Bioactive phytochemicals

Abstract

The present invention provides a process for producing a composition containing (2R,3R,4R,5S)-3,4,5-trihydroxypiperidine-2-carboxylic acid (idoBR1). [Solution] The process includes (a) providing plant material from a plant source including plants of the Cucurbitaceae family; (b) fractionating the plant material to produce an extract concentrated with idoBR1; (c) assaying the extract for (i) inhibitory activity against sialidase or TNF-alpha; or (ii) IL-10 stimulating activity; and (d) formulating the assayed extract with cosmetic, nutritional, or pharmaceutically acceptable excipients or carriers to produce cosmetic, nutritional, or pharmaceutical compositions.

Description

Technical Field

[0001] The present invention relates to a process for the production of compositions comprising (2R,3R,4R,5S)-3,4,5-trihydroxypiperidine-2-carboxylic acid (idoBR1), and various products, compounds, compositions, medical uses and methods based thereon, and their use in the preparation of various compositions for medical use, including the treatment of inflammation, infections, skin disorders, and in vivo inhibition of xyalidase activity, as well as processes for isolating and purifying said compositions from various plant sources.

[0002] The present invention also relates to a method for monitoring the quality of Cucurbitaceae extracts (e.g., Cucumis extracts), a process for producing Cucurbitaceae extracts, and Cucurbitaceae extracts (and particularly Cucumis extracts) obtainable by such processes.

Background Art

[0003] Cucumis sativus is a widely cultivated plant of the family Cucurbitaceae, including pumpkins. Cucumis sativus originated in India and has been cultivated in West Asia for at least 3000 years and is thought to have been introduced to other regions of Europe by the Romans. Records of cucumber cultivation appear in France in the 9th century, in England in the 14th century, and in North America in the mid-16th century.

[0004] Cucumbers and cucumber extracts have long been recognized as having anti-inflammatory properties and have been used topically for various types of skin problems, including swelling under the eyes and sunburn. Cucumbers were very popular in the ancient civilizations of Egypt, Greece, and Rome, and were used not only as food but also for their skin-healing properties.

[0005] Iminosaccharide acids (ISAs) constitute a more widely distributed subclass of phytochemicals known as iminosaccharides. Many known ISAs are phytochemicals and are present in plant tissues as secondary metabolites (they may play a role in defense). While iminosaccharides are widely distributed in plants (Watson, A. et al., 2001, Phytochemistry 56, 265), iminosaccharide acids are far less distributed and more difficult to isolate and identify (Martinez, R., et al., 2019, Amino acids 51, 991).

[0006] The iminosaccharide acid idoBR1 is present in older cucumber varieties but not in the given modern commercial varieties. It has been shown to be a major component of the given cucumber fruit and is the only iminosaccharide acid in cucumbers. It is a minor component in the given pumpkins and gourds.

[0007] WO2013 / 054070 identifies idoBR1 as a key bioactive component in anti-inflammatory herbal medicines based on Cucumis extracts.

[0008] Herbal food additives and therapeutic agents There is currently great interest in the use of herbal remedies and supplements, and there is increasing approval from food manufacturers, healthcare companies, and medical professionals that herbal products are valuable and can complement established formulations and treatments. Herbal food additives and supplements are now widely used.

[0009] However, quality control of herbal food additives is difficult due to the complex properties and inherent heterogeneity of plant materials. The materials used in herbal and plant-based food additives are typically whole plants, parts of plants, or extracts. Since plant materials contain many different chemical components, the materials are complex mixtures. This makes standardizing and controlling the quality of the materials extremely difficult. Furthermore, many herbal food additives are mixtures of two or more plant-based components, and therefore mixtures of mixtures, adding another level of complexity. Moreover, the manufacturing recipes and methods used are often not uniform and may remain undisclosed. These factors make it extremely difficult to ensure that two samples of a given product, obtained from different sources and seemingly identical, actually contain the same mixture of components. This problem, which makes quality control of such materials difficult, even limits the use of certain herbal extracts among herbal experts.

[0010] Furthermore, in the practical application of herbal medicines or as food supplements / nutritional aids, plants are not readily available locally, and therefore require sourcing from distant sources. However, the supply of such plants from remote locations can be unstable and inaccurate, particularly because detailed monographs, including identity and quality standards, do not exist for many of these plants. The complex mixtures of components found in medicinal plants vary considerably in type and concentration depending on many factors, including the plant source, where the plant grows, what other plants or microorganisms grow nearby, the time of year the plant is harvested, the conditions under which the material is stored and processed, and the extraction procedures used.

[0011] Therefore, there is a need for a sophisticated process that can profile herbal products containing idoBR1, thereby establishing standard specifications for plant-derived products that may be related to activity, enabling quality control in the production of herbal medicines, food additives, cosmetics, and nutritional supplements, and allowing for the ideal (structural and / or functional) quantification of bioactive elements.

[0012] The inventors have now discovered that idoBR1 exhibits inhibitory activity against sialidase and TNF-alpha, while also exhibiting IL-10 stimulating activity. This finding means that idoBR1, a related bioactive element, can now be rapidly and easily functionally assayed after fractionation of plant materials, enabling the development of improved processes for formulating cosmetic, nutritional, or pharmaceutical compositions based on extracts from plant materials from plant sources, including plants of the Cucurbitaceae family.

[0013] Therefore, the provision of cosmetic, nutritional, or pharmaceutical compositions conforming to standard specifications is greatly facilitated: the present invention enables rapid assay of the functional quality of extracts. Furthermore, time-consuming and expensive physical characterization (e.g., by GC-MS and / or HPLC) becomes optional. In addition, since the functionality of the extract is assayed, the potential effects of interference / inhibition by co-extracted parts can be monitored. This may be particularly important in cosmetic applications where the extract is formulated for topical application. [Overview of the Initiative]

[0014] Therefore, according to the present invention, a process for producing a composition comprising (2R,3R,4R,5S)-3,4,5-trihydroxypiperidine-2-carboxylic acid (idoBR1), wherein the process is (a) A process of providing plant material from a plant source including plants of the Cucurbitaceae family, (b) A step of fractionating the plant material to produce an extract in which idoBR1 is concentrated, (c) Assaying the extract for (i) inhibitory activity against sialidase or TNF-alpha; or (ii) IL-10 stimulating activity, (d) The process of formulating the assayed extract using cosmetic, nutritional, or pharmaceutically acceptable excipients or carriers to produce cosmetic, nutritional, or pharmaceutical compositions. A method is provided that includes this.

[0015] The plant source may include plants of the genus Cucumis or Cucurbita. The preferred species of the genus Cucumis is Cucumis sativus (cucumber). The preferred species of the genus Cucurbita is Cucurbita melos or Cucurbita moschata. [Modes for carrying out the invention]

[0016] All publications, patents, patent applications, and other references mentioned herein are incorporated herein by reference in their entirety for all purposes as if each individual publication, patent, or patent application were included by reference in its entirety, as specifically and individually indicated to be incorporated by reference and as if its contents were fully described.

[0017] definition As used herein, and unless otherwise specifically indicated, the following terms are intended to have the following meanings in addition to any broader (or narrower) meanings that the terms may enjoy in the art:

[0018] Unless otherwise required by context, the use of the singular form in this specification shall be interpreted as including the plural form, and vice versa. The term "a" or "an" used in relation to an entity shall be interpreted as referring to one or more of that entity. Thus, the terms "a" (or "an"), "one or more," and "at least one" are used interchangeably in this specification.

[0019] As used herein, the term “comprise,” or its variations such as “comprises,” or “comprising,” should be interpreted as indicating the inclusion of any described complete entity (e.g., feature, element, characteristic, property, method / process step, or limitation) or group of complete entities (e.g., feature, element, characteristic, property, method / process step, or limitation), but not the exclusion of any other complete entity or group of complete entities. Thus, as used herein, the term “comprises” is inclusive or open-ended and does not exclude additional undescribed complete entities or methods / process steps.

[0020] The phrase "essentially derived from" is used to require a specific whole or process that does not substantially affect the characteristics or functions of the claimed invention.

[0021] As used herein, the term "consists of" is used to indicate the existence of a complete set described (e.g., a feature, element, characteristic, property, method / process step or limitation) or a single set of complete sets (e.g., a feature, element, characteristic, property, method / process step or limitation).

[0022] As used herein, the term “disease” is used to define any abnormal condition that impairs physiological function and is associated with specific symptoms. The term is used more broadly to encompass any disorder, illness, abnormality, pathology, disease, condition, or syndrome in which physiological function is impaired, regardless of the nature of its etiology (or whether the etiological basis of the disease has been established). Thus, it includes conditions resulting from infection, trauma, injury, surgery, radiation ablation, poisoning, or malnutrition.

[0023] As used herein, the terms “treatment” or “to treat” refer to an intervention (e.g., administration of a drug to a subject) that cures, improves or reduces the symptoms of a disease or eliminates (or reduces the effects of) its cause(s) (e.g., pathological diversification). In this context, the term is used synonymously with the term “therapy.”

[0024] Also, the term "treatment" or "treating" refers to an intervention (e.g., administration of an agent to a subject) that prevents or delays the onset or progression of a disease or reduces (or eradicates) its incidence in a treated population. In this case, the term "treatment" is used synonymously with the term "prevention".

[0025] The term "subject" (which is to be construed as including "individual", "animal", "patient" or "mammal" where the context permits) defines any subject to which treatment is suggested, particularly a mammalian subject. Mammalian subjects include, but are not limited to, humans, domestic animals, farm animals, zoo animals, sports animals and pet animals. In a preferred embodiment, the subject is a human.

[0026] References herein to the treatment of diabetes are to be construed as including the treatment of type 1 and type 2 diabetes per se, as well as prediabetes (impaired glucose tolerance) and insulin resistance. The term "prediabetes" or "impaired glucose tolerance" defines a state in which there is an elevation in the level of glucose or glycosylated hemoglobin in the absence of diabetes.

[0027] As used herein, an effective amount of a compound or composition is an amount that can be administered to a subject without undue toxicity, irritation, allergic response, or other problems or complications and that is commensurate with a reasonable benefit / risk ratio, but is sufficient to provide the treatment or prevention as indicated by a desired effect, e.g., a permanent or temporary amelioration of the subject's condition. The amount will vary for each subject depending on the subject's age and general condition, mode of administration, and other factors. Thus, while the exact effective amount cannot be specified, one of ordinary skill in the art can determine the appropriate "effective" amount in any individual case using routine experimentation and the general knowledge of the background. Treatment results in this context include eradication or alleviation of symptoms, reduction of pain or discomfort, prolongation of survival, improvement of mobility and other markers of clinical improvement. Treatment results need not be a complete cure.

[0028] The term phytochemicals is used herein in a broad sense to encompass any chemical components of plants, including macromolecules and micromolecules. Important examples include alkaloids (e.g., iminosaccharides and iminosaccharide acids, selected from structural classes such as pyrrolidine, piperidine, pyrrolizidine, indolizidine, tropane, and nortopane), carbohydrate analogs, phenolic compounds, terpenoids, enzyme inhibitors, glycosides, nucleotides, amino acids, lipids, and sugars.

[0029] The term "isolated," when applied to a compound of the present invention, is used herein to indicate that the compound exists in a physical environment different from that in nature. For example, an isolated compound may be substantially isolated with respect to the complex cellular environment in which it naturally exists (e.g., it may be concentrated or purified). Thus, an isolated compound may take the form of a concentrated fraction or extract of any of the plant sources described herein.

[0030] When isolated materials are concentrated or purified, the absolute level of concentration or purity is not critical, and those skilled in the art can easily determine an appropriate level depending on the intended use of the material. A purity level of at least 0.1% w / w, 0.2% w / w, 0.3% w / w, 0.4% w / w, 0.5% w / w, 0.6% w / w, 0.7% w / w, 0.8% w / w, 0.9% w / w, 1.0% w / w, 1.1% w / w, 1.2% w / w, 1.3% w / w, 1.4% w / w, 1.5% w / w, 1.6% w / w, 1.7% w / w, 1.8% w / w, 1.9% w / w, or 2.0% w / w is preferred.

[0031] A purity level of at least 0.5–2.0% w / w, for example, at least 0.8–1.5% w / w, or for example, at least about 1.0% w / w, is particularly preferred. If the material is isolated from a natural source, a level of 5–10% w / w can be easily obtained by using suitable concentration techniques such as ion exchange chromatography, if necessary.

[0032] In some situations, the isolated compound may form part of a composition (e.g., a more or less crude extract containing many other substances) or buffer system that may contain other components. In other situations, the isolated compound may be purified to an essentially homogeneous state, as determined by spectrophotometric methods, NMR, or chromatography (e.g., GC-MS of trimethylsilyl derivatives).

