Bioactive phytochemicals of plants in the genus Ziziphus and guarana

Abstract

To provide a process for producing a composition containing 4-hydroxymethyl proline, a pharmaceutical composition, a crude drug, or a method for monitoring the quality of a nutritional supplement, and a method for selecting a breeding line and / or variety of a plant.SOLUTION: A composition contains selected isolated 4-hydroxymethyl-proline or a pharmaceutically acceptable salt or a derivative thereof. Further, provided is novel 4- hydroxymethyl-N-methyl-proline together with a preparation process thereof, a composition containing the same, and use thereof as a medical drug and a medicine. Further, also provided is a process of producing a crude drug including a step of monitoring the quality of the crude drug by detecting the presence or absence of one or more of 4-hydroxymethyl-N-methylproline in a sample of the crude drug or by measuring the amount thereof.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to a composition containing 4-hydroxymethyl-N-methylproline, and energy - Applications include disease control, blood glucose control, inflammation, bacterial infection, skin disorders, and in vivo inhibition of sialidase activity. , metabolic syndrome (e.g., central obesity, elevated triglyceride levels and Any disease related to diabetes (including type 1 diabetes, type 2 diabetes, and insulin resistance) Their use in medicine, including the treatment of disorders, and the composition from various plant sources. Processes for isolating and purifying, as well as various products, compounds, and compositions. This relates to materials, medical applications, and methods based thereon.

[0002] The present invention also modifies the quality of crude drugs, supplements, and extracts of plants of the genus Ziziphus and guarana. Method for producing crude drugs based on extracts of Ziziphus jujuba and guarana Processes, and herbal food additives, foods, and obtained by such processes. Regarding beverages. [Background technology]

[0003] Jujube species The genus Ziziphus belongs to the family Rhamnaceae and includes thorny shrubs and small trees. It is a genus of trees with many species. They are distributed in tropical and subtropical regions worldwide. In fact, it is an edible drupe, yellowish-brown, red, or black, often very sweet and high in sugar, with a pleasant texture and flavor. Its taste is reminiscent of dates. Well-known species include Z. jujuba, Z. spina-christi, Z. lotus, Z. mauritiana, and Z. Joazeiro is one example.

[0004] Zizyphus jujuba, also known as Chinese dates, is a traditional Chinese medicine. Among the fruits used is the jujube. It is believed to nourish the heart and calm the mind. It is used to treat nervousness, insomnia, and palpitations. Other traditional uses of the jujube genus include Treatment for diseases such as digestive disorders, liver disease, urinary tract disorders, diabetes, skin infections, and fever. It is included. It is used as a treatment for eczema (Khiljee at al., 20 11, Journal of Pakistan Association of D Ermatologists 21, 112).

[0005] Zizyphus spina-christi is a wild tree that grows in desert areas. It is abundant throughout the Middle East. Its leaves are a rich source of protein for desert animals, and small The brown fruit is eaten by the locals. All parts of the plant have traditionally been eaten by the locals. I use it to maintain my health. It has antibacterial, antifungal, antioxidant, and anti-inflammatory properties derived from plants. It is caused by the fact that the leaf extract is used to treat wounds and skin diseases. Dried leaf infusion It is used as a skin cleanser and hair treatment. Its root bark is used to treat pain. It is also used in folk medicine.

[0006] The chemical composition of this plant has been well documented (Abdel-Zaher). , A. et al., 2005, J. Ethnopharmacol.101 ,129). Many peptides and cyclopeptide alkaloids, flavonoids, trityl Penoid saponin glycosides and betulinic acid have been identified in varying amounts in various species of the genus. are present (Han, B., et al., 1990, Phytochemist ry 29, 3315、Solati, J. & Soleimani, N., 2010, Acta Diabetol., 47, 219).

[0007] Recently, the composition, antioxidant activity, and hepatoprotective effect of the aqueous extract of Zizyphus jujuba have been evaluated (Liu, N. et al., 2017, RSC Adv.7 , 6511). A more detailed analysis of Z. jujuba fruits was carried out by Villanueva, J.R. & Villanueva, L.R. (2017, P hytotherapy Research, 31, 347).

[0008] The literature includes many scientific studies on the use of Z. spina-christi for the treatment of diabetes (e.g., Nesseem, D., et al., 2 009, Pharmazie 64, 104). The extracts of the plant are reported to have antihyperglycemic activity (e.g., Abdel-Zaher, A. et al., 2005, J. Ethnopharmacol.101,129). The methanol extracts of the roots of Z. spinachristi and Z. jujuba used in the treatment of alloxan-induced diabetic rats have been claimed to have beneficial effects on diabetic rats with reduced hyperglycemia, hyperlipidemia, and lipid peroxidation (Hussein, H., et al., 2006 , International Journal of Pharmacology 2, 563). Hussein et al. believe that the activity is due to the strong antioxidant activity that is not a characteristic of the compounds of the present invention.

[0009] Tanira, M., et al., (1988, Internationa (Journal of Pharmacology 2, 563) used Z in mice. Ethanol extract of Spina christi leaves was tested for anti-inflammatory and moderate effects. It showed a degree of antipyretic effect. Motamedi, H. et al., (2009, A sian Journal of Plant Sciences 8, 187) Ethanol of Z. spina-christi as a potential source of new antimicrobial compounds Leaf extracts of sulfur and methanol have been reported, but currently, common fatty acids of many plants ( It has been revealed that hexadecanoic acid contains potent antibacterial activity. Medi et al. found 4-hydroxymethyl-N-methyl-proline to have potential efficacy It does not specify any particular ingredient or suggest the use of fruit.

[0010] Borgi, W. et al.,(2007, Fitoterapia 78, 16) Aqueous and methanol extracts of the root bark of Z. lotus are used in the acute inflammatory process. It showed a significant anti-inflammatory effect during the period, and it was reported that this was related to antioxidant effects. racts of Z. spina-christi for cosmetics U.S. Patent No. 5,849,302 and EP081, titled "and psoriasis" 5842A2 uses a 10% aqueous extract of Z. spina-christi leaves. The leaf extract was described as having anti-inflammatory activity against the skin of healthy volunteers exposed to UV light. The study showed a 17.5% reduction in redness in all subjects.

