial cells, such as ICAM-1, VCAM-1 and E-Selectin, and their cognate ligands on leukocytes. Therefore, ICAM-1, VCAM-1 and E-Selectin are responsible for the recruitment of inflammatory cells such as neutrophils, eosinophils and T lymphocytes from the circulation to the site of inflammation. These adhesion proteins are normally present at low levels on the surface of endothelial cells, but are strongly induced by various inflammatory cytokines such as TNF-α.

Other inflammatory processes, inflammatory bowel disease (IBD) is a group of processes that cause inflammation of the intestine. The inflammation lasts for a long time and usually recurs. The two main types of IBD are Crohn's disease and ulcerative colitis.

Crohn's disease occurs when the lining and wall of the intestine become inflamed, causing ulcers. Although Crohn's disease can occur in any part of the digestive system, it often occurs in the lower part of the small intestine where it joins the large intestine.

Ulcerative colitis is a chronic inflammatory disease of unknown etiology that affects the colon. The course of the disease can be continuous or relapsing, mild or severe. The earliest lesion is inflammatory infiltration with the formation of abscesses at the base of Lieberktihn's crypts (intestinal crypts). Coalescence of these swollen and ruptured crypts causes separation of the overlying mucosa from its blood supply, leading to ulceration. Signs and symptoms of the disease include cramping, lower abdominal pain, rectal bleeding, and frequent, liquid stools consisting primarily of blood, pus, and mucus with few fecal particles. Acute, severe or chronic persistent ulcerative colitis may require a total colectomy.

Another inflammatory disease is atherosclerosis. Atherosclerosis is a disease that affects arterial blood vessels. It is a chronic inflammatory response in the walls of the arteries, mainly due to the deposition of lipoproteins (plasma proteins that carry cholesterol and triglycerides). It is commonly called hardening or lining of the arteries. It is caused by the formation of more plaques in the arteries, causing inflammation of the arteries.

The most common therapy for the treatment of inflammatory processes involves the use of non-steroidal anti-inflammatory drugs (NSAIDs), e.g. naproxen, diclofenac, ibuprofen to relieve symptoms such as pain. Despite the widespread use of NSAIDs, many individuals do not tolerate well the doses needed to treat the disease for a long time, as NSAIDs are known to cause gastric erosions. In addition, NSAIDs only treat the symptoms of the disease and do not eliminate the cause.

Viruses

The severe acute respiratory syndrome disease associated with the coronavirus (COVID-19) is a syndrome of viral replication coupled with a host inflammatory response. Cytokine storm and viral evasion of cellular immune responses may play equally important roles in the pathogenesis, clinical manifestation, and outcome of COVID-19. Systemic anti-inflammatory cytokines and biomarkers are elevated as the disease progresses to advanced stages and are associated with poorer survival.

SARS-CoV-2 activates the innate immune system and causes the release of a large number of cytokines, including IL-6, which can increase vascular permeability and cause fluid and blood cell leakage into the alveoli, resulting in symptoms such as dyspnea and respiratory failure [33], Higher mortality is associated with the result of worsening ARDS (acute respiratory distress syndrome) and tissue damage that can lead to organ failure and/or death.

Pathogenesis of COVID-19 and a strategy for the use of edible plants

To better understand how nutraceuticals or phytomolecules can effectively act against novel coronaviruses, it is essential to understand the structural features and their associated targets and receptors. In addition, understanding the mechanism of action of common antiviral drugs and the responsible targets for drug development may be useful for designing treatment regimens for

COVID-19 from natural sources. SARS-CoV-2, like other HCoVs, is a positive-sensitive single-stranded RNA virus with two groups of proteins that form characteristic markers: structural protein such as spikes (S), nucleocapsid (N), matrix (M), envelope (E); and nonstructural proteins such as nspl2-RNA-dependent RNA-polymerase (RdRp), nsp3papain-like proteinases, nsp5-3C-like major protease and nsP13 SARS-CoV helicase [33, 6], Primary nsP13 helicase, 3CL protease, nspl2 -RNA-dependent RNA-polymerase (RdRp) become a major target for drug development. In addition to these proteins, the binding and internalization of the viral spike glycoprotein (S) within host cell ACE-2 receptors may also be a target to prevent virus entry into newer host cells [9], SARS-CoV-2 recognizes human angiotensin-converting enzyme-2 (ACE-2), thus demonstrating that it is essential for entry into host cells by invasion of alveolar epithelial cells, subsequent viral replication and primary infection of host lung cells, as ACE-2 is highly expressed in the heart, lungs, intestines, kidneys and blood vessels [128], The expression of ACE-2 is significantly increased in patients with diabetes and hypertension, and the connecting link of the related comorbidity is the receptor for angiotensin-converting enzyme-2 (ACE2), because this receptor is the site of virus replication, therefore, a strategy was devised for the development of newer antiviral drugs, which specifically targeted ACE-2 [69], Different plants produce phytomolecules that can be used for ci targeting these viral targets, which has already been done in the case of other viral diseases such as SARS, HIV, HCV, etc. [108, 49, 94],

At the molecular level, SARS-CoV-2 vims binds to angiotensin-converting enzyme-2 (ACE-2), which is present in the lungs of the human host. The binding of the virus to host cells via its trimeric spike glycoprotein makes this protein a key target for potential therapies and diagnostics. It has been reported that in SARS-CoV-2, the S2 subunit in each spike monomer contains a fusion peptide, a transmembrane domain and a cytoplasmic domain that is highly conserved and could be a potential target for antiviral (anti-S2) compounds [16]. Viral replication occurs, causing a cellular response. Infiltration of large numbers of inflammatory cells occurs, comprising innate immune cells and adaptive immune cells [135, 123]. Neutrophils are mostly innate immune cells that cause lung damage [124], On the other hand, adaptive immune cells are mainly T-cells, i.e. cytotoxic CD8 + T cells that not only destroy vims but further damage the lung [114]. This accelerates the progression of the systemic inflammatory response, leading to a massive increase in various cytokines such as TNFα, IL1, IL6, IL10, etc. This is called a cytokine wave. Due to the increase of various cytokines, inflammation and apoptosis of type 1 and type 2 cells in the alveoli occur. This disrupts oxygen transport functions, causing cell death in the lung alveoli and causing acute respiratory distress syndrome (ARDS) [17]. Figure 2 shows the pathogenesis of the disease leading to a cytokine storm and multi-organ dysfunction and ultimately death.

Among the proposed mechanisms of SARS-CoV-2-induced lung injury is a cytokine storm triggered by an unbalanced type 1 and type 2 T-helper cell response, resulting in an uncontrolled and generalized inflammatory response [96], Elevated inflammatory cytokines (Interferon γ, interleukin (IL-) 1 β, IL-6, IL-12) and chemokines (CXCL10 and CCL2) circulating throughout the body are associated with lung inflammation and the pathogenesis of ARDS due to inflammatory damage to the alveolar membrane, resulting in increased lung permeability and secretion of protein-rich pulmonary edema fluid into the airspaces, resulting in respiratory insufficiency and major causes of complications leading to multiple organ failure [138]. Antiviral herbs with added anti-inflammatory properties that protect the lungs from infection may be explored for synergistic therapy purposes.

Weak immune mechanisms associated with cytokine surges are one of the main causes that ultimately lead to reduced cellular oxygenation at the alveolar level, which is thought to be the main cause of death in COVD-19. In addition to this respiratory impairment, there are also platelet events involving open reading frames (ORFs), particularly ORF8 proteins, which, upon binding to SARSC0V2, lead to the dissociation of iron from the 1-beta chain of hemoglobin, which binds to the surface glycoprotein porphyrin, thereby causing impairment of internal respiration [62],

On average, SARS-CoV2 has a longer incubation period of 2-14 days in the human body, probably due to its immune evasion property, which effectively escapes the host's immune detection in the early stage of infection [91], Herbal preparations that act as immunomodulators can serve as prophylactic treatment, if added to the daily diet, to prevent infection during this critical spread interval at the community level and help control the disease in the community and help speed recovery from infection. Considering the above strategies for the treatment, control and prevention of COVID-19, the search for potential plants with the above properties can help in the development of natural plant antiviral drugs against the pandemic disease.

Therefore, there is a need to develop improved and natural replacement drugs with synergistic properties with reduced side effects for the prevention and treatment of inflammatory diseases caused by increased IL-1 and TNF-a. Herbs are known all over the world and are used to treat many ailments. There is evidence that plant-derived products have potential pharmacological and multiple therapeutic effects in the treatment of many diseases and tend to have less harmful side effects than synthetic drugs.

REVIEW OF SIMILAR PATENTS

Numerous attempts to use the above-mentioned natural plants and synthetic ingredients separately or in a synergistic combination are presented in the patent literature, namely:

In US Patent Publication US7776911B2, author Govindarajan Padmanaban and colleagues, assignee: Indian Institute of Science (Bangalore), presented an antimalarial drug containing a synergistic combination of curcumin and artemisinin.

Korean patent KR 1020090099238A, 2009 presents antiviral agents derived from Curcuma longa that prevent avian influenza, swine influenza and novel influenza.

In WO2014197129A2, the author Ghorbani et al (USA) presented a composition and a method of treating pain. The composition includes turmeric extract, boswellia extract, ginger extract, holy basil extract, rosemary extract, white willow extract and alpha-lipolic acid, each in very small amounts.

In US20030096027A1, Babish et al., assignee: Metaproteomics LLC, USA, presented a curcuminoid composition that exhibits synergistic inhibition of cyclooxygenase-2 (COX2), reduction of inflammation in the stomach and kidney.

Indian patent application IN202021022497 belonging to Rukhmini Enterprises presents a formulation that combines a special form of standardized curcumin, vitamin K2-7 (PureK2TM), vitamin C and L-selenomethionine (RightSelTM) as a prophylaxis for outbreaks of COVID-19. The biggest advantage of this formulation is its two-way properties. While boosting immunity, the virus also attacks by blocking the replication protein. The presence of vitamin C and L-selenomethionine (RightSelTM) helps to strengthen immunity, while curcumin and vitamin K2-7 (PureK2TM) have an antiviral effect.

Sanjeeb Kumar Sahoo in W02011101859A1 (2011) described a water-soluble curcumin-loaded nanoparticulate system for cancer treatment composed of glyceryl monooleate (GSO), polyvinyl alcohol (PVA) and Pluronic F-127, with a uniform particle size that less than 200 nm, which has a high surface charge and high zeta potential, about - 32 mV, which increases the solubility, stability and bioavailability of the entrapped curcumin used for cancer treatment. The curcumin-loaded nanoparticle system is prepared by incorporating curcumin into the GMO liquid phase and then emulsifying this mixture with PVA and pluronic solution F-127. Then, the final emulsion was lyophilized with a lyophilizer to obtain a lyophilized powder. It has been investigated that curcumin exhibits its proliferative activity against cancer cells by inducing apoptosis.

Ranchodbhai Patel, EP2587940A1, presented nanoparticle compositions of hydrophobic phenolic compounds such as curcuminoids or isoflavones and polymethoxylated flavones in combination with hydrophobic polymers such as zein, gliadin, hordein, secalin or combinations of these polymers. This composition showed higher water dispersibility, stability against accumulation and sedimentation, and higher bioavailability. This is due to the nanonization of hydrophobic phenolic compounds and hydrophobic prolamins containing the polymer. They are used as antioxidants, anti-inflammatory and anti-cancer agents.

In US20100179103A1, Ketan Desai presented a combination of curcumin cyclodextrin in the form of microemulsion, solid lipid nanoparticles, solid powder, liquid and microencapsulated oil or gel, which is used for the prevention and treatment of various diseases such as Alzheimer's disease, asthma, rheumatoid arthritis, oncological diseases, etc. , because curcumin has anti-inflammatory and anti-angiogenic properties. Since curcumin is very poorly absorbed and has very low bioavailability, the patent described a method to increase curcumin delivery by complexing it with cyclodextrin, which can increase bioavailability.

Thomas M. DiMauro in US20100292512A1 presented methylated curcumin-methoxy stilbene hybrid molecules used primarily in the treatment of cancer. Curcumin does not readily penetrate the human gastrointestinal tract and is subject to intestinal metabolism and rejection, with less than 1% of oral curcumin entering plasma. The small amount of curcumin that enters the bloodstream is rapidly metabolized by the kidneys and liver. Although curcumin is highly lipophilic (and readily crosses the blood-brain barrier), only very small amounts of orally administered curcumin are detected in serum and brain tissue. High oral doses of curcumin have been reported to cause problems such as rash and diarrhea and headaches, possibly caused by curcumin metabolites. They describe the intranasal use of a formulation by delivering an effective amount of curcumin to the olfactory mucosa across the cribriform plate and into the brain for the treatment of neurodegenerative diseases such as Alzheimer's disease.