[0033] The term herbal medicine is used herein to define a pharmaceutical composition in which at least one active element (e.g., a compound) is not chemically synthesized but is a phytochemical component of a plant. In most cases, this non-synthetic active element is not isolated (as defined herein) but exists together with other phytochemicals associated with the plant source. However, in some cases, the plant-derived bioactive element(s) may be present in concentrated fractions or may be isolated (sometimes including a high degree of purification). However, often, herbal medicines contain more or less crude extracts, infusions or fractions of a plant or even the whole (or part thereof) of an untreated plant, in which case the plant (or part thereof) is usually at least dried and / or ground. Herbal medicines may be in the form of food supplements, food additives, nutritional supplements, or beverages, and may be supplied in unit doses as herbal pharmaceutical kits or packs.

[0034] The term herbal food is used herein to define a composition in which at least one component is not chemically synthesized but is a phytochemical component of a plant. In most cases, this non-synthetic component is unpurified but exists together with other phytochemicals associated with the plant source. However, in some cases, the plant-derived component(s) may be present in concentrated fractions or isolated (sometimes to a high degree of purity). However, often herbal food additives include more or less crude extracts, infusions or fractions of a plant or even the whole (or part thereof) of an untreated plant, in which case the plant (or part thereof) is usually at least dried and / or ground. Thus, the term includes herbal foods in the form of additives and supplements for use with foods and beverages.

[0035] The term "bioactive element" is used herein to define a phytochemical component that is necessary or sufficient for the pharmaceutically effectiveness of the herbal medicine in which it is contained. In the present invention, the bioactive element includes idoBR1.

[0036] The term "nutritional supplement" is used herein to define a food product (or its isolate) that provides physiological benefit or protection against a disease. Preferred nutritional supplements of the present invention are anti-inflammatory.

[0037] The term "standard specification" is used herein to define the properties or phytochemical profile that correlate with the acceptable quality of a herbal medicine, cosmetic, or nutritional supplement. In this context, the term "quality" is used to define the overall suitability of the product for its intended use and includes the activity of idoBR1 at appropriate concentrations.

[0038] The term phytochemical profile is used herein to define a set of characteristics relating to different phytochemical components.

[0039] Functional assay The extract of the present invention is assayed for (i) inhibitory activity against sialidase or TNF-alpha; or (ii) IL-10 stimulating activity. Functional assays may include biological assays. Biological assays may be performed in vivo or in vitro and may include enzyme inhibition assays (e.g., sialidase inhibition). Other biological assays include receptor binding assays, cell assays (including cell replication, cell-pathogen and cell-cell interactions, and cell secretion assays), immunoassays, antimicrobial activity assays (e.g., bacterial and viral cell binding and / or replication), and toxicity assays (e.g., LD 50 (Includes assays.)

[0040] Functional characterization can also be carried out indirectly through forms of characterization that allow for the identification of one or more indices of biological activity.

[0041] Exemplary technologies are described in more detail below.

[0042] sialidase Inhibition of sialidase (neuraminidase) activity by idoBR1 or extracts containing it can be determined by an enzyme assay in which neuraminidase activity is measured, for example, using an enzyme from Clostridium perfringens (Sigma-Aldrich). The assay is based on an enzyme that cleaves the 2'-(4-methylumbelliferyl)-α-DN-acetylneuraminic acid (MUNANA) substrate to release the fluorescent product 4-methylumbelliferone (4-MU). Therefore, the inhibitory effect is (IC 50 The value is determined based on the concentration of idoBR1 or extract required to reduce enzyme activity by 50% (to obtain the value).

[0043] The preferred method may be as follows: 1. Prepare the reaction mixture. Sodium phosphate buffer (pH 4.5), 10 mmol of MUNANA4MU-NeuAc (100 μl in buffer), 0.1 mg of enzyme per 100 μl, 100 μl of plant extract or compound. Incubate at 2.37C for 10-30 minutes. 3.1. The reaction is stopped by adding 25 ml of 0.25 M glycine-NaOH (pH 10.4). 4. The released 4-methylumbelliferone (4-MU) (emission at 448 nm, excitation at 365 nm) is measured by fluorescence analysis.

[0044] TNF-α and IL-10 The reduced TNF-α and increased IL-10 levels due to the extract can be measured using ELISA in either cell cultures (e.g., THP-1 monocytes) or whole blood samples.

[0045] THP-1 cells are commercially available. Cultured cells can be placed in RPMI complete medium in a microtiter plate (e.g., 5 cells per well in a 96-well plate), incubated for 24 hours, and then PMA (10 ng / ml) can be added to the 96-well plate to differentiate THP-1 cells and determine their effect on TNF-α and IL-10 production. Cells should be pre-treated with cucumber extract, for example, 200 μg / ml to 25 μg / ml, followed by 2 hours of LPS (100 ng / ml) stimulation. After incubation, the cell supernatant is aspirated from each well into a sterile microcentrifuge tube and centrifuged at 1000 rpm for 2-3 minutes to settle any cells present. The cell supernatant is then used for assessment of the presence of TNF-α or IL-10 using ELISA. Sandwich ELISA plates coated with suitable antibodies are widely available (e.g., R&D Systems, USA).

[0046] For whole blood analysis, an aliquot (800 μl) of whole blood can be incubated with an extract dissolved in RPMI1640 for a 48-hour pre-incubation period, after which LPS (10 μg / ml) is added and incubation is continued for a further 20 hours at 37°C in a humidified (100%) atmosphere of 5% CO2 in air. At the end of the incubation period, the supernatant consisting of plasma is collected by centrifugation at 10,000 g for 30 seconds at room temperature, and TNF-α and IL-10 levels are measured using a human TNF-α and IL-10 ELISA assay (e.g., a kit available from BioSource Europe SA, Belgium).

[0047] physical characterization The extracts of the present invention may also be physically characterized (though this is not mandatory). This can take the form of quantitative determination of phytochemical components(s) present in any given fraction or at other stages in the process, measurement of component purity, determination of molecular weight (or molecular weight distribution or various statistical functions thereof in the case of fractions containing multiple different phytochemical components), determination of molecular formula(e) (e.g., by nuclear magnetic resonance), and various spectral analyses.

[0048] Particularly useful spectral characteristics include: • Mass spectrum (e.g., mass vs. charge (m / z) value vs. abundance), and / or • Chromatography data (e.g., spectrum, column retention time, elution profile, etc.), and / or • Photodiode array (PDA) spectra (e.g., in both the UV and visible ranges), and / or • Electrochemical detection (ED) or evaporative light scattering (ELSD) detection; and / or • Nuclear magnetic resonance (NMR) spectrum ( 1 H and / or 13 (Includes spectral data obtained via CNMR) It includes.

[0049] Spectral characterization can be combined with fractionation steps. For example, GC-MS and HPLC-PDA-MS-ED-ELSD (as described herein) can be used to combine fractionation with the acquisition of mass spectra, UV-Vis spectra, electrochemical reaction or mass fraction data, and chromatographic spectral data.

[0050] Any or all of the above characteristics can be used to define a “chemical fingerprint” of any given sample (or any fraction or phytochemical component thereof).

[0051] chemical characterization The extracts of the present invention may also be chemically characterized (though this is not essential). This may take the form of measuring, among other things, the chemical reactivity of the phytochemical components(s), their solubility, stability, and melting points.

[0052] Medical Use of the Compounds of the Present Invention neoplasm The compounds of the present invention are sialidase inhibitors and are therefore applicable to the treatment or prevention of diseases and disorders mediated by sialidase activity and / or sialic acid.

[0053] Sialidases are attractive therapeutic targets because they are involved in a variety of pathological processes, including bacterial and viral infections and neoplasms. The expression of sialidases Neu1 and Neu3 appears to be altered in diabetes (e.g., Neu1 activity discussed by Natori, Y., et al., 2013, Biol. Pharm., Bull., 36, 1027). Sialidases are also involved in atherogenesis (Sukhorukov, V.N., et al., 2017, Curr. Pharm. Des., 23, 4696) and osteoarthritis (Katoh, S., et al., 1999, J Immunol., 162, 5058).

[0054] Therefore, the compounds of the present invention are applicable to the treatment or prevention of neoplasms / proliferative disorders, as will be described in more detail below.

[0055] As used herein, the term “neoplasm” is used in its strict sense to define a disease involving the abnormal proliferation of neoplastic cells. The term includes benign, precancerous, and malignant neoplasms (as defined above) and is used synonymously with the term “proliferative disorder.”

[0056] Neoplasms arise from inappropriately high levels of cell division and / or low levels of apoptosis or senescence in neoplastic cells that have acquired genetic or epigenetic changes that release them from normal physiological control (i.e., the cells have been transformed). Neoplasms typically produce structures known as neoplasms (abnormal masses of tissue that grow at a rate exceeding that of normal tissue, do not coordinate with normal tissue, and persist in the same excessive manner even after the stimulus that caused the change has ceased). Most neoplasms form large masses of tissue (solid tumors), while some neoplasms do not form such individual tissue masses. These include cervical intraepithelial neoplasms, anal intraepithelial neoplasms, and leukemia.

[0057] Neoplasms can be benign, potentially malignant, or malignant. Benign neoplasms include uterine fibroids and melanocytic nevi (skin moles), which are non-invasive and do not progress to malignant neoplasms. Potentially malignant (precancerous) neoplasms include carcinoma in situ, which is non-invasive but may eventually develop into a malignant neoplasm.

[0058] Malignant neoplasms are new organisms (tumors) that invade and destroy surrounding tissues, form metastases, and can ultimately kill the host. The terms "malignant neoplasm" and "cancer" are used synonymously in this specification.

[0059] The terms “proliferative disorder” and “neoplasm” may be used herein to define a class of diseases involving the pathological growth of cells in vivo.

[0060] Therefore, proliferative disorders include cancer, cancer metastasis, smooth muscle cell proliferation, systemic sclerosis, cirrhosis of the liver, adult respiratory distress syndrome, idiopathic cardiomyopathy, lupus erythematosus, retinopathy (e.g., diabetic retinopathy), cardiac hypertrophy, benign prostatic hyperplasia, ovarian cysts, pulmonary fibrosis, endometriosis, fibromatosis, hamartoma, lymphangiomatosis, sarcoidosis, and desmoid tumors. Neoplasms involving smooth muscle cell proliferation include excessive cell proliferation in the vascular system (e.g., intima-smooth muscle cell hyperplasia, restenosis, and vascular occlusion (especially biologically or mechanically mediated vascular injury, e.g., stenosis after angioplasty)). Furthermore, intima-smooth muscle cell hyperplasia may include hyperplasia in smooth muscle outside the vascular system (e.g., occlusion in the bile ducts, bronchial airways, and kidneys in patients with renal interstitial fibrosis). Non-cancerous proliferative disorders also include hyperproliferation of cells in the skin, such as psoriasis and its altered clinical forms, Reiter's syndrome, pityriasis rubra pilaris, and hyperproliferative variants of keratinization disorders (including actinic keratosis, senile keratosis, and scleroderma).

[0061] The term “neoplasm” is also used herein in a broad sense to define diseases involving abnormal growth and / or differentiation of cells in vivo, and thus encompasses hyperplasia, metaplasia, and dysplasia.

[0062] Hyperplasia is defined as a condition in which normal (unaltered) cells in an organ or tissue proliferate to an abnormal degree. Therefore, it can result in overall organ enlargement, the formation of benign tumors, or be visible only under a microscope. Hyperplasia is a physiological response to certain stimuli, and hyperplastic cells retain normal regulatory mechanisms (unlike neoplastic growth, where cells proliferate in an abnormal manner unresponsive to normal physiological control). Examples include congenital adrenal hyperplasia, endometrial hyperplasia, benign prostatic hyperplasia (prostatic enlargement), mammary hyperplasia (including ductal hyperplasia), focal epithelial hyperplasia (Heck's disease), sebaceous gland hyperplasia, and hepatic hyperplasia.

[0063] Metaplasia refers to a condition in which one type of mature, differentiated cell is replaced by another type of mature, differentiated cell. Examples include squamous metaplasia of columnar epithelial cells in salivary ducts (when stones are present), squamous metaplasia of transitional epithelium in the bladder (again, when stones are present or associated with infection), glandular metaplasia of the esophagus (Barrett's esophagus) in patients with acid reflux, and ossification in connective tissue.

