MULTI-TARGETED COMPOSITIONS FOR MITIGATING ACUTE RESPIRATORY DISTRESS SYNDROME

Abstract

A method for treatment of viral infections, especially SARS-CoV-2 infections, said method comprising the administration of hydrolysable tannins.

Claims

1. A method of preventing, inhibiting, mitigating and/or treating Covid-19 comprising administering to an individual exposed to or infected with the SARS-CoV-2 virus an effective amount of one or more hydrolysable tannin.

2. The method of claim 1 wherein the hydrolysable tannins are characterized as glucose esterified with gallic-, ellagic-, chebulic-, modified ellagic-, and modified chebulic-acids and combinations thereof.

3. The method of claim 1 wherein the hydrolysable tannins have the following structure: ##STR00011## wherein R.sub.1, R.sub.2, R.sub.3, R.sub.4 and R.sub.5, which may be the same or different, are independently selected from hydroxy and ester moieties of gallic acid, ellagic acid, chebulic acid, dehydrohexahydroxydiphenic acid (DHHDP) and hexahydroxydiphenic acid (HHDP), provided that no more than 4, preferably no more than 3, most preferably no more than 2 of the R.sub.x groups are hydroxyl.

4. The method of claim 1 wherein the hydrolysable tannin is selected from the group consisting of Pentagalloyl glucose, Chebulinic acid, Chebulagic acid, Pedunculagin, Tellimagrandin I, Tellimagrandin II, Geraniin, Corilagin, Casuaricitin, and Nupharin.

5. The method of claim 1 wherein the hydrolysable tannin is administered following exposure or potential exposure to the SARS-CoV-2 virus but before the manifestation of symptoms of Covid-19 in an amount effective to inhibit or prevent RNA replication and/or prevent or inhibit the binding of the SARS-CoV-2 virus to the receptor site.

6. The method of claim 1 wherein the hydrolysable tannin is administered following exposure or infection to the SARS-CoV-2 virus but before the manifestation of symptoms of Covid-19 in an amount effective to prompt or enhance the initial immune response.

7. The method of claim 6 wherein the hydrolysable tannin is administered to an individual with a compromised immune response.

8. The method of claim 6 wherein the administration of the hydrolysable tannin up-regulates interferon alpha and/or interferon gamma.

9. The method of claim 1 wherein the hydrolysable tannin is administered to an individual manifesting symptoms of Covid-19.

10. The method of claim 9 wherein the hydrolysable tannin is administered to an individual manifesting symptoms of acute respiratory distress syndrome.

11. The method of claim 9 wherein the hydrolysable tannin is administered to an individual manifesting symptoms of cytokine storm and the effective amount is sufficient to down-regulate pro-inflammatory cytokines.

12. The method of claim 11 wherein the effective amount is sufficient to down-regulate Interleukin 6 and/or Interleukin 8.

13. A method of preventing, inhibiting, mitigating and/or treating acute respiratory distress syndrome comprising administering an effective amount of one or more hydrolysable tannins to an individual exposed to a substance or organism known to induce inflammation of the respiratory system and/or suffering a disease or other medical condition known to induce inflammation of the respiratory system.

14. The method of claim 13 wherein the organism is an influenza virus or a corona virus.

15. The method of claim 13 wherein the organism is the SARS-CoV-2 virus.

16. The method of claim 13 wherein the hydrolysable tannins are characterized as glucose esterified with gallic-, ellagic-, chebulic-, modified ellagic-, and modified chebulic-acids and combinations thereof.

17. The method of claim 13 wherein the hydrolysable tannins have the following structure: ##STR00012## wherein R.sub.1, R.sub.2, R.sub.3, R.sub.4 and R.sub.5, which may be the same or different, are independently selected from hydroxy and ester moieties of gallic acid, ellagic acid, chebulic acid, dehydrohexahydroxydiphenic acid (DHHDP) and hexahydroxydiphenic acid (HHDP), provided that no more than 4, preferably no more than 3, most preferably no more than 2 of the R.sub.x groups are hydroxyl.

18. The method of claim 13 wherein the hydrolysable tannin is selected from the group consisting of Pentagalloyl glucose, Chebulinic acid, Chebulagic acid, Pedunculagin, Tellimagrandin I, Tellimagrandin II, Geraniin, Corilagin, Casuaricitin, and Nupharin.

19. The method of claim 13 wherein the hydrolysable tannin is administered following exposure or potential exposure to the substance or organism but before the manifestation of symptoms respiratory distress.

20. The method of claim 13 wherein the hydrolysable tannin is administered following exposure to the substance or organism but before the manifestation of symptoms.

21. The method of claim 13 wherein the hydrolysable tannin is administered to an individual manifesting symptoms of acute respiratory distress syndrome.

22. The method of claim 14 wherein the hydrolysable tannin is administered following exposure or potential exposure to the virus but before the manifestation of symptoms of the associated disease in an amount effective to inhibit or prevent RNA replication and/or prevent or inhibit the binding of the virus to the receptor site.

23. The method of claim 14 wherein the hydrolysable tannin is administered following exposure or infection to the virus but before the manifestation of symptoms of associated disease in an amount effective to prompt or enhance the initial immune response.

24. The method of claim 23 wherein the hydrolysable tannin is administered to an individual with a compromised immune response.

25. The method of claim 23 wherein the administration of the hydrolysable tannin up-regulates interferon alpha and/or interferon gamma.

26. The method of claim 14 wherein the hydrolysable tannin is administered to an individual manifesting symptoms of the associated disease.

27. The method of claim 26 wherein the hydrolysable tannin is administered to an individual manifesting symptoms of acute respiratory distress syndrome.

28. The method of claim 26 wherein the hydrolysable tannin is administered to an individual manifesting symptoms of cytokine storm and the effective amount is sufficient to down-regulate pro-inflammatory cytokines.

29. The method of claim 28 wherein the effective amount is sufficient to down-regulate Interleukin 6 and/or Interleukin 8.

30. A method for tailoring the treatment of an individual exposed to and/or infected with a virus known to cause respiratory distress, which treatment involves the administration of a hydrolysable tannins, said method comprising assessing i) the phase of the infection, ii) the viral load, iii) the level of interferon alpha and/or gamma, iv) the level of interleukin 6 and/or 8 and/or v) the manifestation of symptoms of the viral infection and, based thereon selecting the timing for said administration, the selection of the hydrolysable tannin, and the amount of the administration.

Description

DETAILED DESCRIPTION

[0045] For a proper understanding and appreciation for the teachings and associated benefits of the compositions and methods taught in this specification, a sense of how viral infections progress, particularly how Covid-19 progresses, is necessary. For convenience, and since it is the key virus of interest, this discussion will be focused on SARS-CoV-2 virus: though, its applicability applies to most if not all viral infections, particularly those arising from an influenza virus or a coronavirus.

[0046] While some infectious disease specialists have developed significant experience in treating patients with COVID 19, many therapies continue to be used without evidence of benefit and without regard to timing, potentially causing more harm than would have been realized had no treatment been applied at all. Hence, consensus regarding the phases of COVID-19 is critical for understanding the appropriate timing for the study and delivery of therapeutics. In this respect, it is possible and likely that some of the failures in randomized controlled trials seen to date were due to the mis-timing of treatments. The context of disease phases may explain the failure of remdesivir or monoclonal antibody therapies when given late in disease [Daniel O. Griffin et al., The importance of understanding the Stages of COVID-19 in Treatment and Trials, AIDS Rev, 8:23(1):40-47, 2021; G Lippi et al., Coronavirus disease 2019 (COVID-19): the portrait of a perfect storm, Annals of translational medicine, 8(7):497, 2020]. There are three different phases of Covid 19 progression and the fourth one which is related to imbalance in energy metabolism.