[0011] However, 4-hydroxymethyl-N-methyl-proline as described herein (and (Similar compounds) have not been reported from plants of the genus Ziziphus, and these compounds are not found in It has not been suggested that it is involved in any of the research on physical activity. Rather, interest in this point is, The focus had been on structurally unrelated triterpenes, antioxidants, and peptides.

[0012] Guarana As a bioactive phytochemical present in species of the genus Ziziphus, 4-hydroxymethyl Chil-N-methyl-L-proline has not been reported by other groups, and this is surprising. What should be noted is that guarana, Paullinia cupana (Sapindaceae family, Jujube genus) It was also found in the fruits of (unrelated) plants.

[0013] Paullinia cupana is native to the Amazon basin and is particularly common in Brazil. Yes, guarana is best known for the seeds from its fruit, which are almost the same size as coffee beans. It is used as a nutritional supplement and is best known for its high caffeine content. The seeds contain about twice the concentration of caffeine found in coffee seeds: caffeine Compared to 1%-2% of coffee seeds, guarana seeds contain approximately 2%-4.5% of cyanobacteria. It exists in - Bempong, DK, et al., 1993, Int. J. Pharmacog. 31, 175).

[0014] The first report of the use of guarana as a beverage dates back to 1669, when Iez went to the Amazon. During the Sisters' expedition, missionary Joao Felipe Bettendorf, Satellite- The Mawe Indians have a diuretic effect and a therapeutic effect against headaches, fever, and convulsions. It was observed that they were consuming stimulating beverages. Guarana may now be sweetened. Alternatively, carbonated beverages or energy shots, herbal tea ingredients, or capsules. It is used in the ingredients contained in [the product]. Jujubes do not contain caffeine.

[0015] The therapeutic properties of guarana as a stimulant, tonic, and aphrodisiac have been recognized since the first reports of its unique use. It became known all over the world. The seeds contain caffeine (1) due to the stimulating properties of guarana. Due to its content of ,3,7-trimethylxanthine, it is a commercially useful part of the plant. Kofink, M., et al., 2007, European Food Research and Technology 225, 569, Campos, MPD et al., 2011, Journal of Alterna tive and Complementary Medicine 17, 505) .

[0016] In addition to its psychoactive effects, it is believed to possess similar functional properties to green tea, which is also rich in catechins. Therefore, the use of guarana for metabolic disorders is being widely studied. Studies have shown that guarana can help reduce lipid levels. It has a positive effect on metabolism, increases basal energy and weight loss, and may be helpful in treating obesity. This has been shown (for example, Opala, T., et al., 2006, European Journal of Medical Research 11, 343).

[0017] However, other studies have not shown any effect of guarana-containing preparations on body weight (S ale, C., et al., 2006, Int. J. Obes.(Lon d.), 30, 764), this is (for example, the inventors of 4-hydroxymethyl- As first demonstrated with N-methyl-L-proline, in vitro or in vivo To produce quality-controlled extracts that measure compounds shown to have specific activity To highlight the essentials.

[0018] 4-hydroxymethyl-N-methyl-proline (and similar compounds) as described herein None of these have been reported from plants of the genus Guana, and these compounds are not bioactive phytochemicals. It has not been suggested that it is of quality. Rather, interest in this point lies in methylxanthine, catechin N, methylbenzene, cyclic monoterpenes, sesquiterpenes, oleic acid, pau acid, and The focus has been on structurally unrelated chemicals, including methoxyphenylpropene (Ava to, P., et al., 2003, Lipids 38, 773-780 ).

[0019] Herbal food additives and treatments Currently, there is great interest in the use of herbal therapies and supplements, and food manufacturers Healthcare companies and medical professionals recognize the value of herbal products and established formulations. It is increasingly recognized that it can complement treatment. Currently, herbal food additives and Supplements are widely used.

[0020] However, quality control of herbal food additives is challenging due to the complex properties and inherent heterogeneity of plant materials. It is difficult due to its nature. The ingredients used in herb and plant-based food additives are usually, It is the whole plant, a part of it, or an extract. Plant materials contain many different chemical components. Because it is a complex mixture, the material is a complex mixture. This makes it difficult to standardize and control the quality of the material. This becomes extremely difficult. Furthermore, many herbal food additives contain two or more plant-based ingredients. It is a mixture of its components, and therefore a mixture of mixtures, thus requiring a further level of complexity. Furthermore, the recipes and manufacturing methods used are often not uniform and are not disclosed. It may remain unprocessed. Due to these factors, it is obtained from different sources and the table Two samples of a particular product that appear identical on the surface actually contain the same mixture of ingredients. It is extremely difficult to confirm this. It is difficult to control the quality of such materials. This issue, even among herbal experts, is that the use of certain herbal extracts is restricted. He was.

[0021] Another problem is that the plants used in the practice of herbal food additives are often not available locally. Therefore, the fact that it needs to be obtained from a supply source far from the end user However, the supply of such plants from remote locations is particularly difficult for many such plants. Because there is no detailed monograph including identifiability and quality standards, it may be irregular and inaccurate. It is possible. The complex mixture of components found in medicinal plants depends on the plant source, the place where the plant grows, Other plants and microorganisms growing nearby, the time of year when the plants are harvested, and the storage and processing of the materials. Depending on many factors such as the conditions under which it is extracted and the extraction procedure used, the type and concentration can vary greatly. different.

[0022] Therefore, we profile herbal products and medicinal plant materials that may be related to their activity. A highly sensitive process is needed to establish standard specifications for herbal food additives. This enables quality control in the manufacturing process, and enables the production of known or potentially active components. It allows for ideal quantification of minutes.

