Procyanidins for the treatment of endothelial dysfunction triggered by covid-19

Abstract

The invention relates to a natural composition for medical purposes and more specifically to a composition comprising procyanidins, for use in the prevention or treatment of endothelial inflammation and / or endothelial systemic dysfunction triggered by Corona virus disease

Description

[Technical Field]

[0001] This invention relates to a natural composition for medical purposes, and more specifically, to a composition containing proanthocyanidins for the prevention or treatment of endothelial inflammation and / or systemic endothelial dysfunction triggered by COVID-19, including symptomatic post-COVID-19 individuals who have recovered from COVID-19. [Previous Technology]

[0002] Coronavirus disease 2019 (COVID-19) is defined as the disease caused by a novel coronavirus, now known as Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2; formerly known as 2019-nCoV), which was first identified in an outbreak of respiratory illness cases in Wuhan City, Hubei Province, China. It was initially reported to the WHO on December 31, 2019. On January 30, 2020, the WHO declared the COVID-19 outbreak a global health emergency. On March 11, 2020, the WHO declared COVID-19 a global pandemic, the first such designation since the declaration of the H1N1 influenza pandemic in 2009. As of June 9, 2020, more than 7.12 million cases had been reported in 188 countries and territories, resulting in more than 406,000 deaths.

[0003] Common symptoms include fever, cough, fatigue, shortness of breath, and loss of smell and taste. Although most cases result in mild symptoms, some may progress to acute respiratory distress syndrome (ARDS) due to cytokine storm, multiple organ failure, septic shock, and blood clots.

[0004] Complications may include pneumonia, acute respiratory distress syndrome (ARDS), multiple organ failure, septic shock, and death. Cardiovascular complications may include heart failure, arrhythmia, cardiac inflammation, and blood clots. Approximately 20 to 30% of people exhibiting COVID-19 have elevated liver enzymes reflecting liver damage. Neurological manifestations include seizures, stroke, encephalitis, and Guillain-Barré syndrome (which includes loss of motor function). Following infection, children may develop pediatric multisystem inflammatory syndrome with symptoms similar to Kawasaki disease, which can be fatal.

[0005] Varga Zsuzsanna et al., "Endothelial cell infection and endotheliitis in COVID-19," www.thelancet.com, Vol. 395, May 2, 2020, describes the rapid emergence of cardiovascular complications as a key threat of COVID-19, in addition to respiratory illness. However, the underlying mechanisms of the disproportionate effects of SARS-CoV-2 infection on patients with cardiovascular comorbidities remain incompletely understood. SARS-CoV-2 infects the host using the angiotensin-converting enzyme 2 (ACE2) receptor, which is expressed in several organs, including the lungs, heart, kidneys, and intestines. The ACE2 receptor is also expressed by endothelial cells. It is currently unclear whether vascular disturbances in COVID-19 are attributable to viral involvement of endothelial cells. Interestingly, SARS-CoV-2 can directly infect engineered human vascular organoids in vitro. In fact, it has demonstrated endothelial cell involvement across vascular beds in a range of COVID-19 patients. The authors found evidence of direct viral infection of endothelial cells and diffuse endothelial inflammation. Direct viral infection of the endothelium or immune-mediated recruitment of immune cells can lead to widespread endothelial dysfunction associated with apoptosis. These findings demonstrate the presence of viral components within endothelial cells and the accumulation of inflammatory cells, providing evidence of endothelial and inflammatory cell death. This also suggests that SARS-CoV-2 infection, as a direct result of viral involvement and the host's inflammatory response, contributes to the induction of endotheliitis in several organs. Furthermore, the induction of apoptosis and pyroptosis may play an important role in endothelial cell damage in COVID-19 patients. COVID-19 endotheliitis may explain the systemic impaired microcirculatory function and its clinical sequelae in different vascular beds of COVID-19 patients.

[0006] This was also confirmed by Frank Ruschitzka et al. in the Lancet, "Endothelial cell infection and endotheliitis in COVID-19," April 20, 2020. COVID-19 is considered a lung disease. Until now, it has not been clear why patients sustain life-threatening organ failure in organs other than the lungs. An interdisciplinary team from University Hospital Zurich has demonstrated that SARS-CoV-2 directly induces inflammation in the blood vessels, which can lead to organ failure and even death. During the analysis of tissue samples obtained from deceased COVID-19 patients following autopsies, pathologists at University Hospital Zurich found that the patients not only suffered from pneumonia but also from widespread inflammation of all endothelial tissues in all organs. "COVID is a systemic inflammation of the blood vessels, and we can now also call the disease COVID endotheliitis," said Professor Frank, outlining findings from cardiologists, infectious disease physicians, pathologists, and intensive care physicians. Frank Ruschitzka also believes that the treatment of COVID-19 patients must address two points: "We must address viral replication while simultaneously protecting and stabilizing the patient's vascular system. This is primarily applicable to patients with cardiovascular disease who have been diagnosed with impaired endothelial function, and to patients with known risk factors for severe progression of COVID-19."

[0007] Zuhang et al., "Procyanidins and butanol extract of Cinnamomi Cortex inhibit SARS-CoV infection", Antiviral Res. April 2009; 82(1): 73-81, published online on February 11, 2009. doi:10.1016 / j.antiviral.2009.02.001, found that the butanol fraction of cinnamon bark (CC / Fr.2) showed moderate inhibitory activity in wild-type severe acute respiratory syndrome coronavirus (wtSARS-CoV) and HIV / SARS-CoV S pseudovirus infection. Inhibition of pseudoviruses was also observed in cells pretreated with CC and CC / Fr.2 (IC50S, 283.4±16.3 and 149.5±13.5 μg / ml, respectively). However, the highest activity against wtSARS-CoV was observed when the virus was treated with the extract before challenge (IC50S, 43.1±2.8 and 7.8±0.3 μg / ml; SIs, 8.4 and 23.1, respectively). Among the compounds isolated from the CC fraction, proanthocyanidins A2 and B1 showed moderate anti-wtSARS-CoV activity (IC50S, 29.9±3.3 and 41.3±3.4 μM; SIs, 37.35 and 15.69, respectively). The authors also attempted to use transferrin receptor (TfR) as an indicator to determine whether it interfered with the clathrin-dependent endoplasmic pathway. CC / Fr.2 inhibited TfR internalization, but proanthocyanidins did not. In summary, CC / Fr.2 contains unknown substances that can inhibit infection (potentially interfering with endoplasmosis), and it also contains proanthocyanidins that do not inhibit endoplasmosis but do inhibit infection. Therefore, CC extracts contain antiviral activity that acts through different mechanisms depending on the compounds or mixtures.

[0008] JP 2005 314316 A (Kikkoman Corp.) provides (1) a novel anti-SARS coronavirus agent containing proanthocyanidins, catechins or grape extract as active ingredients, (2) an agent containing proanthocyanidins, catechins or grape extract as active ingredients for the prevention or treatment of SARS coronavirus infection, (3) food, beverage, medicine or cosmetic containing the above anti-SARS coronavirus agent or agent for the prevention or treatment of SARS coronavirus infection, and (4) food or beverage containing proanthocyanidins, catecholamines or grape extract as active ingredients and labeled for the prevention or improvement of SARS coronavirus infection.

[0009] JP 2007 217410 A (Yoshida Tsutomu, Yamashita Masako) provides an antiviral composition that eliminates viruses from latently infected cells and is a fundamental therapy for viral infectious diseases; and provides an antiviral agent containing the composition as an active ingredient and an antiviral functional food. The antiviral agent has viral proliferation inhibitory activity by containing fucoidan or proanthocyanidins as active ingredients targeting latently infected cells, or viral induction activity from latently infected cells by containing fucoidan or proanthocyanidins as active ingredients. A pharmaceutical product for treating viruses or an antiviral functional food is obtained by containing the antiviral composition. Particularly preferred is that the fucoidan is a sulfated polysaccharide derived from brown algae, and the proanthocyanidins are either contained in a peanut seed coat extract.

[0010] ISTIFLI ERMAN SALIH et al., "In silico analysis of the interactions of certain flavonoids with the receptor-binding domain of 2019 novel coronavirus and cellular proteases and their pharmacokinetic properties", JOURNAL OF BIOMOLECULAR STRUCTURE & DYNAMICS, December 2013, advance print volume, advance print date October 28, 2020 (2020-10-28), pp. 1-15, XP009525294. This reveals that as of October 2020, COVID-19 had infected more than 35 million people worldwide and caused nearly 1 million deaths. The microorganism causing COVID-19 was named Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2 or 2019-nCoV). The aim of this study was to investigate the interactions of 23 phytochemicals belonging to different flavonoid subgroups with the receptor-binding domain (RBD) of the 2019-nCoV spike glycoprotein and cellular proteases [transmembrane serine protease 2 (TMPRSS2), cathepsin B, and cathepsin L (CatB / L)]. The compounds interacted more strongly with CatB and CatL than with other proteins. Van der Waals bonds and hydrogen bonds play important roles in receptor-ligand interactions. Based on RBCI (relative binding index) analysis to rate the interactions of flavonoids with their target proteins, (-)-epicatechin gallate interacted strongly with all the proteins studied. This was supported by results obtained from molecular dynamics and the Poisson-Boltzmann surface area (MM / PBSA) method. According to Lipinski's five rules, (-)-epicatechin gallate exhibits drug-like properties. Although this molecule cannot cross the blood-brain barrier (BBB), it is concluded that (-)-epicatechin gallate can be evaluated as a candidate molecule in drug development studies against 2019-nCoV because it is not a receptor for P-gp (P-glycoprotein), does not inhibit any of the cytochrome Ps, and has not shown AMES toxicity or hepatotoxicity to eukaryotic cells.

[0011] MAEDA TAKAAKI et al., "Anti SARS-CoV Activity of Extracts from Japanese Pepper (Zanthoxylum piperitum (L.) DC.f.inerme Maki no)", HORTICULTURAL RESEARCH (JAPAN), JP, Vol. 10, No. 2, April 15, 2011 (2011-04-15), pp. 267-272, XP009525292. This study reveals that Japanese pepper (Zanthoxylum piperitum) is native to Japan and has four well-known lineages (Asakura, Takahara, Budou, and Arima), named according to their place of origin or morphology. Restriction site-associated DNA sequencing (RAD-Seq) was used to analyze 93 materials from multiple regions, including these four lineages. Single nucleotide variant analysis was used to divide the plants into eight groups: two groups each for the Asakura and Arima lineages, one group each for the Takahara and Budou lineages, and two additional groups. In one Asakura group and two Arima groups, plants were found in both farmland and mountainous areas, indicating an early stage of cultivation of Japanese pepper. The second Asakura lineage group was closely associated with plants present in multiple regions, representing a second stage of cultivation, as gene-related lineages with the desired traits spread to the periphery after the early cultivation. These results suggest that the cultivation of Japanese pepper is ongoing. Furthermore, this study demonstrates that although the thornless lineage is considered a subspecies of Japanese pepper, the thornless plant is polyphyletic.

[0012] ROH CHANGHYUN et al., "A facile inhibitor screening of SARS coronavirus N protein using nanoparticle-based RNA oligonucleotide", INTERNATIONAL JOURNAL OF NANOMEDICINE, DOVE MEDICAL PRESS, NEW ZEALAND, Vol. 7, January 1, 2012 (2012-01-01), pp. 2173-2179, XP009525291. This study reveals that hundreds of millions of people worldwide have been infected with Severe Acute Respiratory Syndrome (SARS), and the global mortality rate from SARS has increased significantly. Therefore, there is an urgent need to develop effective drug treatments targeting the biological effects of SARS. The authors have previously shown that quantum dot (QD)-bound RNA oligonucleotides are sensitive to the specific recognition of the nucleosheath (N) protein of SARS-related coronavirus (SARS-CoV). In this study, the authors discovered that a designed biochip can be used to analyze inhibitors of the SARS-CoV N protein using nanoparticle-based RNA oligonucleotides. Among the polyphenolic compounds detected, (-)-catechin gallate and (-)-gallocatechin gallate exhibited significant inhibitory activity against the SARS-CoV N protein. (-)-catechin gallate and (-)-gallocatechin gallate weakened binding affinity in a concentrated manner, as demonstrated by QDs-bound RNA oligonucleotides on a designed biochip. At a concentration of 0.05 µg mL⁻¹, (-)-catechin gallate and (-)-gallocatechin gallate showed over 40% inhibitory activity on a nanoparticle-based RNA oligonucleotide biochip system.