[0047] Phase 1: The Pre-Exposure or Incubation Time

[0048] The pre-exposure or incubation time is associated with the risk of developing the associated disease as the viral load increases through viral replication. This is the very first phase, starting with the initial exposure or infection and continuing to the first onset or manifestation of severe symptoms and usually lasts between 2 and 11 days (mean incubation time: 6 days), with patients likely to be infectious 1-3 days before the onset of symptoms. Although the true rate of individuals who will remain asymptomatic, or only mildly symptomatic, until terminal viral shedding is still unknown, some evidence suggests that the number could be as high as 50%. Importantly, this rate may be even underestimated due to under-testing or under-reporting. It now seems reasonable to hypothesize that this pre-symptomatic phase is perhaps the most critical for containment of the outbreak. Interestingly, the viral load of asymptomatic, pre-symptomatic, or mildly symptomatic subjects is comparable to that of patients with overt disease. This highlights the significant risk of viral transmission throughout this first phase. Evidence suggests that 50-80% of all cases may be attributed to transmission from an asymptomatic or pre-symptomatic individual. As such, the relatively long incubation time and considerably high rate of asymptomatic-mild symptomatic individuals explains the rise in number of cases despite public health intervention and public awareness. It is during this phase therapies should be more targeted toward inhibiting viral binding and viral replication. It is also the time in which antivirals, monoclonal antibodies and other therapies that augment innate immune responses such as interferons would have the highest potential for benefits versus harm.

[0049] Phase 2: Early Inflammatory Phase—Progressive Respiratory Involvement

[0050] The viral symptom phase occurs very soon after viral RNA is detectable. For most individuals, that will be the time at which their illness comes to clinical attention. The population in this phase will be predominantly an outpatient population and studying therapeutics in this group will potentially prevent hospitalizations and viral transmission, thereby having a significant impact on resource utilization. The initial clinical manifestations of early inflammatory phase are pulmonary compromise, particularly difficulty or at least the sense of difficulty in breathing, with or without hypoxemia, essentially the early stage of and/or moderate manifestation of acute respiratory distress syndrome, followed by impacts on the cardiac, renal, and other organ systems. Therapeutics that target viral replication and augment the innate immune response such as interferons have a decreasing likelihood of benefit at this later stage of disease since the symptomatic manifestations during this phase are driven by the host's immune responses rather than ongoing viral replication. Here a different type of immunomodulation is required at this stage. The disturbances in the coagulation system also appear to begin during the early inflammatory phase in the 2nd week of illness, but the macrovascular manifestation may not be evident until week three of the illness.

[0051] Phase 3: Late Inflammatory Phase—Cytokine Storm

[0052] The third phase, which develops in around 15% of all SARS-CoV-2 infected subjects, is perhaps the most challenging and intriguing from a physiopathological perspective. In fact, whilst the respiratory phase is mostly attributable to direct cytopathic lung injury caused by viral replication in pulmonary parenchyma, the late pro-inflammatory phase is instead characterized by an abnormal, almost exaggerated, host reaction against the pathogen, either locally (i.e., in the lung) or systemically, thus mimicking the pathogenesis of severe sepsis and severe inflammatory response syndrome (SIRS). Here the individual is suffering fully developed or server acute respiratory distress. Although the precise mechanisms underlying the onset of this disproportionate host response against the virus remain partially elusive, it has now been acknowledged that SARS-CoV-2 infection of dendritic cells and cells of the monocyte/macrophage lineage triggers their activation and active secretion of a vast array of pro-inflammatory cytokines particularly pro-inflammatory interleukins (ILs) such as IL-6, IL-2, IL-7, and IL-8, monocyte chemoattractant protein 1 (MCP-1), macrophage inflammatory protein 1-α (MIP 1-α), granulocyte colony stimulating factor (GSF), C-X-C motif chemokine 10 (CXCL10) and tumor necrosis factor-α (TNF-α). The renin-angiotensin-aldosterone system (RAAS) also plays a very relevant role in this phase. Specifically, the binding of SARS-CoV-2 to its receptor angiotensin-converting enzyme 2 (ACE2) at the surface of host cells may be associated with profound derangement of RAAS, culminating in the increased activity of angiotensin II (Ang II) and decreased activity of angiotensin 1,7 (Ang 1,7), thus fostering vasoconstrictive, inflammatory, oxidative and fibrotic injuries.

[0053] Phase 4: Lack of Energy

[0054] The relatively high proportion of people chronically infected with SARS-CoV-2 (‘the long haulers’), who do not make a straight-forward recovery in the post viral period of their illness, almost certainly reflects damage done by the host response to the initial infection. A severe body response such as a cytokine storm can give rise to oxidative and inflammatory damage and generalized oxidative stress, and this suggests that the antioxidant therapies might be beneficial [E Wood et al., Role of mitochondria, oxidative stress and the response to antioxidants in myalgic encephalomyelitis/chronic fatigue syndrome: A possible approach to SARS-CoV-2 ‘long-haulers’? Chronic Dis Trans' Med, 7(1):14-26, 2021]. Antioxidant therapy is known to improve the levels of the abundant natural antioxidant, glutathione (which is important for redox balance), and to strengthen the immune response [ME Soto et al., Is antioxidant therapy a useful complementary measure for Covid-19 treatment? An algorithm for its application, Medicine, 56:1-29, 2020]. Physiological changes in SARS-CoV-2 that enhance the production of reactive oxygen species could be ameliorated by free radical scavengers [G D Mironova et al., Prospects for the use of regulators of oxidative stress in the comprehensive treatment of the novel Coronavirus Disease (COVID-19) and its complications, Eur Rev Med Pharmacol Sci, 24:8585-8591, 2020]. Oxidative stress and ongoing pathogenesis in SARS-CoV-2 are almost certainly linked [L Delgado-Roche et al., Oxidative stress as key player in severe acute respiratory syndrome coronavirus (SARS-CoV) infection, Arch Med Res, 51:384-387, 2020].

[0055] The tail phase is now appreciated to be a common feature of COVID 19. Growing numbers of individuals are reporting suffering from this aspect of COVID 19 and support groups have formed of “long haulers”. As we learn more about the post exhaustive fatigue of this disease, it appears to be distinct from that described in chronic fatigue syndrome (CFS). SARS-CoV-2 potentially drains the body's energy and ATP reserves through continued aggressive inflammatory response in the airways, thus damaging the airways and the alveoli, in the lungs, and make carbon dioxide exchange difficult. As a result, the metabolism of the patient cannot provide sufficient energy to support the life processes. Like several other health problems, COVID-19 disease is associated with a difficulty in keeping a balance of the energy budget in the body. As the energy budget worsens, the patient's body tries to balance the budget by scavenging building blocks and energy from the healthy tissues. When the budget becomes unmanageable the patient dies. The disease may be more fatal for the elder patients, both because they may have additional health problems or also because they may have a slower energy metabolism, or their metabolism may produce at a slower rate [M. Özilgen and B Yilmaz, COVID-19 disease causes an energy supply deficit in a patient. Int J Energy Res. 2020; 1-4. 10.1002/er.5883; Bayram Yilmaz et al, Energetic and exergetic costs of COVID-19 infection on the body of a patient, International Journal of Exergy, 32(3): 314-327, 2020].

[0056] For purposes of simplicity and a better understanding the present teachings, the following terms have the meanings as presented.