[0023] The inventors have found that the plant distribution of specific 4-hydroxymethyl-N-methyl-proline varies. We discovered a correlation with jujube plants and guarana, which are used to treat certain diseases. It is of great commercial and medical importance, and is therapeutically active 4-hydroxymethyl-N-methyl This enables the synthesis and / or isolation of a new family of til-proline for the first time, and standard specifications This enables the manufacture of improved herbal medicines that meet certain criteria, and a series of therapeutic treatments based on them. [Overview of the project]

[0024] This disclosure includes the following [1] through

[43] . [1] [ka] Alternatively, the composition comprising isolated 4-hydroxymethyl-proline selected from those pharmaceutically acceptable salts or derivatives, wherein R represents an optionally substituted C1-6 alkyl, C1-6 alkenyl, or C1-6 alkynyl. [2] The above 4-hydroxymethyl-proline is [ka] The composition according to [1] above, selected from a pharmaceutically acceptable salt or derivative thereof. [3] The composition according to [1] or [2] above, wherein the isolated 4-hydroxymethyl-proline is synthetic or purified from a plant source selected from (a) plants of the genus Ziziphus and (b) plants of the genus Guarana. [4] The composition according to [3] above, wherein (a) the plant source is selected from plant species selected from Z. jujuba, Z. spina-christi, Z. lotus, Z. mauritiana, and Z. joazeiro, or (b) the plant source is selected from plant species of Paullinia cupana. [5] The composition according to any one of the above [2] to [4], wherein the plant source comprises fruit, fruit part, fruit extract, fruit juice, seeds, bark, root, and / or leaves. [6] A pharmaceutical composition comprising the composition described in any one of the preceding items [1] to [5] above, and a pharmaceutically acceptable excipient. [7] The pharmaceutical composition described in [6] above, in the form of (a) a pharmaceutical pack, kit or patient pack, or (b) a unit dosage form. [8] A composition according to any one of the preceding items [1] to [7] for use in the treatment or prevention. [9] A composition according to any one of the preceding items [1] to [8] for use in a method of treating energy-utilizing diseases.

[10] The above energy utilization disorders include (a) homeostasis disorders; (b) metabolic disorders; (c) glucose metabolism dysfunction; (d) appetite disorders; (e) insulin resistance; (f) diabetes mellitus (e.g., type 1 or type 2 diabetes); (g) prediabetes; (h) metabolic syndrome; (i) obesity; (j) wasting syndromes (e.g., cancer-related cachexia); (k) muscle disorders; (l) gastrointestinal disorders; (m) growth retardation; (n) hypercholesterolemia; (o) atherosclerosis; (p) age-related metabolic dysfunction; (q) hyperglycemia; (r) impaired glucose tolerance; (s) hyperinsulinemia; (t) diabetes mellitus; (u) metabolic acidosis; (v) cataracts; (w) diabetic neuropathy; (x) diabetic nephropathy; (y A composition for use as described in [8] above, selected from (z) diabetic retinopathy; (aa) macular degeneration; (bb) glomerulosclerosis; (cc) diabetic cardiomyopathy; (dd) glucose metabolism disorder; (ee) arthritis; (ff) hyperlipidemia; (gg) osteoporosis; (hh) osteopenia; (ii) bone loss; (jj) brittle bone syndrome; (kk) acute coronary syndrome; (ll) infertility; (mm) short bowel syndrome; (nn) chronic fatigue; (oo) eating disorders; (pp) bowel motility disorder; (qq) glucose metabolism disorder; (rr) fatty liver; (ss) polycystic ovary syndrome; (tt) hemochromatosis; and (uu) acanthosis nigricans.

[11] The composition according to any one of the above [1] to [8] for use in a method of treating an inflammatory disorder.

[12] The above 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) graft rejection; (i) vasculitis; (j) asthma; (k) acne; (l) osteoarthritis; (m) inflammation of the oral mucosa and gastrointestinal tract; (n) inflammation of the eye; (o) nose (p) inflammation of the ear; (q) steroid-responsive inflammatory diseases; (r) inflammatory diseases of the skin (e.g., actinic keratosis, acne vulgaris, acne comedones, acne rosacea, and acne nodularis and cystic acne, allergic contact dermatitis, angioedema, bullous pemifigoids, cutaneous drug reactions, erythema multiforme, scleroderma erythematous, photodermatitis, psoriatic arthritis, scleroderma and urticaria, psoriasis, dermatitis, atopic dermatitis, scleroderma, steroid-responsive dermatitis) (s) skin conditions associated with symptomatic disorders, urinary tract pruritus, radiation, chemotherapy, and exposure to environmental irritants; (s) inflammatory autoimmune diseases (e.g., ankylosing spondylitis, Crohn's disease, ulcerative colitis, Alzheimer's disease, multiple sclerosis, motor neuron disorders, 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) arthritis; (y) graft-versus-host disease; (z) chronic pelvic inflammatory disease; (a') endometriosis; (b') chronic hepatitis; (c') tuberculosis; and (d') inflammation of the skin, e.g., caused by exposure to sunlight, allergens, irritants, or burns.

[13] The composition for use described in

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

[14] The composition for use as described in

[13] above, wherein the autoimmune disease is selected from Graves' disease; rheumatoid arthritis; Hashimoto's thyroiditis; vitiligo; diabetes mellitus (e.g., type 1 diabetes or type 2 diabetes); pernicious anemia; multiple sclerosis; glomerulonephritis; systemic lupus erythematosus 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).

[15] The composition for use as described in

[13] above, wherein the inflammatory disease 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; enteropathic arthritis; reactive arthritis and arthritis associated with inflammatory bowel disease.

[16] The composition for use described in

[13] above, wherein the allergy described above is selected from atopic allergy, allergic rhinitis, allergic conjunctivitis, atopic dermatitis, hypereosinophilia, irritant bowel syndrome, allergen-induced migraine, bacterial allergy, bronchial allergy (asthma), contact allergy (dermatitis), delayed allergy, pollen allergy (hay fever), drug allergy, sting allergy, bite allergy, gastrointestinal allergy; food allergy; and physical allergy, such as cold urticaria, angioedema, cholinergic urticaria and photosensitivity.

[17] A composition according to any one of the above [1] to [8] for use in a method of treating neoplasms.

[18] The composition for use described in

[17] above, wherein the neoplasm is selected from benign, precancerous and malignant neoplasms, hyperplasia, metaplasia and dysplasia.

[19] The composition for use described in

[17] above, wherein the neoplasm is a malignant neoplasm (cancer).

[20] The composition for use described in

[19] above, wherein the malignant neoplasm is selected from (a) carcinoma; (b) blastoma; (c) leukemia; (d) lymphoma; (e) myeloma; (f) sarcoma; and (g) mixed-type cancer.

[21] The composition for use according to

[20] above, wherein the malignant neoplasm is a carcinoma selected from carcinomas of the bladder, breast (e.g., primary breast tumor, lymph node-negative breast cancer, invasive tubular adenocarcinoma of the breast 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 cancer), stomach, cervix, thyroid, prostate, gastrointestinal system (e.g., gastrointestinal stromal tumor), or skin (e.g., squamous cell carcinoma).