[0064] Dysplasia is defined as a condition characterized by the abnormal maturation of cells within a tissue. This typically consists of an proliferation of immature cells, accompanied by a corresponding decrease in the number and location of mature cells. For example, cervical epithelial dysplasia is characterized by an increase in a population of immature cells confined to the mucosal surface. Myelodysplastic syndrome, or hematopoietic dysplasia, exhibits an increase in the number of immature cells in the bone marrow and a decrease in mature, functional cells in the blood. Other examples include neurofibromatosis.

[0065] Hyperplasia, metaplasia, and dysplasia are usually reversible conditions resulting from irritants (e.g., injury or trauma). In contrast, neoplasms are usually irreversible and are associated with cell transformation.

[0066] The compounds of the present invention are generally applicable to the treatment of any neoplasm, including proliferative disorders, benign, precancerous, and malignant neoplasms, hyperplasia, metaplasia, and dysplasia.

[0067] Therefore, the present invention is applicable to the treatment of proliferative disorders including, but not limited to, cancer, cancer metastasis, smooth muscle cell proliferation, systemic sclerosis, cirrhosis of the liver, adult respiratory distress syndrome, idiopathic cardiomyopathy, lupus erythematosus, retinopathy (e.g., diabetic retinopathy), cardiac hypertrophy, benign prostatic hyperplasia, ovarian cysts, pulmonary fibrosis, endometriosis, fibromatosis, hamartoma, lymphangiomatosis, sarcoidosis, and desmoid tumors. Neoplasms including smooth muscle cell proliferation include excessive cell proliferation in the vascular system (e.g., intima-smooth muscle cell hyperplasia, restenosis, and vascular occlusion (especially biologically or mechanically mediated vascular injury, e.g., stenosis after angioplasty)). Furthermore, intima-smooth muscle cell hyperplasia may include hyperplasia in smooth muscle outside the vascular system (e.g., occlusion in the bile ducts, bronchial airways, and kidneys in patients with renal interstitial fibrosis). Non-cancerous proliferative disorders also include hyperproliferation of cells in the skin, such as psoriasis and its altered clinical forms, Reiter's syndrome, pityriasis rubra pilaris, and hyperproliferative variants of keratinization disorders (including actinic keratosis, senile keratosis, and scleroderma).

[0068] Particularly preferred is the treatment of malignant neoplasms (cancer). The present invention is applicable to the treatment of any cancer, including those selected from the following main groups: (a) cancer; (b) blastoma; (c) leukemia; (d) lymphoma; (e) myeloma; (f) sarcoma; and (g) mixed-type cancer.

[0069] Cancer refers to malignant neoplasms of epithelial origin or cancers of the endometrium inside or outside the body. Carcinomas, which are malignant tumors of epithelial tissue, account for 80-90 percent of all cancer cases. Epithelial tissue is found throughout the body. It is present in the skin, as well as in the coverings and endometrium and internal passages of organs, such as the gastrointestinal tract. In preferred embodiments, the cancers treated according to the present invention are selected from cancers of the salivary glands; colon; rectum; appendix; lungs; thymus; breast; cervix; bladder and eye.

[0070] The present invention is applicable to the treatment of all blastomas, including hepatoblastoma (e.g., nephroblastoma, non-epithelial kidney tumor, rhabdoid kidney tumor, renal sarcoma and pPNET of the kidney), medulloblastoma, pancreaticblastoma, pulmonaryblastoma, pleuropulmonaryblastoma, and neuroblastoma (including common peripheral nerve cell tumors as well as ganglioblastoma and retinoblastoma).

[0071] The present invention is applicable to the treatment of all leukemias, myeloproliferative disorders and myelodysplastic disorders, including lymphocytic leukemia (e.g., progenitor cell leukemia, mature B-cell leukemia, mature T-cell leukemia, and NK-cell leukemia); acute myeloid leukemia; chronic myeloproliferative disorders; myelodysplastic syndromes and other myeloproliferative disorders. Therefore, the present invention is applicable to the treatment of various leukemias, including lymphocytic, lymphocytic, or lymphoblastic leukemia (malignancies of lymphocytic and lymphocytic blood cell lineages), and polycythemia vera or erythremia (malignancies of various blood cell products, but with erythrocyte dominance).

[0072] Lymphoma develops in the glands or nodules of the lymphatic system, which is a network of blood vessels, nodules, and organs (particularly the spleen, tonsils, and thymus) that produce white blood cells, or lymphocytes, that purify bodily fluids and fight infection. Unlike leukemia, sometimes called "fluid cancer," lymphoma is a "solid cancer." Lymphoma can also occur in certain organs, such as the stomach, breast, or brain. These lymphomas are called extranodal lymphomas. Lymphoma is further classified into two categories: Hodgkin lymphoma and non-Hodgkin lymphoma. The presence of Reed-Sternberg cells in Hodgkin lymphoma is what diagnostically distinguishes it from non-Hodgkin lymphoma. The present invention is applicable to the treatment of all such lymphomas and reticular neoplasms, including (a) Hodgkin lymphoma; (b) non-Hodgkin lymphoma (e.g., progenitor cell lymphoma, mature B-cell lymphoma, mature T-cell lymphoma and NK-cell lymphoma); (c) Burkitt lymphoma and (d) other lymphoreticular neoplasms, including mantle cell lymphoma.

[0073] Therefore, the present invention is applicable, for example, to the treatment of tumors of glands or nodules of the lymphatic system (including the spleen, tonsils, and thymus), as well as to a wide range of lymphomas, including extranodal lymphomas of the stomach, breast, and brain.

[0074] Myeloma is a cancer that originates from plasma cells in the bone marrow. Therefore, the present invention is applicable to the treatment of hematopoietic malignancies and blood malignancies, including those of the lymphoid system (e.g., leukemia, acute lymphoblastic leukemia, chronic lymphoblastic leukemia, B-cell lymphoma (such as diffuse large B-cell lymphoma), T-cell lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, hairy cell lymphoma, and Burkitt lymphoma) as well as myeloid hematopoietic malignancies (e.g., acute myeloid leukemia, chronic myeloid leukemia, myeloid leukemia, and imatinib-sensitive and refractory chronic myeloid leukemia, myelodysplastic syndrome, bortezomib-sensitive and refractory multiple myeloma, myeloproliferative disorders or promyelocytic leukemia, and follicular thyroid carcinoma).

[0075] The present invention applies to the treatment of all sarcomas. Sarcomas refer to cancers originating from supporting and connective tissues, such as bone, tendons, cartilage, muscle, and fat. The most common sarcomas, usually occurring in young adults, often present as painful lumps on bones. Sarcomatous tumors typically resemble the tissue from which they grow. Exemplary sarcomas for treatment according to the present invention include osteosarcoma (or osteogenic sarcoma); chondrosarcoma; leiomyosarcoma (smooth muscle); rhabdomyosarcoma (skeletal muscle); mesosarcoma or mesothelioma (inner lining of body cavities); fibrosarcoma (fibrous tissue); angiosarcoma or hemangioendothelioma (vascular tissue); liposarcoma; glioma; astrocytoma; myxosarcoma (primitive embryonic connective tissue); and mesenchymal or mixed mesodermal tumors (mixed connective tissue type). Fibrosarcomas include peripheral nerve sheath tumors and other fibrous neoplasms, such as fibroblasts and myofibroblasts, nerve sheath tumors, and other fibromatous neoplasms. Kaposi's sarcoma is also included. Furthermore, soft tissue sarcomas include, for example, Ewing's tumor and Askin's tumor of soft tissue, pPNETs of soft tissue, extrarenal rhabdoid tumors; fibrohistiocytic tumors; synovial sarcomas; bone and chondrocyte neoplasms of soft tissue, as well as alveolar soft tissue sarcomas. Osteosarcoma (malignant bone tumors) include malignant fibrous neoplasms of bone; malignant chordomas and odontogenic malignancies. Gliomas include oligodendrogliomas, mixed and unspecified gliomas, and neuroepithelial glial tumors.

[0076] The present invention is applicable to the treatment of mixed-type cancers, including, for example, adenosquamous carcinoma, mixed mesodermal tumors, carcinosarcoma, and teratocarcinoma. Therefore, the present invention is applicable to the treatment of various CNS, PNS, and mixed intracranial and intramedullary neoplasms, including astrocytoma, neuroblastoma, glioma, schwannoma, ependymoma, and choroid plexus tumor (e.g., ependymoma and choroid plexus tumor); intracranial and intramedullary embryonic tumors (e.g., medulloblastoma, undifferentiated neuroectodermal tumor (PNET), medullary epithelioma, atypical teratomatous / rhabdoid tumor, and other intracranial and intramedullary neoplasms (e.g., pituitary adenoma and sella turcica cancer, tumor (craniopharyngioma), pineal parenchymal tumor, neuronal and mixed neuron-glial tumor, meningioma, and typical intracranial and intramedullary neoplasms).

[0077] Therefore, the present invention is particularly applicable to the treatment of intracranial and intramedullary germ cell tumors; intracranial and intramedullary germ cell tumors; intracranial and intramedullary teratomas; intracranial and intramedullary embryonic carcinomas; intracranial and intramedullary yolk sac tumors; intracranial and intramedullary choriocarcinomas; and mixed-type intracranial and intramedullary tumors.

[0078] The present invention is also applicable to the treatment of various germ cell tumors, trophoblast tumors, and gonadal neoplasms. Therefore, the present invention is applicable to the treatment of malignant extracranial and extragonadal germ cell tumors, including, for example, extracranial and extragonadal malignant germ cell tumors, extracranial and extragonadal malignant teratomas, extracranial and extragonadal embryonic carcinomas, extracranial and extragonadal yolk sac tumors; extracranial and extragonadal choriocarcinomas and typical extracranial and extragonadal malignant mixed germ cell tumors. The present invention is also applicable to the treatment of malignant gonadal germ cell tumors, including, for example, malignant gonadal germ cell tumors, seminomas, malignant gonadal teratomas, gonadal embryonic carcinomas, gonadal yolk sac tumors, gonadal choriocarcinomas, mixed-type malignant gonadal tumors and malignant gonadal adenoblastomas.

[0079] infectious disease The compounds of the present invention are sialidase inhibitors and are therefore applicable to the treatment or prevention of diseases and disorders mediated by sialidase activity and / or sialic acid. Such diseases and disorders include infectious diseases (including bacterial and viral infections).

[0080] The compounds of the present invention may have anti-infective (e.g., pathostatic or pathocidal) activity against any infectious agent. Therefore, the compounds of the present invention can target a wide range of different infectious agents (i.e., have activity against them). Thus, the present invention is widely applicable to the treatment or prevention of any infectious disease or infectious illness, including infectious diseases involving viruses, bacteria, fungi, protozoa, prions, or metazoan organisms.