[0057] “Preventing” or “prevention” refers to reducing the risk of manifesting acute respiratory distress syndrome.

[0058] “Treating” or “treatment” refers to reversing, alleviating, arresting, inhibiting, mitigating or ameliorating at least one of the clinical symptoms associated with acute respiratory distress syndrome, inhibiting the progression of acute respiratory distress syndrome, as well as delaying the onset of at least one or more symptoms of acute respiratory distress syndrome in a patient who has been exposed to or is infected with a microbe, especially a viral agent, that induces or is associated with the manifestation of acute respiratory distress syndrome. In following, treating or treatment also refers to inhibiting acute respiratory distress syndrome, either physically, (e.g., stabilization of a discernible symptom), physiologically, (e.g., stabilization of a physical parameter), or both, and to inhibiting at least one physical parameter that may or may not be discernible to the patient.

[0059] “Improve” or “improvement” is used to convey the fact that the present inventive composition has manifested or effected changes, most notably beneficial changes, in either the characteristics and/or the physical attributes of the tissue to which it is being provided, applied or administered, including, for example, boost the Innate and adaptive immunity through interferons (IFNs), reduce replication of coronavirus for example via RNA-dependent RNA polymerase (RdRp) and reduce inflammation by reducing, for example, IL-6 and/or IL-8, etc. These terms are also used to indicate that the symptoms or physical characteristics associated with the diseased state are diminished, reduced or eliminated.

[0060] “Inhibiting” generally refers to delaying the onset of the symptoms, delaying or stopping the progression of the symptoms, alleviating the symptoms, or eliminating the symptoms associated with acute respiratory distress syndrome.

[0061] Furthermore, again for simplicity, while the present teachings are applicable to addressing or treating acute respiratory distress syndrome generally, whether arising from environmental exposures, e.g., chemical exposure, smoke, etc., allergens, or microorganisms, e.g., fungi, molds, bacteria and viruses, the following description is specifically focused on viral infections, especially influenza and coronavirus infections, most especially exposure and infection by the SARS-CoV-2. Many of the teachings are equally applicable, at least to certain aspects of the acute respiratory distress: though it is also recognized that each type of expose may and does involve different pathways and the development and evolution of acute respiratory distress depending upon the initiator or cause thereof. Similarly, it is recognized that even within the class of viral infections, differences in viral pathways, the development and evolution of the infection, and the disease manifestation are found whereby the exact same response associated with the compositions and methods taught herein may and/or are different.

[0062] According to a first aspect of the present teaching there is provided a method for preventing, inhibiting, mitigating and/or treating bacterial, fungal and/or viral infections, most especially those associated with or known to cause acute respiratory distress syndrome, most especially for preventing and/or mitigating the manifestation of acute respiratory distress syndrome, said method comprising administering to an individual exposed to or infected with such microorganisms and/or manifesting inflammation of the respiratory system or suffering from acute respiratory distress an effective amount of one or more select hydrolysable tannins, In particular, there is provided a method of preventing, inhibiting, mitigating and/or treating acute respiratory distress syndrome associated with or caused by the influenza virus or a coronavirus, most especially the SARS-Cov-2 virus, comprising administering an effective amount of one or more hydrolysable tannins characterized as glucose esterified with gallic-, ellagic-, chebulic-, modified ellagic or modified chebulie-acids and combinations thereof.

[0063] According to a second aspect of the present teaching there is provided a method for preventing and/or inhibiting viral RNA replication and/or the binding of viruses, particularly pathogenic viruses, to their host receptor, said method comprising administering to individuals exposed to and/or infected with said viral microorganisms, especially influenza viruses and coronaviruses, most especially the SARS-CoV-2 virus, an effective amount of select hydrolysable tannins, most especially the hydrolysable tannins, said hydrolysable tannins characterized as glucose esterified with gallic-, ellagic-, chebulic-, modified ellagic- and modified chebulic acids and combinations thereof. In particular, it has now been found that these hydrolysable tannins interfere with and/or inhibit viral RNA replication and/or the ability of the virus to bind to the ACE2 receptor. The prevention of viral RNA replication and/or the prevention of the binding of the virus to the host receptor results in a reduced viral load, particularly as compared to an untreated individual, and the prevention, inhibition and/or mitigation of the symptoms association with said viral infections, particularly acute respiratory distress syndrome, most especially hyperinflammation and/or cytokine storm.

[0064] According to a third aspect of the present teaching there is provided a method for promoting and/or enhancing the immune response in individuals with compromised immune responses and/or to viral infections, particularly infections due to viruses which are known or found to poorly induce or even fail to induce the interferon response, particularly the interferon alpha and interferon gamma responses, most especially the interferon gamma response, especially the coronaviruses such as SARS, SARS-CoV and SARS-CoV-2, most especially SARS-CoV-2. Specifically, it has now been found that the immune response of T cells, NKT cells and/or NK cells and, in particular, the interferon alpha and interferon gamma responses may be induced or upregulated by the administration of select hydrolysable tannins, most especially the hydrolysable tannins to the individual exposed to and/or infected with the virus and/or in individuals with a compromised Type I interferon response, wherein the hydrolysable tannins are characterized as glucose esterified with gallic-, ellagic-, chebulic-, modified ellagic- and modified chebulic-acids and combinations thereof.

[0065] According to a fourth aspect of the present teaching there is provided a method for preventing, inhibiting, mitigating and or treating acute respiratory distress, most notably, the manifestation of hyperinflammation and or a cytokine storm in the respiratory system, said method comprising administering to individuals exposed to or infected with a virus know to induce or elevate the risk for acute respiratory syndrome an effective amount of select hydrolysable tannins, most especially the hydrolysable tannins, said hydrolysable tannins characterized as glucose esterified with gallic-, ellagic-, chebulic-, modified ellagic- and modified chebulic-acids and combinations thereof. Most especially, according to this embodiment, the present method is directed to the administration of said hydrolysable tannins to individuals exposed to and/or infected with influenza viruses and coronaviruses, most especially the SARS-CoV-2 virus. In particular, it has been found that the administration of the hydrolysable tannins down-regulate pro-inflammatory interleukins, especially IL-6 and IL-8, as well as other pro-inflammatory cytokines.

[0066] According to a fifth aspect of the present teaching there is provided a method for tailoring the treatment of an individual exposed to and/or infected with a virus, particularly viruses which are known or found to poorly induce or even fail to induce the interferon response and/or induce or manifest symptoms of acute respiratory distress syndrome, which method comprises administering to said individual one or more of select hydrolysable tannins, the timing, selection of the hydrolysable tannin, and amount of the administration based upon i) the phase of the infection, ii) the viral load, iii) the level of interferon alpha and/or gamma, iv) the level of interleukin 6 and/or 8 and/or v) the manifestation of symptoms of the viral infection, particularly the manifestation of symptoms associated with or a precursor to acute respiratory distress; wherein the hydrolysable tannins are characterized as glucose esterified with gallic-, ellagic-, chebulic-, modified ellagic- and modified chebulic-acids and combinations thereof. This method is especially applicable to the treatment of individuals exposed to and/or infected with an influenza virus or a coronavirus, most especially the SARS-CoV-2 virus, either as an early phase treatment to prevent viral RNA replication and/or binding to its host receptor and/or inducer of the interferon response. Alternatively, it may be used during the course of the infection to prevent, inhibit and/or mitigate symptoms of acute respiratory distress such as hyperinflammation and/or cytokine storms. It may also be used in the late phase of such infections to help bring the immune response back to a more normal, pre-infection, state: thereby addressing hangover or lingering symptoms of the viral infection due to the impact of the infection on the immune and inflammatory responses.