[22] The composition according to

[20] , wherein the malignant neoplasm is a lymphoid leukemia, for example, a leukemia selected from progenitor cell leukemia, mature B-cell leukemia, mature T-cell leukemia and NK-cell leukemia, acute myeloid leukemia, chronic myeloproliferative disorders and myelodysplastic syndromes.

[23] The composition for use according to

[20] above, wherein the malignant neoplasm is (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, such as mantle cell lymphoma.

[24] The composition for use according to

[20] above, 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.

[25] A composition according to any one of the above [1] to [8] for use in a method of treating a bacterial or viral infection.

[26] The composition for use according to

[25] above, wherein the method comprises inhibiting the growth of symbiotic bacteria and / or pathogenic bacteria in vivo.

[27] The composition for use according to

[25] above, wherein the method comprises disrupting host-bacterial cell interactions and / or inhibiting or eliminating bacterial biofilm formation in a mammalian (e.g., human) host.

[28] The composition for use according to

[26] or

[27] above, wherein the method comprises the treatment or prevention of diseases and disorders mediated or characterized by the presence of bacterial biofilms (e.g., subgingival plaque biofilms and fascial biofilms).

[29] A composition according to any one of the above [1] to [8] for use in a method of treating periodontal disease, bacterial vaginosis, and / or diseases caused by infection by Tannerella forsythia, Tannerella denticola, Porphyromonas gingivalis, or Gardnerella vaginalis.

[30] A composition according to any one of the above [1] to [8] for use in a method of treating atheromatosis, for example, atherosclerosis.

[31] The composition according to any one of the above [1] to [8] for use in a method for regulating the growth of symbiotic bacteria in a mammalian host.

[32] The composition for use according to

[31] above, wherein the method comprises modulating the composition of symbiotic bacteria in a mammalian host, such as a human host.

[33] The composition according to any one of the above [1-8] for use in a method of controlling blood glucose in a mammal, for example, a human subject.

[34] The method according to any one of the above items [9] to

[33] , comprising administering an effective amount of the composition described in any one of the above items [1] to [8].

[35] Use of the compositions described in any one of the above [1] to [8] for the treatment of a disease or for use in the method described in any one of the above [9] to

[33] , for the manufacture of a pharmaceutical product.

[36] A cosmetic, nutritional supplement, herbal, or pharmaceutical composition comprising the composition described in any one of the above items [1] to [8], and optionally further comprising a cosmetic, nutritional supplement, or pharmaceutically acceptable excipient or carrier.

[37] A cosmetic method for reducing swelling or erythema of the skin, comprising administering to a subject, for example, by topical application to the skin, one of the compositions described in any one of the above paragraphs [1] to [8] or

[36] .

[38] The composition, composition for use, method, or use described in any one of the preceding [1] to

[37] , wherein the isolated 4-hydroxymethyl-proline is present in the composition at a level of at least 0.5%, 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, or 99% w / w (on a dry weight basis).

[39] A process for producing any one of the compositions described in [1] to

[38] above, (a) A step of providing plant material from any one of the plant sources described in any one of the above [2] to [5], (b) the step of extracting 4-hydroxymethyl-proline as described in [1] above from the plant material, and then (c) The step of formulating the extracted 4-hydroxymethyl-proline together with a pharmaceutically acceptable excipient to produce a pharmaceutical composition, The above process, including.

[40] A process for producing a pharmaceutical composition, herbal medicine, or dietary supplement, comprising the step of monitoring the quality of the pharmaceutical composition, herbal medicine, or dietary supplement by detecting the presence or absence of 4-hydroxymethylproline described in [1] above, or by measuring its amount, in a sample of the pharmaceutical composition, herbal medicine, or dietary supplement.

[41] A method for monitoring the quality of a pharmaceutical composition, herbal medicine, or nutritional supplement, (a) A step of providing a sample of a pharmaceutical composition, herbal medicine, or nutritional supplement, (b) The step of detecting the presence or absence of 4-hydroxymethylproline described in [1] above in the sample, or measuring its amount; The above method, including.

[42] A process for producing a supplemented food or beverage, comprising (a) providing a composition described in any one of the preceding items [1] to

[41] , and (b) adding the composition from step (a) to a food or beverage to produce a supplemented food or beverage.

[43] A method for selecting a breeding line and / or variety of a plant as described in either paragraph [3] or [4] above, (a) A step of providing a material which is derived from the plant described in [3] or [4] above, for example, the plant part described in [5] above, (b) The step of determining the presence or absence of 4-hydroxymethylproline described in [1] or [2] above in the material of step (a) above, and / or measuring the amount thereof, The above method, including. This invention is based, at least in part, on the remarkable discovery that the plant distribution of certain 4-hydroxymethyl-N-methyl-proline compounds correlates with medicinal plants used to treat various diseases. Thus, the family of 4-hydroxymethyl-N-methyl-proline compounds has been identified for the first time as important bioactive components of established herbal medicines.

[0025] Therefore, according to the present invention, isolated 4-hydroxymethyl- selected from the following Proline: [ka] Alternatively, a composition comprising pharmaceutically acceptable salts or derivatives thereof is provided, where, R is optionally substituted with a C1-6 alkyl, C1-6 alkenyl, or C1-6 a This represents Lukinil.

[0026] In a preferred embodiment, 4-hydroxymethyl-proline is [ka] or selected from pharmaceutically acceptable salts or derivatives thereof.

[0027] Other embodiments of the present invention are defined in the claims appended herein.

[0028] The specific 4-hydroxymethyl-N-methyl-proline described herein is novel. According to the present invention, the novel 4-hydroxymethyl-N-methylproline itself is , the preparation processes thereof, compositions containing them, and those as pharmaceuticals and drug products. Provided in conjunction with the use of such 4-hydroxymethyl-N-methyl as described herein. Some of the chyl-prolines, as far as is known, are recognized as having biological activity. Therefore, it is not claimed to be a pharmaceutical product in itself.

[0029] Embodiments of herb quality monitoring according to the present invention According to another aspect of the present invention, one or more 4-hydroxymethyl- To detect the presence or absence of N-methylproline, or to measure its amount. Therefore, a process for manufacturing crude drugs, which includes a step of monitoring the quality of the crude drugs, It will be provided.