[0013] SALMAN SAAD et al.: "Virtual screening of immunomodulatory medicinal compounds as promising anti-SARS-CoV-2 inhibitors", FUTUNK VILOGY, FUTUNK MEDICINE LTD., UK, Vol. 15, No. 5, April 30, 2020 (2020-04-30), pp. 267-275, XP009525290. This study reveals Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2), a malignant viral disease causing acute respiratory distress, leading to global mortality and morbidity. The aim was to screen different immunomodulatory drug compounds to prevent their interaction with SARS-CoV-2 viral proteins. Materials & Methods: Autodock vina was used to analyze a library of immunomodulatory drug compounds with antiviral capabilities targeting the SARS protease, spike protein, and non-structural proteins (NSP-9,15). Results: Among more than 300 pharmaceutical compounds, only six compounds—arzanol, ferulic acid, genistein, resveratrol, rosmarinic acid, and hydroquinone—showed significant interactions with SARS virus proteins by forming low-binding-energy hydrogen bonds with the active site residues. Further ADMET (absorption, distribution, metabolism, excretion, and toxicity) analyses demonstrated favorable pharmacokinetic properties and low acute toxicity in these compounds. Conclusion: This study provides compelling evidence that these pharmaceutical compounds exhibit antiviral activity against SARS-CoV-2 and could be further used to treat this disease.

[0014] All such literature reveals the study of silicon dioxide.

[0015] Specifically, the literature cited by Zuhang et al., JP 2005 314316 A, JP 2007 217410 A, MAEDA TAKAAKI et al., and ROH CHANGHYUN et al. describes the antiviral activity of proanthocyanidins derived from different sources by inhibiting the growth and proliferation of SARS-CoV / SARS-CoV2.

[0016] On the other hand, ISTIFLI ERMAN SALIH et al. and SALMAN SAAD et al. have described compounds containing proanthocyanidins as having a high affinity or binding ability to SARS-CoV2 proteins, thereby also exhibiting antiviral activity. However, the antiviral activity of proanthocyanidins is not within the scope of this invention and prior art is therefore not applicable in this context.

[0017] During the ongoing global pandemic of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), the number of patients recovering from COVID-19 continues to rise. COVID-19 can cause multi-organ disease with a wide range of symptoms, including lung problems, thrombotic complications and cardiovascular dysfunction, renal dysfunction, gastrointestinal symptoms, neurological problems, and more. This pleiotropic clinical presentation is attributed to endothelial dysfunction, coagulopathy, microcirculatory and inflammatory problems. However, many cases of persistent symptoms have been reported even after recovery from acute COVID-19. A significant responsibility now is to find and develop solutions to this growing global problem following the COVID-19 pandemic.

[0018] Following SARS-CoV-2 infection, significant lung symptoms can persist, especially in individuals who have been admitted to a hospital and managed in a hospital ward for at least one week (Nalbandian A, Sehgal K, Gupta A, Madhavan MV, McGroder C, Stevens JS et al., Post-acute COVID-19 syndrome, Nature Medicine, 2021;27(4):601-15).

[0019] Individuals permitted to enter the intensive care unit tend to have greater residual disease, morphological damage, and especially respiratory damage due to a combination of disease, systemic complications, treatment and intubation.

[0020] However, COVID-19 patients with non-severe symptoms also suffer from persistent effects such as fatigue, recurrent headaches, attention deficit disorder, anxiety or depression, and more (Lopez-Leon S, Wegman-Ostrosky T, Perelman C, Sepulveda R, Rebolledo PA, Cupio A, et al. More than 50 Long-term effects of COVID-19: a systematic review and meta-analysis. medRxiv, 2021: 2021.01. 27.21250617).

[0021] Permanent lung damage and scarring can also be seen after significant involvement of the lungs and respiratory tract during viral infection. Post-COVID-19 fibrosis is estimated to be prevalent in one-third of hospitalized patients with SARS-CoV-2 infection (Ahmad Alhiyari M, Ata F, Islam Alghizzawi M, Bint I Bilal A, Salih Abdulhadi A, Yousaf Z., Post COVID-19 fibrosis, an emerging complication of SARS-CoV-2 infection, IDCases, 2020;23:e01041-e). The long-term effects and consequences of viral infection leave most patients symptomatic, weak, sleep-deprived, and unable to lead a normal life or work for extended periods, often exceeding six months.

[0022] Proanthocyanidins represent a group of plant polyphenols found in roots, bark, and fruits, possessing an astringent taste. Proanthocyanidins include subgroups of proanthocyanidins and prodelphin. Proanthocyanidins are biopolymers composed of flavanine subunits. Proanthocyanidins are composed of catechin and epicatechin units (also known as monomeric proanthocyanidins). Proanthocyanidins are extracted from plant materials using solvents such as water, ethanol, acetone, or liquid carbon dioxide by conventional methods. The extract is purified by solvent / solvent extraction, ultrafiltration, or chromatography. The purified extract is concentrated by solvent evaporation, freeze-drying, or spray drying.

[0023] An extract rich in proanthocyanidins from the bark of the French pine tree is marketed by Horphag Research Institute under the trademark Pycnogenol®. The extract contains 70 to 75% by weight of proanthocyanidins and other flavanols, such as catechins, epicatechins, and taxols; see Grimm et al., "Single and multiple dose pharmacokinetics of maritime pine bark extract (Pycnogenol) after oral administration to healthy volunteers," BMC Clinical Pharmacology, August 3, 2006, 6:4 - http: / / www.biomedcentral.com / 1472-6904 / 6 / 4.

[0024] Other extracts rich in proanthocyanidins are available from grape seeds, cypress berries, cocoa beans, or other plant materials. Pycnogenol® pine bark extract has been shown to stimulate endothelial nitric oxide synthase and induce vasodilation (Fitzpatrick, DF, Bing, B., Rohdewald, P., 1998).

[0025] US2004137081A1 (Rohdewald P. et al.) discloses that sexual wellness or sexual fitness is enhanced over time by administering a proanthocyanidin source and an arginine source on a daily basis. The two sources can be incorporated into a composition or taken separately from the package. The arginine source can be a salt or peptide of L-arginine and aspartic acid, such as arginine aspartate. Proanthocyanidins stimulate endothelial NO- synthase, which acts as a catalyst for the synthesis of nitric oxide from the receptor, which is the arginine source. Sufficient nitric oxide is released over time to enhance sexual wellness or sexual fitness. In cases of low levels of androgens in both sexes, the composition may contain sex hormones or sex hormone precursors or sex hormone stimulants or sex hormone bioavailability stimulants as another component.

[0026] The vascular endothelium is an active paracrine, endocrine, and autocrine organ, essential for regulating vascular tone and maintaining vascular homeostasis. Endothelial dysfunction is a major determinant of microvascular dysfunction, shifting vascular balance towards increased vasoconstriction, subsequently leading to local organ ischemia, inflammation accompanied by related tissue edema, and a procoagulant state.

[0027] Currently, the basic principle of the therapy is to stabilize the endothelium while dealing with viral replication. This strategy is particularly relevant to vulnerable patients with pre-existing endothelial dysfunction, which is associated with male sex, smoking, hypertension, diabetes, obesity, and a pre-existing cardiovascular disease, all of which are associated with adverse outcomes of COVID-19.

[0028] Therefore, there is a need for an effective and safe natural composition for the treatment or prevention of endothelial inflammation and / or systemic endothelial dysfunction triggered by COVID-19, including symptomatic post-COVID-19 individuals who have recovered from COVID-19. [Summary of the Invention]

[0029] The applicant has unexpectedly discovered that compositions containing proanthocyanidins show promising potential in the prevention and / or treatment of endothelial inflammation and / or systemic endothelial dysfunction in patients infected with Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). This safe, natural composition is particularly promising in the treatment and prevention of endothelial inflammation and / or systemic endothelial dysfunction triggered by COVID-19.

[0030] Contrary to prior art documents, this invention is solely intended to treat endothelial dysfunction resulting from COVID-19. No antiviral activity or effect against SARS-CoV2 self-infection is produced in this invention, but the invention addresses the consequences of such infection; therefore, the applicant believes that the problems addressed by the cited documents are different from those addressed by this invention.

[0031] However, Weichmann, F. and Rohdewald, P., Projected supportive effects of Pycnogenol® in patients suffering from multi-dimensional health impairments after a SARS-CoV2 infection, Int J Antimicrob Agents, 2020, 56(6): 106191, revealed that SARS-CoV2 strongly affects endothelial cells, triggering inflammation and / or coagulopathy, and leading to microcirculatory dysfunction, accompanied by endothelial problems and prethrombotic symptoms. Symptoms resulting from COVID-19 include endothelial dysfunction, coagulopathy, cytokine storm, microcirculatory problems, and capillary leak syndrome. Data from previous studies using Pycnogenol® provide strong evidence for its potential beneficial effects on improving endothelial function and normalizing and stabilizing microcirculatory function and platelet activity in patients with COVID-19, as well as exerting anti-inflammatory and antioxidant effects.

[0032] It has been shown that the virus SARS-CoV2 that causes COVID-19 strongly affects endothelial cells, leading to endothelial activation, prethrombotic symptoms and microcirculatory dysfunction, increased inflammation and coagulopathy.

[0033] Proanthocyanidins such as Pycnogenol® have been shown to improve endothelial function by stimulating endothelial nitric oxide synthase (eNOS), which amplifies NO production and ultimately leads to increased vascular lumen, adequate tissue perfusion and better blood circulation.

[0034] In addition, Pycnogenol® has been shown to improve the microcirculation perfusion system. In several clinical studies, the levels of O2 and CO2 in subcutaneous tissues, the diameter of microvessels in the nails, and blood flow velocity were increased after Pycnogenol® supplementation.

[0035] In addition, the increase in endothelial NO production induced by Pycnogenol® also causes platelet activation, thereby reducing platelet aggregation and reducing the risk of thrombosis, stroke or heart attack.

[0036] Pycnogenol®’s potent anti-inflammatory activity has been thoroughly studied, showing that the levels of pro-inflammatory cytokines, such as COX-1 and 2, 5-LOX, TNF-α, IL-1β, IL-6 and NF-κB, have been reduced to normal levels.

[0037] During inflammation, various reactive oxygen species are produced, which in turn burn inflammasomes and trigger the secretion of interleukins. The antioxidant activity of Pycnogenol® has been studied in several clinical studies, showing increased plasma antioxidant capacity (expressed as oxygen free radical uptake capacity) and reduced plasma oxidative stress as measured by plasma free radicals.

[0038] In addition, the Pycnogenol® metabolite M1 (δ-(3,4-dihydroxyphenyl)-γ-valerolactone), which promotes endothelial cell absorption, has been shown to exert direct anti-inflammatory activity by reducing iNOS (inducible nitric oxide synthase) expression and excess nitrite production.

[0039] In one embodiment of the present invention, a composition comprising proanthocyanidins and at least one suitable excipient is provided for the prevention or treatment of endothelial inflammation and / or systemic endothelial dysfunction triggered by COVID-19 induced by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection.