AU2015220913B2 by Bombardelli et al., Indena S. p. A (IT) is an invention relating to a composition with analgesic and anti-inflammatory action. A composition with analgesic and anti-inflammatory action, consisting of curcumin in a complex with phospholipids and an extract from the roots and rhizomes of the narrow-leaved American straw.

In US6264995B1, Newmark et al (USA) described an invention that represents an herbal composition that reduces inflammation in bones and joints by inhibiting the cyclooxygenase-2 enzyme, prepared from holy basil, turmeric, ginger, green tea, rosemary.

In W02012168450A1, author Thimmermann et al presented to Queen Mary and Westfield College (GB) grantees an invention designed to treat wounds or bleeding from organ damage and related disorders (especially brain, burn and brain damage) using an antimalarial agent , which is called artemisinin, and its derivatives. The present invention also relates to the treatment of myocardial infarction and coronary heart disease (and related disorders) using an antimalarial substance called artemisinin and its derivatives. The present invention also relates to the use of artemisinin and its derivatives in coronary artery bypass surgery, heart transplantation and ischemia/reperfusion related diseases.

US8232417B1 to Gupta et al., assigned to Bioderm Research, Scotsdale, AZ (USA) discloses an invention relating to certain artemisinin derivatives classified as sesquiterpenes with an endoperoxide group. The compounds of this invention have broad-spectrum antibacterial and antifungal biological activity suitable for topical or oral use in the treatment of infections and skin diseases in mammals, including acne, rosacea, topical wounds, infections, dandruff.

Korean patent KR101682512B1 disclosed a synergistic combination of 3-O-acetyl-l 1-ketobeta-boswellic acid (AKBA), a selectively enriched boswellia extract and a non-acidic boswellia resin extract (BNRE). Nutritional, pharmaceutical and nutritional supplements. The composition(s) can be used to prevent, control and treat diseases associated with inflammation, including inflammation and asthma, arthritis, endothelial dysfunction and the like. The invention also includes the alleviation of inflammatory biomarker proteins or molecules whose expression/formation is altered in the inflammatory disease process.

US9381221B2 by Velez-Rivera et al (MX) discloses a phytocomposition containing, together with other natural plants, boswellia extract for the treatment of pain associated with joint diseases. Rheumatoid arthritis, osteoarthritis, fibromyalgia and gouty arthritis are among the most common joint diseases.

US2009/0209581 A1 to Habash et al discloses a composition and method useful in the treatment or prevention of immunological diseases such as autoimmune disease. As used in this patent, an immunological disease is one that involves appropriate or exaggerated host immune responses. In a preferred embodiment, the agent used to alter the expression pattern of a gene associated with these diseases is a nitroxide antioxidant. Tempol is a stable nitroxide radical characterized by the chemical formula 4-hydroxy2.2.6,6-tetramethylpiperidine-1-oxyl, which has antioxidant properties. The present recipients have discovered that tempol also has the novel property of altering the expression of genes that encode proteins associated with immunological diseases such as autoimmune disease.

In US2010/0196456A1, US 2010/0129453A1, Strasser (MiVital, Switzerland) used gum arabic and/or rosin as natural emulsifiers to emulsify poorly soluble drugs including Q10, curcumin and others. But the lack of stable quality of natural excipients is obvious. These emulsions may only be relevant as a dietary supplement.

In US2015/0072012A1 Sripathy et al and in US2011/0229532A1 Nair et al (Laila Pharmaceuticals Pvt, IN) claimed that nanoformulations of hydrophobic compounds can be used as ayurvedic / dietetic formulations of pharmaceutical / nutraceuticals. But these emulsions refer to microemulsions of separate solubilized mean values of 200 nm and with much more widespread 80% emulsifiers. Furthermore, they also claim that release of the API for more than 24 hours does not affect spontaneous self-emulsification by the method of the present invention.

In patent series US2008/022.0102A1, US 2016/0128939 A9, US2004/0192768A1, US 20200129452 by Behnam, Dariush, assigned to Aquanova AG (Darmstadt, DE), claimed that the different solubilities of the separate active chemicals are mostly at a rather high percentage nonionic surfactants, especially polysorbate 20/polysorbate 80 and often ethanolic solvents and have nothing to do with SNEDDS.

In US2018/0071210A1 and PCT/GB20 19/050009, author Wilkhu et al., assigned to GW Research Limited, Cambridge (UK), presented oil-free cannabinoids mainly in solid and gel form with excipients and with poloxamer as emulsifiers. The solvent can be selected from the group consisting of diacetin, propylene glycol, triacetin, monoacetin, propylene glycol diacetate, triethyl citrate and mixtures thereof. A type IV oral pharmaceutical formulation (OPF) containing at least one cannabinoid, at least one solvent, and at least one poloxamer can be rehydrated by adding 20 mL of water for injection at room temperature or by adding 20 mL of water for injection at 37°C. The formulation may be essentially water-free, alcohol-free and/or oil-free, but this invention does not represent any spontaneous self-nano-emulsification reaction of the aforementioned formulations.

In US205/0232952A1, W02003074027A2 by Lambert et al., assigned to Novagali Pharma SA (FR), a drug delivery system containing one or more poorly water-soluble or water-insoluble therapeutic agents, vitamin E, one cosolvent selected from propylene glycol and ethanol, one or more bile salts, TPGS and a further surfactant derived from hydrogenated castor oil tyloxapol and polyoxyl. Unfortunately, when the drug concentration was higher than 1.5% w/w, the drug easily precipitated from the resulting microemulsions. The grafting time was approximately 2 hours as the concentration was increased to 2.5% w/w.

In WO2012/033478, Murty et al., assigned to Murty Pharmaceuticals, Inc., 518 Codell Drive, Lexington, KY 40509 (USA) used type I, type II and type III cannabinoid preparations of SEDDS.

The most suitable micelle nano-self-emulsifying system (SNEDDS) suitable for the composition in the present invention is described in the SIPO application at the Office of Intellectual Property of the Republic of Slovenia (2021) P202100024 and the patent application at the Patent Office in Russia (2021) W21013724/2021106347 by Victor Bolduev and colleagues (SI). Currently, many poorly soluble natural products on the market claim to have high bioavailability, which is obtained by using phospholipids or vegetable oils, etc. However, none of the previous discoveries investigate a self-nanoemulsifying formulation (SNEDDS) containing the synergistic properties of curcumin, boswellia, artemisinin, vitamin C and optionally cannabinoids and/or nitroxides to achieve spontaneous self-emulsification of APIs without an oil phase.

PURPOSE OF THE INVENTION

The purpose of this invention is therefore to develop an improved and natural alternative medicine with synergistic properties with reduced side effects for the prevention and treatment of inflammatory disorders caused by the increase of IL-1 and TNF-α by cytokine storm and provide antioxidant, anti-inflammatory, immunomodulatory, antiviral properties and anticancer properties, which can be designed for multiple therapeutic applications.

The present invention describes a new herbal composition that includes the synergistic properties of curcumin, boswellia, artemisinin, vitamin C and optionally cannabinoids and/or nitroxides. The composition can be used to treat a variety of inflammatory problems with minimal side effects. In addition, the invented composition is on a nanomicelle support and has self-emulsifying properties.

As far as we know from available sources, there is no report on similar nano self-nano emulsified pharmaceutical synergistic compositions from curcumin (0.1-5%), boswellia (0.1-5%), artemisinin (0.1-3%), vitamin C (0 .1-6%) and optionally cannabinoids (0.1-5%) and nitroxide (0.1-2%). The composition can be on liquid or solid pharmaceutically acceptable carriers, but it can also be used in combination with at least one other anti-inflammatory agent.

BRIEF DESCRIPTION OF IMAGES

FIGURE 1 Flowchart for the synthesis of CimetrA™

FIGURE 2 Weight - male

FIGURE 3 Weight - female

FIGURE 4 Organ weight — male

FIGURE 5 Organ weight - female

FIGURE 6 Group 4M. Animal #42, brain - hippocampus. No pathological and cytotoxic changes.

FIGURE 7 Group 4M. Animal #42, the heart. No pathological and cytotoxic changes.

FIGURE 8 Group 4M. Animal #42, the lungs. No pathological and cytotoxic changes.

FIGURE 9 Group 4M. Animal #42, liver. No pathological and cytotoxic changes.

FIGURE 10 Group 4M. Animal #42, kidneys. No pathological and cytotoxic changes.

FIGURE 11 Group 4M. Animal #42, the spleen. No pathological and cytotoxic changes. The white pulp looks active.

FIGURE 12 Group 4M. Animal #42, spinal cord, cervical segment. No pathological and cytotoxic changes. White matter shows vacuolization due to technical artifact.

FIGURE 13 Group 4M. Animal #42, sciatic nerve. No pathological and cytotoxic changes.

FIGURE 14 Group 4M. Animal #42, the tongue. No pathological and cytotoxic changes. Arrows indicate some mast cells (resident cells) in the striated muscle.

FIGURE 15 Viability of PBMCs

FIGURE 16 TNF-a concentration

FIGURE 17 Concentration of IL-6

FIGURE 18 Concentration of IL-1 β

FIGURE 19 IL-1 Ra concentration

FIGURE 20 FIGURE 21 IL -IRa concentration

FIGURE 21 Daily change of the NEWS-ranking in both considered groups

SUMMARY OF THE INVENTION

The invention relates to a nanomicellar pharmaceutical synergistic composition consisting of curcumin (0.1-5%), boswellia (0.1-5%), artemisinin (0.1-3%), vitamin C (0.1-6% ) and optionally cannabinoids (0.1-5%) and/or nitroxides (0.1-2%), where the percentages given are percentages by weight.

This invention relates to a nanomicelle pharmaceutical synergistic composition with antioxidant, anti-inflammatory, immunomodulating, antiviral, anti-cancer properties and is intended for several therapeutic applications.

This invention relates to a micellar pharmaceutical synergistic composition produced in liquid form (oral mucosal spray, drops) or pharmaceutically acceptable solid form (powder).

DETAILED DESCRIPTION OF THE INVENTION

It should be understood that the detailed description and specific examples illustrating embodiments of the invention are provided by way of illustration only, as various changes and modifications within the spirit and scope of the invention will be understood by those skilled in the art. A person skilled in the art can use this invention to its fullest extent based on the description herein. The following specific implementation examples should be understood as merely illustrative and in no way limiting the rest of the disclosure.

Unless otherwise specified, all technical and scientific terms used herein have the same meaning as is commonly understood by one skilled in the art to which the invention belongs.

The term inflammatory disorder as used herein refers to a disease or condition characterized by chronic inflammation, including (but not limited to) rheumatoid arthritis, osteoarthritis, juvenile rheumatoid arthritis, psoriatic arthritis, refractory rheumatoid arthritis, chronic non-rheumatoid arthritis arthritis, osteoporosis / bone resorption, coronary heart disease, atherosclerosis, vasculitis, ulcerative colitis, psoriasis, Crohn's disease, respiratory distress syndrome in adults, skin delayed hypersensitivity disorders, septic shock syndrome and inflammatory bowel diseases.

The term pharmaceutically acceptable as used herein means that the carrier, diluent, excipient and/or salt must be compatible with the other ingredients of the formulation and must not be harmful to the recipient.

The term pharmaceutically acceptable carrier as used herein means a non-toxic, inert solid, semi-solid, diluent, encapsulating material or excipient of any kind. Some examples of materials that can serve as pharmaceutically acceptable carriers are sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; malt; gelatin; talc; as well as other non-toxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweeteners, flavors and perfumes; at the discretion of the formulator, preservatives and antioxidants may also be present in the composition.

Screened plants according to the present invention that have antioxidant, anti-inflammatory, immunomodulatory, antiviral and anticancer properties

Artemisinin (ART) is isolated from the traditional Chinese medicine Artemisia annua L [4], The plant is also known as qinhao or sweet wormwood. Artemisia annua (qinghao) is a plant of the Asteraceae family that has been used in traditional Chinese medicine for centuries [126], the sesquiterpene lactone artemisinin (ART), the active medicinal ingredient, was discovered in the 1970s. Since then, chemical structural modification studies have been carried out to obtain new compounds with enhanced antimalarial activity and improved pharmacological properties. ART derivatives are well-tolerated drugs for you. This safety is one of the reasons why their effectiveness has been studied in other diseases, not just malaria. ART derivatives are active against other parasites, cancer cells and viruses, albeit with lower potency, with an effective concentration in the micromolar range compared to the nanomolar range as antimalarials [31]. Many studies demonstrate the activity of artemisinin and its derivatives, among others, against various viral diseases such as HCMV, HHV-6A, HSV-1, EPV, HBV, HCV, BVDV, HIV-1 and (R)CMV [30, 98, 23 ].

