AU2021106617A4

AU2021106617A4 - A novel formulation of Backhousia Citriodora and Zingiber extracts with antiviral effects against SARS-CoV-2

Info

Publication number: AU2021106617A4
Application number: AU2021106617A
Authority: AU
Inventors: Kerrie Symons
Original assignee: Individual
Current assignee: Individual
Priority date: 2021-08-23
Filing date: 2021-08-23
Publication date: 2021-11-11
Anticipated expiration: 2029-08-23

Abstract

The present innovations are novel formulations combining Backhousia citriodora oil and Zingiberofficinale, particularly 6-gingerol Citral and Zingibain, that inhibit the attachment and entry of SARS-CoV-2 virus by mediating the Spike protein, ACE2 and TMPRSS and interfering with the fusion of the virus with the host cell. The novel oil in water emulsion, leaf paste and SMEDDS formulations have other bacterial fungal and viral activity specifically human respiratory syncytial virus (RSV) S. aureus, E. coli, P. aeruginosa, C. albicans, MRSA, A. niger, K. pneumoniae and P. acnes and is predicated on the quenching of the sensitization and the toxicity of citral by the 6-gingerol and the formulations have anti-platelets aggregation and vasorelaxing effects.

Description

EDITORIAL NOTE 2021106617

There are 10 pages of description only.

Description

The angiotensin-converting enzyme 2 (ACE2), a host cell receptor, has been found to play a crucial role in virus cell entry; therefore, ACE2 blockers can be a potential target for anti viral intervention.

A composition comprising lipophilic extracts of Zingiber officinale and Backhousia citriodora have potent ACE2 inhibitory effects in epithelial cells anddownregulate ACE2 expression.

Conditions tied to elevated furin levels - including obesity, hypertension and diabetes - are connected with vulnerability to more severe forms of Covid-19.

The Spike glycoprotein of SARS-CoV-19 - the causative pathogen for Covid-19 - contains a furin cleavage complex (FFC). The modified S glycoprotein can interact with the cell surface receptor angiotensin-converting enzyme 2 (ACE2) which is the specific receptor found in human cells, particularly epithelium, that the virus uses to gain entry. The S1 or receptor binding domain contacts with ACE2, which is facilitated by a furin cleavage. Research has shown that the S protein of SARS-CoV2 is between 10 and 20 times more likelyto bind to human ACE2 than the S protein of previous coronaviruses. This means that SARS-CoV-2 is significantly more infectious than both SARS and MERS. SARS-CoV-2 uses endogenous furin to cleave the S protein in the trans-Golgi network right after virion assembly. The mechanism separates furin from other virally hijacked proteases, which increases the pathogenity of SARS-CoV-2.

The presence of FCC allows the virus to spread systematically and cause higher rates of severe disease and death. Furin is present in most tissues and is highlyexpressed in the lungs which may explain how the virus gains entry into the respiratory tract and causes infection. The presence of elevated furin levels in the obese and diabetic population makes them more susceptible to being infected with SARS-CoV-2 and more likelyto experience severe complications as it spreads rapidly throughout the body including coagulopathy. This innovation has synergistic antiviral effects and citral and 6-gingerol are effective in preventing viral entry and replication and based on their binding abilitywith multiple COVID-19 targets, with anti-protease targets.

This innovation is a formulation of Backhousia citriodora oil, 6-gingerol that is a furin inhibitor and has an accepted risk-benefit ratio.

Further citral and 6-gingerol inhibit the action of furin in SARS-CoV-2 the furin enzyme plays a key role to cleave glycoprotein which is the first stepto initiate an infection. SARS-CoV-2 has a furin cleavage site atthe junction of subunits S1 and S2 This innovation disrupts the binding and fusion machinery required by coronaviruses. Further cytokine storm is the exaggerated immune response often observed with viral infections and this innovation is a potent anti-inflammatory combination.

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is a new type of corona virus that is responsible for the COVID-19 pandemic. Not surprisingly, spike protein of SARS-CoV-2 can directly bind to angiotensin converting enzyme 2 receptor of host cells (ACE2) indicating that an ACE2 inhibitor could block SARS-CoV-2 entry.