[0040] Preferably, the present invention relates to an oral composition comprising proanthocyanidins derived from plant extracts and at least one suitable excipient, wherein the plant extracts are selected from the group consisting of: pine bark, grape seed, apple, cocoa bean, peanut skin, cranberry extracts or combinations thereof, wherein the oral composition is a source of an ingredient selected from the group consisting of: δ-(3,4-dihydroxyphenyl)-g-valerolactone, δ-(3-methoxy-4-hydroxy-phenyl)-γ-valerolactone, catechin, epicatechin, ferulic acid, gallic acid, 4-hydroxybenzoic acid, caffeic acid, protocatechuic acid, zeaxanthin and mixtures thereof, wherein the ingredient is responsible for mediating an anti-inflammatory effect on the endothelium, and the composition is used to treat endothelial inflammation and / or systemic endothelial dysfunction triggered by SARS-CoV-2 infection.

[0041] Another object is to provide an oral composition comprising proanthocyanidins derived from plant extracts and at least one suitable excipient, wherein the plant extracts are selected from the group consisting of: pine bark, grape seed, apple, cocoa bean, peanut skin, cranberry extracts or combinations thereof, wherein the oral composition is a source of an ingredient selected from the group consisting of: δ-(3,4-dihydroxyphenyl)-g-valerolactone, δ-(3-methoxy-4-hydroxy-phenyl)-γ-valerolactone, catechin, epicatechin, and arbutin. The composition comprises ferrous acid, gallic acid, 4-hydroxybenzoic acid, caffeic acid, protocatechuic acid, zeaxanthin, and mixtures thereof, wherein the components are responsible for mediating anti-inflammatory effects on the endothelium, and the composition is intended for the treatment of endothelial inflammation and / or systemic endothelial dysfunction triggered by COVID-19 induced by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, characterized in that the oral composition is administered to symptomatic COVID-19 post-recovery individuals.

[0042] In another embodiment, the present invention provides a dietary or food supplement, food preparation, beverage, pharmaceutical nutritional product, or pharmaceutical comprising the composition of the present invention.

[0043] In another embodiment, the present invention provides an oral composition comprising a source of an ingredient selected from the group consisting of: δ-(3,4-dihydroxyphenyl)-g-valerolactone, δ-(3-methoxy-4-hydroxy-phenyl)-γ-valerolactone, catechin, epicatechin, ferulic acid, gallic acid, 4-hydroxybenzoic acid, caffeic acid, protocatechuic acid, zeaxanthin, and mixtures thereof, said composition being used to treat endothelial inflammation and / or systemic endothelial dysfunction triggered by SARS-CoV-2 infection, characterized in that said oral composition is administered from a symptomatic COVID-19 individual who has recovered from COVID-19.

[0044] In another embodiment, the present invention provides a method for treating or preventing endothelial inflammation and / or systemic endothelial dysfunction triggered by COVID-19 induced by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, the method comprising administering to an individual in need an effective amount of the composition used according to the present invention.

[0045] Preferably, the method of treating or preventing endothelial inflammation and / or systemic endothelial dysfunction triggered by SARS-CoV-2 infection comprises orally administering an effective amount of the oral composition of the present invention or the agent of the present invention to an individual in need, wherein the individual in need is a symptomatic individual who has recovered from COVID-19.

[0046] Other objects and advantages of the present invention will become apparent to those skilled in the art after referring to the appended claims and examining the following embodiments.

Implementation Method

[0047] Although similar or equivalent methods and materials described herein can be used to practice or test the invention, suitable methods and materials will be described below. All publications, patent applications, patents and other references mentioned herein are incorporated herein by reference in their entirety. The publications and applications discussed herein provide only those disclosures prior to the filing date of this application. Nothing herein should be construed as an admission that the invention is not entitled to rely on prior inventions prior to such publications. Furthermore, the materials, methods and examples are illustrative only and are not intended to be restrictive.

[0048] In case of conflict, this specification (including its specific definitions) shall prevail.

[0049] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the subject matter belongs. As used herein, the following definitions are provided to facilitate understanding of the invention.

[0050] As used in this specification and the claims, unless the context clearly requires otherwise, the singular forms "a / an" and "the" include the plural references.

[0051] In some cases, the presence of expansive words and phrases, such as “one or more,” “at least,” “but not limited to,” or other similar phrases, should not be construed as implying that a narrower situation is expected or required in the absence of such expansive phrases.

[0052] In addition, unless otherwise stated, the use of "or" means "and / or".

[0053] Similarly, "comprise", "comprises", "comprising", "include", "includes" and "including" are interchangeable and not intended to be restrictive. The term "comprise" is generally used in the sense of inclusion, that is, allowing the existence of one or more features or components.

[0054] It should also be understood that, where the term "comprising" is used in the description of various embodiments, those skilled in the art will understand that in some specific cases, the language "consistent with" or "comprises with" may be used instead to describe the embodiments.

[0055] The terms "preferred" and "preferably" refer to embodiments of the invention that provide particular benefits in certain circumstances. However, other embodiments may also be preferred in the same or other circumstances. Furthermore, the listing of one or more preferred embodiments does not imply that other embodiments are not applicable, nor is it intended to exclude other embodiments from the scope of the invention.

[0056] As used herein, the term "pine bark extract" refers to an extract of the bark of the French sea pine, such as Pycnogenol® (Horphag), which is commercially available. The terms "Pycnogenol®," "pine bark extract," and "French sea pine bark extract" are interchangeable. *P. pinaster* and *P. maritimeensis* should be understood to refer to the same organism commonly known as "French sea pine." Therefore, these terms are interchangeable.

[0057] As used herein, the term "extract" includes any preparation obtained from a plant, fruit or vegetable using extraction methods.

[0058] The term "food cuisine" generally refers to materials of plant or animal or synthetic origin that contain essential nutrients for maintaining growth, repair and important processes in living organisms and providing energy, such as carbohydrates, proteins, fats, vitamins, minerals, etc.

[0059] "Dietary or food supplements" refer to products containing substances such as vitamins, minerals, foods, herbal medicines, and amino acids, intended to supplement the daily intake of such substances. Dietary supplements are available in pill, tablet, capsule, powder, or liquid form and are intended for oral administration.

[0060] The term "medicinal nutritional product" refers to any substance, or a portion thereof, that provides medical or health benefits, including the prevention and treatment of disease. Such products range from individual nutrients, dietary supplements, and specific diets to genetically engineered foods, herbal products, and processed foods such as cereals, soups, and beverages. It also refers to products isolated or purified from food, generally sold in pharmaceutical forms not typically associated with food, and proven to have physiological benefits or provide protection against diseases such as chronic diseases.

[0061] The term "beverage" means a liquid intended for drinking, which may be water, flavored water, soft drink, alcoholic beverage, health drink or concentrated beverage based on dairy products (milk) or juice.

[0062] "Pharmaceutical-acceptable excipients or carriers" are any materials that do not interfere with the pharmacological activity of the active ingredient or degrade its bodily function in an individual to whom it may be administered, but which contribute to the manufacture of dosage forms or administration of compositions. Examples of pharmaceutically acceptable excipients include (but are not limited to) maltodextrin, calcium phosphate, and fused silica. Pharmaceutically acceptable excipients also include flavoring agents and various additives (such as other vitamins and minerals), all solvents, dispersion media, coatings, isotonics and absorption delay agents, sweeteners and the like, non-toxic excipients (such as wetting agents or emulsifiers), pH buffers and the like, such as sodium acetate, sorbitol monolaurate, triethanolamine oleate, and inert components (such as talc and magnesium stearate), which are standard excipients in the manufacture of tablets, capsules, and other dosage forms.

[0063] As used herein, the terms "individual" or "patient" are recognized in this art and are used interchangeably herein to refer to mammals, including dogs, cats, rats, mice, monkeys, cattle, horses, goats, sheep, pigs, camels, and preferably humans. In some embodiments, an individual is an individual requiring treatment or an individual suffering from a disease or condition. However, in other embodiments, an individual may be a normal individual. The terms do not indicate a specific age or sex. Therefore, it is intended to cover male or female adult individuals and newborn individuals.

[0064] The term "effective amount" refers to the amount necessary to achieve a physiological effect. A physiological effect can be achieved by administering a single dose or by repeated administration. Of course, the dose administered will vary depending on known factors such as the physiological characteristics of the particular composition; the individual's age, health status, and weight; the nature and severity of the symptoms; the type of concurrent treatment; the frequency of treatment; and the desired effect, which can be adjusted by those skilled in the art.

[0065] Surprisingly, the administration of compositions containing proanthocyanidins in suitable excipients is particularly promising in the treatment and prevention of endothelial inflammation and / or systemic endothelial dysfunction triggered by COVID-19.

[0066] Endothelium is a single layer of squamous endothelial cells lining the inner surface of blood vessels and lymphatic vessels. Endothelium forms an interface between circulating blood or lymph in the lumen and the rest of the vessel wall. Endothelial cells form a barrier between blood vessels and tissues and control the flow of substances and fluids into and out of tissues. Endothelial cells in direct contact with blood are called vascular endothelial cells, while those in direct contact with lymph are called lymphoendothelial cells. Vascular endothelial cells are arranged throughout the circulatory system, from the heart to the smallest capillaries.

[0067] These cells have unique functions, including fluid filtration, vascular tone, hemostasis, neutrophil recruitment, and hormone transport in glomeruli such as those of the kidney.

[0068] The endothelium lining the inner surface of the heart chambers is called the endocardium. Impaired function can cause serious health problems throughout the body.

[0069] "Endothelial inflammation" is an immune response within the vascular endothelium, in which it becomes inflamed, also known as endotheliitis. Individuals with COVID-19 may suffer from inflammation of all endothelial tissues in a wide range of organs. Endothelial inflammation has been observed to lead to widespread endothelial dysfunction in COVID-19. COVID-19 endotheliitis is a major cause of systemic impaired microcirculatory function and its clinical sequelae in different vascular beds of COVID-19 patients.

[0070] Endothelial activation encompasses a range of endothelial responses to inflammatory signals, including changes in thrombotic resistance, altered vasodilatory tone, and loss of barrier function. Upon activation, the endothelium rapidly facilitates cell transport. Leukocyte activation and migration are essential for normal innate and adaptive immunity. The term endothelial dysfunction can be applied to states where the endothelial cell phenotype imposes a net burden on the host. Endothelial responses to damage can lead to vasoconstriction, vasodilation, vascular leakage, and inflammation. Endothelial activation can transform the internal vascular surface from a non-adhesive barrier into a barrier that replenishes leukocytes, promotes coagulation, and further induces inflammatory processes.

[0071] Vasculitis is a group of conditions in which blood vessels are damaged by inflammation. Myocarditis, also known as inflammatory cardiomyopathy, is an inflammation of the myocardium. Both vasculitis and myocarditis are associated with endothelial activation and have been observed in patients with COVID-19.

[0072] Endothelial dysfunction, or loss of normal endothelial function, is a hallmark of vascular disease and is often considered a key early event in the development of atherosclerosis. Impaired endothelial function leading to hypertension and thrombosis is commonly seen in patients with coronary artery disease, diabetes, hypertension, hypercholesterolemia, and smokers. Endothelial dysfunction has also been shown to predict future adverse cardiovascular events and is present in inflammatory diseases such as rheumatoid arthritis and systemic lupus erythematosus. Endothelial dysfunction is a result of changes in endothelial function. Following fat (lipid) accumulation and under inflammatory stimulation, endothelial cells become activated, characterized by the expression of molecules such as E-selectin, VCAM-1, and ICAM-1, stimulating the adhesion of immune cells. In addition, transcription factors (substances that increase intracellular protein production) become activated; specifically AP-1 and NF-κB, leading to increased expression of cytokines such as IL-1, TNFα, and IFNγ, promoting inflammation. This state of endothelial cells promotes the accumulation of lipids and lipoproteins in the intima, leading to atherosclerosis. It subsequently recruits leukocytes and platelets, as well as promotes the proliferation of smooth muscle cells, resulting in fatty streaks. The lesions and persistent inflammation in the intima lead to endothelial shedding, which interferes with the endothelial barrier, causing damage and subsequent functional impairment.