[0067] Hydrolysable tannins (HTs) occur in nature and are generally characterized as glucose esterified with gallic-, ellagic-, chebulic-, modified ellagic- and modified chebulic-acids and combinations thereof. Other hydrolysable tannins, such as those based upon other polyhydric alcohols, e.g., fructose, xylose, saccharose, and structures like hamamelose, as well as those substituted with other acid groups such as hexahydroxydiphenic acid (HHDP) are also believed suitable for use in the practice of the present teaching and are within the scope of its claims, though focus is on the aforementioned hydrolysable tannins. In following, hydrolysable tannins are readily hydrolyzed by acidic, alkali, or enzymatic (tannase or β-glucosidase) hydrolysis, particularly hydrolysis with sulfuric or hydrochloric acid. Polygalloyl esters are called gallotannins (GTs), which give gallic acid (GA) on hydrolysis whereas ellagic esters are referred to ellagitannins (ET) and give hexahydroxydiphenic acid (which spontaneously dehydrates to ellagic acid (EA) upon hydrolysis.

##STR00001##

[0068] Hydrolysable tannins may contain both galloyl and hexahydroxydiphenoyl functionalities. ETs can be defined in a narrow sense as hexahydroxydiphenoyl esters of carbohydrates or cyclitols, while the definition of ETs in a wider sense cover compounds derives from further oxidative transformations, including oligomerization processes [T Okuda et al., Ellagitannins Renewed the concepts of tannins, Chapter 1, Chemistry and Biology of Ellagitannins, World Scientific Co Pte: td, http://www.worldscibooks.com/chemistry/679.html]. Monomeric and oligomeric HTs contain one or more polyhydroxyphenoyl groups such as HHDP or its oxidized forms (dehydrohexahydroxydiphenoyl, DHHDP, chebuloyl, or neochebuloyl, m- or p-dehydrodigalloyl and valoneoyl (VL), tergalloyl (TG), macaranoyl, or flavogallonoyl as tris-galloyl and/or gallagyl as tetrakis-galloyl group and so on.

[0069] Sources of hydrolysable tannins are well known: preferred hydrolysable tannins are derived from various herbs and plants, including, but not limited to, Occimum gratissmium, Occimum sanctum, Mollugo pentaphylla L, Hypericum triquetrifolium, Ampelopsis brevipedunculata (Maxim.) Trautv. (AB), Withania somnifera, Terminalia chebula, Terminalia bellerica, Terminalia citrina, Terminalia catappa, Euphoria longana, Terminalia macroptera, Terminalia arjuna, Emblica officinalis, Gaila chinensis, teas generally, including Sideritis raseri, particularly from their fruits, leaves, peels and/or roots. Especially preferred sources are the hydrolysable tannins derived from foodstuffs such as, but not limited to, almonds, Terminalia chebula fruit, Terminalia bellerica fruit, Terminalia arjuna fruit, Emblica officinalis fruit, Gaila chinensis fruits, cashew nuts, pistachios, mangos, hazelnuts, persimmons, chestnuts, walnuts, guacas, cloves, pimento, pomegranates, plums, apricots, peaches, bird cherries, strawberries, raspberries, blackberries, black currants, gooseberries, grapes, muscadine grapes, bearberry, and the like. Although natural hydrolysable tannins are preferred, it is also to be appreciated that the hydrolysable tannins for use in the practice of the present teaching can also be synthesized by esterification of polyhydric alcohol with the respective acids, e.g, gallic acid, chebulic acid, ellegic acid and/or HHDP.

[0070] As noted above, the selection of the hydrolysable tannin can be tailored to the specific timing of its administration and its intended purpose or objective, all as taught herein. One aspect of the hydrolysable tannins that affects its selection for use at a particular phase of an infection is its hydrophobicity. As shown in the following schematic, hydrophobicity can be selected based upon the groups or acid esters as well as the polyol base:

##STR00002##

wherein DHHDP=dehydrohexahydroxydiphenoyl, HHDP=hexahydroxydiphenoyl. R can be for instance hydrogen, hydroxyl, galloyl, HHDP or other ET or GT substitute groups [V. Virtanes and M Karonen, Partition Coefficients (IogP) of Hydrolysable Tannins, Molecules, 25(16): 3691, 2020]. In this respect, it has come to be appreciated that hydrophobicity is one of the essential physicochemical properties that affects how a compound interacts with lipids and permeates cell membranes.

[0071] Structures of common phenolic acids present as esters in hydrolysable tannins are as follows:

##STR00003##

[0072] Preferred hydrolysable tannins for use in the practice of the present teaching include those according to the following structure:

##STR00004##

wherein R.sub.1, R.sub.2, R.sub.3, R.sub.4 and R.sub.5, which may be the same or different, are independently selected from hydroxy and ester moieties of gallic acid, ellagic acid, chebulic acid, dehydrohexahydroxydiphenic acid (DHHDP) and hexahydroxydiphenic acid (HHDP), provided that no more than 4, preferably no more than 3, most preferably no more than 2 of the Rx groups are hydroxyl. Exemplary hydrolysable tannins are as follows (“+” means the moiety is bridged across the designated R positions):

[0073] Pentagalloyl glucose: R.sub.1=R.sub.2=R.sub.3=R.sub.4=R.sub.5=G

[0074] Chebulinic acid: R.sub.1=R.sub.3=R.sub.5=G; R.sub.2+R.sub.4=C

[0075] Chebulagic acid: R.sub.1=G; R.sub.2+R.sub.4=C; R.sub.3+R.sub.5=HHDPA

[0076] Pedunculagin: R, =OH; R.sub.2+R.sub.3=HHDPA; R.sub.4+R.sub.5=HHDPA

[0077] Tellimagrandin I: R.sub.1=OH; R.sub.2=R.sub.3=G; R.sub.4+R.sub.5=HHDPA

[0078] Tellimagrandin II: R, =R.sub.2=R.sub.3=G; R.sub.4+R.sub.5=HHDPA

[0079] Geraniin: R.sub.1=G; R.sub.2+R.sub.4=DHHDPA; R.sub.3+R.sub.5=HHDPA

[0080] Corilagin: R.sub.1=G; R.sub.2=R.sub.4=OH; R.sub.3+R.sub.5=HHDPA

[0081] Casuaricitin: R.sub.1=G; R.sub.2+R.sub.3=HHDPA; R.sub.4+R.sub.5=HHDPA

[0082] Nupharin A: R.sub.1=R.sub.2=R.sub.5=G; R.sub.3+R.sub.4=HHDPA

[0083] The structures of the foregoing as well as a couple additional hydrolysable tannins are as follows:

##STR00005## ##STR00006## ##STR00007## ##STR00008## ##STR00009## ##STR00010##

[0084] Each of the treatment methods described above involves the administration of an effective amount of the tannins to the individual exposed to or infected with the virus of concern. An “effective amount” is evidenced by the manifestation of an improvement, inhibition, and/or benefit with respect to the purpose for which the hydrolysable tannin is being applied, which, in turn is dependent upon the timing of its administration. For example, an effective amount in relation to the ability to prevent or delay RNA replication and/or the binding of the virus to the host receptor is evidenced by a lower viral load as compared to what is normally expected or common in individuals to whom the hydrolysable tannin was not administered. Similarly, an effective amount for enhancing or initiating the immune response may be established by an up-regulation in the interferon alpha and/or interferon gamma: particularly in individuals whose immune response is compromised. In the case of individuals manifesting the signs of infection, an effective amount is such as will prevent, delay, inhibit and/or improve or shorten the duration of the manifestation of hyperinflammation of the respiratory system and/or cytokine storms. This may manifest visually or may be evaluated by assessing the level of pro-inflammatory cytokines, especially the pro-inflammatory interleukins, such as IL-6 and/or IL8. Preferably, the methods involve the administration of an amount of one or more hydrolysable tannins which effect at least a 20% down regulation in IL-6 and/or IL-8 and/or their corresponding downstream cytokine/chemokine and/or at least a 20% up regulation in IL-12, IFN-alpha and/or IFN-gamma and/or their corresponding downstream cytokine/chemokine as opposed to the response to the same trigger in the absence of the hydrolysable tannin: down regulation and up regulation being evidenced by a reduction or inhibition or a promotion or enhancement, respectively, in the expression or generation/production of the aforementioned interleukins and/or interferons and/or their corresponding downstream cytokine/chemokine, as appropriate. More preferably, the extent of the modulation, i.e., the down regulation and/or up regulation, is at least a 30%, most preferably at least a 50%, as compared to the same trigger in the absence of the hydrolysable tannin.

[0085] Following on the foregoing, the specific amount of the hydrolysable tannin to be administered to a given patient will vary depending upon the timing and purpose of its administration, the specific hydrolysable tannin to be administered, the delivery method, the specific disease and/or trigger for the event being addressed (e.g., chemical exposure, bacterial infection, viral infection, etc.), the weight of the patient, etc. The comparative efficacy of the various hydrolysable tannins, as well as combinations thereof, can be ascertained by simple trial and error and/or by further in-vitro assessment of gene expression. Again, administration of the hydrolysable tannins prevent, delay, or mitigate the appearance or manifestation of symptoms of the disease, enable patients to recover faster from acute respiratory distress and/or other manifestations of the immune response being addressed, reduce or lessen the severity of the acute respiratory distress and/or other manifestations of the immune response, and reduce the risk of death from acute respiratory distress, especially from that associated with influenza and coronavirus infections, most especially COVID-19.

[0086] The hydrolysable tannins may be administered as a preventative prior to exposure to the pathogen, but, are more likely and preferably administered subsequent to the exposure to the pathogen, but in advance of the manifestation of the symptoms associated with the infection, e.g., following a known exposure, but before diagnostic confirmation. Again, such early administration is difficult absent strict and continual screening tests, hence, the hydrolysable tannins, from a practical perspective, particularly absent clinical symptoms, are more likely to be administered following manifestation of the symptoms of the infection/inflammation. Still, if a person known to exposed also suffers from a compromised immune response and/or there is no indication of a suitable immune response despite the detection of a viral load, it is desirable to administer the hydrolysable tannin as soon as possible to initiate or enhance the immune response, particularly that pertaining to the key interferons, most notably interferon alpha and/or gamma. At the same time, because of the key roles played by cytokines in the immune-response system, it is important not to administer the treatment too early as to interfere with the nature response to the infection or invasion as this may lead to an earlier and faster progression of the disease. For example, it may be desirable to administer the hydrolysable tannins to help initiate and ramp up the immune response; but, to stop the treatment once the immune response is active and allow that response to take its natural path, while continuing to monitor the symptoms and/or level of pro-inflammatory cytokines, particularly the pro-inflammatory interleukins, especially IL-6 and/or IL8. On the other hand, it symptoms worsen or acute respiratory distress syndrome manifests, it is preferable to have initiated administration or, as the case may be, reinitiated, the administration of the hydrolysable tannins once adverse respiratory symptoms are manifesting, particularly that associated with hyperinflammation and/or cytokine storm. The need to administer the hydrolysable tannin is especially warranted if or once other symptoms of the infection or disease are starting to decrease or wane, e.g., if fever is dropping, achiness is less severe, etc, or if viral load is dropping and/or the individual is no longer testing positive, yet, respiratory distress continues as this is indicative of cytokine storm.

[0087] The hydrolysable tannins may be administered as is, but are preferably administered as a therapeutic composition in a proper delivery vehicle. Additionally, the hydrolysable tannins may be used alone or in combination with antimicrobial agents, especially antibiotics and/or antiviral agents, and/or with other therapeutic agents such as plasma treatments, antibody treatments (e.g., Tocilizumab), and the like and/or in combination with other anti-inflammatory agents, antioxidants, vitamins and the like. Indeed, it is believed that the afore mentioned combinations are not only cumulative in their benefits but provide synergy in helping patients recover from acute respiratory distress syndrome, especially from that associated with influenza and coronavirus infections. Selection will depend, in part, upon the particular infection or microbe being addressed. For example, indications are that azithromycin, hydroxychloroquine, chloroquine, remdesivir, several nucleotide analog drugs—including favipiravir, ribavirin, galidesivir, and EIDD-2801, and combinations thereof are effective in the treatment of Covid-19. Hence, their combination with the present hydrolysable tannins of the present teachings are beneficial in boosting the Innate and adaptive immunity through interferons (IFNs), reducing replication of coronavirus for example via RNA-dependent RNA polymerase (RdRp) and reducing inflammation by reducing, for example, IL-6 and/or IL-8, etc.

[0088] As noted above, hydrolysable tannins can be used as is, i.e., as 100% of the composition to be administered; however, the hydrolysable tannins are preferably incorporated into a pharmaceutical composition in which the hydrolysable tannin(s) account for from about 0.01 to about 99 weight percent of the pharmaceutical composition. Preferably the hydrolysable tannin(s) will comprise from about 0.5 to about 30 wt %, more preferably from about 0.5 to about 20 wt %, most preferably from about 1.0 to about 10 wt % of the pharmaceutical composition. Another factor playing into the concentration of the hydrolysable tannin in the pharmaceutical composition is the dose or rate of application of the composition to the patient. Obviously, dosing itself depends upon a number of factors including the concentration and/or purity of the hydrolysable tannin(s), the individual to whom the composition is to be administered, the mode of administration, the form in which the pharmaceutical composition is to be administered, the disease or symptom to be addressed, etc. Generally speaking, an appropriate dose of the hydrolysable tannin(s), or of the pharmaceutical composition comprising the hydrolysable tannin(s), can be determined according to any one of several well-established protocols including in-vitro and/or in-vivo assays and/or model studies as well as clinical trials. For example, animal studies involving mice, rats, dogs, and/or monkeys can be used to determine an appropriate dose of a pharmaceutical compound. Results from animal studies are typically extrapolated to determine appropriate doses for use in other species, such as for example, humans. Similarly, an appropriate oral dosage for a particular pharmaceutical composition containing one or more hydrolysable tannins will depend, at least in part, on the gastrointestinal absorption properties of the compound, the stability of the compound in the gastrointestinal tract, the pharmacokinetics of the compound and the intended therapeutic profile: all of which is readily ascertainable.

[0089] As noted above, the hydrolysable tannins are preferably administered as a therapeutic composition comprising the hydrolysable tannin and a pharmaceutically acceptable vehicle such as a pharmaceutically acceptable diluent, a pharmaceutically acceptable adjuvant, a pharmaceutically acceptable excipient, a pharmaceutically acceptable carrier, or a combination of any of the foregoing with which a pharmacological active agent, including the hydrolysable tannins provided by the present disclosure, can be administered to a patient, which does not destroy or have a marked adverse effect on the activity of the therein contained hydrolysable tannins and which is non-toxic when administered in doses sufficient to provide a therapeutically effective amount. Such vehicles are well known and standard in the pharmacological art. Exemplary carriers include fillers, binders, humectants, disintegrating agents, solution retarders, absorption accelerators, wetting agents, absorbents, or lubricating agents. Other useful excipients include magnesium stearate, calcium stearate, mannitol, xylitol, sweeteners, starch, carboxymethylcellulose, microcrystalline cellulose, silica, gelatin, silicon dioxide, and the like.