[0030] In another embodiment, the present invention relates to a method for monitoring the quality of crude drugs, comprising the following steps. The method provides: (a) the step of providing a sample of crude drug; and (b) the step of providing the sample of the crude drug The presence or absence of one or more 4-hydroxymethyl-N-methylproline in the substance is detected. A step of dispensing or measuring the amount.

[0031] In this context, the term quality refers to the overall suitability of the herb for its intended use. Used to define compatibility, for example, one or more 4-hydroxymethyl This may include the presence or absence of til-N-methyl-proline (at appropriate concentrations), which means that Depending on the use, condition, purity, and undesirable supplemental ingredients and / or contaminants of a particular source It indicates an acceptable or unacceptable level of contamination.

[0032] In a further embodiment, the present invention relates to a supplemented food or beverage comprising the following steps. It provides a process for manufacturing [the product]. (a) Provide plant parts of species of the genus Jujube and / or Paullinia cupana. Steps; (b) a step of monitoring the quality of the plant part by the method of the present invention; and (c) Adding a plant part (or one or more fractions thereof) to food or beverage to supplement A step in manufacturing a food or beverage.

[0033] This aspect of the present invention has broad utility in the manufacture of any supplemented food or beverage. Headings may be used for any food or beverage, including refrigerated and hot foods and beverages. Foods and beverages, sweet foods and beverages, carbonated drinks, alcoholic beverages and non-alcoholic beverages It includes.

[0034] The process and method of the present invention preferably involves one or more of the samples of herbal food additives. The presence or absence of the above 4-hydroxymethyl-N-methylproline is detected, or The method further includes the step of measuring the amount.

[0035] The present invention allows for the detection of the presence of one or more 4-hydroxymethyl-N-methyl-proline lines. In the embodiment, one or more 4-hydroxymethyl-proline selected from the following are , [ka] [In the formula, R is optionally substituted with a C1-6 alkyl, C1-6 alkenyl, or C A process involving assay, detection, or monitoring of 1-6 alkynyls is preferred. .

[0036] The present invention allows for the detection of the presence of one or more 4-hydroxymethyl-N-methyl-proline lines. In other embodiments, one or more 4-hydroxymethylproly selected from the following: N, [ka] A process involving assay, detection, or monitoring is preferred.

[0037] The present invention also aims to provide crude drugs obtained by the methods and processes of the present invention. [Brief explanation of the drawing]

[0038] [Figure 1] Figure 1 shows the effect of compound 1 (μM) on LPS-stimulated TNF-α production in human blood. [Figure 2] Figure 2 shows the effect of extracts from jujube plants on blood glucose levels. [Figure 3] Figure 3 shows the effect of extracts from jujube plants on blood glucose levels. [Figure 4] Figure 4 shows the effect of extracts from jujube plants on blood glucose levels. [Figure 5] Figure 5 shows the effect of extracts from jujube plants on blood glucose levels. [Modes for carrying out the invention]

[0039] All publications, patents, patent applications, and other references referred to herein are the individual publications. It is specifically and individually indicated that a work, patent, or patent application is incorporated by reference, The entire thing is by reference for any purpose as if its contents were completely enumerated. This specification is incorporated herein.

[0040] definition As used herein, unless otherwise indicated, the following terms are used in the same sense as they are used in other contexts. In addition to the broader (or narrower) meanings that may be enjoyed in the technical field, the following meanings It is intended to have

[0041] Unless otherwise specified in the context, the use of the singular form here is to be interpreted as including the plural form, and The reverse is also true. The terms "a" or "an" used in relation to an entity are... It is interpreted as referring to one or more entities. In such cases, the term "a" (or "an") "One or more" and "at least one" are used interchangeably in this specification.

[0042] When used herein, the term "includes" or "includes" or "including" Such variations include any enumerated whole (integer) (e.g., features, elements, Characteristics, features, steps in a method / process, or limitations) or complete entities (e.g., features, Read to indicate the inclusion of a group of elements, characteristics, properties, steps in a method / process, or limitations. This should be included, but it does not exclude any other perfect field or group of perfect fields. Therefore, this When used in a specification, the term "includes" can mean both ends or open ends. Yes, and do not exclude additional, unlisted, complete or method / process steps.

[0043] The phrase "essentially consisting of" in this specification refers to a specific complete entity (plural). or steps, and not substantially affect the characteristics or functions of the claimed invention It is used when potatoes are needed.

[0044] When used herein, the term "consisting of" means the enumerated complete entities (e.g., For example, a feature, element, characteristic, property, step or limitation of a method / process) or a complete body (For example, only groups of features, elements, characteristics, properties, steps or limitations of a method / process) It is used to indicate existence.

[0045] As used herein, the term “disease” means a condition that impairs physiological function and is characterized by a specific symptom. It is used to define any abnormal condition related to the state. This term refers to the nature of the pathogenesis ( Regardless of whether the etiological basis of the disease has actually been established, physiological function To broadly encompass impaired disabilities, illnesses, abnormalities, conditions, diseases, states, or syndromes It is used in cases of infection, trauma, injury, surgery, radiation ablation, poisoning, or malnutrition. This includes conditions arising from scarcity.

[0046] When used herein, the terms “treatment” or “to treat” refer to the symptoms of a disease. To cure, improve, or alleviate the condition, or the cause(s) (e.g., pathological multiplicity) This refers to an intervention (for example, administering medication to the subject) that eliminates (or reduces the impact of) a condition. In this case, this term is used synonymously with the term "therapy."

[0047] Furthermore, the terms "treatment" or "to treat" imply the onset or progression of a disease. To prevent or delay the disease, or to reduce (or eradicate) its incidence within the treated population. This refers to an intervention (for example, administering medication to the subject) that aims to prevent It is used synonymously with the term "".

[0048] The term "subject" (where the context allows, "individual," "animal," "patient," or "mammalian") (Should be read as including "thing") refers to any subject requiring treatment, especially suckling animals Define the object. Mammal objects include humans, domesticated animals, livestock, zoo animals, and sports This includes, but is not limited to, live animals and pet animals. In a preferred embodiment, Elephants are humans.