ACE2, a trans-membrane protein that is mainly involved in vasodilation, and is an important target for controlling hypertension. ACE2 receptor targets are those which have been used for diabetics and heart disease, additionally ACE2 receptors are present in the intestines, kidneys, oral mucosa and heart endothelium.

COVID-19, the disease that SARS-CoV-2 causes, can spread to the lungs, causing pneumonia. Ventilator acquired pneumonia is a common nosocomial infection in such patients.

The main function of proteases in any living organism is the cleavage of proteins resulting in the degradation of damaged, misfolded and potentially harmful proteins and therefore providing the cell with amino acids essential forthe synthesis of new proteins. Plant proteases can also play a part as signal molecules and in contrast to human-encoded enzymes possess exceptional features such as higher stability, unique substrate specificity and a wide pH range for enzymatic activity. These valuable features make plant-derived proteolytic enzymes suitable for Covid19 therapies.

The innate immune system provides the first barrier to infection and disease through various processes including activation of inflammation. Inflammation is a biological process whereby immune cells, including macrophages, respond to, and work together to eliminate damage from injury and disease. Chronic or sustained inflammation can be detrimental to the health and growth of humans and animals. Immune modulating functional plant compounds extracted from Backhousia citriodora and Zingiber officinale are a safe approach to controlling inflammation in Covid19. 6-gingerol modifies macrophage function and has a significant effect on mRNA expression of antibacterial and cytokine genes COX2, TNFa, IL-1B, analysed by real time quantitative polymerase chain reaction (qPCR).

Studies also point out that treatment with transmembrane protease shrine 2 (TMPRSS2) inhibitor significantly blocked COVID 19 entry, therefore either ACE2 or TMPRSS2 blockers are a potential targets for anti-viral intervention. Other studies reported that the Covid-19 receptor binding domain was capable of entering cells expressing human ACE2 while other receptors are ineffective, confirming that human ACE2 is the prime receptor for Covid-19 targeting the precise receptor ACE2.

ACE2 overexpression was frequently observed in gastrointestinal tissues and colon cell lines which is comparatively higher than that of other tissues including lung tissues. The development of new drugs for targeting ACE2 and treating Covid19 could be time consuming making the safetyand efficacyof plant proteases pivotal. The traditional medicine systems from many geographical areas use gingeras the primary treatment of viral infections including Covid19. In addition, natural products including diterpenoids sesquiterpenoids and triterpenoids have been shown to inhibit SARS-CoV-2. Citral has been shown to be capable of inhibiting ACE2.

As the entry of SARS-CoV-2 is mediated by ACE2 receptor, people undergoing drug therapies for diabetes and hypertension are more susceptible to SARS-CoV-2 as they would be inherently overexpressing ACE2 receptors.

The Covid-19 genome does not have the processing protease for the viral membrane fusion glycoprotein precursors, entry of this virus into cells is determined primarily by host cellular, trypsin-type, processing proteases that proteolytically activate the fusion glycoprotein precursors of Covid-19.

At leastfive different processing proteases have been identified in the airwaves of humans and animals. The proteases determine the infectious organ tropism of Covid-19 as well as the efficiency of viral multiplication in the airway, and sometimes in the brain. Proteases in the upper respiratory tract are suppressed by secretory leukoprotease inhibitor, and those in the lower respiratory tract are suppressed by pulmonary surfactant which, by adsorption, inhibits the interaction between proteases and viral membrane proteins. Since protease activities predominate over those of endogenous inhibitory compounds under normal airway conditions, administration of protease inhibitors in the early stage of infection significantly suppresses viral entry and viral multiplication.

SARS-CoV-2, the virus that causes COVID-19, enlists the help of two enzymes on the surface of human cells in order to invade them. This innovation is a compound that inhibits both enzymes. Viral enzymes can only invade cells and replicate with the assistance of the cells own molecular machinery. Proteases on the cells own surface must split open the protein spikes that give the virus its characteristic crown-like appearance.