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Turmeric (Curcuma longa L.) belongs to the ginger family (Zingiberaceae). Plant roots contain several secondary metabolites, including curcuminoids, sesquiterpenes, and steroids [82]; with the curcuminoid curcumin being the main component of the yellow pigment and the main bioactive substance. Chemically, curcumin is diferuloylmethane, a diarylheptanoid that belongs to the class of natural phenols. Curcuma longa L. and its polyphenolic compound curcumin have been the subject of many antimicrobial investigations due to their extensive traditional use and low side effects. Antimicrobial activities of curcumin extract and C. longa rhizomes against nucleotides are proposed as a therapeutic target for antiviral and anticancer compounds. Among 15 different polyphenols, curcumin is proposed as a potent antiviral compound by this process with inhibitory activity against the effect of IMPDH either in a non-competitive or competitive manner [24], The study of various bioconjugates of curcumin against various viruses including various bacteria, viruses, fungi and parasites. The promising results of the antimicrobial activity of curcumin were a good example to enhance the inhibitory effect of existing antimicrobial agents through synergism [75]. As a plant derivative, curcumin has been shown to have a broad spectrum of antiviral activity against various viruses.

Boswellia, or frankincense, is a resinous herbal extract of the boswellia tree that has been used in natural medicine practices for centuries. Its anti-inflammatory effects mean it can help with inflammatory conditions such as rheumatoid arthritis, inflammatory bowel disease and asthma. Treatment for these conditions usually focuses on reducing inflammation. By helping to control inflammation, boswellia can reduce the symptoms of chronic inflammatory conditions [112]. The resin compounds with true anti-inflammatory effects are pentacyclic triterpenes of the boswellic acid type.

Boswellic acids inhibit leukotriene biosynthesis in neutrophil granulocytes. The effect is triggered by boswellic acid, which binds to the enzyme. In clinical trials, promising results have been observed in patients with rheumatoid arthritis, chronic colitis, ulcerative colitis, Crohn's disease, bronchial asthma, and peritumoral brain edema [1]. Its medicinal properties are also widely recognized, especially for the treatment of inflammatory conditions, but also for some cancer diseases, wound healing and its antimicrobial action. Scientific research is already beginning to support the benefits of boswellia, but most studies to date have used cell or animal models. Scientists do not know the in-depth effects of this substance on humans, so many more clinical trials on humans are needed before doctors can recommend this treatment [112]. In vitro studies and animal models show that boswellic acids have been found to inhibit the synthesis of specific anti-inflammatory enzymes that cause cellular saturation and optimal reduction of the risk of heart disease, stroke and cancer in healthy individuals. As an antioxidant and antiatherogen, ascorbic acid is credited with many health benefits. There is compelling evidence of lipid antioxidant protection by ascorbic acid both with and without iron supplementation in animals and humans [1], Prolonged bronchoconstriction, chemotaxis and increased vascular permeability have occurred [2, 3].

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Ascorbic acid (vitamin C) is one of the most important and main vitamins for human health. It is required for many physiological functions in human biology. Based on the available biochemical, clinical and epidemiological studies, the current recommended daily dose for ascorbic acid should be 80-90 mg/day [32] to discuss the role of ascorbic acid in enhancing immunity during cold infections. Ascorbic acid has been shown to stimulate the immune system by increasing T-cell proliferation in response to infection. These cells can destroy infected targets by cytolysis by producing large amounts of cytokines and helping B cells to synthesize immunoglobulins to control inflammatory reactions. Furthermore, ascorbic acid has been shown to block pathways that lead to T-cell apoptosis, thereby promoting or sustaining the proliferation of T-cells to attack infection. This mechanism is proposed for the enhanced immune response observed after vitamin C administration during cold infections [11],

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Cannabinoids are a group of compounds that mediate their effects via cannabinoid receptors. The discovery of A9-tetrahydrocannabinol (THC) as the main psychoactive principle in marijuana and the identification of cannabinoid receptors and their endogenous ligands led to significant developments in research aimed at understanding the physiological functions of cannabinoids. Cannabinoid receptors include CB1, which is predominantly expressed in the brain, and CB2, which is found primarily on cells of the immune system. The fact that CB1 and CB2 receptors have been found on immune cells suggests that cannabinoids play an important role in regulating the immune system. Recent studies have shown that THC administration in mice induced marked apoptosis in T cells and dendritic cells, resulting in immunosuppression. In addition, several studies have shown that cannabinoids downregulate cytokine and chemokine production and, in some models, upregulate T-regulatory cells (Tregs) as a mechanism to suppress inflammatory responses [78],

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Cannabidiol CBD /Π\ <5 2) OH \6 1/ 1 — 'λ ΙΙβ'^ΪΙ H 0^ 0 Tetrahydrocannabinol and cannabinol THC, CBN . 1 °°\ < _k m) /σΓο\ _o JT/ / to \« o/ O en ) ' T 0 Cannabielsoin CBE /X®\5a <? /79 \ O / 5 \ __/ 9bp 4Π In JA? H about ^x —0 Mo-tetrahydrocannabinol zso-THC 7 ¹⁰ H 0^ Cannabicyclol CBL /Tal/ Evil TKAo 0^ ***0 Cannabicitran CBT **0

One of the main mechanisms of immune suppression by cannabinoids is the induction of cell death or apoptosis in immune cell populations. Under normal conditions, maintenance of homeostasis requires apoptosis, which involves morphological changes (i.e., cell shrinkage, nuclear fragmentation, and membrane blebbing) and molecular changes (i.e., induction of caspases and cytochrome c leakage). The extrinsic pathway of apoptosis is initiated by ligation of death receptors (i.e., CD95) on the cell surface, leading to the activation of major caspases such as caspase 3, 8, and 10. The intrinsic pathway of apoptosis is initiated via mitochondria and caspase 9; cytochrome c and caspase 3 are the main factors in the induction of cell death [78],

Cannabinoid action on cytokines

Cytokines are signaling proteins that are synthesized and secreted by immune cells upon stimulation. They are modulatory factors that balance the initiation and resolution of inflammation. One of the possible mechanisms of cannabinoid-mediated immune control during inflammation is dysregulation of cytokine production by immune cells and disruption of a well-regulated immune response. In addition, cannabinoids can affect the immune response and host resistance by disrupting the balance between cytokines produced by the T-helper subsets, Thl and Th2. In vitro studies were performed to compare the effect of THC and cannabinol on cytokine production by human T, B, CD8 + , NK and eosinophil cell lines. However, the results were variable depending on the cell lines and concentrations used. Both anti-inflammatory and anti-inflammatory effects of THC were demonstrated in this study, suggesting that different cell populations have different thresholds for response to cannabinoids. In general, TNF-α, GM-CSF, and IFN-γ levels decreased with drug treatment. Interestingly, the levels of the anti-inflammatory cytokine IL-10 decreased after THC treatment, but the levels of the inflammatory cytokine IL-8 increased. In other studies, the cannabinoid CP55,940 has been shown to have a stimulatory effect on several cytokines in the human promyelocytic cell line HL-60 at nanomolar concentrations. At the molecular level, THC has also been shown to inhibit LPS-stimulated mRNA expression of IL1α, IL-Ιβ, IL-6 and TNF-α in cultured rat microglial cells; however, the effect was independent of cannabinoid receptors. In another study, mice were fed Corynebacterium parvum in vivo, followed by administration of the synthetic cannabinoids WIN55,212-2 and HU210. Animals were then challenged with LPS. The results showed a decrease in TNF-α and IL-12 levels, but an increase in serum IL-10 levels. It has been shown that this effect depends on CB1 receptors. During chronic inflammation, suppression of IL-6 can reduce tissue damage. AjA has been reported to prevent joint tissue damage in animal models of adjuvant arthritis. Recent studies have shown that addition of AjA to human monocyte-derived macrophages in vitro reduces the secretion of IL-6 from the activated cells, suggesting that AjA is suitable for the treatment of joint inflammation in patients with systemic lupus erythematosus (SLE). , rheumatoid arthritis (RA) and osteoarthritis.

They observed that the CB2 agonist HU-308 attenuated liver ischemia/reperfusion injury by decreasing the levels of TNF-α, ΜΙΡ-1α and MIP-2 in serum and liver homogenates. Recent in vitro studies have also shown a potent anti-inflammatory effect of synthetic cannabinoids (CP55,940 and WIN55,212-2). Both CP55,940 and WIN55,212-2 downregulated IL-6 and IL-8 cytokine production from IL-1β-stimulated fibroblast-like rheumatoid synoviocytes (FLS) through a non-CB1/CB2-mediated mechanism [78] ,

Cannabinoids and multiple sclerosis

Multiple sclerosis (MS) is an autoimmune disorder mediated by myelin-specific self-reactive T cells, macrophages/microglial cells, and astrocytes. The action of these cells leads to demyelination of nerve fibers and axons in the CNS in humans and causes many signs and symptoms such as muscle spasms, tremors, ataxia, weakness or paralysis, constipation and loss of bladder control. There is both anecdotal and clinical evidence demonstrating the effectiveness of cannabinoids in the treatment of MS. In 1994, a study was conducted with 112 MS patients (57 men and 55 women) from the United States and Great Britain; all patients treated themselves with the help of cannabis. The results of the research showed that the use of cannabis in more than 90% of patients improves the symptoms of their disease, such as spasticity, pain, tremors and depression. In eight different clinical studies, patients with MS also reported benefits of THC (used by ingestion, inhalation, injection, or rectal suppositories), cannabis (used by ingestion or inhalation), and the cannabinoid receptor agonist Nabilone TM (used by ingestion) for the treatment of spasticity, pain, tremors and ataxia. Cannabinoid use also improved objective test scores such as handwriting tests and bladder control tests. Cannabinoids are generally beneficial in the treatment of MS because of their neuroprotective and immunosuppressive properties. In this chapter, we will focus on the latter and discuss the actions of endogenous, natural and synthetic cannabinoids on immune cells in the CNS during MS [78].

Cannabinoids and colitis

During inflammation, several different cellular pathways are activated in the intestinal tract, leading to a pathological state [132], It has been shown that the functional CB1 receptor is expressed in the human ileum and colon, and the number of CB1-expressing cells increases significantly after inflammation increased [72, 86]. The protective role of these CB1 receptors during inflammation was demonstrated in a study analyzing the role of the endogenous cannabinoid system in the development of experimental colitis in mice induced by intrarectal treatment with 2,4-dinitrobenzene sulfonic acid (DNBS) or oral dextran sodium sulfate (DSS) administration. [72]. The DSS model, originally reported by Okayasu and colleagues, was used to investigate the role of leukocytes in the development of colitis [81]. Oral administration of 5% DSS in drinking water can cause acute colitis due to chemical damage to the colon. In addition, long-term administration of DSS induces colorectal carcinoma, which resembles the sequence of dysplasia and carcinoma observed during cancer development in human ulcerative colitis [136], On the other hand, intestinal inflammation induced by intrarectal administration of DNBS has many characteristic features of Crohn's disease in humans, including the induction of IL-12-mediated inflammation with a large Thl-mediated response [80]. The involvement of the endogenous cannabinoid system in the modulation of the acute phase of DNBS-induced colitis was further supported by higher levels of CB1-encoding transcripts in wild-type mice after induction of inflammation. They observed that genetic ablation of CB1 receptors made mice more susceptible to inflammatory damage. Furthermore, similar to CB1-deficient mice, pharmacological blockade of CB1 with the specific antagonist SR141716A resulted in exacerbation of colitis [72], A protective role of the endogenous cannabinoid system was observed 24 h after DNBS treatment, becoming more evident on days 2 and 3 . However, increased spontaneous proliferative activity of the smooth muscle cell membrane in DNBS-treated colons of CB1 − / mice was observed as early as 8 h after DNBS treatment, indicating that inflammation-induced smooth muscle irritation occurred at an earlier stage than in wild mice. This further supports the idea that the endogenous cannabinoid system is protected from inflammatory changes. These data suggest that activation of CB1 and the endogenous cannabinoid system provides an early and important physiological step in colonic self-protection against inflammation.