[0049] As used herein, the term “energy utilization disorder” refers to an abnormal energy This term encompasses any disease or disorder resulting from use. Therefore, this term refers to homeostatic disorders. This includes harm and disease, metabolic disorders, glucose metabolism dysfunction, and appetite disorders. This term includes insulin resistance, various forms of diabetes, and metabolic syndrome. Obesity, wasting syndrome (e.g., cancer-related cachexia), muscle disorders, gastrointestinal disorders, growth retardation, high blood pressure This includes lesterolemia, atherosclerosis, and age-related metabolic dysfunction. The term also includes conditions associated with metabolic syndrome, obesity, and / or diabetes. Including, for example, hyperglycemia, impaired glucose tolerance, hyperinsulinemia, diabetes, metabolic acidosis, white blood sugar Internal organ failure, diabetic neuropathy, diabetic nephropathy, diabetic retinopathy, macular degeneration, glomerular sclerosis Diabetic cardiomyopathy, insulin resistance, glucose metabolism disorders, arthritis, hypertension, hyperlipidemia bloodemia, osteoporosis, osteopenia, bone loss, brittle bone syndrome, acute coronary syndrome, infertility, short bowel syndrome This group includes chronic fatigue, eating disorders, and bowel motility disorders.

[0050] The term "metabolic syndrome" as used herein refers to a condition characterized by three or more of the following symptoms: Used to define a condition characterized by its presence: central obesity (101 in men). 6 centimeters (40 inches) or more; for women, 88.9 centimeters (35 inches) or more. Waist measurement of 150 mg / dL or higher); high levels of triglycerides (150 mg / dL or higher); low levels Bell's HDL (less than 40 mg / dL for men, less than 50 mg / dL for women), and hypertension Pressure (130 / 85 mmHg or higher).

[0051] Therefore, this term follows the World Health Organization's definition of metabolic syndrome. The following conditions are included, as defined by this: (a) Fasting plasma glucose levels exceeding 6.1 mmol / L (b) blood pressure greater than 140 / 90 mmHg; and (c) one or more of the following: (i) Plasma triglycerides greater than 1.7 mmol / L; (ii) 0.9 and 1.0 mmol / L HDL less than L (for men and women, respectively); (iii) 30 kg / m 2 A body that surpasses Case index.

[0052] References to the treatment of metabolic syndrome in this specification refer specifically to obesity (e.g., middle-aged). Cardiac obesity, as well as elevated serum triglycerides and diabetes (type 1 and type 2 diabetes and insulin Any of the disorders associated with metabolic syndrome, including phosphorus resistance, or This should be interpreted as including all treatments.

[0053] References in this specification to the treatment of type 1 or type 2 diabetes refer to type 1 and type 2 diabetes itself. This should be interpreted as including treatment for prediabetes (early-stage diabetes) and insulin resistance. ru.

[0054] The terms "prediabetes" or "early-stage diabetes" refer to elevated levels in individuals without diabetes. This defines the state in which glucose or glycosylated hemoglobin is present.

[0055] When used herein, the effective amount of a compound or composition is a reasonable benefit / risk ratio. Appropriately, without excessive toxicity, irritation, allergic reactions, or other problems or complications. The desired effect that can be administered to elephants is, for example, a permanent or temporary improvement of the condition in question. This defines a sufficient amount to provide treatment or prevention for the symptoms that appear. This amount depends on the individual's age and The results may vary among subjects depending on their overall condition, administration mode, and other factors. Therefore, it is impossible to specify the exact effective amount, but those skilled in the art can determine it through routine experiments and backtesting. Using general knowledge of grounds, determine the appropriate "effective" amount in each individual case. This may be possible. In this context, treatment outcomes include the complete elimination or reduction of symptoms, pain or discomfort. These include reduced discomfort, extended survival time, improved mobility, and other markers of clinical improvement. The treatment outcome does not need to be a complete cure.

[0056] As used herein, the term “medicine kit” is used in all cases, at the discretion of the user. The side packaging contains a means of administration (e.g., a measuring device) and / or a means of delivery (e.g., an inhaler). Defines various pharmaceutical compositions of one or more unit doses, accompanied by an injector or syringe. In a pharmaceutical kit containing the above compound / drug combination, each compound / drug is a single It may be one or more formulations. The unit dose(s) are contained in the blister pack. The drug kit may optionally include instructions for use.

[0057] As used herein, the term “medicine pack” may optionally refer to a common outer packaging. Defines various pharmaceutical compositions in one or more unit doses contained within the package. Two or more compounds In a pharmaceutical pack containing a combination of drugs, the individual compounds / drugs may be single or non-single. It may be a single formulation. The unit dose(s) may be contained in a blister pack. The medication pack may optionally include an instruction manual.

[0058] As used herein, the term “patient pack” refers to a medical pack for the entire course of treatment. Define the package prescribed to the patient, which contains the composition. A patient pack typically contains one or more components. It includes blister packs (multiple packs are possible). The patient pack is used by the pharmacist to prepare the patient's medication. It is superior to the conventional prescription of splitting the supply from the bulk supply, and the patient is always, usually the patient You can access the package insert included in the patient pack, which is not included in the prescription. This has been shown to improve patient compliance with physician's instructions.

[0059] The term "pharmaceutically acceptable derivative" as applied to the compounds of the present invention refers to the parental compounds of the present invention. This defines compounds obtained (or that can be obtained) by the chemical derivatization of substances. Therefore, pharmaceutically acceptable derivatives are those that do not cause excessive toxicity, irritation, or allergic reactions. (That is, administration to or contact with human tissue that is commensurate with a reasonable benefit / risk ratio) Suitable for use. Preferred derivatives are alkylation, esterification, or acidation of the parent compound. This is obtained (or can be obtained) by the chemical reaction. Therefore, the chemical reaction of the present invention pharmaceutically acceptable derivatives of the compound include N-oxides and their esters.

[0060] The pharmaceutically acceptable derivatives of the present invention possess some or all of the biological activities described herein. It can be retained. In some cases, biological activity is increased by derivatization. Derivatives are prodra It may act as a bug, and one or more of the biological activities described herein are inhibitory. This can only occur after voltification. Particularly preferred prodrugs are those with one or more free hydroxyls. It is an ester derivative that is esterified and activated by in vivo hydrolysis. Conductivity also affects other biological activities of the compound, such as bioavailability and / or globulin activity. The lycosidase inhibition profile may be enhanced. For example, derivatization can enhance CNS penetration (e.g., It may increase the penetration of the blood-brain barrier.