[0073] In vascular diseases, endothelial dysfunction is a systemic pathological state of the endothelium. In addition to acting as a semipermeable membrane, the endothelium is also responsible for maintaining vascular tone and regulating oxidative stress by releasing mediators such as nitric oxide, prostacyclin and endothelin and controlling local angiotensin-II activity.

[0074] Proanthocyanidins represent a group of flavonoids, including proanthocyanidins, prodelphinidin, and propelin subgroups. Proanthocyanidins are homogeneous or heterogeneous polymers composed of monomeric units of catechin or epicatechin, linked by 4-8 or 4-6 bonds, resulting in a large number of isomers of proanthocyanidins. Typically, proanthocyanidin oligomers have a chain length of 2-12 monomeric units. Proanthocyanidins can be synthesized or extracted from plant materials. Non-limiting examples of plant material sources for proanthocyanidins include grape seeds, grape skins, pine bark, ginkgo leaves, cocoa beans, tamarind fruit, tomatoes, peanut shells, almonds, apples, cranberries, blueberries, and tea leaves.

[0075] Proanthocyanidins represent a group of plant polyphenols found in roots, bark, and fruits, and have an astringent taste. Proanthocyanidins include subgroups of proanthocyanidins and prodelphin. Proanthocyanidins are biopolymers composed of flavanane subunits.

[0076] Proanthocyanidins are composed of catechin and epicatechin units (also known as monomeric proanthocyanidins). Proanthocyanidins are members of the flavonoid proanthocyanidin (or condensed tannin) class. They are oligomeric compounds formed by catechin and epicatechin molecules. In addition to catechin and epicatechin, proanthocyanidins also contain gallic acid.

[0077] Proanthocyanidins, comprising smaller bioactive / bioavailable polymers (four or more catechins), represent a group of condensed flavan-3-ols found in many plants, most notably apples, pine bark, cinnamon, cherifolia, cocoa beans, grape seeds, grape skins, peanut skins, and red wine from grapes (Vitis vinifera, common grape). However, raspberries, cranberries, blackcurrants, green tea, black tea, and other plants also contain these flavonoids, as does cocoa beans. Proanthocyanidins can also be isolated from the heartwood of Quercus petraea and Quercus robur (barrel oak). Acai oil, obtained from the fruit of the açaí palm (Acai berry (Euterpe oleracea)), is rich in many proanthocyanidin oligomers.

[0078] On average, each serving of apple contains about eight times more proanthocyanidins than wine, with some varieties, such as Red Delicious and Granny Smith, having the highest levels.

[0079] A well-known product containing proanthocyanidins is an extract of the bark of the French sea pine (Pinus pinaster), which is commercially available under the trade name Pycnogenol® as a food supplement. See also US Patents 3,436,407 (MASQUELIER JACQUES); US 5,720,956 (ROHDEWALD, PETER); and US 6,372,266 (SUZUKI NOBUTAKA et al., Horphag Research Ltd.), all of which are incorporated herein by reference. Pycnogenol® is a standardized bark extract of the French sea pine, Aiton, Atlantica des Villar subspecies. The quality of this extract is described in detail in the United States Pharmacopeia (USP 28) (Maritime Pine Extract. In: United States Pharmacopeia. Rockville: United States Pharmacopeial Convention, Inc.; 2005, pp. 2115-2116). The extract consists of a polyphenol concentrate, which is also found in fruits and vegetables, but at lower concentrations. Polyphenols are composed of flavonoids (especially proanthocyanidins) and phenolic acids. All of these components have the ability to inactivate free radicals. Rohdewald P., A review of the French maritime pine bark extract (Pycnogenol®), a herbal medication with a diverse pharmacology, Int J Clin Pharmacol Ther 2002; 40(4): 158-168. Pycnogenol®, which is between 65-75%, is a proanthocyanidin containing catechin and epicatechin subunits with different chain lengths (Rohdewald P., A review of the French maritime pine bark extract (Pycnogenol®), an herbal medication with a diverse clinical pharmacology, Int J Clin Pharmacol Ther 2002; 40: 158-168). Other components are polyphenol monomers, phenolic acids or cinnamic acid and their glycosides (Id.).According to USP 28, the standardized Pycnogenol® extract contains between 65% and 75% proanthocyanidins (70 + / - 5% proanthocyanidins), and the compound is known for its relatively significant antioxidant and anti-inflammatory activities as well as other effects (Rohdewald P. "Pycnogenol®, French Maritime Pine Bark extract", Encyclopedia of Dietary Supplements, 2005, pp. 545-553).

[0080] In one embodiment of the present invention, a composition comprising proanthocyanidins and at least one suitable excipient or thereof is provided for the prevention or treatment of endothelial inflammation and / or systemic endothelial dysfunction triggered by COVID-19 induced by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection.

[0081] The compositions of the present invention contain 20% w / w to 95% w / w of proanthocyanidins and a suitable excipient (qsp) in sufficient quantity per 100% total volume. Preferably, the compositions of the present invention contain about 30% w / w to 80% w / w of proanthocyanidins, more preferably about 40% w / w to 80% w / w of proanthocyanidins, and even more preferably about 60% w / w to 80% of proanthocyanidins and a suitable excipient (qsp).

[0082] According to a preferred embodiment, the composition of the present invention comprises 65% to 75% w / w proanthocyanidins. Pycnogenol® extract is standardized to contain between 65% and 75% proanthocyanidins (70 + / - 5% proanthocyanidins). Thus, for example, a 100 mg tablet of the composition of the present invention contains between 65 mg and 75 mg of proanthocyanidins.

[0083] According to one embodiment of the present invention, endothelial inflammation is selected from the group consisting of: endotheliitis, myocarditis or vasculitis.

[0084] According to a preferred embodiment, the endothelial inflammation consists of severe endotheliitis.

[0085] Endothelial tissue is a layer of cells that acts as a protective barrier in blood vessels and regulates and balances various processes in the microvessels. Disruption of this regulatory process can, for example, cause circulatory disorders in organs and body tissues, leading to cell necrosis and thus the death of such organs or tissues.

[0086] "Endodermatitis" is an immune response within the endothelium of blood vessels, in which the blood vessels become inflamed. This condition can lead to edema of the surrounding tissues (including the matrix) and can cause irritation and pain.

[0087] SARS-CoV-2 has been shown not only to trigger lung inflammation, which subsequently causes other complications, but also to directly lead to systemic endotheliitis, an inflammation of all endothelial tissues in the body, affecting all vascular beds—in the blood vessels of the heart, brain, lungs, kidneys, and intestines. The consequences are fatal: this leads to severe microcirculatory disturbances that damage the heart, trigger pulmonary embolisms and vascular occlusion in the brain and intestines, and can also lead to multiple organ failure and even death. The endothelial tissues of younger patients are generally able to cope well with the viral attack. The situation is different for patients with hypertension, diabetes, heart failure, or coronary artery disease, all of whom share a common characteristic—significantly weakened endothelial function. If such patients are infected with SARS-CoV-2, they will be particularly at risk, as their already weakened endothelial function will be further impaired, especially during periods of peak viral replication.

[0088] "Myocarditis," also known as inflammatory cardiomyopathy, is an inflammation of the heart muscle. Symptoms may include shortness of breath, chest pain, decreased exercise capacity, and irregular heartbeat. The duration of the problem can vary from hours to months. Complications may include heart failure due to dilated cardiomyopathy or cardiac arrest. Myocarditis is most commonly attributed to viral infections.

[0089] "Vasculitis" is a group of conditions caused by inflammation that damages blood vessels. Both arteries and veins are affected. Lymphangitis (lymphangiitis obliterans) is sometimes considered a type of vasculitis. Vasculitis is mainly caused by leukocyte migration and the damage it causes. Although both occur in vasculitis, inflammation of veins (phlebitis) or arteries (arteritis) are separate entities.

[0090] According to a specific embodiment of the present invention, the vasculitis is Kawasaki-like disease.

[0091] Kawasaki disease is a syndrome of unknown cause that causes fever and primarily affects children under 5 years of age. It is a form of vasculitis in which blood vessels become inflamed throughout the body. Fever usually lasts more than five days and is not affected by commonly used medications. Other common symptoms include large lymph nodes in the neck, a rash in the genital area, and redness of the eyes, lips, palms, or soles of the feet. The skin on the hands and feet may peel within three weeks of onset, after which it usually heals. In some children, coronary artery aneurysms may form in the heart. Although the cause is unclear, it is likely due to an autoimmune response triggered by an infection in genetically susceptible individuals. It is not transmitted from person to person. Diagnosis is usually based on individual signs and symptoms. Other conditions that may be similar include scarlet fever, juvenile rheumatoid arthritis, and pediatric multisystem inflammatory syndrome associated with COVID-19.

[0092] According to Russell M Viner et al., The Lancet, "Kawasaki-like disease: emerging complication during the COVID-19 pandemic", Vol. 395, No. 10239, pp. 1741-1743, June 6, 2020, studies from several countries have confirmed that severe illness and death from COVID-19 in children are rare, and accurate estimates are unavailable due to the lack of a true population denominator. However, attention has now shifted to children's susceptibility to infection for two reasons. First, the extent to which children can transmit COVID-19 is key to how countries reopen communities after lockdowns. Second, emerging concerns about a new, severe Kawasaki-like disease in children associated with COVID-19, including the description of the outbreak in Italy by Lucio Verdoni and colleagues in The Lancet, are altering our understanding of this disease in children. Verdoni and colleagues described ten cases of Kawasaki-like disease (seven boys and three girls; age 7.5 years [SD 3.5]) in Bergamo, Italy, during the peak of the pandemic in the country (February 18 to April 20, 2020), with a monthly incidence rate approximately 30 times higher than the Kawasaki disease incidence rate observed in the past five years.

[0093] The compositions of the present invention contain proanthocyanidins derived from plant extracts or synthetic materials (i.e., synthetic proanthocyanidins).

[0094] The plant extract may be selected from the group consisting of proanthocyanidins containing the extract, said extract being selected from pine bark, cypress fruit, grape seed, apple, peanut skin, walnut, pomegranate, tomato, almond, tea, hawthorn, cocoa bean extract or combinations thereof.

[0095] The extract rich in proanthocyanidins is natural and preferably a plant extract having more than 50% by weight (dehydrated extract) of proanthocyanidins, more preferably more than 70% by weight, and even more preferably more than 75% by weight of proanthocyanidins. Preferably, the plant extract according to the invention is derived from pine bark, and more preferably, the plant extract is Pycnogenol®.

[0096] In a preferred embodiment, the formulation containing proanthocyanidins may be a pine bark extract. The pine bark may be from the coastal pine, such as from Pycnogenol®. In a preferred embodiment, the composition may contain proanthocyanidins at a concentration of 10% to 100% of the total weight. For example, the Pycnogenol® composition may be diluted or concentrated to contain 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 90%, or 95% proanthocyanidins. Concentration may be performed using known methods such as column chromatography or affinity chromatography.

[0097] According to an embodiment of the present invention, the composition containing proanthocyanidins is a source of an ingredient selected from the group consisting of: δ-(3,4-dihydroxyphenyl)-g-valerolactone, δ-(3-methoxy-4-hydroxy-phenyl)-γ-valerolactone, catechin, epicatechin, ferulic acid, gallic acid, 4-hydroxybenzoic acid, caffeic acid, protocatechuic acid, zeaxanthin, and mixtures thereof, wherein the ingredient is responsible for mediating the anti-inflammatory effect on the endothelium.

[0098] Unbound by theory, it is believed that the components described herein mediate anti-inflammatory effects by reducing nitrite production in LPS-stimulated macrophages through direct NO quenching and downregulation of iNOS (inducible NO synthase).