[0090] The hydrolysable tannin(s), more appropriately, pharmaceutical compositions comprising the hydrolysable tannins, can be administered through any conventional method. The specific mode of application or administration is, in part, dependent upon the form of the pharmaceutical composition, the primary purpose or target of its application (e.g., the application may be oral if intending to address the disease generally or by nasal application or inhalation if intending to address primarily the symptom of acute respiratory distress syndrome. Suitable modes of administration include, for example, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, oral, sublingual, intracerebral, intravaginal, transdermal, rectal, nasal or inhalation. The preferred modes of administration are oral, by nasal application, or inhalation. The former allows for absorption through epithelial or mucous linings of the gastrointestinal tract (e.g., oral mucosa, rectal, and intestinal mucosa, etc.) while the latter allows direct application to the tissue of the respiratory tract that is manifesting the symptoms of respiratory distress. Furthermore, again, depending in part upon the form of the administration, the pharmaceutical compositions of the present disclosure can be administered systemically and/or locally. Finally, the form of the pharmaceutical composition containing the hydrolysable tannin(s) and its delivery system varies depending upon the parameters already noted. For example, orally administered pharmaceutical compositions of the present teaching can be in encapsulated form, e.g., encapsulated in liposomes, or as microparticles, microcapsules, capsules, etc.

[0091] For preparing pharmaceutical compositions containing the hydrolysable tannin(s) for use in the present methods, pharmaceutically acceptable carriers can be either solid or liquid. Solid form preparations include powders, tablets, pills, capsules, cachets, suppositories, lozenges, and dispersible granules. A solid carrier can be one or more substances which may also act as diluents, flavoring agents, solubilizers, lubricants, suspending agents, binders, preservatives, tablet disintegrating agents, or an encapsulating material including, for example, magnesium carbonate, magnesium state, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, chewing gum, methylcellulose, sodium carboxy-methlycellulose, a low melting wax, cocoa butter, and the like. In powders, the carrier is a finely divided solid, which is in a mixture with the finely divided active component. In tablets, the active component is mixed with the carrier having the necessary binding capacity in suitable proportions and compacted in the shape and size desired.

[0092] Liquid preparations include solutions, suspensions, and emulsions, for example, water or water-propylene glycol solutions. For example, parenteral injection liquid preparations can be formulated as solutions in aqueous polyethylene glycol solution. The hydrolysable tannin(s) may thus be formulated for parenteral administration (e.g. by injection, for example bolus injection or continuous infusion) and may be presented in unit dose for in ampoules, pre-filled syringes, small volume infusion or in multi-dose containers with an added preservative. The compositions may take such forms as suspensions, solutions, or emulsions in oily or aqueous vehicles, and may contain formulation agents such as suspending, stabilizing and/or dispersing agents. Alternatively, the active ingredient may be in powder form, obtained by aseptic isolation of sterile solid or by lyophilization from solution, for constitution with a suitable vehicle, e.g. sterile, pyrogen-free water, before use.

[0093] Aqueous solutions suitable for oral or inhalation use can be prepared by dissolving or suspending the hydrolysable tannin(s) in water and adding suitable colorants, flavors, stabilizing and thickening agents, as desired. Aqueous suspensions suitable for oral use can be made by dispersing the finely divided active component in water with viscous material, such as natural or synthetic gums, resins, methylcellulose, sodium carboxy-methylcellulose, or other well-known suspending agents. Compositions suitable for oral administration in the mouth includes lozenges comprising the active agent in a flavored base, usually sucrose and acacia or tragacanth; pastilles comprising the active ingredient in an inert base such as gelatin and glycerin or sucrose and acacia; and mouthwashes comprising the active ingredient in suitable liquid carrier.

[0094] Finally, solutions or suspensions may be applied directly to the nasal cavity by conventional means, for example with a dropper, pipette, or spray. Similarly, solutions or suspensions may be applied directly to the respiratory tract by conventional means, for example, by a spray, nebulizer, or inhaler. The compositions may be provided in single or multi-dose form. In compositions intended for administration to the respiratory tract, including intranasal compositions. The suspension or solutions or active will generally have a small particle size for example of the order of 5 microns or less. Such a particle size may be obtained by means known in the art, for example by micronization, atomization, etc.

[0095] Following upon the foregoing, the therapeutic compositions provided by the present disclosure can be formulated in a unit dosage form. A unit dosage form refers to a physically discrete unit suitable as a unitary dose for patients undergoing treatment, with each unit containing a predetermined quantity of the hydrolysable tannin compositions. A unit dosage form can be for a single daily dose, for administration 2 times per day, or one of multiple daily doses, e.g., 3 or more times per day. When multiple daily doses are used, a unit dosage form can be the same or different for each dose. One or more dosage forms typically comprise a dose, which can be administered to a patient at a single point in time or during a time interval.

[0096] Of course, one may vary the dosing with time if the desired outcome for the treatment fails to manifest. For example, if viral load increases rapidly or the manifestation of symptoms rapidly advances, particularly in immune compromised individuals or if it appears that a proper or normal immune response is not initiate, it would be desirable to initiate or increase the dosage to promote the immune response. Similarly, if there is a marked worsening of acute respiratory distress, particularly with a subsiding in other factors or symptoms of the disease, it would be appropriate to initiate administration or increased the dose of administration of the hydrolysable tannin(s). For example, one may monitor the status of a patient and adjust the dosage, its frequency, etc. to either drop their levels of pro-inflammatory interleukins to normal levels or to a more controlled, moderate level sufficient to maintain an immune response to the pathogen. Or, if the immune response seems to be lacking, one may want to administer the hydrolysable tannin(s) to boost the innate and adaptive immunity through interferons (IFNs). Furthermore, if exposure is known with viral load increasing, though still asymptomatic, it would be desirable to initiate or increase the dosage to reduce replication of coronavirus via, for example, RNA-dependent RNA polymerase (RdRp) or interference with the binding site of the host. Similarly, it may be desirable to administer a large initial dose to boost the Innate and adaptive immunity through interferons (IFNs) and/or to reduce replication of coronavirus and guard against subsequent hyperinflammation and/or another cytokine storm. Furthermore, one may increase the dose or issue a large dose if the patient's symptoms worsen after treatment has begun.

[0097] The compositions containing the hydrolysable tannin(s) (also referred to as the “active” or “actives” hereinafter) can be formulated for immediate release or for delayed or controlled release. In this latter regard, certain embodiments, e.g., an orally administered product, can be adapted for controlled release. Controlled delivery technologies can improve the absorption of an active agent in a particular region, or regions, of the gastrointestinal tract in the case of orally administered doses or in the respiratory tract in the case of nasal or inhalation administered doses. Controlled delivery systems are designed to deliver the active in such a way that its level is maintained within a therapeutically effective window and effective and safe blood levels are maintained for a period as long as the delivery system continues to deliver the active with a particular release profile. Controlled delivery of orally administered actives typically and preferably produces substantially constant blood levels of the active over a period of time as compared to fluctuations observed with immediate release dosage forms. Controlled delivery of inhalation administered actives typically and preferably produces substantially constant levels of the active in the tissue of the respiratory tract over a period of time as compared to fluctuations observed with immediate release dosage forms. For some actives, maintaining a constant blood and/or tissue concentration of the active throughout the course of treatment is the most desirable mode of treatment as immediate release of the active may cause the blood or tissue level of the active to peak above that level required to elicit the most desired response. This results in waste of the active and/or may cause or exacerbate toxic side effects. In contrast, the controlled delivery of the active can result in optimum therapy; not only reducing the frequency of dosing, but also reducing the severity of side effects. Examples of controlled release dosage forms include dissolution-controlled systems, diffusion-controlled systems, ion exchange resins, osmotically controlled systems, erodable matrix systems, pH independent formulations, and gastric retention systems.