[0061] The term "pharmaceutically acceptable salt" means that a salt is pharmaceutically acceptable to humans without excessive toxicity, irritation, or allergic reactions. Suitable for use in contact with the tissues of lower animals, and the benefit / risk ratio is reasonable. Defines non-toxic organic or inorganic acid addition salts of free bases. Appropriate pharmaceutically acceptable salts This is well known in the relevant technical field. Examples include inorganic acids (e.g., hydrochloric acid, hydrobromic acid, Sulfuric acid and phosphoric acid), organic carboxylic acids (e.g., acetic acid, propionic acid, glycolic acid, milk Acids, pyruvic acid, malonic acid, succinic acid, fumaric acid, malic acid, tartaric acid, citric acid, ascorbic acid Rubic acid, maleic acid, hydroxymaleic acid, dihydroxymaleic acid, benzoic acid, f Phenylacetic acid, 4-aminobenzoic acid, 4-hydroxybenzoic acid, anthranilic acid, cinnamic acid, (Salicylic acid, 2-phenoxybenzoic acid, 2-acetoxybenzoic acid, and mandelic acid) Salts of organic sulfonic acids (e.g., methanesulfonic acid and p-toluenesulfonic acid) are also mentioned. It can be done.

[0062] These salts and free base compounds exist either in a hydrated form or substantially anhydrous form. It may exist. Crystal forms including all polymorphic forms of the compound of the present invention are also intended, and generally, chemical The acid addition salts of the compound are soluble in water and various hydrophilic organic solvents, and their free base forms and In comparison, it exhibits a higher melting point and increased solubility.

[0063] In this specification, the term "alkyl" refers to a linear or branched saturated hydrocarbon chain. Definition: The term "C1-C6 alkyl" refers to a linear molecule having 1 to 6 carbon atoms. This refers to branched saturated hydrocarbon chains. Examples include methyl, ethyl, n-propyl, and ethyl. Examples include sopropyl, t-butyl, and n-hexyl. The alkyl group of the present invention is optional. Therefore, it can be substituted with one or more halogen atoms.

[0064] In this specification, the term "alkenyl" means a compound containing at least one carbon-carbon double bond. Defines linear or branched hydrocarbon chains. The term "C1-C6 alkenyl" is used. This refers to a linear or branched unsaturated hydrocarbon chain having 1 to 6 carbon atoms. Examples include ethenyl, 2-propenyl, and 3-hexenyl. The Kenyl group can be optionally substituted with one or more halogen atoms.

[0065] In this specification, the term "alkynyl" means a compound containing at least one carbon-carbon triple bond. Defines linear or branched hydrocarbon chains. The term "C1-C6 alkynyl" is used. This refers to a linear or branched unsaturated hydrocarbon chain having 1 to 6 carbon atoms. Examples include ethinyl, 2-propynyl, and 3-hexynyl. The quinyl group can be optionally substituted with one or more halogen atoms.

[0066] The term phytochemicals encompasses any chemical components of plants, including macromolecules and small molecules. Therefore, it is used in a broad sense in this specification. An important example is alkaloids (for example) For example, imino sugars and imino sugar acids, such as the structural classes pyrrolidine, piperidine, and pyrrolidine. (Selected from gin, indolizidine, tropane, and nortropane), carbohydrate analogs phenol compounds, terpenoids, enzyme inhibitors, glycosides, nucleotides, amino acids Lipids and sugars are examples.

[0067] When applied to the compounds of the present invention, the term isolated refers to the naturally occurring rings of the compound. It exists in a physical environment different from the boundary (or, in the case of synthetic compounds, is purified to some extent). This is used herein to indicate that an isolated compound is present in nature. For example, an isolated compound is present in nature. For the complex cellular environment (or the starting products, intermediates, and slow processes involved in its synthesis) (For some or all of the cinder, solvent, reactants and / or by-products), substantially isolated ( For example, it can be purified. Therefore, the isolated compound can be used as a plant source as described herein. It can take the form of any of the concentrated fractions or extracts.

[0068] When an isolated material (e.g., a synthetic, non-naturally occurring compound) is purified, the purity is... The absolute level is not important; a person skilled in the art can determine an appropriate level of purity depending on the application in which the material will be used. The degree can be easily determined. In the case of synthetic materials, the purity level is 85-99% w / w. It is within the range and may exceed 99% w / w. However, especially if the material is not from a natural source If separated, 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 A purity level of 0.9% w / w or 2.0% w / w is preferred.

[0069] Particularly preferable is at least 0.5-2.0% w / w, for example, at least 0.8-1 This represents a purity level of 0.5% w / w, for example, at least about 1.0% w / w. By using appropriate concentration techniques such as ion exchange chromatography, the material can be naturally concentrated. When isolated from the source, levels of 5-10% w / w can be easily obtained.

[0070] Depending on the circumstances, isolated compounds may be cruder in composition (for example, containing many other substances). (or a less coarse extract) or forms part of a buffer system, which is, for example, It may contain other components. In other situations, isolated compounds may be measured spectrophotometrically, for example, by NMR. Alternatively, it can be determined by chromatography (e.g., GC-MS of trimethylsilyl derivatives). When defined, it can be refined to an essential level of uniformity.

[0071] The term "crude drug" as used herein means a drug in which at least one active ingredient (e.g., a compound) is chemically transformed. To define pharmaceutical compositions that are not scientifically synthesized, but rather consist of phytochemical components of plants It is used. In most cases, this non-synthetic active ingredient is not isolated (as defined here). It is present (as is the case) along with other phytochemicals associated with the source plant. However, in some cases, plant-derived bioactive components may be included in the concentrated fraction. They may be isolated (often involving high levels of purification). However, in many cases, they are raw. The medicine is a coarser or less coarser extract, infusion, or fraction of the plant, or unprocessed. This includes the entire plant (or part thereof) of the treatment, but in such cases, the plant (or part of the plant) ) are usually at least dried and / or ground. Herbal medicines are available in the form of dietary supplements and beverages. Alternatively, it may be provided as a herbal medicine kit or herbal medicine pack in multiple single doses.