[0099] According to Grimm et al., "Single and multiple dose pharmacokinetics of maritime pine bark extract (Pycnogenol) after oral administration to healthy volunteers," BMC Clinical Pharmacology, August 3, 2006, 6:4- http: / / www.biomedcentral.com / 1472-6904 / 6 / 4, the first systematic pharmacokinetic analysis of the components and metabolites of standardized maritime pine bark extract (USP quality) was performed on human volunteers after single and repeated administration. The components of the extract were bioavailable and detectable in the plasma of all individuals. The pharmacokinetic parameters calculated for the compounds identified so far are similar to those from other studies. Furthermore, the steady-state concentrations of catechins, caffeic acid, ferulic acid, and M1 (δ-(3,4-dihydroxyphenyl)-γ-valerate) were described for the first time, and the plasma concentrations of taxanes in humans were presented for the first time. The detection of ten previously unknown bioavailable components and metabolites by Pycnogenol reveals the potential to discover active compounds with anti-inflammatory biological effects.

[0100] According to the present invention, the composition also comprises at least one suitable excipient, wherein the suitable excipient is preferably a pharmaceutically acceptable excipient.

[0101] Examples of suitable excipients for the present invention include (but are not limited to) anti-adhesives, adhesives (e.g., crude cellulose, tragacanth gum or gelatin), coatings, disintegrants, fillers, diluents, softeners, emulsifiers, flavorings, colorants, adjuvants, lubricants, functional agents (e.g., nutrients), viscosity modifiers, build-up agents, slip agents (e.g., colloidal silica), surfactants, penetrants, diluents or any other inactive ingredients or combinations thereof.

[0102] For example, the compositions of the present invention may include excipient materials selected from the group consisting of: calcium carbonate, colorants, whitening agents, preservatives and flavoring agents, triacetin, magnesium stearate, sterotes, natural or artificial flavorings, essential oils, plant extracts, fruit flavorings, gelatin or combinations thereof.

[0103] Where appropriate, the formulations of the present invention may include other artificial or natural sweeteners, bulk sweeteners, or combinations thereof. Bulk sweeteners include both caloric and non-caloric compounds. Non-limiting examples of bulk sweeteners include sucrose, dextrose, maltose, dextrin, dehydrated invert sugar, fructose, high-fructose corn syrup, levulose, galactose, corn syrup solids, tagatose, polyols (e.g., sorbitol, mannitol, xylitol, lactitol, erythritol, and maltitol), hydrogenated starch hydrolysate, isomalt, trehalose, and combinations thereof.

[0104] Where appropriate, the compositions of the present invention may further comprise nonsteroidal anti-inflammatory drugs, such as acetylsalicylic acid (aspirin).

[0105] According to a preferred embodiment of the present invention, the composition is adapted for oral administration.

[0106] Preferably, the oral administration is in the form of food preparation, dietary supplement, pharmaceutical nutritional product or beverage.

[0107] Alternatively, the present invention provides a pharmaceutical preparation comprising a composition as defined above.

[0108] Therefore, the present invention further provides food preparations, dietary or food supplements, pharmaceutical-like nutritional products, beverages, and pharmaceuticals comprising the compositions of the present invention. As described above, the pharmaceuticals may further comprise acetylsalicylic acid (aspirin) and pharmaceutically acceptable excipients.

[0109] Preferably, the dietary supplement, pharmaceutical-like nutritional product, or pharmaceutical preparation of the present invention is administered at a dose of 25 mg / day to 500 mg / day, more preferably 25 mg / day to 300 mg / day. The dietary supplement, pharmaceutical-like nutritional product, or pharmaceutical preparation of the present invention contains 20% w / w to 80% w / w proanthocyanidins and suitable excipients qsp.

[0110] Preferably, the composition of the present invention comprises a Pycnogenol® extract, which is standardized to contain between 65% and 75% proanthocyanidins. Thus, a 100 mg tablet may contain between 65 mg and 75 mg of proanthocyanidins, more preferably 70 mg of proanthocyanidins.

[0111] The present invention also provides a method for treating or preventing endothelial inflammation and / or systemic endothelial dysfunction triggered by COVID-19 induced by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, the method comprising administering an effective amount of the composition or agent according to the present invention to an individual in need.

[0112] Preferably, the composition or pharmaceutical preparation is administered orally.

[0113] More preferably, the composition or agent is administered at a dose of 25 mg / day to 500 mg / day, more preferably 25 mg / day to 300 mg / day.

[0114] The individual in need is an animal, preferably a mammal, and more preferably a human.

[0115] If intended for oral administration, the composition or agent of the present invention may be in the following forms: for example, tablets, capsule tablets, pills, hard capsules or soft capsules, lozenges, flat capsules, dispensable powders, granules, suspensions, elixirs, dispersions or any other form reasonably adapted for such administration.

[0116] In a preferred embodiment, the composition according to the invention comprises proanthocyanidins as the sole active ingredient administered to an individual.

[0117] Also covers compositions comprising sources of ingredients selected from the group consisting of: δ-(3,4-dihydroxyphenyl)-g-valerolactone, δ-(3-methoxy-4-hydroxy-phenyl)-γ-valerolactone, catechin, epicatechin, ferulic acid, gallic acid, 4-hydroxybenzoic acid, caffeic acid, protocatechuic acid, zeaxanthin, and mixtures thereof, said compositions being used for the prevention or treatment of endothelial inflammation and / or systemic endothelial dysfunction triggered by COVID-19 induced by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection.

[0118] Preferably, the ingredient is derived from a formulation containing 20% ​​to 95% w / w proanthocyanidins. More preferably, the ingredient is derived from a formulation containing 65% to 75% w / w proanthocyanidins.

[0119] According to another embodiment, the present invention provides an oral composition comprising proanthocyanidins derived from plant extracts and at least one suitable excipient, wherein the plant extracts are selected from the group consisting of: pine bark, grape seed, apple, cocoa bean, peanut skin, cranberry extracts or combinations thereof, wherein the oral composition is a source of an ingredient selected from the group consisting of: δ-(3,4-dihydroxyphenyl)-g-valerolactone, δ-(3-methoxy-4-hydroxy-phenyl)-γ-valerolactone, catechin, epicatechin, ferulic acid, gallic acid, 4-hydroxybenzoic acid, caffeic acid, protocatechuic acid, zeaxanthin and mixtures thereof, wherein the ingredient is responsible for mediating anti-inflammatory effects on the endothelium, and the composition is used to treat endothelial inflammation and / or systemic endothelial dysfunction triggered by SARS-CoV-2 infection.

[0120] In a preferred embodiment, the present invention provides an oral composition comprising proanthocyanidins derived from plant extracts and at least one suitable excipient, wherein the plant extracts are selected from the group consisting of: pine bark, grape seed, apple, cocoa bean, peanut skin, cranberry extracts or combinations thereof, wherein the oral composition is a source of ingredients selected from the group consisting of: δ-(3,4-dihydroxyphenyl)-g-valerolactone, δ-(3-methoxy-4-hydroxy-phenyl)-γ-valerolactone, catechins, epicatechin, etc. The composition comprises catechin, ferulic acid, gallic acid, 4-hydroxybenzoic acid, caffeic acid, protocatechuic acid, zeaxanthin, and mixtures thereof, wherein the components are responsible for mediating anti-inflammatory effects on the endothelium, and the composition is intended for the treatment of endothelial inflammation and / or systemic endothelial dysfunction triggered by COVID-19 induced by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, characterized in that the oral composition is administered to symptomatic COVID-19 post-recovery individuals.

[0121] Endothelial dysfunction triggered by COVID-19 has been observed to persist for months after SARS-CoV-2 infection in post-COVID patients (Riou M, et al., Reduced Flow-Mediated Dilatation Is Not Related to COVID-19 Severity Three Months after Hospitalization for SARS-CoV-2 Infection, Journal of Clinical Medicine, 2021; 10(6):1318, https: / / doi.org / 10.3390 / jcm10061318). The value of assessing endothelial function (using flow-mediated dilatation (FMD)) has been proposed, such as for risk stratification and early detection of vascular sequelae, as well as long-term cardiovascular complications in COVID-19 patients. (Evans, PC et al., Endothelial Dysfunction in COVID-19: A Position Paper of the ESC Working Group for Atherosclerosis and Vascular Biology, and the ESC Council of Basic Cardiovascular Science, Cardiovasc. Res. 2020).

[0122] Therefore, it has been observed that endothelial dysfunction triggered by COVID-19 can persist for months in patients who have recovered from COVID following SARS-CoV-2 infection. Therefore, the oral composition according to the invention can be used indiscriminately to treat endothelial inflammation and / or systemic endothelial dysfunction in patients with COVID infection, including patients post-COVID-19 or individuals who have recovered from COVID-19.

[0123] Preferably, the oral composition of the present invention comprises 20% to 95% w / w proanthocyanidins and is administered at a dose of 25 mg / day to 300 mg / day.

[0124] More preferably, the oral composition contains 65% to 75% w / w proanthocyanidins.

[0125] According to one embodiment, systemic endothelial dysfunction is selected from the group comprising: endothelial blood flow and microcirculation problems or endothelial coagulation problems.

[0126] In particular, the endothelial blood flow and microcirculation function problems are selected from the group including: renal function problems, pulmonary function problems, liver function problems, cognitive function problems, blood pressure problems related to endothelial dysfunction, and blood velocity problems related to endothelial dysfunction.

[0127] Specifically, endothelial coagulation problems are selected from the group that includes thrombosis and platelet aggregation.

[0128] In particular, endothelial inflammation is selected from the group that includes: endotheliitis, myocarditis or vasculitis.

[0129] Preferably, vasculitis is Kawasaki-like disease.

[0130] According to another embodiment of the present invention, the endotheliitis is severe endotheliitis.

[0131] Pycnogenol® is a preferred pine bark extract.

[0132] According to one embodiment, the suitable excipient is a pharmaceutically acceptable excipient.

[0133] Advantageously, the oral composition of the present invention is in the form of food preparation, dietary supplement, pharmaceutical nutritional product or beverage.

[0134] According to one embodiment, the present invention provides a pharmaceutical preparation comprising the oral composition of the present invention.

[0135] Preferably, the pharmaceutical preparation or dietary supplement of the present invention is administered at a dose of 25 mg / day to 300 mg / day.

[0136] The present invention also provides a method for treating or preventing endothelial inflammation and / or systemic endothelial dysfunction triggered by COVID-19 induced by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, comprising orally administering an effective amount of the oral composition or agent of the present invention to an individual in need, wherein the individual in need is a symptomatic COVID-19 individual who has recovered from COVID-19.

[0137] Preferably, the oral composition or agent of the present invention is administered at a dose of 25 mg / day to 300 mg / day / patient.

[0138] Another object of the present invention is to provide an oral composition comprising a source of an ingredient selected from the group consisting of: δ-(3,4-dihydroxyphenyl)-g-valerolactone, δ-(3-methoxy-4-hydroxy-phenyl)-γ-valerolactone, catechin, epicatechin, ferulic acid, gallic acid, 4-hydroxybenzoic acid, caffeic acid, protocatechuic acid, zeaxanthin, and mixtures thereof, said oral composition for treating endothelial inflammation and / or systemic endothelial dysfunction triggered by SARS-CoV-2 infection.

[0139] According to another embodiment, an oral composition comprising a source of an ingredient selected from the group consisting of: δ-(3,4-dihydroxyphenyl)-g-valerolactone, δ-(3-methoxy-4-hydroxy-phenyl)-γ-valerolactone, catechin, epicatechin, ferulic acid, gallic acid, 4-hydroxybenzoic acid, caffeic acid, protocatechuic acid, zeaxanthin, and mixtures thereof, wherein the oral composition is administered from a symptomatic post-COVID-19 individual who has recovered from COVID-19.

[0140] Preferably, the source of the ingredient is a formulation containing 20% ​​to 95% w / w proanthocyanidins.

[0141] More preferably, the source of said ingredient is derived from a formulation containing 65 to 75 w / w% proanthocyanidins.

[0142] In Example 1, the applicant did evaluate the effect of Pycnogenol® on improving endothelial function, microcirculation and the inflammatory marker IL-6 in symptomatic individuals with COVID-19 over a period of 3 weeks compared with a control group.