[0098] An appropriate controlled release oral dosage and ultimate form of a pharmaceutical composition containing the hydrolysable tannin(s) will also depend upon a number of factors. For example, gastric retention oral dosage forms may be appropriate for compounds absorbed primarily from the upper gastrointestinal tract, and sustained release oral dosage forms may be appropriate for compounds absorbed primarily from the lower gastrointestinal tract. Again, it is to be expected that certain hydrolysable tannins are absorbed primarily from the small intestine whereas others are absorbed primarily through the large intestine. It is also to be appreciated that while it is generally accepted that compounds traverse the length of the small intestine in about 3 to 5 hours, there are compounds that are not easily absorbed by the small intestine or that do not dissolve readily. Thus, in these instances, the window for active agent absorption in the small intestine may be too short to provide a desired therapeutic effect in which case large intestinal absorption must be channeled and/or alternate routes of administration pursued.

[0099] Where additional pharmacological actives may be and preferably are also present in the compositions according to the present teaching, the amount by which they are present and/or the dosage amount will typically be consistent with their conventional concentration and rates of application. For example, such other actives will be present in an amount of from about 0.5 to about 30 wt. %, more preferably from about 0.5 to about 20 wt. %, most preferably from about 1.0 to about 10 wt. % of the pharmaceutical composition. Of course, as noted, the combination of these other pharmacological actives with the hydrolysable tannin(s) also provide enhanced performance and/or synergy whereby the amounts of each and/or the dose of each is generally less than required for the use of the individual active compounds on their own.

[0100] Without further elaboration, it is believed that one skilled in the art can, using the preceding description, utilize the present invention to its fullest extent. The following preferred specific embodiments are, therefore, to be construed as merely illustrative, and not limitative of the remainder of the disclosure in any way whatsoever.

EXAMPLES

Example 1—Inhibition of RNA-Dependent RNA Polymerase (RdRp)

[0101] A study was undertaken in which the impact of select hydrolysable tannins in accordance with the present teaching on the enzymatic activity of SARS-CoV-2 RNA Polymerase (RdRp), in vitro was evaluated. The experiment was performed using the SARS-CoV-2 RNA Polymerase (RdRp) Assay Kit cat. #S2RPA100KE from Profoldin (Hudson, Mass.), according to the manufacturer's instructions provided with kit. The test materials, whose identity and concentration are presented in Table 1, were solubilized and diluted in sterile distilled water and samples were added to the plate according to the plate design. Fluorescent signal proportional to the RNA polymerase activity was quantified at ex/em 485/530 nm with Applied Biosystem, (Foster City, Calif.) multi-well plate fluorometer Cytofluor 4000. Statistical significance was assessed with paired Student test. Deviations of ≥20% as compared to water control with p values below 0.05 were considered statistically significant. Results are summarized in Tables 1 and 2.

TABLE-US-00001 TABLE 1 Inhibition of RNA-dependent RNA polymerase (RdRp) Test Material % of Control p value Water (control) 100 1 Chebulinic acid (0.01045 μM) 10 μg 25 0.000 Chebulinic acid (0.0522 μM) 50 μg 14 0.000 Chebulinic acid (0.20905 μM) 100 μg 7 0.000 Chebulagic acid (0.01047 μM) 10 μg 35 0.000 Chebulagic acid (0.05237 μM) 50 μg 10 0.000 Chebulagic acid (0.2094 μM) 100 μg 8 0.000 Chebulic acid (0.02807 μM) 10 μg 105 0.463 Chebulic acid (0.1403 μM) 50 μg 96 0.530 Pentagallolyl glucose (0.01063 μM) 10 μg 32 0.000 Pentagallolyl glucose (0.0531 μM) 50 μg 12 0.000 Pentagallolyl glucose (0.2126 μM) 100 μg 4 0.000 Lopinavir (10 μM) 91 0.237 Lopinavir (50 μM) 80 0.028

TABLE-US-00002 TABLE 2 EC.sub.50 values EC50 Statistical significance/ Test Material values Comments Chebulinic acid <0.01 μM 0.000/Highly significant Chebulagic acid <0.01 μM 0.000/Highly significant Pentagallolyl glucose <0.01 μM 0.000/Highly significant Remdesivir*  0.77 μM Statistically significant Lopinavir   >50 μM 0.028/Marginally significant *Wang et al., RNA-dependent RNA polymerase of SARS CoV-2 as therapeutic target J Medical Virology, 2021 93:300-310,

[0102] As shown in Tables 1 and 2, neither Lopinavir, a commercial inhibitor of viral RNA replication, nor chebulic acid manifested effective inhibition of SARS-CoV-2 RNA replication. In stark contrast, the hydrolysable tannins, namely Chebulinic acid, Chebulagic acid and Pentagallolyl glucose, all exhibited a marked inhibition of SARS-CoV-2 RNA replication. Additionally, each of the hydrolysable tannins demonstrated a markedly lower EC.sub.50, even as compared to Remdesivir, currently a key antiviral agent used in the treatment of COVID-19, the disease associated with SARS-CoV-2 infections.

Example 2—Inhibition of Interleukins

[0103] A second study was undertaken to assess the ability of the select hydrolysable tannins to inhibit select pro-inflammatory interleukins, namely IL-6 and IL-8. The study was conducted using reconstituted human ciliated airway tissue. Specifically, Ninety Six 3D ciliated airway tissues (cat. #34839) were obtained from Mattek (Ashland, Mass.) and were cultured in Assay Media (cat. #031921JRD, MatTek). The tissue samples were rinsed twice before incubation with the MTT reagents. The test materials themselves were solubilized and diluted in sterile distilled water and added to the feeder chamber medium contacting the basal side of the tissues on Day 1. The test material was also applied topically on Day 2 together with LPS (added where indicated at 1 μg/ml, from Escherichia coli 0113, Cayman Chemical Company, Ann Arbor, Mich.) for total contact time of 72 h. The experiments were tested in triplicates or duplicates.

[0104] Following the exposures to the test materials, 1L-6 and IL-8 were quantified in the tissue culture—conditioned medium by sandwich ELISA (Table 4A and 4B). The effect of the test materials on mitochondrial metabolism was measured by the MTT assay, which quantifies the activity of mitochondrial dehydrogenases, such as succinate dehydrogenase, implicated in the respiratory electron transport chain in mitochondria. The MTT conversion values were also used to standardize the IL-6, IL-8 output data to tissue viability. The results are presented in Tables 3 and 4.