This splitting changes the shape of the spikes, exposing the binding sites that allow the virus to gain entry to the cell. The spikes of coronaviruses contain three "cleavage sites" where particular proteases can splitthe proteins, the cleavage sites and their respective proteases help determine how pathogenic the virus is. This invention is a compound that inhibits two particular proteases and protects cells from invasion by SARS-CoV-2.

One of the proteases, called furin, is used by some of the most pathogenic coronaviruses and is one factor that helps SARS-CoV-2 spread so easily.

Over the past several decades, the frequency of antimicrobial resistance and its association with serious infectious diseases has increased at alarming rates. VAP and Mucormycosis (blackfungus) deaths are soaring with Covid-19. With the alarming increase in antibiotic resistance in various microbial pathogens, identification of an alternative strategy and novel agent to counter bacterial resistance and that is generally safe for human consumption (GRAS). 6-gingerol has good oral availabilityand Citral is considered a potent antifungal and antibiotic.

SARS-CoV-2 viral entry factors such as ACE2 and TMPRSS members were broadly enriched in epithelial cells of the glands and oral mucosae. Saliva from SARS-CoV-2 infected individuals harbours epithelial cells exhibiting ACE2 and TMPRESS expression and sustains SARS-CoV-2 infections.

Periodontal Disease and Mucositis

Periodontal disease is an inflammatory condition that affects the supportive tissue of the teeth and various studies report the prevalence and severity of periodontal disease is greater among diabetic patients. In a double-blinded clinical trial, diabetic patients received ginger supplementation for 2 months. The results showed that the intervention significantly reduced the levels of TNF-A, IL-6 and hs-CRP ingingergroup post-intervention. It is indicated that ginger may inhibit the synthesis of both prostaglandin and leukotriene. Such dual inhibition of cyclooxygenase (COX) and lipoxygenase (LOX) means a more effective anti inflammatory agent.

Proinflammatory cytokines such as IL1-B.IL-6ANDTNF-a are activated through activation of reactive oxygen species activityinT2DM patients with CP. There are several compounds in gingeracting as a blocker of serotonin receptors and serotonin receptors may be correlated with the reduction of TNF-a IL-1b IL-6 IL-2 and prostaglandins.

In this innovation the citral in B. citriodora and Z.officinale shows appreciable antimicrobial activity against Gram-positive and Gram-negative bacteria as well as fungi. Media composition and inoculum size had no observable effect on activity but alkaline pH increased citral activity. The growth rates of Escherichia coli cultures were reduced at concentrations of citral<0.01% v/v, while concentrations <0.03% v/v produced rapid reduction in viable cells followed by limited regrowth. In a non-growth medium, 0.08% and 0.1% v/v showed rapid bactericidal effects.

The availability of adequate supplies of monoterpenes aldehydes, particularly citral, is an issue of great importance to a number of industries, especiallythe pharmaceutical industry. Production with new or improved processes employing readily available source of starting material, lemon myrtle. The present invention is directed to this very important end of mass supply of citral from lemon myrtle starting material.

The present invention represents a potent immune modulator and stimulates and cleans the lymphatic system.

Oral mucositis also called stomatitis, is a common, debilitating complication of cancer chemotherapy and radiography occurring in about 40% of patients. It results from the systemic effects of cytotoxic chemotherapy agents and from the local effects of radiation to the oral mucosa. Oral mucositis is inflammation of the mouth the pain and discomfit may limit the patient's ability to tolerate either chemotherapy or radiotherapy.

This innovation extracts the bioactive compounds 6-gingerol and Zingibain from the Zingiber officinale rhizome to quench the toxicity of the citral in Backhousia citriodora oil and paste to enable their use in nasal and throat sprays.

The present invention relates to compositions consisting of an association of lipophilic extracts of Zingiber officinale (Zo) and Backhousia citriodora (Bc) which the prevents and treat of Covid-19. The combination of Bc and 6-gingerol provides a multifaced attack on

Covid-19 and inhibits the bacterial and fungal infections associated with the pathogens of the virus. Backhousia citriodora and Zingiber officinale strongly reduced the human ACE2 levels in HT 29 cells from 17.68ng/mL (control) 0,1% DMSO to 1.43 ng/mL and 4.34ng/L without displaying cytotoxicity. ACE2 Protein level was significantly reduced by Bc and Zo essential oils and significantlydownregulated ACE2.