[0081] Therefore, the present invention is broadly applicable to the treatment or prevention of viral infections; bacterial infections; protozoan infections; fungal infections; prion infections; and / or metazoan (e.g., parasitic helminth) infections or invasions. The compounds of the present invention may also be applied to the treatment or prevention of chronic, dormant, or latent viral, bacterial, protozoan, fungal, prion, or metazoan (e.g., parasitic helminth) infections or invasions. • Viral targets include, but are not limited to, the following viruses (or virus classes): Retroviridae (e.g., human immunodeficiency viruses, including HIV-1); Picornaviridae (e.g., poliovirus, hepatitis A virus; enterovirus, human coxsackievirus, rhinovirus, echovirus); Calciviridae (e.g., gastroenteritis strains); Togaviridae (e.g., equine encephalitis virus, rubella virus); Flaviridae (e.g., dengue virus, encephalitis virus, yellow fever virus); Coronaviridae (e.g., coronavirus); Rhabdoviradae (e.g., vesicular stomatitis virus, rabies virus); Filoviridae (e.g., Ebola virus); Paramyxoviridae (e.g., parainfluenza virus, mumps virus, measles virus, respiratory syncytial virus); Orthomyxoviridae (e.g., influenza virus); Bungaviridae (e.g., Hantan virus, Bunya virus, phlebovirus, and nairovirus); Arena viridae (hemorrhagic fever viruses); Reoviridae (e.g., reovirus, orbivirus and rotavirus); Birnaviridae; Hepadnaviridae (hepatitis B virus); Parvoviridae (parvovirus); Papovaviridae (papillomavirus, polyomavirus); Adenoviridae (most adenoviruses); Herpesviridae (herpes simplex virus (HSV) 1 and 2, varicella-zoster virus, cytomegalovirus (CMV), herpesviruses); Poxviridae (smallpox virus, vaccinia virus, poxvirus); and Iridoviridae (e.g., African swine fever virus); and unclassified viruses (e.g., virulence factors of spongiform encephalopathy, factors of hepatitis delta (considered a defective satellite of hepatitis B virus), HCV virus (causing non-A, non-B hepatitis); Norwalk and related viruses, and astroviruses).Of the aforementioned, those particularly preferred are HIV, hepatitis A, hepatitis B, hepatitis C, rabies virus, poliovirus, influenza virus, meningitis virus, measles virus, mumps virus, rubella, pertussis, encephalitis virus, papillomavirus, yellow fever virus, respiratory syncytial virus, parvovirus, chikungunya fever virus, hemorrhagic fever virus, and herpesvirus, especially varicella, cytomegalovirus, and Epstein-Barr virus. • Bacterial targets include, but are not limited to, Gram-negative and Gram-positive bacteria. Examples of bacteria that can be targeted by the compounds of the present invention include Helicobacter pylori, Borelia burgdorferi, Legionella pneumophilia, Mycobacterium spp. (e.g., M. tuberculosis, M. leprae, M. avium, M. intracellulare, M. kansaii, and M. gordonae), Staphylococcus aureus, Neisseria gonorrhoeae, Neisseria meningitidis, Listeria monocytogenes, Streptococcus pyogenes (Group A Streptococcus), Streptococcus agalactiae (Group B Streptococcus), Streptococcus viridans, Streptococcus faecalis, Streptococcus bovis, any anaerobic species of the genus Streptococcus, Streptococcus pneumoniae, Campylobacter spp., Enterococcus spp., Haemophilus influenzae, and Bacillus. This includes, but is not limited to, Anthracis, Corynebacterium spp. (including C. diphtheriae), Erysipelothrix rhusiopathiae, Clostridium perfringens, Clostridium tetani, Enterobacter aerogenes, Klebsiella spp. (including K. pneumoniae), Pasturella multocida, Bacteroides spp., Fusobacterium nucleatum, Streptobacillus monilijormis, Treponema pallidium, Treponema pertenue, Leptospira spp., Rickettsia spp., and Actinomyces spp. (including A. israelii).Bacteria that form biological membranes in vivo are specific targets of the compounds of the present invention, and these include Tannerella forsythia, Tannerella denticola, Porphyromonas gingivalis, and Gardnerella vaginalis. • Fungal targets include, but are not limited to, Cryptococcus neoformans, Histoplasma capsulatum, Coccidioides immitis, Blastomyces dermatitidis, Chlamydia trachomatis, and Candida albicans. Protozoan targets include, but are not limited to, Plasmodium spp. (including Plasmodium falciparum, Plasmodium malariae, Plasmodium ovale, and Plasmodium vivax), Toxoplasma spp. (including T. gondii and T. cruzii), Leishmania spp., Cryptosporidium spp. (including C. parvum), Cyclospora spp. (including C. cayetanensis), Entamoeba (including E. histolytica), and Giardia spp. (including G. lamblia). Metazoan targets include parasites or pathogens, such as parasitic worms (e.g., Schistosoma spp.).

[0082] Inhibition of bacterial growth in vivo Sialidase activity is key to the utilization of sialoconjugate sugars and is involved in host-pathogen interactions with bacteria. Glycoprotein-associated sialic acids have been proposed as essential in vivo nutrient sources for Tannerella forsythia when growing in biofilms (Roy, S., 2011, Microbiology, 157, 3195). The sialidase inhibitory properties of the compounds of the present invention are also applicable to the inhibition of commensalism and / or pathogenic bacterial growth in vivo, particularly to the disruption of host-bacterial cell interactions, including the inhibition or elimination of bacterial biofilms in mammalian (e.g., human) hosts.

[0083] Therefore, the compounds are applied to the treatment or prevention of diseases and disorders mediated or characterized by the presence of bacterial biofilms (e.g., subgingival plaque biofilms and mucosal biofilms).

[0084] Such diseases include periodontal disease, bacterial vaginosis, and other diseases caused by Tannerella forsythia, Tannerella denticola, Porphyromonas gingivalis, and Gardnerella vaginalis (the latter species being associated with bacterial vaginosis and premature birth).

[0085] Regulation of commensal bacterial growth The sialidase inhibitory properties of the compounds of the present invention are also applicable to the regulation of the composition of the host microbiome (and particularly commensal bacteria), for example, the regulation of the composition of commensal bacteria in mammalian (e.g., human) hosts. Regulation of the gut microbiome is particularly preferred.

[0086] Atheroma formation The compounds of the present invention are sialidase inhibitors and are therefore applicable to the treatment or prevention of atheroma formation, because sialidase is involved in this process (Sukhorukov, VN, et al., 2017, Curr. Pharm. Des., 23, 4696) and osteoarthritis (Katoh, S., et al., 1999, J Immunol., 162, 5058). Thus, the compounds of the present invention are applicable to the treatment and prevention of atherosclerosis.

[0087] inflammation The compounds of the present invention inhibit sialidase, which is thought to be involved in the TNF-α-induced inflammatory process in osteoarthritis (Gee, K. et al., 2003, J Biol Chem. 278, 37275). Furthermore, the compounds of the present invention may suppress or inhibit TNF-α activity. Thus, they can be applied to any disorder in which inflammation plays a role in the impairment of physiological function and / or symptoms and / or pain. For example, the compounds of the present invention can be used as anti-inflammatory agents to reduce or eliminate acute, chronic, local or systemic inflammation, for example.

[0088] Inflammation occurs when tissue is damaged by microorganisms, trauma, chemicals, heat, cold, sunburn, or any other adverse event. Endogenous chemicals (e.g., bradykinin, histamine, and serotonin) are released during injury or wound, and such chemicals activate and attract tissue macrophages and other white blood cells. During this process, chemical mediators such as TNF-α are released, causing inflammation.

[0089] Inflammatory disorders are characterized by persistent or chronic inflammation. In such conditions, prolonged inflammation leads to tissue destruction, resulting in widespread damage and ultimate malaise of the affected tissues and / or organs.

[0090] Therefore, the compounds of the present invention are applicable to the treatment of delocalized inflammatory disorders, such as those affecting multiple organs. Such disorders include those resulting from immune dysfunction (and thus potentially containing autoimmune components). Such conditions include systemic lupus erythematosus (SLE), scleroderma, and hypersensitivity reactions.

[0091] There is growing evidence that inflammation is linked to the development of type 2 diabetes.

[0092] The compounds of the present invention are also applicable to the treatment of localized inflammatory disorders, including skin inflammation and chronic prostatitis, glomerulonephritis, inflammatory bowel disease, pelvic inflammatory disease, reperfusion injury, rheumatoid arthritis, transplant rejection, vasculitis, asthma, acne, osteoarthritis, oral mucositis, gastroenteritis, ocular, nasal and otorhinolaryngeal inflammation, and other steroid-responsive inflammatory disorders.

[0093] In particular, the compounds of the present invention are applicable to the treatment of inflammatory skin diseases. These include, for example, actinic keratosis, acne (including acne vulgaris, comedones, rosacea, and cystic nodular acne), allergic contact dermatitis, angioedema, bullous pemphigoid, skin drug reactions, erythema multiforme, lupus erythematosus, actidermy, psoriatic arthritis, scleroderma and urticaria, psoriasis, dermatitis (e.g., atopic dermatitis), scleroderma, steroid-responsive inflammatory skin disorders (e.g., uremic pruritus), and skin conditions associated with exposure to sunlight, radiation, chemotherapy, and environmental irritants.

[0094] The compounds of the present invention are also applicable to the treatment of inflammatory autoimmune diseases. Such diseases may include certain tissues or organs (such as musculoskeletal tissue, as in rheumatoid arthritis and ankylosing spondylitis), GI ducts (e.g., as in Crohn's disease and ulcerative colitis), CNS (e.g., as in Alzheimer's disease, multiple sclerosis, motor neuron disease, Parkinson's disease and chronic fatigue syndrome), pancreatic beta cells (e.g., insulin-dependent diabetes mellitus), adrenal glands (e.g., Addison's disease), kidneys (e.g., Goodpasture syndrome, IgA nephropathy and interstitial nephritis), exocrine glands (e.g., Sjögren's syndrome and autoimmune pancreatitis), and skin (e.g., psoriasis and atopic dermatitis).

[0095] Other inflammatory disorders treatable according to the present invention include conditions such as osteoarthritis, periodontal disease, diabetes (including type 2 diabetes and diabetic nephropathy), chronic obstructive pulmonary disease, atherosclerosis, graft-versus-host disease, chronic pelvic inflammatory disease, endometriosis, chronic hepatitis A, and tuberculosis.

[0096] Pharmacology The compositions and compounds of the present invention may be administered topically, or orally, or parenterally, including intravenous, intramuscular, intraperitoneal, subcutaneous, transdermal, respiratory (aerosol), rectal, vaginal, and topical (including oral and sublingual) administration.

[0097] The dosage administered can vary significantly depending on the specific dosage unit used, the duration of treatment, the age and sex of the patient being treated, the nature and severity of the disorder being treated, and the specific compound selected.

[0098] Typically, the effective dose of the compound administered is in the range of approximately 0.01 mg / kg to 500 mg / kg per day. A unit dose may contain 0.05 to 500 mg of the compound and may be taken once or more times per day. The compound may be administered with a pharmaceutical carrier using any conventional dosing unit form, whether oral, parenteral, or topical, as described below.

[0099] The preferred route of administration is oral administration. Typically, the preferred dose is in the range of 0.01 to 500 mg per kilogram of body weight of the recipient per day, preferably in the range of 0.1 to 50 mg per kilogram of body weight per day, and most preferably in the range of 1 to 5 mg per kilogram of body weight per day.

[0100] The desired dose is preferably provided as a single dose for daily administration. However, 2, 3, 4, 5, or 6 or more secondary doses may be used, administered at appropriate intervals throughout the day. These secondary doses may be administered, for example, in unit dosing forms containing 0.001 to 100 mg, preferably 0.01 to 10 mg, most preferably 0.5 to 1.0 mg of the active ingredient per unit dosing form.

[0101] formulation When isolated from natural sources, idoBR1 may be purified. However, the compositions of the present invention may take the form of unit doses as herbal medicines, food supplements, food additives, nutritional supplements, beverages, or as herbal pharmaceutical kits or packs as defined above. Such herbal medicines are preferably analyzed before use to determine whether they meet standard specifications.

[0102] The herbal medicine for use according to the present invention may be a dried plant material. Alternatively, the herbal medicine may be a processed plant material, the processing of which includes physical or chemical pretreatment, e.g., powdering, grinding, freezing, evaporation, filtration, pressing, spray drying, extrusion, supercritical solvent extraction, and tincture production. If the herbal medicine is administered or sold in the form of a whole plant (or part thereof), the plant material may be dried before use. Any convenient drying form may be used, including freeze-drying, spray-drying, or air-drying.

[0103] The compounds of the present invention can be separated from high molecular weight components such as proteins and polysaccharides by extraction in a polar solvent (ethanol / water mixture, e.g., ≥50% v / v (e.g., up to about 70% v / v) ethanol / water mixture, etc.). Other suitable techniques include various membrane techniques, including microfiltration, ultrafiltration, and nanofiltration. Alternatively or additionally, electrodialysis may also be used to concentrate charged compounds. These methods use membranes with pore sizes that allow only molecules smaller than a certain size to pass through, or depend on the charge on the molecules that are allowed to pass through or not through the membrane. Anion and cation exchange resins may also be used to concentrate compounds.

[0104] When isolated from natural sources, compounds for use according to the present invention may be purified. In embodiments in which the compounds are formulated with pharmaceutically acceptable excipients, any suitable excipients may be used, including, for example, inert diluents, disintegrants, binders, lubricants, sweeteners, flavorings, colorants, and preservatives. Suitable inert diluents include sodium carbonate and calcium, sodium phosphate and calcium, and lactose, while corn starch and alginic acid are suitable disintegrants. Binders may include starch and gelatin, while lubricants, if present, are usually magnesium stearate, stearic acid, or talc.

[0105] The pharmaceutical composition may take any suitable form, for example, tablets, elixirs, capsules, solutions, suspensions, powders, granules, and aerosols.

[0106] The pharmaceutical composition may take the form of a kit of components, which may include the composition of the present invention along with instructions for use and / or a number of different components in unit dosage forms.

[0107] Tablets for oral use may contain the compound for use according to the present invention, mixed with pharmaceutically acceptable excipients such as inert diluents, disintegrants, binders, lubricants, sweeteners, flavorings, colorants, and preservatives. Suitable inert diluents include sodium carbonate and calcium, sodium phosphate and calcium, and lactose, while corn starch and alginic acid are suitable disintegrants. Binders may include starch and gelatin, while lubricants, if present, are usually magnesium stearate, stearic acid, or talc. If desired, tablets may be coated with a substance such as glyceryl monostearate or glyceryl distearate to slow absorption in the gastrointestinal tract. Capsules for oral use include hard gelatin capsules in which the compound for use according to the present invention is mixed with a solid diluent, and soft gelatin capsules in which the active ingredient is mixed with water or an oil such as peanut oil, liquid paraffin, or olive oil.