The cannabinoid system and liver injury

Over the past few years, awareness of the cannabinoid system in the pathophysiology of liver disease has increased. CB1 and CB2 receptors have been shown to be elevated in the early stages of liver damage [14]. Although it has been proven that the embryonic liver expresses receptor mRNA

CB2, adult liver hepatocytes and endothelial cells showed only weak physiological levels of CB1 receptor expression and were shown to produce low levels of endocannabinoids. CB1 receptors have been found to be upregulated in vasculature endothelium and in myofibroblasts found in fibrotic bands of cirrhotic livers in humans and rodents [121], CB2 receptors are also expressed in myofibroblasts, inflammatory cells and biliary epithelial cells [58] . In recent years, there is increasing evidence that endocannabinoids may regulate the pathophysiology of liver diseases, including acute forms of liver injury, liver fibrosis, and cirrhosis. In normal liver, endocannabinoids are low, which may be due to the high level of expression of FAAH, which is responsible for the degradation of AEA [21], AEA levels in liver and serum have been shown to increase in acute hepatitis and fatty liver [83]. In fatty liver, the increase in AEA is due to the decreased ability of FAAH to degrade AEA. Taken together, the above studies suggest that endocannabinoids and their receptors may play a key role in regulating liver fibrogenesis; therefore, targeting cannabinoid receptors may be a new tool for the prevention and treatment of liver damage.

Cannabinoids and RA

Rheumatoid arthritis is a chronic inflammatory disease that affects approximately 1% of the human population and is characterized by joint destruction, deformity, and loss of function associated with joint stiffness, pain, swelling, and tenderness [54], Major populations of immune cells involved in joint damage , are macrophages, T-cells, fibroblast-like synoviocytes and DCs. The main cytokines are TNF-α and IL-1 [54, 87]. Cannabinoids and their anti-inflammatory properties have been studied in animal models of RA and in human cells from RA patients, and these studies demonstrate the anti-arthritic properties of these natural plant compounds [84,142, 42, 5]. Interestingly, most studies on RA and cannabinoids have focused on the use of non-psychoactive cannabinoids. CBD is the main non-psychoactive component of cannabis, and its protective effect has been demonstrated in murine collagen-induced arthritis [70]. In this study, the authors showed that daily oral (5 mg/kg) or intra-peritoneal (25 mg/kg) administration of CBD inhibited disease progression. In addition, the study showed that CBD-treated mice had less proliferation in ex vivo activated draining lymph node cells, reduced levels of IFN-γ secreted by activated lymph node cells, and decreased synovial TNF-α production cells in the knee. Sumariwalla and colleagues used another synthetic non-psychoactive cannabinoid, HU-320, and showed that this compound improved pre-existing arthritis in mice [119]. Lymph node cells from HU-320-treated mice showed a reduced proliferative response when mouse cells were incubated with collagen II 7 days after inflammation.

Cannabinoids and cancers with inflammatory components

Inflammation strongly influences the development of certain types of cancer, which is thought to account for 15-20% of all cancer deaths worldwide [5]. The hallmarks of cancer-related inflammation include the presence of inflammatory cells in tumor tissue and the regulation of tumor growth, metastasis, and angiogenesis by inflammatory mediators (eg, chemokines, cytokines, and prostaglandins). The link between inflammation and cancer is now widely accepted, and NSAIDs have been shown to reduce the risk of various cancers. The use of these drugs reduces the risk of colon cancer by 40-50% and is thought to be preventive for lung, esophagus and stomach cancer [71]. Therefore, inflammation can be considered a therapeutic opportunity in some types of cancer. Cannabinoids have also recently become widely used as antitumor agents [46, 104] based on their ability to inhibit tumor angiogenesis [15] or induce direct apoptosis or cell cycle arrest in neoplastic cells [78]. The proposal focused on the antiproliferative effects of these compounds on various tumors such as breast and prostate cancer, pheochromocytoma and malignant gliomas [46, 78]. Our laboratory has reported that in vitro THC and other cannabinoids can induce apoptosis in transformed murine and human T-cells [65], including primary acute lymphoblastic human leukemia cells. Furthermore, treatment of mice with T-cell leukemia with THC could cure about 25% of the mice [65]. We further showed that THC treatment resulted in disruption of the MAPK/ERK kinase/ERK signaling module, which was required for apoptotic death [57, 74]. The role of endocannabinoids as potential inhibitors of endogenous tumor growth was suggested in a study where AEA and 2-AG levels were found to be higher in precancerous polyps than in fully developed colon carcinomas [63]. Recent in vivo studies have shown that selective targeting of CB2 receptors resulted in inhibition of colorectal tumor growth through ceramide stimulation-mediated apoptosis [63]. In a xenograft model of thyroid cancer, substances that blocked the degradation of endocannabinoids also increased tissue levels of AEA and 2-AG and reduced tumor growth [20], Various attempts have been made to inactivate cannabinoid-degrading enzymes, thereby increasing local concentration of endocannabinoids on the surface of tumor cells. This leads to the anti-tumor effects of CB-receptors, which are indicators in various types of cancer, such as thyroid, brain and prostate cancer [78], Although most of the effects of cannabinoids are mediated by CB-receptors, AEA has been shown to have an effect on cancer cells by interacting with the TRPV1 receptor [78] or with cholesterol-rich lipid rafts [29]. Cannabinoids have also been reported to differentially regulate signaling pathways in normal cells compared to cancer cells. In malignant diseases such as thyroid cancer, lymphoma, melanoma, pancreatic and breast cancer, the levels of cannabinoid receptors in the tumor are often higher compared to normal cells of the same origin, which in turn increases the sensitivity to cannabinoids in malignant diseases [15, 78]. Furthermore, many animal studies have reported antiproliferative and proapoptotic effects of cannabinoids on tumor cells but not on normal tissue [ 15 , 40 ]. Thus, the role of the cannabinoid system in cancer suggests that this system is involved in the regulation of many functions essential for cancer development.

TEMPO (or tempol) is a small molecule permeating superoxide dismutase (SOD) mimetic that can act as SODI, SOD2, SOD3 and catalase in vivo (NOT to be confused with the brand name paracetamol Tempol, also known as acetaminophen, an analgesic that is available without a prescription and is a generic form of Tylenol®).

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It attenuates superoxide anion- and peroxynitrite-induced inflammation, lowers blood pressure in various models, and exhibits neuroprotective effects. It restores mitochondrial and cardiac functions in TNFα-induced oxidative stress and reduces cardiac hypertrophy in chronic hypoxic rats.

dr. In the article “Effects of tempol and redox-cyclic nitroxides in models of oxidative stress” [131], Christopher S. Wilcox explains: Tempol is a redox-cyclic nitroxide that promotes the metabolism of many reactive oxygen species (ROS) and improves the bioavailability of nitric oxide. It has been extensively studied in animal models of oxidative stress. Tempol has been shown to protect mitochondria from oxidative damage and improve tissue oxygenation. Tempol improved insulin responsiveness in models of type II diabetes and improved dyslipidemia, reduced weight gain, and prevented diastolic dysfunction and heart failure in high-fat models of metabolic syndrome. Tempol protected many organs, including the heart and brain, from ischemia/reperfusion injury. Tempol prevented podocyte damage, glomerulosclerosis, proteinuria, and progressive loss of renal function in salt-overload and mineralocorticosteroid models. It reduced damage to the brain or spinal cord after ischemia or trauma and had an analgesic effect on the spine. Tempol improved survival in several models of shock. It protected normal cells from radiation, but at the same time preserved the sensitivity of tumor cells to radiation. Its paradoxical pro-oxidative action in tumor cells resulted in a decrease in spontaneous tumor formation. Tempol has been effective in some models of neurodegeneration. Thus, tempol effectively prevented many of the harmful effects of oxidative stress and inflammation that underlie radiation damage and many aging-related diseases.

Treatment of Covid-19 with tempol

While the subject of over 2000 publications, the SOD-mimetic nitroxide label tempol has been shown to be effective in the experimental treatment of several human and animal diseases while exhibiting minimal toxicity [90, 133]. By supporting the role of redox signaling in its mechanism of action, tempol modulates many redox-dependent cellular signaling systems. These include e.g. BRCA1, CtBPl, p53, PARP, HIF-Ια, HIF-2a, VEGF, IL-6, BARDI, RAD51, uPAR, NF-κΒ and mGluRs, several of which have been detected in the pathogenesis of Covid-19. It is important that the treatment of alopecia in the initial stage was associated with the therapeutic use of the drug tempol, which for many years promoted other aids. One of the inspirations for this application was the discovery in the mid-1970s that orgotheine, a pharmacological formulation of bovine liver SODI, inhibits hair loss (effluvium) in experimental diabetes in rodents. This implies a role for redox signaling in diabetes and in the laser cycle, a general model for redox modulation of cyclic cellular processes [89, 88], Tempol also improves radiation-induced hair loss in humans, a form of anagen effluvium [36] . Here we report the apparent efficacy of tempol in the treatment of early Covid-19 (SARS-CoV-2 infection), supporting a key role of oxidative stress and redox signaling in Covid-19. This may also explain the common association of Covid-19 with alopecia and the alleged efficacy of some hair loss treatments in the experimental treatment of this disease.

The term therapeutically effective amount, as used herein, means an amount of a compound or composition (eg, curcumin (0.1-5%), boswellia (0.1-5%), artemisinin (0.1-3%), vitamin C (0.1-6%) and optional CBD/CBG (0.1-5%) and TEMPO nitroxides (0.1-2%) sufficient to produce a significant positive change in the condition to be managed or treated , but low enough to avoid potential side effects within the scope of sound medical judgment (with a reasonable benefit/risk ratio).The therapeutically effective amount of a compound or composition will depend on the particular condition being treated, the age and physical condition of the end user, the severity the condition being treated/prevented, the duration of treatment, the nature of the concomitant therapy, the specific compound or composition used, the particular pharmaceutically acceptable carrier used, and similar factors As used herein, all percentages are by weight unless otherwise specified.

The term active pharmaceutical ingredient as used herein refers to curcumin, boswellia, artemisinin, vitamin C, and optionally the cannabinoids CBD/CBG and TEMPO nitroxides.

The actual doses of the active ingredient, curcumin, boswellia, artemisinin, vitamin C and optionally the cannabinoids CBD/CBG and TEMPO nitroxides within the scope of the invention can be varied to obtain the amount of active ingredient effective to achieve the desired therapeutic response for a particular patient, composition and method of use.

The dosage level selected will depend on a number of factors, including the activity of the particular active ingredient, curcumin, boswellia, artemisinin, vitamin C and optional cannabinoids CBD/CBG and TEMPO nitroxides, route of administration, time of administration, rate of elimination of the particular composition being used, duration treatment, use in combination with other extracts, age, sex, weight, condition, general health and previous medical history of the patient being treated, and factors well known in the medical profession.

The compositions of the presented invention are suitable for the treatment of both acute and chronic forms of inflammatory disorders caused by TNF-α, interleukins (IL-1, IL-6, IL-8) and ICAM-1, VCAM1 and E-selectin, in particular rheumatoid arthritis, juvenile rheumatoid arthritis, psoriatic arthritis, osteoarthritis, refractory rheumatoid arthritis, chronic non-rheumatoid arthritis, osteoporosis/bone resorption, coronary heart disease, vasculitis, ulcerative colitis, psoriasis, adult respiratory distress syndrome, Alzheimer's disease in humans. Also, the compositions of this invention can be used to treat inflammation in diseases such as inflammatory bowel disease, Crohn's disease, septic shock syndrome, atherosclerosis, and various autoimmune diseases, among other clinical conditions. The present invention also relates to a method for the treatment of inflammatory disorders, which comprises administering the compositions selectively orally.

The following examples illustrate, but do not limit, the scope of the invention. Those skilled in the art should understand that the present discussion is of a few exemplary embodiments only and is not intended to limit the broader aspects of the present invention, the broader aspects of which are included in the exemplary construction.