The nasal or oral suspension or vapour spray inhaler containing a combination of Bc and Zo will deliver the active ingredients specifically in the host cell receptor's active site (Furin TMPRSS2) to block the attachment of viral S protein via hindering the S protein cleavage activation and membrane fusion.

Citral has been shown to exhibit both antiviral and antifungal activities. Aldehydes and particularly citral have long been considered to contain anti-tumour properties. Citral's germicidal properties were measured using the Ridley-Walker test devised by Penfold and Grant in which the RW co-efficient of citral is a high 19.5 (19 times higher than the power of phenols) though lower than that of thymol. The pure oil of Backhousia citriodora has a coefficient of 16, considerably higher than that of Eucalyptus and tea tree oils.

Quenching

This innovation quenches Citral sensitization with 6-gingerol.

In 1976, RIFM published a brief report indicating that human sensitization to citral could be reduced by the presence of a-pinene, or (+) - limonene. Seven subsequent and more rigorous studies have demonstrated similar quenching effects, three in guinea pigs and four in humans. Citral induced sensitization reactions in guinea pigs at concentrations above 0.5% were reduced by the co-presence of equal quantity of (+) -limonene. This innovation claims that the quenching effect of 6-gingerol was reduced by the co-presence of an equal quantity of 6-gingerol.

In a study carried out in the 1950's Backhousia citriodora were tested for antibacterial action against human pathogens such as Staphylococcus aureus and Staphylococcus typhi while 15 of the 34 oils passed the test, the most potent of these was Bc. Tincture 1: 45% alcohol Dose 2-4 ml

Citral is contained in a number of leave on, rinse off cosmetic products in very low concentrations (0.0001%-0.0002%).

Zingiber Officinale

The main pungent compound in fresh Australian gingerare gingerols whereas the pungency of dry ginger is due to shogaols. 6-shogaol is a dehydrated form of 6-gingerol. The most abundant gingerol is 6-gingerol while other gingerols with different chain length are present in small quantities. Jolad et al have identified 51 compounds on organically grown ginger including citral.

6-gingerol and Zingibain are the most bioactive compounds isolated from ginger rhizome. Gingerol is believed to inhibit the anti-serotonin 3 receptor function leading to anti-emetic properties. Additionally, it has been shown to effect gastric motility and potentially have an antispasmodic effect on the gastrointestinal system. 6-gingerol has poor water solubility and oral biological availability.

6-gingerol is implicated in apoptosis, cell cycle regulation, cytotoxicity and inhibition of angiogenesis. It prevents serotonin release and platelet aggregation. 6-gingerol has immunomodulatory, antiapoptotic, antihyperglycemic and antiemetic properties. 6-gingerol is a bioactive substance down regulates proinflammatory cytokine release by macrophages. The innovation Bc oil as an effective drug carrier which improved the water solubilityof 6 gingerol while sustaining the drug release or antigen as well as prolonging in vivo acting time of the drug coupled with oral bioavailability enhancement.

The present innovation is an oil in water emulsion consisting of Backhousia citriodora oil and fractions of Zingiberofficinale, predominantly 6-gingerol and Zingibain, that quenches the toxicity of citral the predominate bioactive in Bc and prevents and treats Covid-19 and other fungal and bacterial diseases.

The present innovation acts on different stages of viral infection cycle for prevention and treatment as follows: 1 Stimulates the innate immunity for killing the virus via T cells or NK cell mediated immune modulation by protease inhibition 2 Blocks the angiotensin converting-enzyme 2 (Ace2) receptor a host cell receptor has been found to play a crucial role in virus cell entry. 3 This innovation interferes with the interaction of the virus with the host cell entry receptors to prevent their cellular endocytosis, replication and infection 4 This innovation protects the vital organs by executing the anti-inflammatory against cytokine storm induced organ damage. 5 The novel combination is an immunomodulatory and anti-inflammatory agent inhibit 1L -10 production and 1L-1B as the key mediator of the inflammatory response and essential forthe host response. 6 The nebuliser helps the combination of Bc and Zo reach their active ingredients to the lungs and the oesophagus rapidly to enable the combinatorial formulation to protect the patient from virus driven lung injury fibrosis and infection by local action 7 This innovation makes possible large-scale production of citral protease inhibitors from large Lemon Myrtle plantations and 6-gingerol from ginger farms around the world. 8 The increasing use of antibodies and enzymes preparations has caused a corresponding need for agents capable of controlling the activity of enzymatic processes. Previously enzyme inhibitors and, in particular, inhibitors of proteolytic enzymes have been prepared from mammalian tissue which limits the amount of material which can be produced to such an extent that commercial production is not feasible economically to treat the world population.