[0143] Individuals in both groups were similar at baseline. Progressive improvement was observed in both groups under standard management (SM) and SM combined with supplementation. Patients supplemented with Pycnogenol® showed significantly better improvement compared to the control group. No side effects of the supplementation were observed; tolerability was optimal. Progressive evolution over time was observed in all target measurements.

[0144] At the time of inclusion, flow-mediated dilatation (FMD) was low in all individuals. After 2 weeks, FMD was significantly higher in the Pycnogenol® group compared with the control group (p < 0.05 vs. control group), and after 3 weeks, it continued to improve compared with the control group (p < 0.05 vs. control group).

[0145] This improvement was also observed in reactive hyperemia measured by increased skin flow (laser Doppler measurement) after the release of the occluded hypersystolic cuff. The difference from the control group was statistically significant after 1 week and continued to increase after 2 and 3 weeks. This not only confirms the improvement in endothelial function but also the improvement in microcirculation.

[0146] Regarding the inflammatory marker IL-6, which was elevated at baseline, it gradually decreased over 3 weeks. After 3 weeks, the IL-6 level in the Pycnogenol® group was significantly lower than that in the control group (p < 0.05). The difference from the control group was statistically significant after 1 week.

[0147] In summary, Pycnogenol® provides an important solution for managing key parameters associated with symptomatic COVID-19 syndrome.

[0148] In Example 2, the applicant did evaluate, within 3 months, the effects of Pycnogenol® on symptoms of post-COVID-19 syndrome in symptomatic individuals who had recovered from COVID-19, as well as on improving endothelial function, microcirculation, inflammatory markers, and oxidative stress, compared with a control group.

[0149] Individuals in both groups were similar at baseline. Progressive improvement was observed in both groups under standard management (SM) and in the combination of SM and supplementation. Patients supplemented with Pycnogenol® showed significantly better improvement compared to the control group. No side effects of the supplementation were observed; tolerability was optimal. Progressive evolution over time was observed in all target measurements.

[0150] Endothelial dysfunction in all individuals at enrollment was assessed by flow-mediated dilation (FMD) and reactive hyperemia of the fingers in the microcirculation (laser Doppler measurement) following release of the occlusive hyperconstriction cuff. Endothelial function was significantly improved in the Pycnogenol® group at 1 month and 3 months (p < 0.05 compared to the control group). Ankle swelling rate (RAS) measured by strain gauge was significantly reduced in the supplementation group (p < 0.05) compared to the control group, which showed improved capillary filtration rate. At enrollment, renal cortical flow velocity indicated reduced perfusion in all patients (lower systolic and diastolic flow velocities). Renal cortical flow velocity was significantly increased by the supplementation compared to the control group (p < 0.05), with improvements in systolic velocity and diastolic components. High-sensitivity CRP (hs-CRP) and Il-6 plasma levels gradually decreased over 3 months, with a significantly greater decrease in the supplementation group (p < 0.05). The number of patients with normal plasma IL-6 levels at the end of the study was higher due to supplementation (p < 0.05). ESR followed the same pattern, with a gradual and more significant decrease in supplemented individuals (p < 0.02). Oxidative stress was significantly lower in the supplementation group compared to the control group (p < 0.05). Blood pressure and heart rate were normalized in all individuals in the supplementation group; systolic blood pressure was significantly lower in the supplementation group at the end of the study (p < 0.05). At the end of the study, all other blood parameters (including platelets and coagulation factors) were within normal ranges.

[0151] In summary, oral administration of Pycnogenol® provides an important option for treating some of the symptoms and signs associated with post-COVID-19 syndrome. This assessment provides some basic principles for the use of Pycnogenol® in this condition, which will be of great significance in the coming years.

[0152] Those skilled in the art will understand that changes and modifications are permitted in the invention described herein, except for those specifically described. It should be understood that the invention includes all such changes and modifications without departing from its spirit or essential characteristics. The invention also includes all steps, features, compositions, and compounds individually or collectively mentioned or indicated in this specification, and any and all combinations of said steps or features, or any two or more thereof. Therefore, the invention should be considered as encompassing all aspects described without limitation, and its scope is determined by the appended claims, which are intended to cover all changes occurring within the meaning and scope of equivalents.

[0153] Multiple references are cited throughout this manual, and each reference is incorporated into this document in its entirety.

[0154] The foregoing description will be more fully understood with reference to the following examples. However, such examples are exemplary methods for implementing the invention and are not intended to limit the scope of the invention.

[0155] Example 1: Pycnogenol® reduces endothelial and microcirculatory dysfunction and inflammation in symptomatic individuals with COVID-19:

[0156] The goal of this controlled study is to evaluate the effects of Pycnogenol® on endothelial function, microcirculation, and the inflammatory marker IL-6 in patients with COVID-19 compared to a control group.

[0157] Individual, Method

[0158] Methods: Ten individuals with symptomatic COVID-19 were enrolled on the day of their doctor / hospital consultation. One group of five followed standard rehabilitation management, while five similar individuals received a daily supplement of 150 mg Pycnogenol® (at three 50 mg doses) in addition to standard treatment for four weeks.

[0159] COVID-19 individuals aged 35-70 years who had no significant history of COVID-19 before contracting the disease and were willing to participate were included in the registration.

[0160] COVID-19 is diagnosed by detecting SARS-CoV-2 RNA using nasopharyngeal samples via reverse transcription polymerase chain reaction (RT-PCR).

[0161] Exclusion criteria were any acute or systemic illness, medication, or other supplement intake. During the 3-week enrollment, 10 individuals with COVID-19 received standard management (SM) or a combination of Pycnogenol® and SM. One group of 5 followed common recovery management, while 5 similar individuals received 150 mg Pycnogenol® daily (in three 50 mg doses) in addition to standard management. Follow-up lasted 3 weeks.

[0162] Study parameters were assessed in all patients at baseline and at week 1, week 2 and week 3 at the end of enrollment.

[0163] The supplement study is open and comparative.

[0164] Research Endpoint

[0165] All study parameters were assessed before 10 am at room temperature (20°C) after 20 minutes of acclimatization.

[0166] Consider the following research endpoints:

[0167] 1. Endothelial function & microcirculation.

[0168] a. Flow-mediated dilation: Flow-mediated dilation (FMD) of the brachial artery is a non-invasive technique for assessing the establishment of endothelial function. The technique is performed as previously described by measuring brachial artery dilation after hypersystolic occlusion (Enseleit F, Sudano I, Periat D, Winnik S, Wolfrum M, Flammer AJ et al., Effects of Pycnogenol on endothelial function in patients with stable coronary artery disease: a double-blind, randomized, placebo-controlled, cross-over study, Eur Heart J., 2012;33(13):1589-97); arterial size was measured using high-resolution ultrasound before and 1 minute after brachial cuff release.

[0169] b. Reactive hyperemia: Reactive hyperemia is a non-invasive assessment of peripheral microvascular function and endothelial function. Laser Doppler flowmeter (LDF) non-invasively measures skin flow (specified LDF units) within minutes after arterial occlusion. Flow is measured after occlusion during the same procedure described for assessing the brachial artery (within the same test period and within the same time range). Finger flow is measured at rest before occlusion as an average of one-minute continuous recordings. As previously described, increased distal, fingertip laser-Doppler finger flow is measured after occlusion (Freccero C, Holmlund F, Bornmyr S, Castenfors J, Johansson AM, Sundkvist G et al., Laser Doppler perfusion monitoring of skin blood flow at different depths in finger and arm upon local heating, Microvasc Res., 2003;66(3):183-9). This increase in flow is considered a measure of reactive hyperemia in the microcirculation, and it is reduced or eliminated in patients with severe vascular disease or diabetic microangiopathy. It is measured as an increase in skin flow after occlusion (laser Doppler flow increase %).

[0170] 2. Inflammatory markers

[0171] Interleukin-6 plasma levels in pg / mL: Elevated IL-6 levels can indicate an ongoing inflammatory response and may be consistent with systemic infection, local infection, or chronic inflammatory disease. IL-6 is considered a nonspecific marker associated with inflammatory responses; it is not a diagnosis of any specific disease or disease process (including COVID-19).

[0172] The primary assessments during the study were endothelial function and the inflammatory marker IL-6.

[0173] Result

[0174] Ten individuals with moderate / severe COVID-19 symptoms were included in the study. Two groups were formed. One group of five patients followed standard management without supplementation (control group), while five similar individuals received Pycnogenol® in addition to standard management. Both groups were similar at inclusion. No one withdrew from the study.

[0175] No side effects of the supplement were observed; best tolerability.

[0176] At the end of the study, all individuals tested positive for antibodies against SARS-CoV-2.

[0177] The results of the evaluation parameters of the study are shown in Table 1.

[0178] Table 1: Overview including all assessment parameters. *=p<0.05 relative to control. PY=Pycnogenol®. CON=Control group. Visit 1 Baseline Visit 2 Week 1 Visit 3 Week 2 Visit 4 Week 3 1. Endothelial function & microcirculation a. FMD [%] PY 6.9±0.9 7.1±0.7 8.46±1.1* 11.14±1.1* Con 7.0±0.84 6.7±0.9 6.9±0.8 7.0±0.7 b. Post-occlusion reactive hyperemia of finger skin flow [increase %] PY 10.6±1.6 13.6±2.0* 16.4±1.6* 18.5±1.5* Con 10.6±1.4 10.8±1.3 11.1±1.4 12.5±1.4 2. Inflammatory markers b. IL-6 [PG / ML] PY 9.3±1.7 5.1±2.0* 2.6±0.6* 1.8±0.7* CON 9.2±2.3 8.5±2.4 7.4±2.3 6.6±2.2

[0179] At the time of inclusion, vascular screening in all included individuals did not reveal significant vascular problems (plaques, intimal-medial thickening, aneurysms). It is important to note that vascular atherosclerotic lesions can alter endothelial function.

[0180] Endothelial function and microcirculation

[0181] At inclusion, flow-mediated dilatation (FMD) was low in all individuals. After 2 weeks, the Pycnogenol® group (8.46±1.1%) had significantly higher FMD compared to the control group (6.9±0.8%) (p<0.05 vs. control group), and after 3 weeks, the Pycnogenol® group continued to improve compared to the control group (7.0±0.7%) (11.14±1.1%) (p<0.05 vs. control group).

[0182] This improvement was also observed in reactive hyperemia measured by increased skin flow (laser Doppler measurement) after the release of the occluded hypersystolic cuff. The difference between the control group and the control group was statistically significant at 1 week (13.6±2.0% vs. 10.8±1.3%), and continued to increase at 2 weeks (16.4±1.6% vs. 10.8±1.3%) and 3 weeks (18.5±1.5% vs. 12.5±1.4%). This not only confirms the improvement in endothelial function but also the improvement in microcirculation.

[0183] Regarding the elevated inflammatory marker IL-6 at baseline, it gradually decreased over 3 weeks. After 3 weeks, IL-6 levels were significantly lower in the Pycnogenol® group compared to the control group (p < 0.05). After one week, plasma IL-6 levels in the Pycnogenol® group decreased significantly from 9.3 ± 1.7 pg / mL to 5.1 ± 2.0 pg / mL, and further decreased to 1.8 ± 0.7 pg / mL after 3 weeks. The difference with the control group was statistically significant after 1 week. In the control group, plasma IL-6 levels decreased from 9.2 ± 2.3 pg / mL to 8.5 ± 2.4 pg / mL after 1 week, and further decreased to 6.6 ± 2.2 pg / mL after 3 weeks.

[0184] In summary, Pycnogenol® provides an important solution for managing key parameters associated with symptomatic COVID-19 syndrome.

[0185] This assessment provides some interesting basic principles for the use of Pycnogenol® in this condition, which will be of great significance in the coming years.

[0186] Example 2: Preventive effects of Pycnogenol® on cardiovascular risk factors (including endothelial function) and microcirculation in individuals who have recovered from COVID-19.