[0108] Formulations for rectal administration may be provided as suppositories having a suitable base material, for example, cocoa butter or salicylate. Formulations suitable for vaginal administration may be provided as pessaries, tampons, creams, gels, pastes, foams or spray formulations containing the active ingredient in addition to a suitable carrier known in the art.

[0109] For intramuscular, intraperitoneal, subcutaneous, and intravenous applications, the compounds of the present invention are typically supplied as sterile aqueous solutions or suspensions buffered to an appropriate pH and isotonicity. Suitable aqueous vehicles include Ringer's solution and isotonic sodium chloride. The aqueous suspension according to the present invention may contain suspending agents, such as cellulose derivatives, sodium alginate, polyvinylpyrrolidone, and tragacanth gum, as well as wetting agents, such as lecithin. Suitable preservatives for aqueous suspensions include ethyl and n-propyl p-hydroxybenzoate.

[0110] The compounds of the present invention may also be provided as liposome formulations.

[0111] For oral administration, one or more compounds may be formulated into solid or liquid preparations such as capsules, pills, tablets, lozenges, melts, powders, granules, solutions, suspensions, dispersions, or emulsions (solutions, suspensions, dispersions, or emulsions may be aqueous or non-aqueous). Solid unit dosage forms may be capsules of the usual hard or soft shell gelatin type, containing, for example, surfactants, lubricants, and inert fillers, such as lactose, sucrose, calcium phosphate, and corn starch.

[0112] In another embodiment, the compounds of the present invention are tableted using conventional tablet base materials, such as lactose, sucrose, and corn starch, combined with a binder, such as acacia, corn starch, or gelatin; a disintegrant intended to aid in the breakdown and dissolution of the tablet after administration, such as potato starch, alginic acid, corn starch, and guar gum; a lubricant intended to improve the flow of tablet granulation and prevent the tablet material from adhering to the surface of the tablet die and punch, such as talc, stearic acid, or magnesium stearate, calcium, or zinc; a pigment, a coloring agent; and a flavoring agent intended to improve the aesthetic qualities of the tablet and make it more palatable to patients.

[0113] Suitable excipients for use in oral liquid medications include diluents with or without pharmaceutically acceptable surfactants, suspensions, or emulsifiers, such as water and alcohols, such as ethanol, benzyl alcohol, and polyethylene alcohol.

[0114] The compounds of the present invention may also be administered parenterally, i.e., subcutaneously, intravenously, intramuscularly, or intraperitoneally.

[0115] In such embodiments, the compound may be provided as an injectable dose in a physiologically acceptable diluent together with a pharmaceutical carrier (which may be a sterile liquid or a mixture of liquids). Suitable liquids include pharmaceutically acceptable surfactants (e.g., soaps or detergents), suspending agents (e.g., pectin, carbomer, methylcellulose, hydroxypropylmethylcellulose, or carboxymethylcellulose), or water, physiological saline, aqueous dextrose and related sugar solutions with or without emulsifiers and other pharmaceutical adjuvants, alcohols (e.g., ethanol, isopropanol, or hexadecyl alcohol), glycols (e.g., propylene glycol or polyethylene glycol), glycerol ketals (e.g., 2,2-dimethyl-1,3-dioxolane-4-methanol), ethers (e.g., poly(ethylene-glycol) 400), oils, fatty acids, fatty acid esters or glycerides, or acetylated fatty acid glycerides. Suitable oils that may be used in the parenteral formulation of the present invention are of petroleum, animal, plant, or synthetic origin, such as peanut oil, soybean oil, sesame oil, cottonseed oil, corn oil, olive oil, petrolatum, and mineral oil. Suitable fatty acids include oleic acid, stearic acid, and isostearic acid. Suitable fatty acid esters are, for example, ethyl oleate and isopropyl myristate.

[0116] Suitable soaps include fatty alkali metals, ammonium, and triethanolamine salts; suitable detergents include cationic detergents, e.g., dimethyldialkylammonium halides, alkylpyridinium halides, and alkylamine acetates; anionic detergents, e.g., alkyl, aryl, and olefin sulfonates, alkyl, olefin, ethers, and monoglyceride sulfates, and sulfosuccinates; nonionic detergents, e.g., fatty amine oxides, fatty acid alkanolamides, and polyoxyethylene polypropylene copolymers; and amphoteric detergents, e.g., alkyl-beta-aminopropionates, and 2-alkylimidazoline quaternary ammonium salts, as well as mixtures.

[0117] The parenteral compositions of the present invention typically contain about 0.5 to about 25% by weight of the compounds for use according to the present invention in solution. Preservatives and buffers may also be used. To minimize or eliminate irritation at the injection site, such compositions may contain a nonionic surfactant having a hydrophilic-lipophilic balance (HLB) of about 12 to about 17. The amount of surfactant in such formulations is in the range of about 5 to about 15% by weight. The surfactant may be a single component having the above HLB, or a mixture of two or more components having the desired HLB. Examples of surfactants used in parenteral formulations are the class of polyethylene sorbitan fatty acid esters, e.g., sorbitan monooleate and high molecular weight adducts of hydrophobic bases and ethylene oxide formed by the condensation of propylene oxide and propylene glycol.

[0118] One or more compounds for use as herein may also be administered topically, in which case the carrier may preferably comprise a solution, ointment, or gel substrate. The substrate may comprise, for example, one or more of the following: petrolatum, lanolin, polyethylene glycol, beeswax, mineral oil, diluents such as water and alcohol, and emulsifiers and stabilizers. Topical formulations may contain concentrations of the compounds of about 0.1 to about 10% w / v (weight per unit volume).

[0119] When used adjunct, one or more compounds for use as herein may be formulated for use with one or more other drugs. Thus, adjunct use may be reflected in specific unit doses designed to be compatible with (or synergistic with) other drugs, or in formulations in which one or more compounds are mixed with one or more enzymes. Adjunct use may also be reflected in the composition of the pharmaceutical kit of the present invention, in which case the compounds of the present invention are co-packaged with enzymes (e.g., as part of an array of unit doses). Adjunct use may also be reflected in information and / or instructions regarding the co-administration of one or more compounds and / or enzymes.

[0120] Cosmetic formulations The cosmetic composition of the present invention may be selected from, for example, moisturizing compositions, cleansing compositions, or any composition that may provide benefits to the skin. The cosmetic composition of the present invention may include, for example, cosmetically acceptable excipients or carriers selected from those described below.

[0121] In one embodiment, the cosmetic composition is a cleansing composition. A suitable cleansing composition is solid or semi-solid at room temperature. Examples of useful cleansing compositions include, but are not limited to, fatty acid soaps containing glycerin soap, synthetic detergents, and mixtures thereof. Solid cleansing compositions are extensively taught in Soap Technology for the 1990s, the contents of which are incorporated herein by reference. It is desirable that the cleansing composition be fluid.

[0122] In one embodiment of the present invention, the cleansing composition comprises a glycerin soap. Examples of glycerin soaps useful for the present invention include, but are not limited to, those disclosed in U.S. Patent Nos. 4,405,492 and 4,879,063 (the disclosures of which are incorporated herein by reference).

[0123] Suitable examples of fatty acid soaps include soaps derived from hydrocarbon chains with a length of approximately 10 to 22 (including carboxyl carbons), and may be saturated or unsaturated. The soaps may be, for example, sodium salts, potassium salts, ammonium salts, triethanolammonium salts, and mixtures thereof.

[0124] Suitable synthetic detergents include those known in the art for the desired purpose. Examples of detergents useful for personal cleansing include isethionates, sarcosinates, and those derived from commercially available oils such as glyceryl ether sulfonates or coconut oil, which may be pure chain length variants. Other suitable detergents include anionic acyl sarcosinates, methyl acyl taurates, N-acyl glutamates, alkyl sulfosuccinates, alkyl phosphate esters, ethoxylated alkyl phosphate esters, trideceth sulfates, protein condensates, mixtures of ethoxylated alkyl sulfates and alkylamine oxides, betaines, sultaines, and mixtures thereof. Alkyl ether sulfates having 1 to 12 ethoxy groups are also included, particularly ammonium and sodium lauryl ether sulfates.

[0125] The cosmetic composition may be a moisturizing composition.

[0126] Other optional components of the cosmetic composition of the present invention include, but are not limited to, fragrances, air fresheners, preservatives, colorants, dyes, anticoagulants, and personal care ingredients (including, but not limited to, skin and hair care ingredients).

[0127] Examples of suitable personal care ingredients useful in the present invention include, but are not limited to, safe and effective amounts of moisturizers, sunscreens, skin soothing agents, anti-irritants, anti-inflammatory agents, emollients, conditioning agents, wetting agents, deodorants, antiperspirants, artificial sunscreens, antibacterial agents, anti-acne agents, anti-wrinkle agents, anti-skin atrophy agents, skin tightening agents, anti-pruritic agents, antifungal agents, topical anesthetics, skin tone smoothing agents, active natural ingredients, agents for minimizing the appearance of unwanted hair or delaying the regrowth of unwanted hair, skin texture modifiers, and further cleansing agents.

[0128] In one embodiment, the compound may be used from a water or alcoholic water extract by using a water-in-oil (w / o) emulsion, such as those used in the treatment and emollient applications of dry skin.

[0129] Skin emollients function by their ability to remain on the skin surface or within the stratum corneum, acting as lubricants, reducing skin peeling, and improving skin appearance. Typical skin emollients include fatty esters, fatty alcohols, mineral oils, and polyethersiloxane copolymers. Examples of suitable skin emollients include, but are not limited to, polypropylene glycol ("PPG")-15 stearyl ether, PPG-10 cetyl ether, steareth-10, oleth-8, PPG-4 lauryl ether, vitamin E acetate, PEG-7 glyceryl cocoate, lanolin, and combinations thereof. Vitamin E acetate, PEG-7 glyceryl cocoate, and combinations thereof are preferred.

[0130] Examples of suitable humectants include polyhydric alcohols. Suitable polyhydric alcohols include, but are not limited to, glycerol (also known as glycerin), polyalkylene glycols, alkylene polyols and their derivatives (including propylene glycol, dipropylene glycol, polypropylene glycol, polyethylene glycol and their derivatives), sorbitol, hydroxypropyl sorbitol, hexylene glycol, 1,3-dibutylene glycol, 1,2,6-hexanetriol, ethoxylated glycerol, propoxylated glycerol and mixtures thereof.

[0131] Suitable skin soothing agents include, but are not limited to, panthenol, bisabolol, allantoin, aloe, and combinations thereof.

[0132] Suitable conditioning agents include, but are not limited to, dimethicone propyl PG-betaine, dimethicone copolyol, polyquaternium-10, guar, guar derivatives, and combinations thereof. Suitable anti-acne active ingredients include, but are not limited to, salicylic acid, sulfur, lactic acid, glycolic acid, pyruvate, urea, resorcinol, N-acetylcysteine, retinoic acid, benzoyl peroxide, octopirox, triclosan, azelaic acid, phenoxyethanol, phenoxypropanol, flavonoids, derivatives thereof, and combinations thereof. Salicylic acid and benzoyl peroxide are preferred. [Examples]

[0133] The present invention will now be described with reference to specific examples. These are merely illustrative and for illustrative purposes only; they are not intended to limit in any way the scope of the claimed exclusivity or the invention described. These examples constitute the best mode currently contemplated for carrying out the present invention.

[0134] Example 1: Sialidase inhibition by idoBR1 Introduction Sialidases, or neuraminidases, are enzymes that catalyze the cleavage of terminal sialic acids from oligosaccharides and complex carbohydrates. They play a crucial role in regulating the metabolism of sialic acid-containing molecules in biological systems. They are also virulence factors for many viruses and pathogenic bacteria, such as Tannerella forsythia. Human neutrophil sialidase activity has been reported to play a vital role in host inflammatory responses (Glanz, VY, 2019, European J. Pharmacol. 842, 345).

[0135] Methods. The sialidase assay was performed using 2.8 mM and 0.28 mM inhibitors (or water without the inhibitor) and 2.5 nM sialidase (containing NanH from T. forsythia) incubated in the presence of 0.1 mM methylumbelliferyl-N-acetylneuraminic acid in 20 mM sodium phosphate buffer, pH 7.2. The reaction was stopped at 30 and 60 seconds by adding 60 mM sodium carbonate buffer at pH 10.5. The release of fluorescent methylumbelliferone (MU) was quantified by measuring fluorescence emission at 450 nm and excitation at 350 nm. The sialidase activity percentage was expressed as the change in fluorescence between 30 and 60 seconds compared to the reaction without the inhibitor. The reaction was performed in triple succession.