The cytokine storm associated with Covid-19

Treatment of SARS-CoV-2 with a composition within the scope of the present invention

Severe acute respiratory syndrome disease associated with coronavirus 2019 (COVID19) is a syndrome of viral replication coupled with a host inflammatory response. Cytokine storm and viral evasion of cellular immune responses may play equally important roles in the pathogenesis, clinical manifestation, and outcome of COVID-19. Systemic proinflammatory cytokines and biomarkers are elevated as the disease progresses to advanced stages and are associated with poorer survival [95],

SARS-CoV-2 activates the innate immune system and causes the release of a large number of cytokines, including IL-6, which can increase vascular permeability and cause fluid and blood cell leakage into the alveoli, resulting in symptoms such as dyspnea and respiratory failure [39], Higher mortality is associated with the result of exacerbation of ARDS (acute respiratory distress syndrome) and tissue damage that can lead to organ failure and/or death [92]. According to studies published in October 2020, ARDS is the cause of death in 70% of deaths due to COVID-19 [50]. In an analysis of cytokine levels in the plasma of those with severe Sars-CoV-2, the levels of several interleukins and cytokines are remarkably elevated, suggesting a cytokine storm in those most affected [92], In addition, postmortem examination of patients with COVID-19 demonstrated a large accumulation of inflammatory cells in lung tissues, including macrophages and Thelper cells [120], Covid-19 caused by SARS-CoV-2 is characterized by heterogeneous symptoms, ranging from mild fatigue to life-threatening pneumonia, cytokine storm and multi-organ failure [38], A cytokine storm has also been reported in SARS patients, which has been associated with poor outcomes [52]. Although the mechanisms of lung injury and multiorgan failure in Covid-19 are still under investigation, [112] reports of hemophagocytosis and elevated cytokine levels—as well as beneficial effects of immunosuppressants—in affected patients, particularly those who are most severely ill, suggest , that the cytokine storm may contribute to the pathogenesis of Covid-19 [24—8] [25—8]. Serum levels of cytokines that are elevated in patients with Covid-19-associated cytokine storm include interleukin-ΐβ, interleukin-6, IP10, TNF, interferon-γ, macrophage inflammatory protein (MIP) la and 1β, and VEGF [51 , 141], Higher interleukin-6 levels are strongly associated with shorter survival [28], The relative frequency of circulating activated CD4+ and CD8+ T cells and plasma blasts is increased in Covid-19 [73]. In addition to elevated systemic levels of cytokines and activated immune cells, several clinical and laboratory abnormalities are also observed in Covid-19, such as elevated levels of CRP and dymer, hypoalbuminemia, renal dysfunction and effusion, as there are storm disturbances in cytokines. Laboratory test results reflecting hyperinflammation and tissue damage have been found to predict worsening outcomes in Covid-19 [14]. Although immunological dysregulation has been observed in severe cases of Covid-19 [67], it is unknown whether immune hyperactivity or failure to resolve the inflammatory response in severe cases is due to ongoing viral replication or immune dysregulation. The association between nasopharyngeal viral load and cytokine levels (eg, interferon-α, interferon-γ and TNF) and declining viral load in moderate but not severe cases suggests that the immune response is positively associated with viral load [67], Other findings of type I innate immune system defects and type I interferon autoantibodies in the most severe cases of Covid-19 suggest that some patients with Covid-19 may have an inadequate antiviral response [139, 7, 61], Host immune responses and associated symptoms with the immune system, are very different between asymptomatic patients (who have effective control of SARS-CoV-2) and patients with severe Covid-19 (who cannot control the virus), suggesting that host immune dysregulation contributes in some cases to pathogenesis. Another hypothesized mechanism involves autoimmunity due to molecular mimicry between SARS-CoV-2 and a self-antigen. These mechanisms may be involved in subgroups of patients such as children with postinfectious multisystem inflammatory syndrome, a condition that appears to improve with immunomodulatory therapies such as intravenous immune globulin, glucocorticoids, and anti-interleukin-1 and anti-interleukin- 6 therapies. Patients with multisystem inflammatory syndrome very clearly fit the definition of a cytokine storm, as SARS-CoV-2 is no longer present; however, it is not clear whether the cytokine storm is the driving force behind Covid-19 or whether it is a secondary process. In addition, it is now clear that patients with SARS-CoV2 infection can be asymptomatic or have acute Covid-19 with heterogeneous severity, chronic course of Covid-19 or multisystem inflammatory syndrome. A critical question concerns the factors that contribute to the severe cytokine-like phenotype observed in a small proportion of patients. Co-existing conditions such as hypertension, diabetes and obesity are associated with more severe cases of Covid-19, possibly due to a pre-existing chronic inflammatory condition or a lower threshold for developing organ dysfunction due to the immune response.

Some important differences in therapeutic aspects between the cytokine storm associated with Covid-19 and many other cytokine storm disorders should be noted. First, the cytokine storm triggered by SARS-CoV-2 infection may require different therapies from those used for cytokine storms due to other causes. Cytokines may be a key component of the cytokine storm and an essential factor in the antimicrobial response. Thus, blocking cytokine signaling may actually impair SARS-CoV-2 clearance, increase the risk of secondary infections, and result in worse outcomes as seen with the influenza virus [61], as interleukin-6 and other cytokines are potentially critical for both a healthy response to SARS. -CoV-2 as a harmful cytokine storm, it is especially important that the right subgroups of patients with Covid-19 are selected for treatment in a timely manner. Despite positive anecdotal reports, two large, randomized, controlled trials of anti-interleukin-6 antibody therapies failed to demonstrate a survival benefit in hospitalized patients with Covid-19 [48, 118].

Second, the primary site of infection and disease most likely contributes to differences in immune responses and the mechanisms underlying the cytokine storm, with implications for treatment. For example, selective elimination of the primary viral reservoir is beneficial in patients with multicentric Castleman disease associated with HHV-8, but not in patients with Covid-19.

Third, lymphopenia is not commonly seen in cytokine storm disorders, but is a feature of severe Covid-19 disease. It is currently unclear whether the lymphopenia seen in Covid-19 is due to tissue infiltration or lymphocyte destruction.

Fourth, coagulation problems can occur in cytokine storm disorders, but thromboembolic events are more common in cytokine storms associated with Covid-19 [60]. Finally, although cytokine panels were not measured simultaneously on the same platform through the cytokine storms associated with Covid-19 and other cytokine storm disorders, preliminary results suggest that circulating levels of several cytokines such as interleukin-6 and other inflammatory markers such as ferritin, less elevated in Covid-19 than in some other cytokine storm disorders [67]. The levels of inflammatory mediators in lung tissue during SARSCoV-2 infection remain unknown.

Despite many unknowns, a recent randomized controlled trial showing that dexamethasone reduces mortality among the most severe cases of Covid-19, which is characterized by elevated CRP levels and supplemental oxygen requirements and may worsen outcomes in milder cases, suggests that late-stage inflammation phase contributes to mortality [122]. A meta-analysis of seven randomized trials found that 28-day all-cause mortality in critically ill patients with Covid-19 was lower in those treated with glucocorticoids than in those receiving usual care or placebo [116] . An observational study showing that patients with Covid-19 have a good response to glucocorticoids when the CRP level is high, but a poor response when the level is low, is consistent with these findings [59]. Further support comes from positive reports of targeted antagonists against interleukin-1, granulocyte-macrophage colony-stimulating factor, and JAKI and JAK2 in patients with Covid-19 [37, 26, 97]. Similarly, the finding that inflammatory agents such as inhaled interferon-β have a positive effect when given early in the course of the disease is consistent with a model in which immune stimulation that enhances antiviral activity is beneficial early (and likely detrimental late). , while immunosuppression is beneficial late and harmful early. As with dexamethasone, the timing of treatment and the selection of patient subgroups included in the studies are likely to influence the results.

Despite the unknowns regarding the role of immune dysregulation and cytokine storm in Covid-19, hundreds of immunomodulatory drugs are currently being investigated [37]. Many of these treatments have been used in other cytokine storm disorders. Canakinumab, an anti-interleukin-ΐβ monoclonal antibody, and anakinra are being studied for ARDS caused by Covid-19.

Acalabrutinib, a selective Bruton tyrosine kinase inhibitor that regulates B-cell and macrophage signaling and activation, may hold promise for dampening the hyperinflammatory response in Covid-19 [100]. JAKI and JAK2 inhibitors, which are approved for the treatment of many autoimmune and neoplastic conditions, can inhibit signaling by type I interferon, interleukin-6 (and other gpl30 family receptors), interferon and interleukin-2, among other cytokines [140]. Similar to anti-interleukin-6 antibody therapy, Bruton tyrosine kinase and JAK inhibition may prove harmful or ineffective if administered too early, when the immune response to SARS-CoV-2 is critical in controlling viral replication and clearance.

Cytokine storm treatment

The general treatment strategy for cytokine storm includes supportive care to maintain critical organ function, control of the underlying disease and elimination of triggers for abnormal immune system activation, and targeted immunomodulation or nonspecific immunosuppression to limit collateral damage of the activated immune system. As discussed in this review, many drugs are effective in many disorders under the cytokine storm umbrella, and may still be effective in several conditions that have not yet been investigated.

Given the growing number of new therapeutics targeting various aspects of the immune system and our ability to probe the biological mechanisms of disease, further research should focus on identifying drugs that can be used in cytokine storm disorders and accurate diagnostics to select the right drug for the right patients, regardless of the underlying condition [8, 25]. A study of patients with systemic juvenile idiopathic arthritis revealed subgroups of patients with cytokine profiles dominated by interleukin-6 and interleukin-18, suggesting available therapeutic approaches [110]. Similarly, biomarkers have recently been shown to effectively predict which adult patients with Still's disease will respond to anakinra or tocilizumab [127]. Advances in precision oncology suggest that similar efforts are warranted in cytokine storm disorders to identify specific therapeutic targets and signatures of response to certain drugs that transcend disease boundaries. JAK signaling is an interesting target in the cytokine storm because multiple cytokine-receptor pairs can be targeted simultaneously, which can be effective in multiple diseases caused by different cytokines. In addition, both plasma exchange and plasma filtration columns are evaluated in cytokine storm disorders.

Prevention of cytokine storm

However, a major question remains as to why some patients are more prone to cytokine storm than others. Different gene mutations can also be a risk factor for a more severe course of the disease and the occurrence of cytokine storms in COVID-19. Data obtained from global populations indicate that allelic variation in cytokine genes is also strongly influenced by latitude [115, 27]. Latitude is a major environmental factor influencing our evolutionary history in terms of environmental selection. Latitude is therefore related to various factors that include genetic background, biometeorological factors and socio-economic influences. Considering the role of biometeorological factors, sunlight plays a central role in the synthesis of vitamin D, which plays a key role in maintaining immune homeostasis. Genetic factors are known to account for up to 28% of inter-individual variability in serum 25(OH)D concentrations [109]. Genetic and individual differences in vitamin D status have been reported in different populations [64]. Based on this, it can be hypothesized that there is a possibility that vitamin D status has an impact on the geographic variance of COVID-19 [22].

In addition, vitamin D deficiency can lead to increased autoimmunity and increased susceptibility to infections. Vitamin D actually inhibits the production of inflammatory cytokines (i.e., TNF-α and IFN-γ) and promotes the release of anti-inflammatory cytokines. Vitamin D reduces the risk of microbial infection and death through various mechanisms. In a recent review, these mechanisms were classified into three groups, including physical barrier and innate and adaptive immunity [99], COVID-19 viruses disrupt the integrity of the junction, which increases susceptibility to infection by the virus and other microorganisms [101], while vitamin D supports the maintenance of cell junction integrity [ 105 ]. Vitamin D may be valuable in controlling the cytokine storm and disease course in patients with COVID-2019. Deficiency of this vitamin is a cause of increased risk, so vitamin D supplementation could potentially be used [45] Cytokine regulation depends on various upstream regulators, such as toll-like receptors (TLRs), and these are linked to other components of the innate immune system , such as complement elements. TLRs are a family of innate immune sensor proteins that have a key function in the processes of infection, inflammation and immunity [103]; the TLR pathway may be significantly involved in the cytokine storm that occurs during COVID-19 infection. To date, there are no studies on the role of TLR signaling in SARS-CoV-2 infection. However, previous studies suggest that genetic changes within TLRs or TLR signaling affected SARS-CoV infection [109, 103, 125, 43],

In an embodiment, the composition within the scope of the present invention may contain additives that are usually used for the preparation of pharmaceutical formulations. These may include pH buffers, desirable substances and stabilizing components.