This innovation provides a method of preventing or inhibiting replication of a virus in a cell and comprises a method of contacting the cell with a compound according to the various embodiments and provides a novel adjuvant and a highly efficient process for the formulation, characterization and pharmacokinetic 6-gingerol loaded nano structured oil in water emulsion.

Coronavirus disease (Covid-19) caused by novel severe respiratory syndrome coronavirus (SARS-CoV-2) is an ongoing pandemic. The present innovation provides a safe pharmaceutical composition for preventing or treating Covid19. This innovation is a therapeutic strategy which relies on suppressing infectivity and inflammation, along with immune modulation and anticoagulant activity. The angiotensin-converting enzyme 2 (ACE2) receptor, a host cell receptor, has been found to play a crucial role in virus cell entry; therefore, ACE2 cell blockers can be a potential target for antiviral intervention. The ACE 2 inhibitory effects of Bc and Zo display significant ACE2 inhibitory effects in epithelial cells. Immunoblotting and qPCR analysis also confirm that this innovation possess ACE2 inhibitory effects.

The Active Substances

Citral

Citral has been studied in the pharmaceutical industry and has shown antimicibial activity. Citral showed antimicrobial activity against MRSA isolates with minimum inhibitory concentration (MIC) values between 5mg/mL (0.5%) and 40mg/mL (4%), and minimum bacterial concentration (MBC) values between 10mg/mL (1%) and 40mg/mL (4%). The sub inhibitory dose was 2.5mg/mL (0.25%).

The antibacterial and antifungal properties of citral have been proven. Citral at 10% (v/v) dilution showed antibacterial against Staphylococcus aurus, Bacillus subtilus, Escherichia coli and Pseudomonas aeruginosa. At 500 ugmL it completely killed Escherichia coli and Salmonella typhimurium and could inhibit 14 bacteria including Gram-positive cocci and rods and has been proven an effective antifungal compound for 12 fungi, three of which were yeast like and nine filamentous. Further citral can inhibit biofilm formation of both methicillin susceptible S. aureus (MSSA) and methicillin resistant S.aureus (MRSA). Dual species biofilms have increase resistance compared to mono-species biofilm. Citral can induce irritation and sensitisation on human skin.

In this innovation the optimal concentration can be 1% higher when quenched with 6 gingerol.

6-Gingerol

6-gingerol is a bioactive phenolic phytocompound found in fresh ginger rhizome, is an anti inflammatory and has been proven to target viral proteins in in-silico studies. 6-gingerol is one of the main compounds identified in Zingiberofficinale, approved by the TGA as available, accessible and well-studiedfor its therapeutic benefits. Rathinavel reports drug likeness calculation forthe phytocompound gingerol was made in Swiss ADME server and revealed that 6-gingerol possess a molecularweight of 294.39 g/mol, the number of hydrogen bond acceptor and donor are 4 and 6 respectively. Gingerol also possesses excellentTPSA (topological polar surface area) lipophilicities (iLog P) and water solubility (Log S ESOLvalues of 66.76, 3.48 and -2.96 respectively and zero violation for drug likeliness rules such as Lipinski Ghose Veber Egan and Muegge essential for better drug likeness properties.

Citral has been shown to modulate numerous signalling pathways and inhibits inflammatory mediators including cytokines, chemokines, adhesion molecules, prostanoids and eicosanoids. It is claimed that a combination of citral and 6-gingerol have immunomodulatory, anti-inflammatory and antiviral properties.