[0136] result idoBR1 resulted in sialidase inhibition, which was over 30% at both concentrations used (2.8 mM [36%] and 0.28 mM [42%]) and was not clearly dose-dependent. The similar inhibition observed at both concentrations suggests that the inhibition is not competitive.

[0137] Example 2: Inhibition assay of endogenous sialidase activity in THP-1 cells Introduction The purpose of this study was to determine whether cucumber extract containing idoBR1 or more than 1% idoBR1 (Q-actin batch B17CF001) can affect sialidase activity in human THP-1 (monocyte-like) cell cultures. The results of this study may be a combination of decreased sialidase expression or inhibition of the enzyme by idoBR1.

[0138] method Treatment of THP-1 cell lines for sialidase activity testing THP-1 cells were cultured in RPMI medium supplemented with mercaptoethanol and glutamine in a culture flask until 80% confluence, then aspirated and centrifuged at 1500 rpm for 5 minutes. The cell pellet was then resuspended in 1 ml of RPMI complete medium and conventionally counted using a hemocytometer. Cells (5 × 10⁶) were incubated with PMA (10 ng / ml) in a separate dish to differentiate THP-1 cells. To determine sialidase activity, THP-1 cells were pretreated for 1 hour with idoBR1 and cucumber extract-Q-actin (batch number B17CF001) at concentrations of 100 μg / ml to 12.5 μg / ml and 200 μg / ml to 25 μg / ml, respectively, followed by stimulation with LPS (1 μg / ml) for 24 hours. After incubation, sialidase activity was determined using the cells.

[0139] THP-1 cells were washed with phosphate-buffered saline (PBS) and resuspended in ice-cold buffer containing 0.25 M sucrose, 1 mM EDTA, and 0.2 mM phenylmethylsulfonyl fluoride. The cell suspension was sonicated on ice for 15 seconds at a low setting (6% amplitude) (Vibracell™; Sonics and Materials Inc., Newtown, CT), followed by centrifugation at 25,000 g for 15 minutes at 4°C. Lysosomal sialidase activity was determined using the obtained supernatant. Protein quantification of the supernatant was performed using the Bio-Rad protein determination kit as described above. To determine lysosomal sialidase activity, 200 μg of total protein was mixed with 40 nmol of 4-methylumbelliferyl-α-N-acetyl-D-neuraminic acid (Sigma), a lysosomal sialidase-specific substrate, 10 μmol of sodium acetate buffer, pH 4.6, and 200 μg of bovine serum albumin (total volume 200 μl). The sialidase reaction was allowed to proceed at 37°C for 1 hour and stopped by the addition of 0.25 M glycine NaOH, pH 10.4. The released 4-methylumbelliferyl was measured by fluorescence at an excitation wavelength of 365 nm and an emission wavelength of 448 nm (Synergy2 multimode plate reader). Sialidase activity was found to be maximum at 16 hours of cell incubation.

[0140] result [Table 1]

[0141] [Table 2]

[0142] Sialidase activity was found to be highest at 16 hours after LPS (1 μg / mL) treatment; therefore, this incubation period was used for further evaluation of the effects of idoBR1 and cucumber extract on sialidase activity in THP-1 cells.

[0143] [Table 3]

[0144] [Table 4]

[0145] Standard idoBR1 tested at 50 μg / ml and 100 μg / ml showed a maximum relative decrease in sialidase activity of 0.63 and 0.55, respectively, compared to the LPS control. Q-actin at 100 μg / ml and 200 μg / ml showed a maximum relative decrease in sialidase activity of 0.7 and 0.62, respectively, compared to the control (LPS).

[0146] Example 3: Treatment of THP-1 cell lines for CD44-HA (hyaluronic acid) binding activity in the presence of idoBR1 using ELISA. Introduction CD44 has been shown to be involved in hematopoiesis, homing to mucosal lymphoid tissue, and lymphocyte infiltration into inflammatory tissue. Hyaluronic acid (HA) interactions with CD44 and CD168 (RHAMM) can induce numerous cellular behaviors, including the activation of tyrosine kinases, protein kinase C, FAK, and PI3K, MAPK, NFκB, and RAS, as well as cytoskeletal components required for inflammation and cancer. While most cells express several forms of CD44, not all cells constitutively bind to HA (Kryworuchko, M. et al., 1999, Cellular Immunol., 194, 54; Nandi et al., 2000, J. Biol. Chem., 275, 14939). Functionally active HA-adhering CD44 is produced by induction of sialidase via MAPK activation. Studies conducted to understand the role of MAPK in LPS-induced inflammatory responses have shown that MAPKp42 / 44-mediated TNF-α generation and subsequent TNF-α-mediated p38 activation lead to the production of HA-adhering CD44 via sialidase activity (Gee, K. et al., 2003, J Biol Chem. 278, 37275).

[0147] method THP-1 cells were cultured in RPMI medium supplemented with mercaptoethanol and glutamine in a culture flask until 80% confluence, then aspirated and centrifuged at 1500 rpm for 5 minutes. The cell pellet was then resuspended in 1 ml of RPMI complete medium and conventionally counted using a hemocytometer. Cells (5 x 10⁶) were incubated in a separate dish with phorbol 12-myristate 13-acetate PMA) (10 ng / ml) to induce THP-1 cell differentiation. To determine CD44-HA binding activity, THP-1 cells were pretreated for 1 hour with idoBR1 or cucumber extract-Q-actin (batch number B17CF001) at concentrations of 100 μg / ml to 12.5 μg / ml and 200 μg / ml to 25 μg / ml, respectively, followed by stimulation with LPS (1 μg / ml) for 24 hours. After incubation, cell lysates were obtained for further analysis.

[0148] Anti-CD44 monoclonal antibody (Invitrogen, 2 μg) was coated into each well of a 96-well plate in 50 mM carbonate / bicarbonate buffer (pH 9.6) and incubated overnight at 4°C. Unbound antibody was removed using PBS with 0.05% Tween 20 (PBS-T wash solution). The wells were blocked with 1% BSA and incubated at 37°C for 1 hour. The wells were thoroughly washed three times by adding 200 μl of PBS-T wash solution to each well. 50 μl of cell lysate was added to the wells and incubated at 37°C for 1 hour. The wells were then washed three times by adding 200 μl of PBS-T by immersing the wells for 30 seconds before each wash. Biotinylated hyaluronic acid (HA) antibody, followed by streptavidin-HRP, was added to form an immune complex and incubated at 37°C for 60 minutes. The solution was aspirated, and the wells were washed three times by immersing them in 200 μl of washing solution for 30 seconds each. 50 μl each of chromogen A and chromogen B was added to each well. The plate was incubated at 37°C for 15 minutes, away from light. The reaction was stopped by adding 50 μl of stop solution, and the absorbance was read at 450 nm.

[0149] result [Table 5]

[0150] [Table 6]

[0151] CD44-bound HA was found to be 110.45 ng / ml in LPS-stimulated (1 μg / ml) THP-1 cells. ido-BR1 at 100 μg / ml showed the highest reduction in CD44-HA levels (26.62%) in the LPS-induced inflammatory response in THP-1 cells compared to LPS controls. Cucumber extract-Q-actin at 200 μg / ml showed the highest reduction in CD44-HA levels (30.60%) in the LPS-induced inflammatory response in THP-1 cells compared to LPS controls.

[0152] Example 4: Reduction of TNF-alpha production in human blood by idoBR1 and Q-actin cucumber extract. Introduction TNF-α, a cytokine produced by monocytes (macrophages) and T lymphocytes, is a crucial component in the cascade of factors that drive inflammatory responses and has many multifaceted effects as a major orchestrator of disease states (Beutler, B. et al., 1989, Annual Review of Immunology, 7, 625). The biological effects of TNF-α depend on its concentration and site of production: at low concentrations, TNF-α can provide desired homeostasis and protective functions, but at high concentrations, systemically or in specific tissues, TNF-α can exacerbate many inflammatory responses in synergy with other cytokines, particularly interleukin-1 (IL-1). The aim of this study was to evaluate the anti-inflammatory activity of idoBR1 or cucumber extract containing idoBR1 in relation to its ability to modulate TNF- levels in human whole blood.

[0153] method Blood and buffy coat fractions were supplied by the Scottish National Blood Transfusion Service (SNBTS), Glasgow, UK. Ficoll histopaque (1.077 g / l) and lipopolysaccharide (derived from Salmonella abortus equi) were purchased from Sigma-Aldrich Co. Ltd. (UK). PGE was from Cayman Chemical Co. (Ann Arbor, MI). Human TNF-α antibody pairs for the TNF-α ELISA assay were purchased from Invitrogen / Life Sciences Europe. All drugs were dissolved in RPMI1640 from Gibco BRL, UK.

[0154] The blood was used without any further processing after donation. It was kindly supplied from normal, healthy donors (tested by the Scottish National Blood Transfusion Service) by ensuring that all were negative for HIV, hepatitis B and C, CMV, and parasitic diseases, such as malaria. They were also confirmed by our laboratory to be free of acute inflammatory diseases at the time of collection by measuring baseline TNF-α levels, which were always <50 pg / ml.

[0155] Cellular stimulation and measurement of TNF-α Aliquots (800 μl) of whole blood were incubated with the compound dissolved in RPMI1640 for an appropriate pre-incubation period, as shown in the results. LPS was then added, and incubation was continued for a further 20 hours at 37°C in a humidified (100%) atmosphere of 5% CO2 in air. At the end of the incubation period, the supernatant from either the plasma or the culture medium was collected by centrifugation at 10,000 g for 30 seconds at room temperature, and TNF-α levels were measured using a human TNF-α ELISA system (supplied by BioSource Europe SA, Belgium, Invitrogen).

[0156] result The potent activity of cucumber extract and idoBR1 against LPS-induced TNF-α in human blood is shown in the following two tables. Cucumber extract containing 0.09% idoBR1 (prototype Q-actin) reduced TNF-α, while idoBR1 was shown to be effective even at much lower concentrations than 10 μM, confirming that idoBR1 alone may be responsible for the anti-inflammatory effect of cucumber extract. Q-actin contains 10 to 100 times more idoBR1 than the prototype extract used here. Q-actin extract with 1 / 10th the amount of idoBR1 had 1 / 10th the effect on TNF-α (data not shown).

[0157] The second study showed even higher activity of idoBR1 (significant at 0.01 μM) in human blood using pre-incubation. IC of idoBR1 in 48-hour pre-incubation. 50The concentration was calculated as 182 nM for blood and as 27 nM for inhibition of TNF-α production by the human monocyte cell line THP-1. idoBR1 did not significantly alter the viability of THP-1 cells when measured by trypan blue uptake or MTT dye conversion. The inhibitory effect of idoBR1 (10 μM) was equivalent to that of the same pretreatment with dexamethasone (50 μM), inhibiting >50% and >65%, respectively. Mifepristone (a glucocorticoid receptor antagonist) with LPS alone significantly amplified TNF-α production from THP-1 cells, but in the presence of dexamethasone, it reversed the dexamethasone inhibitory effect, whereas idoBR1 did not. The data clearly indicate that idoBR1 can inhibit TNF-α production in human blood. Therefore, it appears to be a potent anti-inflammatory agent. Furthermore, it appears to act through a novel mechanism distinct from the steroid receptor pathway.

[0158] This table shows the effects of various idoBR1 concentrations on LPS-stimulated TNF-α production in human blood. Whole blood was pre-incubated for 48 hours with various concentrations of idoBR1, then LPS (10 μg / ml) was added, and incubation was continued for another 20 hours. After incubation at 37°C (5% CO2, 100% humidity), plasma was collected from the blood by centrifugation, and the level of TNF-α in the plasma samples was measured by ELISA.