In the embodiment, the pH buffer is selected from the group consisting of acetic acid, glacial acetic acid, lactic acid, citric acid, phosphoric acid, carbonic acid, histidine, glycine, barbital, phthalic acid, adipic acid, ascorbic acid, maleic acid, succinic acid, tartaric acid, glutamic acid, benzoic acid, aspartic acid and salts (eg potassium, sodium, etc.) or combinations thereof. The antioxidant is included in an appropriate amount to oxidize excess ions in the formulation. In a preferred formulation, the antioxidant contains less than about 10% by weight. % of the total formulation and, even more preferably, from about 0.05 wt. % to about 6 wt. % of the total formulation.

In an embodiment, the gelling agent is selected from the group consisting of xanthan gum, carrageenan, locust bean gum, guar gum, modified celluloses, low-esterified pectins, and colloidal silica.

In an embodiment, the stabilizing component of the formulation is selected from the group consisting of vitamin-E α-tocopherol, ascorbyl palmitate, BHT (butyl hydroxytoluene), BHA (butyl hydroxy anisole), propyl gallate or malic acid.

If desired, the formulation may further include conventional pharmaceutical additives. Examples of pharmaceutical additives that may be included are co-surfactants (eg, sodium lauryl sulfate), colorants, flavors, preservatives, stabilizers, and/or thickeners. The formulation can have a liquid or semi-solid form and can be filled into a gelatin capsule if desired. After administration, the capsule bursts and releases the formulation. When the formulation comes into contact with an aqueous environment, for example in the gastrointestinal tract, the formulation spontaneously forms an emulsion. One advantage of the invention is that active agents with poor water solubility can be dissolved and formulated into a useful therapeutic formulation.

Certain embodiments disclosed herein relate to free-flowing solid powders prepared from the present invention of self-nano formulations e SNEDDS. Powders can be prepared by subjecting the nano-formulation to encapsulation, nano-spray drying, thin-layer drying or lyophilization; or by combining the nano-formulation with a carrier selected from the group consisting of microcrystalline cellulose, precipitated silica, anhydrous dibasic calcium phosphate, mannitol, hydroxypropyl methylcellulose, cellulose, and mixtures thereof.

Various embodiments relate to the treatment of diseases selected from the group consisting of various viruses, inflammation, immunomodulatory, osteoarthritis, allergy, obesity, neurodegenerative disorders, diabetes, cancer, cardiovascular disorders, and microbiological disorders by administering to subjects the nanoemulsifying (SNEDDS) formulations described herein. , who need it.

Various formulations have been developed using one or a combination of hydrophobic compounds along with single or combinations of surfactants illustrated herein. After describing the disclosed subject matter by reference to certain embodiments, other embodiments will become apparent to one skilled in the art from consideration of the specification.

The content is further described with reference to the following examples. Those skilled in the art will appreciate that many modifications of the materials and methods may be made without departing from the scope of the subject matter presented.

EXAMPLES

This invention is further explained in the form of the following examples. However, it should be understood that the above examples are merely illustrative and should not be taken as limiting the scope of the invention. Experts will be able to recognize the various changes and modifications of the presented implementations. Such changes and modifications may be made without departing from the scope of the invention.

Example 1

ArtemiC ™ / CimetrA ™ are proprietary formulations targeting the treatment of symptoms of COVID-19 developed by MGC Pharma (UK) Ltd., Registration Number: 09750155, Central Working Ecclestone Yards, 25 Ecclestone Plache, London, United Kingdom, SW1W 9NF

ArtemiC™ is a nutritional supplement in the form of micelles, which is supported by the MyCell™ technology, used for the composition of the present invention, is presented in patents US2010/0196456A1 and US 2010/0129453A1 (MiVital, SWISS).

CimetrA™ - pharmaceutical composition based on nano-self-emulsifying drug delivery systems (SNEDDS), which is supported by GraftBio™ technology, suitable for the composition according to the present invention and is described in the SIPO application at the Office of the Republic of Slovenia for Intellectual Property (2021) P202100024 and the patent application at the Russian Patent Office (2021) W21013724/2021106347 by Victor Bolduev et al (SI).

CimetrA™ is an oral self-nanoemulsifying drug system (SNEDDS) according to the present invention containing all API components (curcumin, boswellia, artemisinin) as a lipophilic micelle core and emulsifiers as a micelle shell, vitamin C is a SNEDDS stabilizer and regulator pH-aqueous phases.

The following SNEDDS CimetrA™ formulation is most suitable (for drops, sprays).

Table 1. Formulation of CimetrA™

CimetrA™ formulation m/m % content artemisinin 0.6% API boswellia 1.5% API Curcumin 95% 2.0% API ascorbic acid 6.0% excipient cremaphor CH-40 20% emulsifier PEG-400 5% co-solvent another optional excipient distilled water 64.9%

Curcumin: The curcumin product used was Turmeric Oleoresin Curcumin Powder 95% which has the code EP-5001 from Green Leaf Extraction Pvt Limited, Kerala, India. Curcumin powder is CAS no. 458-37-7 and is a natural product obtained by solvent extraction of the rhizome of Curcuma Longa. According to the manufacturer, the curcumin content in the powder is at least 95%. This curcumin content is determined according to the ASTA 18.0 method.

Boswellia: The term boswellia primarily refers to an extract of the resin of the frankincense plant. In particular: alpha-boswellic acid (CAS number 471-66-9), beta-boswellic acid (CAS number 631-69-6) and their derivatives, 3-O-acetyl-alpha-boswellic acid (CAS number 89913 -60-0), 3-O-acetyl-beta-boswellic acid (CAS number 5968-70-7), 11-keto-beta-boswellic acid (KBA, CAS number 17019-92-0), and 3-O-acetyl -ll-keto-beta-boswellic acid (AKBA, CAS number 67416-61-9).

Artemisinin, also called qinghaosu, is an antimalarial drug derived from the sweet wormwood plant, Artemisia annua. Artemisinin is a sesquiterpene lactone (a compound consisting of three isoprene units attached to cyclic organic esters) distilled from the dried leaves or flower clusters of Artemisia annua.

Self-emulsification depends on the nature of the oil/surfactant pair, the surfactant concentration and oil/surfactant ratio, and the temperature at which self-emulsification occurs. Only very specific combinations of excipients lead to effective self-emulsifying systems (SNEDDS). The effectiveness of drug inclusion in SNEDDS depends on the specific physico-chemical compatibility of the drug / system. So solubility and phase diagram studies are required before formulation to get optimal formulation design.

Preparation of SNEDDS CimetrA™

1. Prepare and sterilize a glass container for mixing the ingredients (3 containers) and for storing the finished product.

2. Container no. 1

2.1. At a room temperature of 22-25 °C, mix 24% LINCOLORH 40/CG and 6% Kollisolv PEG 400. Mix for 15 minutes. Heat the mixture to 60 °C.

Visual inspection of quality. Result: light yellow homogeneous transparent mixture.

2.2. Inject 1.5% boswellia. Dissolve the boswellia completely while stirring for 15 minutes. Heat the mixture to 90 °C.

Visual inspection of quality. Result: dark yellow homogeneous transparent mixture.

2.3. Inject 2.0% curcumin. Dissolve the boswellia completely while stirring for 15 minutes. Heat the mixture to 95-98 °C.

Visual inspection of quality. Result: dark red (almost black) homogeneous mixture.

2.4. Turn off the heating. Allow the mixture to cool to a temperature of 60 while stirring

OC

3. Container no. 2

3.1. Degas (vacuum degassing with stirring) distilled water.

3.2. In container no. 2 pour 53.4% distilled water. Heat to 50 °C.

3.3. Add 6% ascorbic acid to heated distilled water. Dissolve the ascorbic acid for 10 minutes with constant stirring and maintaining a temperature of 50 °C.

Visual inspection of quality. The result: a clear mixture.

3.4. Add 0.5% Kollidon 17PF to heated distilled water. Dissolve Kollidon 17PF minutes while stirring constantly and maintaining a temperature of 50 °C.

Visual inspection of quality. The result: a clear mixture.

4. In container no. 2 slowly pour the contents of container no. 1. Mix thoroughly for 20 minutes and maintain the temperature at 50 °C.

Visual inspection of quality. Result: dark red homogeneous non-viscous transparent mixture.

5. Cool the emulsion to room temperature (20-25 °C).

6. Container no. 3

6.1. Inject 0.6% artemisinin into 2.4% acetone and dissolve completely while stirring for 5 minutes at room temperature.

Visual inspection of quality. The result: a clear mixture.

6.2. In container no. 3 add 4.8% Tween 80 and 1.2% Kollisolv PEG 400. Mix for 10 minutes.

Visual inspection of quality. Result: light yellow homogeneous transparent mixture.

6.3. Put the final mixture from container no. 3 in container no. 2. Mix for 20 minutes at room temperature. Acetone evaporates during mixing.

7. Vacuum the resulting emulsion for 10 minutes.

8. Pour the obtained nanoemulsion into the prepared container for storage.

The resulting emulsion contains the following APIs (in % by weight):

• artemisinin - 0.6% • boswellia - 1.5% • curcumin - 2.0% • ascorbic acid - 6.0% • distilled water - 53.4%

Characteristics of the emulsion:

• solubility in water (SNEDDS technology), • transparency (micelle size is less than 50 nm), • low viscosity, high fluidity (very suitable for use in sprays), • stability during storage (maintenance of all properties, even without formation of sediment, turbidity or phase separation of components).

A flow chart for the production of CimetrA™ is shown in the schematic (FIGURE 1).

Example 2

Solid self-emulsifying / microemulsifying drug delivery systems (S-SNEDDS)

Self-emulsifying / microemulsifying drug delivery systems require incorporation into capsules directly or conversion to granules, pellets and powders for dry-filled capsules as well as tablet preparations. The latter are possible through innovative adaptations of conventional equipment with relative ease and simplicity of process using methods such as melt granulation, melt extrusion, solid support adsorption, spray cooling, spray drying, liquid-based supercritical methods, and high-pressure homogenization. Recently, pellets containing self-emulsifying mixtures have been prepared using the extrusion and circulation technique.

A high content of liquid formulation can be loaded (up to 70%) on the carrier, which not only maintains good flowability, but also enables the production of tablets with good cohesive properties and good content uniformity in both capsules and tablets. This clearly expands the options available to the formulator. In addition to providing the obvious advantages of the in vivo tablet dosage delivery system (improved drug absorption, etc.), the development of solid dosage forms also has the advantage that a high content of liquid formulation can be loaded onto the container and the process provides good content uniformity. In terms of functionality and efficiency, neither the adsorption of the liquid formulation onto the carrier nor the state of the drug in the lipid formulation and the solubility properties of the final solid dosage form should be affected.

Self-emulsification depends on the nature of the oil/surfactant pair, the surfactant concentration and oil/surfactant ratio, and the temperature at which self-emulsification occurs. Only very specific combinations of excipients lead to effective self-emulsifying systems (SNEDDS). The effectiveness of drug inclusion in SNEDDS depends on the specific physico-chemical compatibility of the drug / system. So solubility and phase diagram studies are required before formulation to get optimal formulation design.

Development and characterization of CimetrA™ solid SNEDDS

The ratio of adsorbent and SNEDDS

A proper ratio of Aerosil 200 adsorbent to CimetrA™ liquid SNEDDS can produce the free-flowing properties of solid SNEDDS. In this case, an equal amount of Aerosil 200 and liquid SNEDDS CimetrA™ (50/50% w/w) produced free-flowing powders suitable for immediate tablet compression. On the other hand, a higher and lower amount of Aerosil200 with liquid SNEDDS CimetrA™ (33/67% w/w) produced significant dusting and greasy dust properties.

Free-flowing powders can be obtained in a simple way by adding the optimal amount of liquid lipid formulation to the selected carrier. In the example, liquid SNEDDS m Aerosil 200 50/50 (% w/w) ratios produced a dry free-flowing powder. Significant changes were observed between these ratios due to the high adsorption capacity of the adsorbent (Aerosil200) and retention of liquid SNEDDS in their internal pores. By visual inspection, all SNEDDS powders were dry and free-flowing at this point, indicating the ideal choice of a 50/50 (% w/w) ratio.

Example 3

Experimental and clinical investigations for COVID-19

Data on the correlation between COVID-19 and cytokine/chemokine dysregulation are still limited, but the current in vitro and clinical studies that are available suggest similarities with the reports of SARS and MERS infections.