[0187] The goal of this controlled study is to evaluate the effects of Pycnogenol® on symptoms of post-COVID-19 syndrome and on improvement of endothelial function and microcirculation compared with the control group; inflammatory markers and plasma reactive oxygen metabolites were investigated in symptomatic individuals who had recovered from COVID-19 in this 3-month registry study.

[0188] Individual, Method

[0189] Methods: Sixty individuals who had recovered from COVID-19 with self-diagnostic symptoms were included. One group of 30 followed standard recovery management, while 30 similar individuals received 150 mg Pycnogenol® supplements daily (at three 50 mg doses) in addition to standard management.

[0190] Individuals aged 35-70 who have recovered from COVID-19, have no significant medical history prior to COVID-19, and are willing to participate will be included in the registration. No medication will be used except for symptomatic and occasional pain treatment, and appropriate vitamins and diet.

[0191] Individuals were included at least 2 months after viral infection. COVID-19 was diagnosed using nasopharyngeal samples by detecting SARS-CoV-2 RNA via reverse transcription polymerase chain reaction (RT-PCR).

[0192] Exclusion criteria were any acute or systemic illness, medication, or other supplement intake. During the 90-day study, 60 individuals who had recovered from COVID-19 received standard management (SM) or a combination of Pycnogenol® and SM. One group of 30 followed standard recovery management, while another 30 similar individuals supplemented with 150 mg Pycnogenol® daily (at three 50 mg doses) in addition to standard management. Follow-up lasted for 3 months.

[0193] Study parameters were assessed in all patients at baseline, at week 2, month 1, and month 3 at the end of enrollment.

[0194] The supplement study is open and comparative.

[0195] Study Endpoint

[0196] All study parameters were assessed before 10 am at room temperature (20°C) after 20 minutes of acclimatization.

[0197] Consider the following research endpoints:

[0198] 1. Endothelial function & microcirculation:

[0199] a. Flow-mediated dilation: Flow-mediated dilation of the brachial artery (FMD) is a non-invasive technique for assessing the establishment of endothelial function. The technique is performed as previously described by measuring brachial artery dilation after hypersystolic occlusion (Enseleit F, Sudano I, Periat D, Winnik S, Wolfrum M, Flammer AJ et al., Effects of Pycnogenol on endothelial function in patients with stable coronary artery disease: a double-blind, randomized, placebo-controlled, cross-over study, Eur Heart J., 2012;33(13):1589-97); arterial size was measured 1 minute before and after brachial cuff release using high-resolution ultrasound.

[0200] b. Reactive hyperemia: Reactive hyperemia is a non-invasive assessment of peripheral microvascular function and endothelial function. Laser Doppler flowmeter (LDF) non-invasively measures skin flow (specified LDF units) within minutes after arterial occlusion. Flow is measured post-occlusion during the same procedure described for assessing the brachial artery (within the same test period and within the same time range). Finger flow is measured at rest before occlusion as an average of one-minute continuous recordings. As previously described, increased distal, fingertip laser-Doppler finger flow is measured after occlusion (Hu S, Belcaro G, Cornelli U, Luzzi R, Cesarone M, Dugall M et al., Effects of Pycnogenol(R) on endothelial dysfunction in borderline hypertensive, hyperlipidemic, and hyperglycemic individuals: the borderline study, Int Angiol., 2015;34(1):43-52). This increase in flow is considered a measure of reactive hyperemia in the microcirculation, and it is reduced or eliminated in patients with severe vascular disease or diabetic microangiopathy. It is measured as an increase in skin flow after occlusion (laser Doppler flow increase %).

[0201] c. Ankle Swelling Rate (RAS): This test quantifies capillary filtration at the ankle joint. RAS is measured using a strain gauge volume change recorder (SPG16, Hokanson, USA), where the strain gauge is placed at the minimum ankle circumference while the patient remains still for 30 minutes. The patient is then asked to move to a standing position. RAS is measured by considering the volume of tissue per 100 cm³ in mL / min after supine and standing positions (10 and 20 minutes) (Belcaro G BA, Hoffman U, Nicolaides AN., Laser Doppler. Med Orion., 2006).

[0202] d. Renal cortical blood flow is measured as arterial velocity (in cm / sec) using high-resolution color duplex measurement (Preirus, Hitachi, Japan) (Cesarone MR, De Sanctis MT, Laurora G, Ambrosoli L, Marelli C, Belcaro G, Effects of trandolapril on 24-h ambulatory blood pressure in patients with mild-to-moderate essential hypertension, J Cardiovasc Pharmacol., 1994;23 Supplement 4:S65-72).

[0203] 2. Inflammatory markers & oxidative stress:

[0204] a. Blood high-sensitivity C-reactive protein (hs-CRP) (25): The criteria for hs-CRP levels used in this study were: less than 1.0 mg / L (low risk of cardiovascular disease); hs-CRP between 1.0 mg / L and 3.0 mg / L (intermediate risk of CVD); and hs-CRP levels greater than 3.0 mg / L (high risk of CVD).

[0205] b. Interleukin-6 plasma levels in pg / mL: Elevated IL-6 levels can indicate an ongoing inflammatory response and may be consistent with systemic infection, local infection, or chronic inflammatory disease. IL-6 is considered a nonspecific marker associated with inflammatory responses; it is not a diagnosis of any specific disease or disease process (including COVID-19).

[0206] c. Interleukin-6 (IL-6): The proportion of patients with normal IL-6 (value ≤1.8 pg / mL).

[0207] d. Erythrocyte sedimentation rate (ESR) in mm / hr: Plasma ESR has long been used as a laboratory test to assess acute-phase responses to inflammation. ESR is slightly slower and less sensitive than hs-CRP, but it provides further accurate and valuable information about a patient's inflammatory status. (Lapić I, Padoan A, Bozzato D, Plebani M, Erythrocyte Sedimentation Rate and C-Reactive Protein in Acute Inflammation, Am J Clin Pathol. , 2020;153(1):14-29). The criteria used in this study were as follows: men >50 years, normal ESR value <20; men <50 years, normal ESR value <15; women >50 years, ESR value <30; women <50 years, normal ESR value <20.

[0208] e. Oxidative stress is assessed by measuring plasma free radicals in a drop of blood obtained from the fingertip and is expressed in calories (Cesarone MR, Belcaro G, Carratelli M, Cornelli U, De Sanctis MT, Incandela L et al., A simple test to monitor oxidative stress, Int Angiol., 1999;18(2):127-30).

[0209] 3. Blood pressure (systolic SBP and diastolic DBP) and heart rate (HR).

[0210] The primary assessments during the study were endothelial function and microcirculation parameters at each visit. Secondary clinical outcomes—all of which changed at inclusion—included inflammatory markers such as hs-CRP and IL-6, oxidative stress, and blood pressure / heart rate.

[0211] Statistical analysis: At least 20 individuals in each group (SM and SM+ supplement) were considered to require evaluation of differences in target parameters over 12 weeks. All results and data were considered nonparametric; the Mann-Whitney U test and ANOVA were used for primary symptoms / discomfort and testing.

[0212] Result

[0213] Sixty individuals who had recovered from COVID-19 with self-diagnostic symptoms were included in the study. Two groups were formed. One group of 30 patients followed standard management (SM, control group), while the other 30 similar individuals received Pycnogenol® in addition to SM. The two groups were similar at inclusion. No one withdrew from the study.

[0214] No side effects of the supplement were observed; best tolerability.

[0215] At the end of the study, all individuals tested positive for antibodies against SARS-CoV-2.

[0216] The results of the evaluation parameters of the study are shown in Table 2.

[0217] Table 2: Overview including all assessment parameters*=p<0.05 relative to control. PY=Pycnogenol®. CON=Control group. ----------------------------------------------------------------------------------------------------------------------------------------- Quantity PY 30(14F) 30 30 CON 30(13F) 30 30 Visit 1 Baseline Visit 2 Week 2 Visit 3 Month 1 Visit 4 3rd month 1. Endothelial function & microcirculation A. FMD [%] PY 6.5±1.2 6.6±1.0 12.6±0.9* 18.8±2.8* CON 7.2±1.0 7.3±2,1 8.0±0.9 8.8±1.4 B. Reactive congestion PY 11.2±2.0 16.0±1.0* 18.0±0.8* 24.2±2.3* Increase in skin flow to the fingers after occlusion [%] CON 10.4±2.0 11.0±0.9 13.0±1.0 15.0±1.2 C. RAS ankle swelling rate. PY 2.22±0.01 1.23±0.08* 1.22±0.02* [per 100cm] 3 [The amount of tissue in ml / min] CON 2.26±0.02 2.02±0.04 2.03±0.01 D-renal systolic cortical flow velocity [CM / SEC] PY 20.2±2.0 21.0±1.5 23.0±1.4* 23.2±2.2* CON 19.8±1.6 19.7±0.7 19.6±0.8 20.2±1.0 diastolic component [%] PY 6.1±1 8.0±0.9 11.0±0.4* 14.0±0.9* CON 6.4±0.9 7.0±0.9 8.0±0.8 9.2±0.7 2. Inflammatory markers & oxidative stress A. Plasma HS-CRP PY 3.3±0.5 3.0±0.6 1.2±0.6* 1.2±0.3* [MG / L] CON 3.2±0.4 3.2±0.4 2.7±0.5 2.4±0.2 B. IL-6 [PG / ML] PY 3.0±0.7 1.6±0.5* 1.2±0.3* CON 2.8±0.4 2.3±0.3 2.2±0.8 c. Patients with normal IL-6 levels ≤1.8 pg / mL PY 2 / 30 25 / 30* 26 / 30* CON 3 / 30 11 / 30 16 / 30 D. ESR [MM / HR] PY 26.6±2.2 18.0±1.1 13.0±1.0* 11.0±3.0* CON 27.3±3.0 26.0±0.9 23.0±1.1 19.4±2.2 E. Oxidative stress [CARR unit] PY 411±16 365±11* 358±9* 362±8* CON 418±13 399±22 384±19 387±22 3. SBP [mmHg] PY 138±3.1 135±3.0 133±3.0 131±2.2* CON 139±2.5 138±2.5 137±2.2 137±2.0

[0218] The parameters in both groups gradually improved, and all measured parameters were in the case of SM and SM combined with supplements.

[0219] At the time of inclusion, vascular screening in all included individuals did not reveal significant vascular problems (plaques, intimal-medial thickening, aneurysms). It is important to note that vascular atherosclerotic lesions can alter endothelial function.

[0220] At inclusion, all individuals had low levels of endothelial function and flow-mediated dilation (FMD) in the microcirculation. These significantly improved in both groups. After 1 month, the FMD in the Pycnogenol® group was significantly higher (12.6±0.9%) compared to the control group (8.0±0.9%) (p<0.05 vs. control group), and after 3 months, the FMD in the Pycnogenol® group was even higher (18.8±2.8%) compared to the control group (8.8±1.4%) (p<0.05 vs. control group).

[0221] This improvement was also observed in reactive hyperemia measured by increased skin flow (laser Doppler measurement) after the release of the occluded hypersystolic pressure cuff. The difference between the control group and the control group was statistically significant at 2 weeks (16.0±1.0 vs. 11.0±0.9) and continued to increase at 1 month (18.0±0.8% vs. 13.0±1.0%) and 3 months (24.2±2.3% vs. 15.0±1.2%). This not only confirms the improvement in endothelial function but also the improvement in microcirculation.

[0222] The mean ankle swelling (RAS) rate, measured per 100 cm³ of tissue in mL / min, was significantly lower in the supplementation group compared to the control group (p < 0.05). The differences from the control group were statistically significant after 1 month (1.23 ± 0.08 vs. 2.02 ± 0.04) and after 3 months (1.22 ± 0.02 vs. 2.03 ± 0.01). This demonstrates a significant improvement in capillary filtration rate, an important parameter of microcirculation.