[0159] [Table 7]

[0160] [Table 8]

[0161] Example 5: Effects of idoBR1 and cucumber extract on cytokines IL-10, IL-12, and IL-1β in LPS-stimulated THP-1 cells Introduction IL-10 is an important negative regulator of anti-inflammatory and inflammatory cytokines. Diverse cell types, including T cells, B cells, and monocytes / macrophages, secrete IL-10 under different conditions of immune activation (Moore, K. et al., 1993, Annu. Rev. Immunol., 11, 165). In vitro studies have shown that IL-10 suppresses the release and function of IL-1β, IL-6, TNF-α, granulocyte-macrophage colony-stimulating factor, and IL-12 (Casatella, M. et al., 1993, J. Exp. Med., 178, 2207; de Waal Malefyt, R. et al., 1991, J. Exp. Med., 174, 1209; Fiorentino, D. et al., 1991, J. Immunol., 147, 3815), suggesting a normal endogenous feedback mechanism for controlling immune responses and inflammation (Asadullah, K. et al., 1998, J. Clin. Invest. 101, 783; Joosten, L. et al., 1997, Arthritis Rheum., 40, 249). Studies have demonstrated that IL-10 exerts its repressive effects on IL-12 p40 and p35, as well as on TNF-α gene expression, primarily at the transcriptional level (Aste-Amezaga, M. et al., 1998, J.Immunol., 160, 5936). Among the inflammatory cytokines involved in the pathogenesis of several autoimmune diseases, IL-12 is a major stimulant of IFN-γ production and the development of T helper (Th)1 autoimmune responses (Paunovic, V. et al., 2008, Rheumatology, 47, 771). IL-12 has been shown to synergistically induce the production of IFN-γ and inflammatory cytokines in conjunction with a variety of cytokines. Monocytes / macrophages produce IL-1β along with TNF-α, which mediates inflammation during infection or upon LPS stimulation. It induces inflammatory and catabolic responses independently and in combination with other mediators. The biological activation of cells by IL-1β is mediated by interaction with a membrane receptor, namely IL-1R1 (IL-1RI, CD121a), which can also bind to another IL-1 group receptor, IL-1α.

[0162] method ELISA assay for THP-1 monocytes Sample preparation for ELISA assays Cells were aspirated from an 80% confluent culture flask and centrifuged at 1500 rpm for 5 minutes. The cell pellet was then resuspended in 1 ml of RPMI complete medium and seeded at 1 × 10⁵ cells / well in each well of a 96-well microtiter plate. After 24 hours of incubation, PMA (10 ng / ml) was added to the 96-well plate to differentiate THP-1 cells and determine TNF-α production. THP-1 cells were pretreated for 1 hour with idoBR1 or cucumber extract-Q-actin (batch number B17CF001) at concentrations ranging from 200 μg / ml to 25 μg / ml, sequentially diluted 2-fold, followed by stimulation with LPS (100 ng / ml) for 2 hours. After incubation, the cell supernatant from each well was aspirated into a sterile microcentrifuge tube and centrifuged at 1000 rpm for 2-3 minutes. The cell supernatant was then used to assess the presence of cytokines using ELISA.

[0163] Sandwich ELISA assay ELISA plates coated with antibodies against IL-12, IL-1β, or IL-10 (R&D Systems, USA) were used for the following studies. 50 μL of assay diluent RD1F was thoroughly mixed and added to each well. 200 μL of sample (idoBR1 or extract) or control was added per well and covered with an adhesive strip. After incubation at room temperature for 2 hours, each well was aspirated and washed four times with wash buffer (400 μL). After the final wash, any remaining wash buffer was removed by aspirate or decantation. The plate was inverted and wiped with a clean paper towel. 200 μL of appropriate human conjugate was added to each well, then covered with a new adhesive strip and incubated at room temperature for 1 hour. Aspirate / wash was then repeated. 200 μL of substrate solution was then added to each well and incubated for a further 20 minutes at room temperature in the dark. 50 μL of stop solution was added to each well. The well color changed from blue to yellow. OD was measured at 450 nm within 30 minutes.

[0164] result [Table 9]

[0165] [Table 10]

[0166] [Table 11]

[0167] [Table 12]

[0168] [Table 13]

[0169] [Table 14]

[0170] Both idoBR1 and cucumber extract increased the anti-inflammatory marker IL-10 by 3.04 and 3.65 times, respectively. The IL-10 result indicates an anti-inflammatory effect. idoBR1 at 100 μg / mL showed a 24.53% reduction in IL-12 levels in the LPS-induced inflammatory response in THP-1 cells compared to LPS controls. Cucumber extract Q-actin at 200 μg / mL showed a 25.88% reduction in IL-12 levels in the LPS-induced inflammatory response in THP-1 cells compared to LPS controls. idoBR1 at 100 μg / mL gave a 24.07% reduction in IL-1β levels in the LPS-induced inflammatory response in THP-1 cells compared to LPS controls. Cucumber extract Q-actin at 200 μg / mL showed the highest reduction in IL-1β levels (22.53%) in the LPS-induced inflammatory response in THP-1 cells.

[0171] Example 6: Gym workouts using Q-actin increase IL-10 as measured in human blood. Introduction We demonstrated the modulation of the anti-inflammatory cytokine IL-10 in THP-1 cells by idoBR1 and cucumber extract (Q-actin). Here, we examine cytokine modulation in individuals ingesting Q-actin while under a given strenuous exercise regime that naturally induces muscle inflammation.

[0172] method In an exercise recovery experiment, IL-10 levels in blood samples were measured using 7 placebo subjects and 10 Q-actin subjects. Subjects received 10 mg of either Q-actin or placebo (both in capsule form) twice daily during a 4-day period of strenuous exercise, from day 0 through 1, 2, 3, and on day 4 (the recovery day). Blood samples were collected before and after exercise on days 1, 2, and 3, and at the end of the recovery period on day 4. IL-10 levels were measured by ELISA assay.

[0173] result Subjects given Q-actin tended to show a significant increase in IL-10 upon initiation of exercise, supporting the results obtained in LPS-stimulated THP-1 cells.

[0174] [Table 15]

[0175] Example 7: MAPK signaling effect of idoBR1 and cucumber extract containing idoBR1 Introduction The MAPK signaling cascade plays a role in the initiation of inflammatory responses. Induction of inflammatory cytokine genes requires MAPK activation, and stimulation of the extracellular regulatory protein kinase / mitogen-activated protein kinase (ERK / MAPK) pathway is essential for downstream inflammatory responses (Kaminska, B., 2005, Biochim. Biophys. Acta, 1754, 253; Buchholz, K. et al., 2007, Infection and Immunity, 75, 5924). The MAPK pathway is also required for the expression of inflammatory mediator genes, including COX-2, iNOS, IL-1β, and TNF-α. ERK and / or p38 MAPK have been reported to be involved in the upregulation of IL-1β (Baldassare, J. et al., 1999, J. Immunol., 162, 5367).

[0176] method THP-1 cells were cultured in RPMI medium supplemented with mercaptoethanol and glutamine in a culture flask until 80% confluence, then aspirated and centrifuged at 1500 rpm for 5 minutes. The cell pellet was then resuspended in 1 ml of RPMI complete medium and conventionally counted using a hemocytometer. Cells (5 × 10⁶) were incubated with PMA (10 ng / ml) in a separate dish to differentiate THP-1 cells. To determine protein expression p38 and p42 / 44, THP-1 cells were pretreated for 1 hour with idoBR1-100 μg / ml and 50 μg / ml and cucumber extract (Q-actin batch number B17CF001)-200 μg / ml and 100 μg / ml, followed by stimulation with LPS (1 μg / ml) for 2 hours. After incubation, cells were harvested and whole proteins were isolated.

[0177] Western blot procedure Cell pellets were lysed, and protein concentrations were determined using the Bio-Rad protein determination assay (Bio-Rad). Total cellular proteins were subjected to 8% polyacrylamide SDS gel electrophoresis and subsequently transferred to a polyvinylidene difluoride membrane (Thermoscientific). The membranes were probed with either mouse anti-phospho p38 mAb (Thermoscientific) or mouse anti-phospho p42 / 44 mAb (Thermoscientific), followed by a goat anti-mouse polyclonal antibody (Thermoscientific) conjugated with horseradish peroxidase. All immunoblots were visualized by ECL (Amersham Biosciences). Test samples of Q-actin tested at 100 μg / ml and 200 μg / ml showed relative decreases of 0.92 and 0.83 in phosphorylated p38 expression compared to the LPS control, respectively. IdoBR1 at 50 μg / ml and 100 μg / ml showed a decrease in phosphorylated p38 expression of 0.88 and 0.80, respectively, compared to the LPS control. Q-actin tested at 100 μg / ml and 200 μg / ml showed a relative decrease in phosphorylated ERK42 / 44 expression of 0.81 and 0.78, respectively, compared to the LPS control. IdoBR1 at 50 μg / ml and 100 μg / ml showed a decrease in phosphorylated ERK42 / 44 expression of 0.80 and 0.76, respectively, compared to the LPS control.

[0178] [Table 16]

[0179] [Table 17]

[0180] Therefore, both idoBR1 and cucumber extract containing idoBR1 (Q-actin) have been shown to reduce the MAPK signaling cascade, which plays an essential role in inflammatory responses.

[0181] Example 8: Oral availability and in vivo stability of idoBR1 Introduction The purpose of this study was to investigate the oral availability of idoBR1 from ingested cucumbers / gherkins by measuring its availability in urine. This study not only suggested the oral availability and potential systemic activity of idoBR1, but also confirmed the compound's ability to cross membranes unchanged, thus supporting its local availability.

[0182] method Parisien pickling cucumbers (seeds purchased from Lidl 2013) were organically grown, and one male and one female volunteer each consumed three cucumbers at noon. The consumed raw weight was 260g in each case, including a 30g comparison weight removed from all consumed cucumbers, and this was maintained for analysis. Volunteers did not consume any Cucurbitaceae foods for 15 hours prior to the experiment. Pre-consumption urine samples were collected over 3 hours with t=0, and then samples were collected over 9 hours for females and 15 hours for males. 30g of cucumber samples were homogenized in 50% ethanol (aq), filtered after 15 hours of extraction, and the idoBR1 fraction was bonded to an H+ type cation exchange resin IR120. After washing the column with water, the substance was purged with 2M ammonia solution, dried (52.3mg), and analyzed by GC-MS after trimethylsilylation using Pierce TriSil. Next, 0.2 mg of castanospermine was added to the remaining 51 mg of the substance for comparative quantification purposes. The entire urine sample was treated similarly using cation exchange resin, except that the substance, which had been replaced with ammonia solution, was subjected to a second pass with the same cation exchange resin (ammonium type in this case) to reduce the strong base (which binds to IR120 of the ammonium type), and only the substance that was not retained was preserved. One fraction of urine idoBR was dried and diluted with water to 20 ml. 500 ul of each was sampled and 0.025 mg of castanospermine was added.

[0183] result GC-MS analysis of cucumbers This was performed using Perkin Elmer Turbomass Gold GCMS. The spectrum of the major peak at 10.33 min matched the GCMS spectrum of genuine idoBR1 (900288 PhytoQuest Ltd, UK). The relative response factor between genuine BR1 (900125, PhytoQuest Ltd) and castanospermine was calculated to be 1:2. Assuming the same response factor, the amount of idoBR1 in a 30g sample was estimated to be 1.5mg, which means that the volunteer consumed approximately 260 / 30 × 1.5mg = 13mg of idoBR1.

[0184] Urine results Urine samples collected before cucumber consumption did not show a significant peak in the retention time (10.33 minutes) for idoBR1. After 15 hours, men showed excretion of approximately 2.4 mg of idoBR1 compared to the castanospermine reference peak area, but more accurate measurements of intake and excretion would be needed for a definitive mass balance. Women excreted approximately 2.1 mg of idoBR1. This study confirmed that idoBR1 is orally available and measurable in urine, meaning that it can enter the bloodstream from oral ingestion and be excreted in urine unchanged in at least a significant proportion. This indicates that the compound can cross membranes in the gastrointestinal tract and be significantly present in urine. No obvious conjugation was observed in urine analysis.

[0185] The residual idoBR1 may have remained in the body for a longer period. Therefore, it appears to be a powerful anti-inflammatory agent with long-lasting effects.

[0186] Example 9: Effects of idoBR1 on microglia cells Introduction Microglia are resident macrophages of the central nervous system (CNS). These cells are the primary form of active immune defense in the CNS. In neurodegenerative disorders such as Alzheimer's disease and Parkinson's disease, microglia are chronically activated, promoting the release of inflammatory cytokines that further disrupt normal CNS activity. There is considerable interest in testing the extent to which bioactive food components can mitigate inflammatory effects by reducing oxidative stress and / or reducing inflammatory gene expression.

[0187] method IdoBR1 was applied to cell cultures of the murine microglia cell line BV-2 at suboptimal LPS concentrations of 0, 20, 40, and 80 μg / ml, both with and without LPS. TNF-α and nitrite production were measured after 24 hours. Interestingly, idoBR1 derived from Q-actin was found to be effective in reducing TNF-α and nitrite production by stimulated microglia cells.