Few studies of SARS-CoV-2 infection have been reported so far. One interesting study compared the behavior of SARS-CoV-2 and SARS-CoV in lung tissue. The research team inoculated the viruses into ex vivo human lung tissue samples and reported that SARS-CoV2 was more efficient than SARS-CoV at both replicating and infecting human lung tissues. Furthermore, infection with SARS-CoV-2 was less competent to induce the expression of any IFNs, suggesting that SARS-CoV and SARS-CoV-2 may differ in their ability to control the release of inflammatory cytokines and chemokines. Indeed, SARS-CoV infection increased 11 of the 13 inflammatory factors tested in this study, while SARS-CoV-2 upregulated only five of them (i.e., CXCL10, IL6, CCL2, CXCL1, and CXCL5), although he reproduced more efficiently. The expression of 12 of the 19 IFN and cytokine/chemokine genes tested was significantly lower in human samples infected with SARS-CoV-2 than in samples infected with SARS-CoV. Notably, CXCL8 transcription was increased only by SARS-CoV but not by SARS-CoV-2 infection, while the opposite was detected for CXCL10 [19],

Another research group isolated SARS-CoV-2 from a patient with already developed COVID19 and compared viral tropism and replication capacity with SARS, MERS and 2009 pandemic influenza H1N1 (HlNlpdm) in ex vivo human lung and bronchus samples. To assess extrapulmonary infection, the authors used ex vivo cultures of human conjunctival epithelium (potential portals of infection for SARS-CoV-2) and human colorectal adenocarcinoma cell lines [135], SARS-CoV-2 was able to infect mucosa, cilia, and rods of bronchial pneumocyte cells. type 1 epithelium in the lung and conjunctival mucosa. In the bronchus, rephkation of SARS-CoV-2 was higher than SARS and similar to MERS and lower than HlNlpdm. In the lungs, SARS-CoV-2 replication was comparable to SARS and HlNlpdm, but lower than MERS. In the conjunctiva, rephkation of SARS-CoV-2 was better than that of SARS-CoV. SARS-CoV-2 was less efficient at inducing inflammatory cytokines than H1N1 and MERS. Both SARS-CoV and SARS-CoV-2 thus replicate comparably in the alveolar epithelium; SARS-CoV-2 reproduces in the bronchus more widely than SARS-CoV. These findings provide valuable insight into the transmissibility of SARS-CoV-2 infection and differences between other respiratory pathogens [53],

In a retrospective study, the clinical and immunological characteristics of 21 patients (17 men and four women) affected by COVID-19 were evaluated. These patients were classified into different severity levels according to the guidelines of the National Health Commission of China. In particular, 11 patients with severe form showed a significantly increased value of IL-6, IL-10 and TNF-α in serum compared to reduced absolute numbers of T-lymphocytes, CD4 + T cells and CD8 + T cells compared to moderate cases. This retrospective observational study suggests that SARS-CoV-2 infection may primarily involve T lymphocytes, especially CD4 + and CD8 + T cells, leading to a decrease in T lymphocytes and IFN-γ production by CD4 + T cells. These potential immunological markers may be important due to their association with the severity of the disease of COVID19 [18],

To characterize the transcriptional signatures of the host inflammatory response to SARS-CoV2, Xiong and colleagues performed transcriptomic sequencing of various inflammatory genes from RNA isolated from bronchoalveolar lavage fluid and peripheral blood mononuclear cells of patients with COVID-19. This analysis revealed distinct host inflammatory cytokine profiles to SARS-CoV-2 infection and supports a link between the pathogenesis of COVID-19 and aberrant cytokine release; in particular, CXCL10 was upregulated in peripheral blood mononuclear cells, but no upregulation of the CXCL10 gene was detected in bronchoalveolar lavage fluid. In addition, SARS-CoV-2 induced the activation of many genes involved in apoptosis and P53 in lymphocytes, leading to the suggestion that this activity may be the main cause of the lymphopenia often observed in cases of COVID-19. Transcriptome sequencing analysis of patients with COVID-19 represents an important resource for clinical guidelines on anti-inflammatory treatment and for understanding the molecular mechanisms of the host response [134],

Another study involving 65 patients positive for SARS-CoV-2 showed that the absolute number of CD4 + and CD8 + T cells and B cells gradually decreased with increasing disease severity [129]. In addition, Yang and colleagues analyzed 48 circulating cytokines in 53 patients with COVID-19 (34 severe cases), and 14 had higher scores in patients with a severe clinical history of COVID-19. Among them, CXCL10, CCL7 and IL-1 receptor antagonists were strongly associated with more severe disease and, more importantly, CXCL10 levels were the only ones that had a positive and significant correlation with viral load [137].

Of the 70 patients who survived severe COVID-19 pneumonia, 66 showed significant damage as shown by a CT scan performed before hospital discharge. The lesions varied: from dense lumps of tissue obstructing the blood vessels of the alveoli to tissue lesions. Tissue lesions may represent signs of chronic lung disease and may be irreversible, leaving the patient vulnerable [130]. In addition, survivors of ARDS due to COVID19 may have permanent lung scarring [114]. If the lung tissues are replaced with scars, they no longer function like normal lung tissues, which can lead to poor gas exchange. Similar damage has also been noted in survivors of MERS and SARS, even when these diseases only affected one wing of the lung.

It is important to consider several factors when managing a cytokine storm. Neutralization of a particular cytokine whose level is elevated in the circulation with an existing agent (anti-interleukin-6, anti-TNF, anti-interferon-γ, or anti-interleukin-ΐβ antibody) will not always be effective, and blocking a cytokine with a low or normal level in the circulation is may be effective if it is a key component of the hyperinflammatory circuit or if its tissue level is potentially elevated.

In addition, the various therapies mentioned in this review have distinctive side effect and risk profiles. All targeted drugs have target-specific risks, and combination therapy has more potential risks than single-drug therapy. In addition, pathological hyperinflammation is itself an immunodeficiency that can put patients at risk for infections, and immunosuppressants most likely increase the risk. In this era of cytokine profiling and personalized medicine, patients should be monitored and given appropriate prophylaxis when treated empirically, and randomized, controlled trials should always be performed to assess efficacy and safety.

To advance cytokine storm research and treatment, it will be necessary to pool samples for omics studies and collaborate among experts under different conditions. The introduction of the International Classification of Diseases, 10th Revision, cytokine release syndrome code in 2021 should facilitate electronic research of natural history, pathogenesis, and treatment based on medical records. Once sufficient scientific progress has been made in individualized biomarker-guided treatment of cytokine storms, reliable, rapid, and accessible assays to measure soluble mediators of inflammation in plasma and tissues will be needed.

CimeterA

CimetrA is composed of natural active ingredients and is formulated in microscopic structures known as micelles. The elements that form the structure of these micelles are also exclusively of natural origin. The unique formulation has highly desirable pharmacological properties that ensure an otherwise unattainable high bioavailability of the active ingredients for which it is intended.

Test drug doses are defined according to the following:

• Group 1: CimetrA with a full dose containing a combination of artemisinin 12 mg, curcumin 40 mg, boswellia 30 mg and vitamin C 120 mg in spray delivery divided into 4 separate doses, including doses at 2 a.m. and 2 p.m. afternoon within 48 hours.

• Group 2: CimetrA, with a total dose containing a combination of artemisinin 15.6 mg, curcumin 52 mg, boswellia 39 mg and vitamin C 156 mg in a spray divided into 4 separate doses, including doses at 2 am and 14 .hours in the afternoon within 48 hours.

Example 4

Preclinical data

4.1. Acute toxin study

The study was conducted at the preclinical facility SIA - Science in Action, Ness Ziona, Israel. Science in Action is obliged to comply with the OECD Principles of Good Laboratory Practice ENV/MC/CHEM (98) 17 for toxicity studies; however, this study does not follow all GLP regulations and is therefore considered a non-GLP study. The study follows this protocol and Science in Action SOPs.

The aim of this study was to evaluate the safety and toxicity of low-dose CimetrA spray by oral spray.

Animal handling was performed in accordance with National Institutes of Health (NIH) and Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC) guidelines. Animals were housed in polyethylene cages (3 per cage), size 35 x 30 x 15 cm, with a stainless steel top, where pelleted food and drinking water were provided in a plastic bottle; bedding: clean, steam-sterilized paddy bran was used, and the bedding was changed with the cage at least twice a week. The study was conducted after approval by the Israel Committee for Animal Experiments in accordance with the Israel Animal Protection Law and the Ethics Committee.

Each group of 3 + 3 rats was treated by spraying the oral cavity with experimental substances on the first day.

Each animal was weighed before treatment.

Group 1 (n=3M + n=3 F): 50 μΐ saline per rat

Group 2 (n=3M + n = 3F): 48 pg CimetrA / per kg rat

Group 3 (n=3M + n = 3F): 96 μg CimetrA / per kg rat Group 4 (n=3M + n = 3F): 192 pg CimetrA / per kg rat M means male rat, F means female rat

During all 7 days of the experiment, the animals were monitored and weight was observed to detect the appearance of abnormal clinical signs.

After 7 days, blood samples were taken from all rats for a complete hematological and chemistry panel. Blood 0.2 for unbound EDTA test for hematology panel and 0.5ml for detachable gel tube, up to 0.25ml serum for blood chemistry panel.

After blood collection, the animals were sacrificed, the organs: brain, lungs, heart, liver, spleen and kidneys were removed, weighed and stored in 4% formalin and sent for pathological examination.

4.1.1. The results

The results are shown in FIGURES (FIGURES 2, 3, 4, 5).

Histopathology results

Samples (n = 168) of liver, heart, brain, spleen, spinal cord (uterine, thoracic and lumbar), sciatic nerve, kidney (L + R), lung and tongue of 24 rats fixed in 4% formaldehyde were taken. arrived at Patho-Logica in fixative and was kept in fixative for 48 hours for further fixation. The tissues were then trimmed, placed in embedding cassettes, and routinely processed for paraffin embedding. Seven cassettes were prepared for each animal. Paraffin blocks were cut approximately 4 microns thick. Sections were placed on glass slides and stained with hematoxylin and eosin (HinE). Images were taken with an Olympus microscope (ΒΧ60, serial no. 7D04032) at X4 and XI0 objective magnification and a microscope camera (Olympus DP73, serial NO. OH05504).

Histological evaluation

HinE-stained slides were examined, described and graded using a semi-quantitative classification of five levels (0—4) for the severity of pathological changes (Schafer et al.):

Score 0: The tissue looks normal, without any changes.

Level 1: minimal pathological changes.

Level 2: mild pathological changes.

Level 3: moderate pathological changes.

Level 4: severe pathological changes.

Histopathological evaluation included comparison between treated and control or naïve animals. Pathological findings were described, graded and shown on representative histological images. At the customer's request and only in specific ΤΟΧ studies, NOAEL and LOAEL values were determined after review of all samples from control and treatment groups and comparison between all treatment groups (control, low; intermediate and high doses).

The results

Histopathology • In general, HinE-stained sections showed no pathological changes in all animal samples tested.

• The spleen of all tested animals was reactive and showed a marked proliferation of white pulp lymphocytes. However, this is not considered a pathologic finding and is most likely unrelated to the treatment being tested.

• A population of mast cells was observed in the tongue muscles of all samples. These cells are usually in this area; therefore, this finding is not considered pathological. In all spinal cords, the white matter appeared diffusely vacuolated. This change is considered an artifact due to the decalcification process. We did not observe these changes in brain sections.

• We also studied the pathological evaluation, including grading, of individual animals in different groups (FIGURES 6-14).

Summary and conclusions

All samples appeared normal and did not show any pathological changes.

4.2. An in vitro study

The aim of the study was to examine the effect of CimetrA and their components on the viability of human peripheral blood mononuclear cells (PBMC) and their ability to attenuate the inflammatory response upon stimulation with E-coli-derived lipopolysaccharide.

Preparation of dilutions of CimetrA and their components in culture medium as indicated in Table 2.

Table 2. Solutions of CimetrA and its components in the culture medium

Name Stock concentration (%) Stock conc. (mg/ml) Target concn. in culture media (μg/ml) Dilution factor full composition CimeterA TM artemisinin 0.6 6 3,704 1620 M vitamin C 1.5 15 9,259 1620 dissolved vitamin C 4.5 45 27,778 1620 incense boswellia resin (boswellia) 1.5 15 9,259 1620 curcumin 2 20 12,346 1620

artemisinin 2 20 3,704 5400 mixture vitamin M vitamin C + 1.5 15 9,259 1620 Mono components C 6% dissolved vitamin C 4.5 45 27,778 1620 incense boswellia resin (boswellia) 2.5 25 9,259 2700 curcumin 6 - 60 12,346 4860 ---

Process

PBMCs were thawed and seeded at a density of 5x105 cells/well in 100 μΐ culture medium.