[0223] At enrollment, renal cortical flow velocity was low in all patients, indicating significantly reduced perfusion (lower peak systolic flow and lower diastolic flow component). The supplementation group showed a significant increase compared to the control group, with a rise from 19.8±1.6 cm / sec in the control group to 20.2±1.0 cm / sec, and a significant increase in systolic velocity from 20.2±2.0 cm / sec to 23.2±2.2 cm / sec in the supplementation group. The diastolic component in the Pycnogenol® group was more than double that in the control group (from 6.1±1 to 14±0.9%), which was an increase from 6.4±0.9% to 9.2±0.7%. The diastolic component is expressed as a percentage of diastolic flow velocity to systolic flow velocity.

[0224] Regarding inflammatory markers, plasma levels of hs-CRP and IL-6 were high at baseline and gradually decreased over 3 months. After 3 months, compared with the control group, the levels of hs-CRP and IL-6 in the Pycnogenol® group were significantly lower (p < 0.05). For hs-CRP, the difference was significant after 1 month compared with the control group, which decreased from 3.2 ± 0.4 to 2.4 ± 0.2 mg / L, decreasing from 3.3 ± 0.5 mg / L to 1.2 ± 0.5 mg / L, and remained at 1.2 ± 0.3 mg / L after 3 months. Plasma IL-6 levels in the Pycnogenol® group also decreased significantly after 1 month from 3.0 ± 0.7 to 1.6 ± 0.5, and further decreased to 1.2 ± 0.3 pg / mL after 3 months. The differences with the control group were statistically significant at both 1 month and 3 months. In the control group, plasma IL-6 levels decreased from 2.8±0.4 to 2.3±0.3 after 1 month and to 2.2±0.8 pg / mL after 3 months.

[0225] Compared with the control group, the proportion of patients with IL-6 levels within the normal range (i.e., ≤1.8 pg / mL) was also higher in the supplementation groups after 1 month and 3 months (p<0.05) (25 / 30 and 26 / 30 vs 11 / 30 and 16 / 30).

[0226] After 3 months, ESR followed the same pattern, with a more gradual and significant (p<0.02) reduction in supplemental individuals (from 26.6±2.2 to 11.0±3.0 mm / hr) compared to control patients (from 27.3±3.0 to 19.4±2.2 mm / hr).

[0227] Plasma oxidative stress was assessed by measuring plasma free radicals (PFR). Compared with the control group (from 418±13 to 387±22 cal units), the PFR content expressed in cal units was significantly lower in the supplementation group (from 411±16 to 362±8 cal units) (p<0.05), showing a lower and slower rate of improvement over time.

[0228] At the end of the study, all other blood parameters (including platelets and coagulation factors) were within the normal range.

[0229] Physiological tests: Blood pressure and heart rate were monitored. Blood pressure and heart rate were normalized in all individuals in the supplementation group; at the end of the study, systolic blood pressure (SBP) was significantly lower in the supplementation group (p < 0.05) (from 138 ± 3.1 to 131 ± 2.2 in the supplementation group, compared to 139 ± 2.5 to 137 ± 2.0 in the control group).

[0230] Discussion

[0231] COVID-19 has numerous immediate and long-term sequelae, including neurological symptoms such as loss of smell and taste, headache, anxiety and depression, muscle symptoms such as weakness and fatigue, vasculitis, kidney dysfunction, coagulopathy and even pulmonary fibrosis, which are ongoing research questions.

[0232] The clinical condition of patients after COVID-19 includes common symptoms during the recovery period.

[0233] A more severe condition is termed "prolonged-COVID," characterized by severe clinical symptoms and signs, abnormalities in blood tests, and altered Karnofsky Performance Scale indices, all lasting for several months. In such cases, clinical presentation impairs normal lifestyle and standard activity levels. The incidence of long-term recovery from COVID-19 for at least 3 months with one or more persistent COVID-19 symptoms varied considerably between 32% and 96%.

[0234] Management of clinical situations has not yet been clearly established, and there are no real guidelines. Alleviating recovery symptoms can be a significant problem that needs to be addressed in a short period of time. Specific assessment methods are still needed, and these methods are under development.

[0235] Most physicians attempt to develop treatment plans that are primarily related to the individual and symptom control.

[0236] Over a long period of time, high levels of inflammation may be found in recovered patients.

[0237] Pycnogenol® is a 'soft', safe, natural anti-inflammatory and antioxidant agent studied in several preventative and clinical conditions. This natural remedy, with its high level of safety and highly standardized composition, is used to control inflammation. Natural derivatives can provide a safe solution, where possible, to avoid the continued use of drugs with adverse effects, such as NSAIDs or corticosteroids.

[0238] Pycnogenol® contributes to these beneficial effects through its proven anti-inflammatory activity against the endothelium. Pycnogenol® is primarily composed of proanthocyanidins and small molecules such as catechins, ferulic acid, caffeic acid, and taxol. The proanthocyanidins of Pycnogenol® are metabolized by intestinal bacteria into smaller molecules, including the metabolite M1 (δ-(3,4-dihydroxyphenyl)-γ-pentanolide). These compounds can be detected in the plasma of volunteers supplemented with Pycnogenol®. Metabolite M1 was found to selectively infiltrate blood cells and endothelial cells, where it is highly enriched by promoting uptake, demonstrating intracellular anti-inflammatory effects. Since COVID-19 is described as an "endothelial disease," Pycnogenol® has shown support for recovery from SARS-CoV-2 infection through its anti-inflammatory properties applied directly to the endothelium.

[0239] In addition, some flavonoids found in plasma after ingestion of Pycnogenol® have been shown to be potential inhibitors of angiotensin-converting enzyme 2 (ACE2), a receptor protein essential for SARS-CoV-2 infection.

[0240] This study shows that patients who have recovered from COVID-19 and are supplemented with Pycnogenol® have improved endothelial and microcirculatory function and lower levels of inflammation in the blood compared to the control group.

[0241] Based on these various beneficial effects on health, Pycnogenol® is proposed as a valuable tool for physicians in cases where there are currently no obvious or important solutions.

[0242] In summary, Pycnogenol® provides an important solution for treating some of the symptoms and signs associated with post-COVID-19 syndrome.

[0243] This assessment provides some interesting basic principles for the use of Pycnogenol® in this condition, which will be of great significance in the coming years. [Simplified Explanation of the Diagram]

[0245] None

Claims

1. An oral composition comprising a proanthocyanidin derived from a plant extract and at least one suitable excipient, said plant extract being selected from the group consisting of: pine bark, grape seed, apple, cocoa bean, peanut skin, cranberry extract or combinations thereof, wherein said oral composition is a source of an ingredient selected from the group consisting of: δ-(3,4-dihydroxyphenyl)-g-valerolactone, δ-(3-methoxy-4-hydroxy-phenyl)-γ-valerolactone, catechin, epicatechin, ferulic acid, gallic acid, 4-hydroxybenzoic acid, caffeic acid, protocatechuic acid, zeaxanthin and mixtures thereof, said ingredient being responsible for mediating an anti-inflammatory effect on the endothelium, said oral composition being used to treat endothelial inflammation and / or systemic endothelial dysfunction triggered by COVID-19 induced by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection.

2. An oral composition comprising proanthocyanidins derived from a plant extract and at least one suitable excipient, said plant extract being selected from the group consisting of: pine bark, grape seed, apple, cocoa bean, peanut skin, cranberry extracts or combinations thereof, wherein said oral composition is a source of an ingredient selected from the group consisting of: δ-(3,4-dihydroxyphenyl)-g-valerolactone, δ-(3-methoxy-4-hydroxy-phenyl)-γ-valerolactone, catechin, epicatechin, ferulic acid, gallnut The oral composition comprises 4-hydroxybenzoic acid, caffeic acid, protocatechuic acid, zeaxanthin, and mixtures thereof, wherein the components are responsible for mediating anti-inflammatory effects on the endothelium, and the oral composition is used to treat endothelial inflammation and / or systemic endothelial dysfunction triggered by COVID-19 induced by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, characterized in that the oral composition is administered to symptomatic COVID-19 post-recovery individuals.

3. The oral composition for use as claimed in claim 1 or 2, wherein the oral composition comprises 20% to 95% w / w proanthocyanidins and is administered at a dose of 25 mg / day to 300 mg / day.

4. The oral composition for use as claimed in any one of claims 1 to 3, wherein the oral composition comprises 65% w / w to 75% w / w proanthocyanidins.

5. The oral composition for use as claimed in any one of claims 1 to 4, wherein the systemic endothelial dysfunction is selected from the group comprising problems of endothelial blood flow and microcirculation or problems of endothelial coagulation.

6. The oral composition for use as claimed in claim 5, wherein the endothelial blood flow and microcirculation problems are selected from the group consisting of: renal function problems, pulmonary function problems, liver function problems, cognitive function problems, blood pressure problems related to endothelial dysfunction, and blood velocity problems due to endothelial dysfunction.

7. The oral composition for use as claimed in claim 5, wherein the endothelial coagulation problem is selected from the group including thrombosis and platelet aggregation.

8. The oral composition for use as claimed in any one of claims 1 to 7, wherein the endothelial inflammation is selected from the group comprising endotheliitis, myocarditis or vasculitis.

9. The oral composition for use as claimed in claim 8, wherein the vasculitis is Kawasaki-like disease.

10. The oral composition for use as claimed in claim 8, wherein the endotheliitis is severe endotheliitis.

11. The oral composition for use as claimed in any one of claims 1 to 10, wherein the pine bark extract is Pycnogenol®.

12. An oral composition for use as claimed in any one of claims 1 to 11, wherein the suitable excipient is a pharmaceutically acceptable excipient.

13. The oral composition for use as claimed in any one of claims 1 to 12, wherein the oral composition is in the form of a food preparation, dietary supplement, pharmaceutical nutritional product, or beverage.

14. A pharmaceutical preparation comprising an oral composition for use as described in any one of claims 1 to 12.

15. The pharmaceutical preparation as claimed in claim 14 or the dietary supplement as claimed in claim 13, characterized in that the pharmaceutical preparation or dietary supplement is administered at a dose of 25 mg / day to 300 mg / day.

16. A method for treating or preventing endothelial inflammation and / or systemic endothelial dysfunction triggered by SARS-CoV-2 infection, the method comprising orally administering to an individual in need an effective amount of the oral composition as described in any one of claims 1 to 13 or the agent as described in claim 14, wherein the individual in need is a symptomatic post-COVID-19 individual who has recovered from COVID-19.

17. The method of claim 16, wherein the oral composition for use as described in any one of claims 1 to 13 or the agent as described in claim 14 is administered at a dose of 25 mg / day to 300 mg / day.

18. An oral composition comprising a source of an ingredient selected from the group consisting of: δ-(3,4-dihydroxyphenyl)-g-valerolactone, δ-(3-methoxy-4-hydroxy-phenyl)-γ-valerolactone, catechin, epicatechin, ferulic acid, gallic acid, 4-hydroxybenzoic acid, caffeic acid, protocatechuic acid, zeaxanthin, and mixtures thereof, said oral composition being intended for the treatment of endothelial inflammation and / or systemic endothelial dysfunction triggered by COVID-19 induced by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection.

19. An oral composition comprising a source of an ingredient selected from the group consisting of: δ-(3,4-dihydroxyphenyl)-g-valerolactone, δ-(3-methoxy-4-hydroxy-phenyl)-γ-valerolactone, catechin, epicatechin, ferulic acid, gallic acid, 4-hydroxybenzoic acid, caffeic acid, protocatechuic acid, zeaxanthin, and mixtures thereof, said oral composition being intended for the treatment of endothelial inflammation and / or systemic endothelial dysfunction triggered by SARS-CoV-2 infection, characterized in that said oral composition is administered to a symptomatic COVID-19 individual who has recovered from COVID-19.

20. An oral composition for use as claimed in claim 18 or 19, wherein the source of said ingredient is derived from a formulation containing 20% ​​to 95% w / w proanthocyanidins.

21. An oral composition for use as claimed in claim 18 or 19, wherein the source of said ingredient is derived from a formulation containing 65% to 75% w / w proanthocyanidins.