[0188] [Table 18]

[0189] Equal portions The foregoing description details current preferred embodiments of the present invention. Those skilled in the art will anticipate, in consideration of these descriptions, that numerous modifications and types will arise in its implementation. These modifications and types are intended to be covered within the claims appended herein. The invention described in the claims of the original application of this application is listed below. [1] A process for producing a composition comprising (2R,3R,4R,5S)-3,4,5-trihydroxypiperidine-2-carboxylic acid (idoBR1), wherein the process is: (e) A process of providing plant material from a plant source including plants of the Cucurbitaceae family. (f) A step of fractionating the plant material to produce an extract in which idoBR1 is concentrated, (g) The step of assaying the extract for (i) inhibitory activity against sialidase, (ii) inhibitory activity against TNF-alpha, or (iii) IL-10 stimulating activity, and (h) The process of formulating the assayed extract using cosmetic, nutritional, or pharmaceutically acceptable excipients or carriers to produce a cosmetic, nutritional, or pharmaceutical composition. The process including the process described above. [2] The process described in [1], wherein the plant source includes a plant of the genus Cucumis, for example, the species Cucumis sativus. [3] The process according to [1], wherein the plant source comprises a plant of the genus Cucurbita, for example, a species of Cucurbita melos or Cucurbita moschata. [4] The process according to any one of the prior claims, wherein the plant material comprises a fruit, a fruit part, a fruit extract, a fruit juice, a seed and / or a leaf. [5] The process according to any one of the prior claims, comprising a cucumber, wherein the plant material optionally comprises a cucumber from a plant of the species Cucumis sativus. [6] The process according to any one of the prior claims, wherein idoBR1 is isolated and present in the composition at levels of, for example, at least 1% w / w, 5% w / w, 10% w / w; 15% w / w; 20% w / w; 25% w / w; 30% w / w; 35% w / w; 40% w / w; 45% w / w; 50% w / w, 60% w / w, 70% w / w, 80% w / w, 90% w / w, and 99% w / w (on a dry weight basis). [7] The process described in any one of the preceding [1] to [5], wherein idoBR1 is present in the composition at a maximum level of 5% w / w (on a dry weight basis). [8] The process described in [7], wherein idoBR1 is present in the composition at a maximum level of 1% w / w (on a dry weight basis). [9] The process according to any one of the prior claims, wherein the composition of step (d) is a cosmetic composition.

[10] The process according to any one of [1] to [8], wherein the composition of step (d) is a nutritional supplement composition.

[11] The process according to any one of [1] to [8], wherein the composition of step (d) is a pharmaceutical composition.

[12] The process according to

[11] , wherein the composition of step (d) is in the form of (i) a pharmaceutical pack, kit or patient pack, or (ii) a unit dose.

[13] The formulation step comprises mixing the assayed extract with a pharmaceutically acceptable excipient, according to the process in

[11] or 12.

[14] The process according to any one of the prior claims, wherein in step (c), the extract is assayed for its inhibitory activity against sialidase.

[15] The process according to any one of the prior claims, wherein in step (c), the extract is assayed for inhibitory activity against TNF-alpha.

[16] The process according to any one of the prior claims, wherein in step (c), the extract is assayed for IL-10 stimulating activity.

[17] A composition that can be obtained or produced by the process described in any one of the prior claims, for use in therapy or prevention.

[18] A composition for use as described in

[17] for use in a method for treating inflammatory disorders.

[19] The aforementioned inflammatory disorders include: (a) delocalized inflammatory disorders (e.g., systemic lupus erythematosus (SLE), scleroderma, and hypersensitivity); (b) chronic prostatitis; (c) glomerulonephritis; (d) inflammatory bowel disease; (e) pelvic inflammatory disease; (f) reperfusion injury; (g) rheumatoid arthritis; (h) transplant rejection; (i) vasculitis; (j) asthma; (k) acne; (l) osteoarthritis; (m) oral, mucosal, or gastrointestinal inflammation; (n) ocular inflammation; (o) rhinitis (p) Otitis; (q) Steroid-responsive inflammatory disorders; (r) Skin inflammatory diseases (e.g., actinic keratosis, acne vulgaris, acne comedones, rosacea, acne cystoides, allergic contact dermatitis, angioedema, bullous pemphigoid, skin drug reactions, erythema multiforme, lupus erythematosus, actinic dermatitis, psoriatic arthritis, scleroderma and urticaria, psoriasis, dermatitis, atopic dermatitis, scleroderma, steroid-responsive inflammatory skin disorders, urinary tract infections) (s) toxic pruritus and skin conditions associated with exposure to radiation, chemotherapy and environmental irritants; (s) inflammatory autoimmune diseases (e.g., ankylosing spondylitis, Crohn's disease, ulcerative colitis, Alzheimer's disease, multiple sclerosis, motor neuron disease, Parkinson's disease, chronic fatigue syndrome, insulin-dependent diabetes mellitus, Addison's disease, Goodpasture syndrome, IgA nephropathy, interstitial nephritis, Sjögren's syndrome and autoimmune pancreatitis); (t) osteoarthritis; (u) periodontal disease; (v) diabetic nephropathy; (w) chronic obstructive pulmonary disease; (x) atherosclerosis; (y) graft-versus-host disease; (z) chronic pelvic inflammatory disease; (a') endometriosis; (b') chronic hepatitis; (c') tuberculosis and (d') skin inflammation, e.g., skin inflammation caused by exposure to sunlight, allergens, irritants or burns, as described in

[18] .

[20] The composition for use described in

[18] , wherein the inflammatory disorder is an autoimmune disease, asthma, or allergy.

[21] The inflammatory disorder is an autoimmune disease selected from Graves' disease; rheumatoid arthritis; Hashimoto's thyroiditis; vitiligo; diabetes mellitus (e.g., type 1 or type 2 diabetes mellitus); pernicious anemia; multiple sclerosis; glomerulonephritis; systemic lupus E (SLE, lupus); Sjögren's syndrome; scleroderma; psoriasis; ankylosing spondylitis; myasthenia gravis; pemphigus; polymyositis; dermatomyositis; uveitis; Guillain-Barré syndrome; Crohn's disease; ulcerative colitis; and inflammatory bowel disease (IBD), the composition for use described in

[20] .

[22] The inflammatory disorder is an allergy selected from atopic allergies, allergic rhinitis, allergic conjunctivitis, atopic dermatitis, hypereosinophilia, irritable bowel syndrome, allergen-induced migraine, bacterial allergy, bronchial allergy (asthma), contact allergy (dermatitis), delayed-type allergy, hay fever, drug allergy, puncture allergy, bite allergy, gastrointestinal allergy; food allergy; and physical allergy, such as cold urticaria, angioedema, cholinergic urticaria and photosensitivity, according to the composition for use described in

[20] .

[23] The inflammatory disorder is selected from graft-versus-host disease; sarcoidosis; vascular inflammatory diseases including disseminated intravascular coagulation, atherosclerosis, and Kawasaki disease; vasculitis; Sjögren's syndrome; psoriatic arthritis; enteric arthritis; reactive arthritis and arthritis associated with inflammatory bowel disease; the composition for use according to

[18] .

[24] The composition for use described in

[17] for use in a method of treating neoplasms.

[25] The neoplasm is selected from benign, precancerous, and malignant neoplasms, hyperplasia, metaplasia, and dysplasia, and the composition for use according to

[24] .

[26] The neoplasm is a malignant neoplasm (cancer), the composition for use according to

[25] .

[27] The malignant neoplasm is selected from (a) cancer; (b) blastoma; (c) leukemia; (d) lymphoma; (e) myeloma; (f) sarcoma and (g) mixed-type cancer, and is the composition for use described in

[26] .

[28] The malignant neoplasm is a cancer selected from cancers of the bladder, breast (e.g., primary breast tumor, breast cancer without lymph node metastasis, invasive ductal adenocarcinoma and non-endometrioid breast cancer), colon (e.g., colorectal cancer such as colon adenocarcinoma and colon adenoma), kidney, epidermis (e.g., malignant melanoma), liver, lung (e.g., adenocarcinoma, adrenal cortical, nasopharyngeal, small cell lung cancer and non-small cell lung cancer), esophagus, gallbladder, ovary, pancreas (e.g., exocrine pancreatic adenocarcinoma), stomach, cervix, thyroid, prostate, gastrointestinal system (e.g., gastrointestinal stromal tumor), or skin (e.g., squamous cell carcinoma), according to the composition for use described in

[26] .

[29] The malignant neoplasm is a leukemia selected from lymphoid leukemia, for example, progenitor cell leukemia, mature B-cell leukemia, mature T-cell leukemia and NK-cell leukemia, acute myeloid leukemia, chronic myeloproliferative disorders and myelodysplastic syndromes, the composition for use according to

[26] .

[30] The composition for use according to

[26] , wherein the malignant neoplasm is a lymphoma selected from (a) Hodgkin lymphoma; (b) non-Hodgkin lymphoma, e.g., progenitor cell lymphoma, mature B-cell lymphoma, mature T-cell lymphoma, and NK-cell lymphoma; (c) Burkitt lymphoma; and (d) lymphoreticular neoplasm, e.g., mantle cell lymphoma.

[31] The composition for use described in

[26] , wherein the malignant neoplasm is a sarcoma selected from osteosarcoma; chondrosarcoma; leiomyosarcoma; rhabdomyosarcoma; mesothelioma; fibrosarcoma; angiosarcoma or hemangioendothelioma; liposarcoma; glioma; astrocytoma; myxosarcoma and mesenchymal and mixed mesodermal tumors.

[32] A composition for use as described in

[17] for use in a method for treating a viral infection.

[33] A composition for use as described in

[17] for use in a method for treating a bacterial infection.

[34] The composition for use according to

[33] comprises the method of inhibiting commensalism and / or pathogenic bacterial growth in vivo.

[35] The composition for use according to

[34] comprises inhibiting host-bacterial cell interactions and / or inhibiting or eliminating bacterial biofilm formation in a mammalian (e.g., human) host.

[36] The composition for use according to

[34] or 35, comprising the method for treating or preventing diseases and disorders mediated or characterized by the presence of bacterial biofilms (e.g., subgingival plaque biofilms and mucosal biofilms).

[37] The composition for use described in

[17] for use in a method of treating periodontal disease, bacterial vaginosis and / or disease caused by infection of Tannerella forsythia, Tannerella denticola, Porphyromonas gingivalis or Gardnerella vaginalis.

[38] The composition for use described in

[17] for use in a method for treating atheromatosis, for example, atherosclerosis.

[39] A composition for use as described in

[17] for use in a method for regulating the growth of commensal bacteria in a mammalian host.

[40] The method comprises the modification of the composition for use described in

[39] in a mammalian host, such as a human host.

[41] Use of compositions as defined in

[17] for the treatment of a disease or for use in the manufacture of a drug in any one of the provisions of

[18] to

[40] .

[42] A cosmetic, nutritional, herbal, or pharmaceutical composition comprising a composition that can be obtained or produced by a process described in any one of [1] to

[16] , further comprising optionally cosmetic, nutritional, or pharmaceutically acceptable excipients or carriers.

[43] A cosmetic method for reducing swelling or erythema of the skin, comprising, for example, administering to a subject a composition defined in

[42] by topical application to the skin.

[44] A process for producing a supplemented food or beverage, comprising the steps of (a) providing a composition as defined in

[42] ; and (b) adding the composition of step (a) to a food or beverage to produce a supplemented food or beverage.

[45] A method for monitoring the quality of a cosmetic, nutritional supplement, or pharmaceutical composition, comprising the steps of (a) providing a sample of the composition; and (b) assaying the sample for (i) inhibitory activity against sialidase; (ii) inhibitory activity against TNF-alpha; or (iii) IL-10 stimulating activity.

[46] The cosmetic, nutritional supplement, or pharmaceutical composition is the method according to

[45] , comprising idoBR1.

[47] The method according to

[45] or 46, wherein the cosmetic, nutritional supplement, or pharmaceutical composition comprises plant material from a plant source including plants of the family Cucurbitaceae.

[48] The method according to

[47] , wherein the plant source includes a plant of the genus Cucumis, for example, the species Cucumis sativus.

[49] The method according to

[47] or

[48] , wherein the plant material comprises fruits, fruit parts, fruit extracts, fruit juices, seeds and / or leaves.

[50] The method according to

[49] , wherein the plant material includes a cucumber.

[51] The method according to

[50] , wherein the plant material comprises a cucumber from a plant of the species Cucumis sativus.

[52] idoBR1 for use in any of the methods defined in any one of

[24] to

[40] .

[53] A treatment method as defined in any one of

[24] to

[40] , comprising administering an effective amount of idoBR1 to a subject in need thereof.

Claims

[Claim 1] The invention described in the present specification.

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