PBMCs were incubated for 18 hours at 370°C in 5% CO2.

For the viability test, the cells were centrifuged at 300g for 8 minutes after an incubation period and the medium was cultured with media containing the test products alone or in combination as shown in Table 3.

Table 3: Treatment groups

Group Treatment 1 CimetrA™

2 artemisinin 3 curcumin

4 boswellia 5 vitamin C blend 6 artemisinin + curcumin 7 artemisinin + boswellia 8 artemisinin mixture + vitamin C 9 artemisinin + curcumin + vitamin C blend Group Treatment 10 artemisinin + curcumin + boswellia 11 boswellia + curcumin + vitamin C blend 12 boswellia + artemisinin + vitamin C blend 13 L___L curcumin + boswellia ----

14 curcumin + vitamin C blend 15 boswellia + vitamin C blend 16 solvent —

Processing was performed in triplicate.

Wells with cells in media containing cell culture water (vehicle) served as controls. PBMCs were incubated with subjects/controls for 24 hours at 37°C in 5% CO2. At the end of the incubation period, PBMCs were assessed for viability using the Real Time-Glo (TM) MT cell viability assay according to the manufacturer's instructions. To study the anti-inflammatory effect, PBMCs were pretreated with the subjects individually or in combinations as indicated in the table above and incubated for 3 hours at 37°C in 5% CO2. After the incubation period, PBMCs were stimulated with 100 ng/ml LPS with or without test elements and incubated for 24 h at 37 °C in 5% CO2. An additional set of PBMCs pretreated with the CimetrA formulation were co-stimulated with 10 ng/ml LPS with the CimetrA formulation and incubated for 24 h at 370 °C in 5% CO2. PBMCs treated with 10 ng/ml LPS alone served as control. At the end of the incubation period, conditioned media were collected and subjected to cytokine analysis using a magnetic luminex assay kit for TNFα, IL-IRα, IL-Ιβ, IL-6, and IL-2 according to the manufacturer's instructions.

4.2.1. The results

The results are shown in the figure (FIGURE 15).

Cytokine secretion

The concentration of TNF-α, IL-IRA, IL-Ιβ, IL-6, and IL-2 in conditioned media with PBMCs after pretreatment with test subjects for 3 h and co-stimulation with LPS for 24 h was evaluated using the Human Magnetic Luminex assay (FIGS. 16-20).

Conclusions

In an in vitro study on induced cell culture, CimetrA confirmed its ability to prevent cytokine storms.

Example 5

Clinical data

A randomized controlled clinical trial in moderately hospitalized patients with COVID-19 was conducted in Israel and India. Fifty patients were included in the study, 33 in the treatment group and 17 in the placebo group. 40 patients were included in the study at three hospitals in Israel and 10 at one hospital in India.

Purpose of Study: This study was designed to evaluate the safety and efficacy of CimetrA in patients diagnosed with COVID-19.

Study endpoints

Primary Results • Time to clinical improvement, defined as a National Early Warning Score 2 (NEWS2) of <1= 2, was 24 hours longer compared to routine treatment.

• Percentage of participants with definite or probable drug-related adverse events.

Results in seconds:

• Time to negative PCR.

• Proportion of participants with normalization of fever and oxygen saturation by day 14 from symptom onset.

• Controlled clinical study III. phase aimed at evaluating the effect of CimetrA in patients diagnosed with COVID-19.

• Survival related to COVID-19.

• Frequency and duration of mechanical ventilation.

• Incidence of intensive care unit stay.

• Length of stay in the intensive care unit.

• Duration of supplemental oxygen delivery.

Sl • Additional data will be recorded to supplement the basic set of results.

Population covered by the study

Inclusion criteria

1. Confirmed infection with SARS-CoV-2.

2. A hospitalized patient with moderately stable or worsening COVID-19 who does not require admission to an intensive care unit and, on the other hand, does not improve clinically with continued standard care.

3. Age is 18 years and older.

4. Subjects must be under supervision or admitted to a supervised facility or hospital (home quarantine is not sufficient).

5. They have the possibility of treatment by spraying into the oral cavity.

Exclusion criteria

1. Tube feeding or parenteral nutrition.

2. A patient who needs oxygen supply in addition to the use of nozzles or a simple mask according to assessment 4.

3. Respiratory decompensation requiring mechanical ventilation.

4. Uncontrolled type 2 diabetes.

5. Autoimmune diseases

6. Pregnant or nursing mothers.

7. The need for admission to the intensive care unit during the current hospitalization at any time before the end of participation in the study.

8. Any condition that, in the opinion of the principal investigator, would prevent full participation in this trial or interfere with the assessment of trial endpoints.

Methodology

A multicenter controlled study. 50 adult patients suffering from infection with COVID19. Safety was assessed by collecting and analyzing adverse events, blood and urine laboratory tests, and vital signs. After the screening visit, they received the drug twice a day in the morning and in the evening (every 12 hours) between (Day 1 and Day 2). Patients were randomly assigned in a 2:1 ratio to test drug and standard care or placebo and standard care. The study lasted 2 weeks until completion on day 15 or hospital discharge, whichever occurred later. In case of discharge from the hospital during the study period, the protocol was continued until day 15, regardless of where the subject was. In case of prolonged hospitalization of more than 15 days, subjects continued to be monitored for safety and endpoints until discharge. The basic characteristics of the patients are presented in Table 4.

Table 4. Baseline characteristics of drug and placebo groups.

Cimeter A™ Placebo P Old age 52±14 53 ± 14 0.857 Gender: men 17(52) 8(47) female 15 (46) 9(53) 0.708 Race: Asian 9(27) 1(6) white 23 (70) 16(94) 0.139 Smoking: Currently 5(15) 3(18) in of the past 1(3) 3(18) never 26 (79) 11(65) 0.277

Alcoholic drink: occasionally 2(6) 0 weekly 0 1(6) never 30(91) 16(94) 0.321

Performance data

NEWS analysis of results

All 33 patients in the treatment group had NEWS scores less than or equal to 2 at their last measurement, compared with only 12 of 17 in the placebo group; P=0.015.

There was no difference in the average NEWS score at the first visit, while at the last measurement the score was significantly lower. See Table 5.

Table 5. NEWS Score change in both considered groups

Group statistics Groups St. Average Standard deviation Standard error mean P total NEWS 1 CimeterA 33 1.5152 2.00189 .34848 placebo 17 1.8824 2.05798 .49913 0.546 final NEWS CimeterA 33 .5152 .66714 .11613 placebo 17 2.2353 3.19236 .77426 0.042

In the ITT analysis using imputation, a significant difference appeared between the groups in the last part of the observation session (FIGURE 21).

Room air or supplemental 02 in patients is presented in Table 6.

Table 6. Room air or supplemental 02 by patients

Visit 1 During the study Last seen CimeterA n=33 4 patients 4 patients 0 patients Placebo n=17 3 patients 5 patients 4 patients

Security information

No adverse events or drug-related adverse events were reported during the study.

Conclusions

A phase I clinical study showed a complete safety and efficacy profile of CimetrA, supported by preclinical study results.

Example 6

Packaging and labeling of the medicine

CimetrA formulations will be stored in opaque spray bottles at room temperature and labeled in accordance with GCP and local MoH requirements.

CimetrA formulations will be packaged and labeled with two-part labels. The first part will be firmly attached to the bottle and the second part will be detachable and will be attached to the medication liability record in the CRF when CimetrA is administered. The sticker will contain the following information:

• protocol number, • dose, • batch number, • production date, • retest date.

• name of the sponsor, • instructions for use and storage, • statement "Only for use in tests".

6.1. Supply, distribution and dispatch

MGC and/or its designee will deliver the tested products to the clinical site. The tested product will be manufactured by MGC under EU-GMP manufacturing conditions and will be sufficient to complete the trial.

Each shipment submitted for study will include a shipment form describing the contents of the shipment. This form will help keep current and accurate information in the records. Once the shipment is received, the investigator, coordinator, or pharmacist will confirm its contents and confirm receipt of the trial product stock by signing the shipment form and faxing it to the MGC representative. If, after arrival at the investigation site, the stock of the tested product is damaged or missing, the MGC should be contacted immediately.

6.2. Storage, distribution and return of investigational products

All investigational products sent to the study center must be stored under certain conditions in a secure area accessible only to the investigator and authorized personnel. All investigational products must be stored and inventoried in accordance with applicable government regulations and study procedures. The temperature should be maintained between 20-25 °C.

Empty bottles must be returned to Sponsor for destruction or disposal as chemically hazardous waste in accordance with local regulations.

6.3. Liability and compliance of investigated products

Subject compliance with the trial drug dosing regimen will be assessed by study staff administering the trial drugs.

A medication accountability log will be used at the study center to maintain accurate records of the center's investigational medication supplies (date and quantity received by the investigator, dates of administration to subjects, dates when unused medication is returned to the sponsor or disposed of).

The investigator or designee will be responsible for maintaining accurate records of the quantity and dates of all test product supplies received, handled, and returned. The amount of lost, missing, destroyed, etc. should also be considered and documented. of the product under test. At the end of the study, it should be possible to reconcile the supply records with the records of use and returned stock. In case of discrepancies, invoices must be provided.

Government regulations require that all test materials not used in clinical trials be returned to the sponsor before or at the conclusion of the study. The investigator will return certain copies of the completed distribution and inventory record as indicated on the form.

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Claims (12)

1. A new nanomicelle pharmaceutical synergistic composition that includes curcumin, boswellia, artemisinin, vitamin C and optionally cannabinoids and TEMPO nitroxides. 2. A new nanomicelle pharmaceutical synergistic composition according to claim 1, characterized in that it includes a therapeutically effective amount a) curcumin from 0.1-5 wt. %, boswellia from 0.1-5 wt. %, artemisinin from 0.1-3 wt. %, vitamin C from 0.1-6 wt. % and b) optional cannabinoids from 0.1-5 wt. % and/or TEMPO nitroxides from 0.1-2 wt. %. 3. New nanomicella pharmaceutical synergistic composition according to claim 1, characterized in that the active pharmaceutical ingredients are natural products, synthetically obtained products or their mixture. 4. New nanomicelle pharmaceutical synergistic composition according to claim 1, characterized by the fact that it is encapsulated in micelles a pharmaceutically acceptable nanocarrier of a drug delivery system with self-emulsifying properties - SNEDDS. 5. New nanomicelle pharmaceutical synergistic composition according to claim 1, characterized in that it is in liquid form, such as oral spray or drops for the mucous membrane. 6. New nanomicella pharmaceutical synergistic composition according to claim 1, characterized in that it is in solid form, such as powder, wherein suitable materials as pharmaceutically acceptable carriers are sugars, such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; malt; gelatin; talc; as well as other non-toxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweeteners, flavors and perfumes. 7. New nanomicella pharmaceutical synergistic composition according to claim 1, characterized in that it optionally includes preservatives and antioxidants. 8. New nanomicella pharmaceutical synergistic composition according to claim 1 with antioxidant, anti-inflammatory, immunomodulating, antiviral and anticancer properties for the treatment of several therapeutic indications. 9. A novel nanomicelle pharmaceutical synergistic composition according to claim 1 for the treatment of an inflammatory disorder relating to a disease or condition characterized by chronic inflammation, including, but not limited to, rheumatoid arthritis, osteoarthritis, juvenile rheumatoid arthritis, psoriatic arthritis, unresponsive rheumatoid arthritis, chronic non-rheumatoid arthritis, osteoporosis/bone resorption, coronary heart disease, atherosclerosis, vasculitis, ulcerative colitis, psoriasis, Crohn's disease, adult respiratory distress syndrome, skin delayed hypersensitivity disorders, septic shock syndrome and inflammatory bowel disease. 10. New nanomicella pharmaceutical synergistic composition according to claim 1 for immunomodulation by regulating the circadian rhythm in the body, increasing the immune response to bacterial, viral, fungal and parasitic infections, replacing or transforming damaged tissue by stimulating the growth of fibroblasts and destroying certain tumors. 11. A novel nanomicelle pharmaceutical synergistic composition according to claim 1 for the treatment of severe acute respiratory syndrome of the coronavirus, associated with the coronavirus COVID-19, severe influenza, cytokine storm and alopecia after COVID. 12. New nanomicella pharmaceutical synergistic composition according to claim 1 for the treatment of various forms of cancer by accelerating the metabolism of many reactive oxygen species and improving the biological availability of nitric oxide and protecting normal cells from radiation, while simultaneously maintaining the sensitivity of tumor cells to radiation.