Backhousia citriodora oil and 6-gingerol can to block the virus from entering host cells, prevent viral replication and/or attenuate the exacerbation of the host's immune response.

Zingibain

Zingibain, zingipain or ginger protease (EC 3.4.22.67) (https://enzyme.expasy.org/EC/3.4.22.67) is a cysteine protease enzyme found in ginger rhizome first isolated, purified and reported in 1973 by Ichik et al at Japan Women's University and exists as two isozymes GP-1 and GP11, which were isolated by chromatography, with molecular weights of approximately 22,500 Da.

The binding affinity of S-protein of SARS-CoV-2 with ACE2 is about ten times higher than other Corona viruses including SARS-CoV-1 indicating that CoV-2infection depends on the expression of host cell ACE2 receptor. Furthermore, transmembrane protease serine 2(TMPRSS2) fuse and activate the S-protein=ACE2 complex into the host cell endosome.

Covid-19 initiates infection in upper respiratory tract where commensal bacterial flora exists. The synergism between coronavirus and bacteria has been documented to past outbreaks.

Description of the finished medicinal product

An effective method for isolating 6-gingerol with an optimal formulation 6-gingerol-loaded SMEDDS and effectively improved the oral bioavailability of 6-gingerol. The unit oral dose at a dose of 0.0025 to 50mg/kg of the human body weight. The unit oral dose may comprise 0.01 to about 50mg preferably 0.1mg to 10 mg of the compound.

The preparations can include muco-adhesive mouthwash, gargling agents throat lozenges syrups and nasal/eyedrops for the preclusion/mitigation of COVID-19. 300ug per kg 0.6 % solution.

In addition to the compound as a raw extract, the compounds of the invention may be combined in a pharmaceutical preparation including suitable pharmaceutical carriers excipients where one of these excipients is certified organic ginger starch.

The pharmaceutical composition of the invention may be administered to an animal which may experience the beneficial effects of the compound. The animal being a horse, cow, pig chicken, and the like.

As used herein, the term "prevention" refers to any action that inhibits or delays the onset of Covid-19 by administration of a composition comprising a Backhousia citriodora and Zingiberofficinale extract or a fraction thereof.

As used herein the term treatment refers to any action that improves or advantageously alters symptoms of the disease by administrating of a composition comprising a Bc and Zo or fractions thereof.

The pharmaceutical composition for preventing and treating Covid-19 and its bacterial and anti-fungal of the present invention using methods well known in the art to provide rapid or sustained or delayed release of Bc and Zo extract and all active fractions thereof after administration to a human. In the preparation of the formulation it is preferred that the extracts and active fractions thereof are mixed or diluted with the carrier or enclosed in the carrier in the form of a container.

Accordingly, the pharmaceutical composition for preventing and treating Covid-19 and its bacterial and fungal complications of the present invention is a pharmaceutical formulation using methods well known in the art to provide rapid sustained or delayed release of Bc and Zo and all active fractions thereof after administration to a human.

Oral formulation such as powders, granules, tablets, suspensions, emulsions, syrups, aerosols, external preparations, suppositories and sterile injectable solutions according to conventional methods. It may be used in the form of a formulation and may further included suitable carriers, excipients and diluents commonly used in the preparation of the composition.

For example carriers, excipients and diluents which may be included in the pharmaceutical compositions of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, certified organic ginger starch, acacia gum, Siam benzoin, alginate, gelatine, calcium phosphate, calcium silicate, cellulose, microcrystalline, cellulose, water, magnesium stearate and squalene. When formulated diluents and excipients such as fillers, extenders, binders, wetting agents, lubricants such as magnesium stearate and organic ginger starch are also used.

Liquid preparations for oral use include suspensions, solvents, emulsions and syrups and may include various excipients such as wetting agents, sweeteners and preservatives in addition to commonly used simple diluents such as water and liquid paraffin.

Formulations for parenteral administration include sterile, aqueous, non-aqueous, suspensions, emulsions, lyophilized formulations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, olive oil, injectable esters, ethyl oleate and the like can be used. As the base of the suppository, witepsol, macrogol, Tween 61, cacao butter, glycerogelatine and the like can be used.