TABLE-US-00003 TABLE 3 Inhibition of IL-6 Test Material % of Control p-value Water (control) 100 1.000 Chebulinic acid (10 μgm/ml) 131 0.375 Chebulinic acid (50 μgm/ml) 96 0.945 Chebulinic acid (200 μgm/ml) 18 0.029 Chebulagic acid (10 μgm/ml) 70 N/A Chebulagic acid (50 μgm/ml) 40 0.098 Chebulagic acid (200 μgm/ml) 5 0.020 Pentagallolyl glucose (10 μgm/ml) 91 0.769 Pentagallolyl glucose (50 μgm/ml) 64 0.201 Pentagallolyl glucose (200 μgm/ml) 0 0.017 Chebulic acid (10 μgm/ml) 68 0.559 Chebulic acid (50 μgm/ml) 106 0.892 Chebulic acid (100 μgm/ml) 78 0.568 Lopinavir (10 μM) 73 0.624 Lopinavir (50 μM) 81 0.572

TABLE-US-00004 TABLE4 Inhibition of IL-8 Test Material % of Control p value Water (control) 100 1.000 Chebulinic acid (10 μg/ml) 114 0.014 Chebulinic acid (50 μg/ml) 109 0.220 Chebulinic acid (200 μg/ml) 101 0.837 Chebulagic acid (10 μg/ml) 112 0.131 Chebulagic acid (50 μg/ml) 100 0.986 Chebulagic acid (200 μg/ml) 66 0.000 Pentagallolyl glucose (10 μg/ml) 100 0.693 Pentagallolyl glucose (50 μgm/ml) 104 0.008 Pentagallolyl glucose (200 μg/ml) 0 0.000 Chebulic acid (10 μg/ml) 96 0.703 Chebulic acid (50 μg/ml) 108 0.131 Chebulic acid (100 μg/ml) 111 0.001 Lopinavir (10 μM) 103 0.667 Lopinavir (50 μM) 104 0.006

[0105] The results demonstrate an significant inhibition of the pro-inflammatory cytokines IL-6 and IL-8, most notably IL-6, by the select hydrolysable tannins in, what appears as, a dose dependent relationship. This in view of the results of Example 1 above, demonstrates the ability to tailor treatment based upon the dose amount and the timing of its application. Specifically, the lower doses have demonstrated a marked effect on inhibition of viral RNA replication without an adverse effect on the pro-inflammatory cytokines: thereby allowing the immune response to the viral attack that is manifested, to proceed.

Example 3—Receptor Binding

[0106] As noted in the Background, the receptor binding domain on spike protein S1 of SARS-CoV-2 is the first point of contact between the host and the virus and plays a key role in the interaction with ACE2 that then lead to S2 domain-mediated membrane fusion and incorporation of viral RNA into host cells. Accordingly, as study was undertaken to assess the impact of the hydrolysable tannins on the binding of the SARS-CoV-2 at the ACE receptor.

[0107] Two docking packages were used to generate docking scores for the SARS-CoV-2. The first is the Vina score from Autodock vina [O Trott and A J Olson, AutoDock Vina: improving the speed and accuracy of docking with a new scoring function, efficient optimization, and multithreading. J Comput Chem 2010; 31:455-618], the second is Glide score from Schrodinger package [R A Friesner et al. Extra precision glide: Docking and scoring incorporating a model of hydrophobic enclosure for protein-ligand complexes, J Med Chem, 49:6177-96, 2006; T A Halgren et al., Glide: A New Approach for Rapid, Accurate Docking and Scoring. 2. Enrichment Factors in Database Screening, J Med Chem, 47:1750-1759, 2000]. The binding scores (presented as Kcal/mole) for the hydrolysable tannins versus other established treatments for Covid-19 are presented in Tables 5A thru 5C.

[0108] As evident from the results presented in Tables 5A thru 5C, the select hydrolysable tannins showed significant if not marked lower binding energies (i.e., a tighter binding) as compared to the conventional or currently pursued pharceutical treatments for Covid-19.

TABLE-US-00005 TABLE 5A Binding energy (BE) results for RBD-ACE2 Test Material VinaScore GlideScore Chebulagic acid −7.3 −4.37 Chebulinic acid −7.5 −4.36 Chloroquine −5.3 −3.07 Hydroxychloroquine −5 −3.71 Dexamethasone −6.4 −1.75 Remdesivir −6.2 −3.73

TABLE-US-00006 TABLE 5B Binding energy results for RdRp Test Material VinaScore GlideScore Chebulagic acid −10.1 −6.24 Chebulinic acid −9.6 −7.13 Chloroquine −5.1 −2.75 Hydroxychloroquine −5.2 −3.51 Dexamethasone −7.6 −4.44 Remdesivir −7.0 −4.62

TABLE-US-00007 TABLE 5C Binding energy results for RdRp (cofactor) Test Material VinaScore GlideScore Chebulagic acid −10.1 −6.31 Chebulinic acid −9.7 −6.91 Chloroquine −5.3 −2.68 Hydroxychloroquine −5.2 −3.42 Dexamethasone −7.6 −4.50 Remdesivir −6.8 −4.37

Example 4—Impact on Interferons (IFNα and IFNγ)

[0109] A study has been designed, though not yet completed, to assess the impact of the hydrolysable tannins on interferon production and/or activation. In this study, different doses of Chebulinic acid, Chebulagic acid and Pentagalloyl Glucose are being evaluated to demonstrate their efficacy in boosting interferons using Lipopolysaccharide or viral proteins in Human bronchial epithelial cells 3D model system. In this experiment, 3D ciliated airway tissues (cat. #502-3D-24) obtained from Cell Applications (San Diego, Calif.) are cultured in maintenance medium (cat. #511M-3D-100, Cell Applications). The respective test materials are solubilized in sterile water, with and/or without LPS, Syn TC, EMB, and DMSO for the dexamethasone (DEX) control, at 20 mg/ml: all further dilutions are made in sterile distilled water. All test materials, with the exception of LPS, are assayed at different concentrations in μg/ml (LPS is added topically at 1 μg/ml on Day 2). The test materials are added to the feeder chamber medium contacting the basal side of the tissues on Day 1, then also topically treated on Day 2, together with LPS. Following 24 hours of pre-incubation of the tissue samples with the test substances, the tissue samples are then exposed to 1 μg/ml endotoxin LPS and incubation continued for 48 additional hours. At the end of the experiment tissue culture—conditioned medium is stored at −20° C. until further processing. IFN-α and IFN-γ levels are quantified in the tissue culture—conditioned medium by sandwich ELISA. Colorimetric measurements are performed using Molecular Devices microplate reader MAX190 and SoftMax3.1.2PRO software.

[0110] Though the current study is yet to be completed, based on the results obtained with extracts of Terminalia chebula, especially high tannin content extracts, as set forth in co-pending U.S. patent application Ser. No. 17/035,405, filed on Sep. 28, 2020, the entire contents of which are incorporated herein by reference, it is expected that the tannins themselves, as now claimed, will demonstrate a marked upregulation of Interferon alpha and Interferon gamma.

[0111] Without further elaboration, it is believed that one skilled in the art, using the preceding description, can utilize the present invention to its fullest extent. Furthermore, while the present invention has been described with respect to aforementioned specific embodiments and examples, it should be appreciated that other embodiments, changes and modifications utilizing the concept of the present invention are possible, and within the skill of one in the art, without departing from the spirit and scope of the invention. Furthermore, although not detailed as explicitly above, it is expected that the use of the hydrolysable tannins in the treatment of viral infections, notably, influenza and coronavirus infections, most notably, Covid-19, will help reduce the risk for development of viral drug resistance during therapy, especially with commonly used nucleoside analogues as well as address chronic fatigue associated with the tail phase of COVID 19 disease during the recovery period. In closing, the teachings above, including the preferred specific embodiments, are to be construed as merely illustrative, and not limitative of the remainder of the disclosure and the appended claims in any way whatsoever.