The composition of the present invention is administered in pharmaceutical effective amount.

The term "pharmaceutical effective amount" means an amount sufficientto treat a diseases at a reasonable benefit risk ratio applicable to medical treatment, with an effective dose level of individual type and severity, age, sex, type, type of virus infected type of fungus infected, drug activity, sensitivityto drug, time of administration, route of administration and rate of release, duration of treatment, factors including concurrently used drugs and other factors well known in the medical arts. In a specific embodiment of the present invention. The effective concentration of the Backhousia citriodora and Zingiberofficinale and the active fraction showing an antiviral antibiotic and antifungal effectwas treated at a concentration of 120-240/ml.

The composition of the present invention may be administered as individual therapeutic agents or in combination with other therapeutic agents and may be administered sequentiallyor simultaneously with conventional therapeutic agents and single or multiple administrations.

Taking all of the above factors into consideration it is important to administer an amount that can obtain the maximum effect in the minimum amount without side effect amount and can be easily determined by those skilled in the art.

In another aspect the present invention relates to a food composition for improving or preventing Nonovirus, Salmonella, Clostridium perfringens, Campylobacter and Staphylococcus aureus, comprising Backhousia citriodora and Zingiberofficinale or fractions thereof as an active ingredient.

Food compositions functional foods and beverages fruits canned fruit yoghurt canned foods jams and marmalade fish meat ham sausage dairy products margarine noodles frozen food can be prepared by adding the combination of Backhousia citriodora and Zingiber officinale of the current invention and fractions thereof.

When the present invention is used as a preservative and antimicrobial the mixed amount of the active ingredient may be suitably determined on the purpose of use (prevention health therapeutic treatment). Preferably the antimicrobial may be used for the preparation of preservatives and antimicrobials wet-wipes disinfectant cleaners detergent soaps humidifier fillers masks ointment hand washes or coatings.

The constituents of the excipients, their nature, quantity used, their colouring matter, preservatives, adjuvants, stabiliser, emulsifiers, flavouring, thickeners.

Constituents intended to be ingested and the outer covering includes hard capsules, soft capsules, rectal capsules, coated tablets and film coated tablets.

EDITORIAL NOTE 2021106617

There is 1 page of claims only.

Claims

Claims

What is claimed is:

Claim 1 A novel pharmaceutical composition comprising Backhousia citriodora oil and bioactive 6-gingerol and Zingibain extracted from Zingiberofficinale where 6-gingerol quenches sensitization or harmful effects of the citral on skin and the formulation inhibits entry of the SARS- CoV-2 virus into the host cell and inhibits the crucial steps of pathogenesis mediated by Spike protein , ACE2 and TMPRSS2 in the tissues of the gastrointestinal tract.

Claim 2 the combination in claim 1 a novel oil in water emulsion that exerts anti inflammatory effects against different cytokines and inflammatory mediators and can attenuate cytokine storm when used in conjunction with standard antiviral therapy and blocks LPS-induced activation of NF-KB by inhibiting phosphorylation of IkB and inhibits COX 2 expression.

Claim 3 The combination in claim 1 blocks the binding and attachment of SARS-CoV2 to ACE2-expressing cells thus inhibiting their infection to host cells as an inhibitor in SARS-CoV 2 Viral Spike Protein/ACE2 and uses a nasal spray or throat spray. The combination in claim 1 synergistically inhibits the main protease site ACE2 and binds strongly to the Spike protein ACE 2 and inhibits SARS-CoV-2 3CL protease and benefits from first establishing robust interactions within the S1 and S2 sites while reducing viral shedding of saliva and salivary viral burden by lozenges and mouthwash containing citral quenched by 6-gingerol.

Claim 4 The combination in claim 1 modulates virulence factors in methicillin-resistant Staphylococcus aureus and modifies the biofilm produced by MRSA in the initial stage biofilm formation. This innovation has a synergistic antibiotic effect against Staphylococcus pneumonia and inhibits biofilm formation.

Claim 5 The combination in claim 1 is a new class of anti-platelet agent and the synergistic effects have pharmaceutical activities including anti platelet aggregation and vasorelaxing effects.