[0072] The term "herbal food" as used herein refers to a food in which at least one ingredient is chemically synthesized. It is not used to define the phytochemical components of plants, but rather to define the compositions of plants. In this case, this non-synthetic component is not purified, but other plant compounds associated with the source plant are present. It exists together with chemical substances. However, in some cases, plant-derived components (multiple) are present. It may be contained in the concentrated fraction or (in some cases, isolated in high purity). However, in many cases, herbal food additives are coarser or coarser than the plant. It does not contain extracts, decoctions or fractions, or the whole (or part thereof) of the untreated plant. In such cases, the plant (or part of the plant) is usually at least dried and / or crushed. Therefore, this term includes additives and supplements used in food and beverages. It contains herbal foods in various forms.

[0073] The term "bioactive ingredient" as used herein means that it is necessary or sufficient for the medicinal effect of the herbal medicine containing it. Used to define a phytochemical that is a component. In the present invention, it is a bioactive component (multiple). (OK) contains one or more of the 4-hydroxymethyl-N-methyl-proline compounds of the present invention. nothing.

[0074] The term "nutritional supplement" is used herein to mean a food that provides physiological benefits or treats a disease. Used to define foods (or their isolates) that protect against [unclear]. Preferred nutrition of the present invention Nutritional supplements possess blood glucose regulating activity and are found to have applications in the treatment of energy utilization disorders. Other nutritional supplements of the present invention are anti-inflammatory.

[0075] The term "standard specification" as used herein refers to the acceptable specifications for herbal medicines, cosmetics, or dietary supplements. Used to define properties or phytochemical profiles that correlate with quality. In this context, the term quality defines the overall suitability of a product for its intended use. Used to provide one or more of the present invention at appropriate concentrations This includes the presence of chill-proline.

[0076] The term "phytochemical profile" in this specification refers to a profile related to different phytochemical components. It is used to define the characteristics of a chain.

[0077] In its broadest aspect, the present invention relates to the 4-hydroxymethyl-N-methyl- We intend to explore all optical isomers, racemic forms, and diastereomers of proline. Therefore, the reference to 4-hydroxymethyl-N-methyl-proline in this invention is to diastereoma - as a mixture, as individual diastereomers, as a mixture of enantiomers, Furthermore, 4-hydroxymethyl-N-methyl-proline in the form of individual enantiomers To include.

[0078] chemical synthesis The 4-hydroxymethyl-N-methyl-proline described herein is obtained by conventional methods. It can be prepared by [method]. Methods for preparing heteroaromatic ring systems are well known in the art. In particular, the synthesis method is Comprehensive Heterocyclic Chemistry, Vol. 1 (Eds.: AR Katritzky, CW Rees), Pergamon Press, Oxford, 1984, and Comprehensive Heterocyclic Chemistry II : A Review of the Literature 1982-1995 T he Structure, Reactions, Synthesis, and Uses of Heterocyclic Compounds, Alan R. Katritzky (Editor), Charles W. Rees (Editor) tor), EFV Scriven (Editor), Pergamon It is described in Pr, June 1996. Others that support the synthesis of the target compound Common resources include March's Advanced Organic Chem. istry: Reactions, Mechanisms, and Structs ure, Wiley-Interscience;5th edition (Jan This includes (July 15, 2001).

[0079] Extraction from plant sources / detection there The 4-hydroxymethyl-N-methyl-proline of the present invention as described herein is a naturally supplied material. It can be isolated from its source.

[0080] For example, the compounds of the present invention are extracted and / or purified from plant sources selected from the following. It is possible to: (a) plants of the genus Ziziphus; and (b) plants of the genus Guarana. In a preferred embodiment The plant source will be selected from the following plant species: Z. jujuba, Z. . spina-christi, Z. lotus, Z. mauritiana, Z . joazeiro, and P. cupana.

[0081] The plant material from the above-mentioned plant sources is used in accordance with the present invention to produce 4-hydroxymethyl It can be used as a starting material for the isolation and purification of tyl-N-methyl-proline. The 4-hydroxymethyl-N-methyl-proline of the present invention is water-soluble and has an anionic interaction. It can be concentrated by cation exchange chromatography or cation exchange chromatography. It is possible. They can also be concentrated using size exclusion. Therefore, those skilled in the art Using standard techniques, the 4-hydroxymethyl-N-methyl-proline of the present invention is used It will be understood that it can be easily purified and isolated.

[0082] Any suitable part of the plant source can be used, and in a preferred embodiment, the source This includes or is essentially derived from plant parts selected from fruits, seeds, and leaves. It is possible to use other parts such as roots, stems, and bark.

[0083] Processes suitable for extraction and / or purification include processes that include the following steps: ru: (a) Step of providing materials, for example, plant parts selected from: plant source From, fruit, fruit parts, fruit extracts, fruit juice, seeds, bark, roots and / or leaves; (b) Extract the first sample of the material with a polar solvent to produce a polar extract and a non-polar residue. Steps to achieve; (c) The polar extract from step (b) is subjected to ion exchange chromatography to obtain ion A step of producing an extract concentrated with ionic compounds and nonionic residues; (d) The concentrated extract from step (c) is fractionated by chromatography to obtain the ionic compounds The step of concentrating the substance to produce one or more polar fractions containing one or more compounds of the present invention. .

[0084] Methods of quality control for herbs sample The herb samples used in the method of the present invention are dried plant materials or are used in food and beverages. It may be an aliquot of herbal food additive in the form added to. Alternatively, the sample is Before characterization, the samples can be pre-treated using one of several methods. Pre-treatment can be physical Pretreatment by photochemical means, e.g., powdering, grinding, freezing, evaporation, filtration, pressing, spray drying. This may include extrusion, supercritical solvent extraction, and tincture production.

[0085] Preferably, food additive samples are fractionated before characterization. Solvent extraction (multiple fractions possible) Any suitable sorting method can be used, including the sample. In a preferred embodiment, The compound is fractionated by ion exchange chromatography to separate the polar compound from the nonpolar residue. A concentrated extract is produced. In such embodiments, characterization is preferably performed by air This includes liquid chromatography (GC), such as GC-MS. When using GC, concentration is required. The extracted material is derivatized before chromatography.

[0086] When herbal food additives are managed or sold in the form of the whole plant (or part thereof), The material may be dried before use. Any method including freeze-drying, spray-drying, or air-drying may be used. A convenient form of drying can be used.

[0087] Detection of 4-hydroxymethyl-N-methylproline Any appropriate form of characterization of food additive samples can be used, including, Detection of the presence or absence of 4-hydroxymethyl-N-methyl-proline in the sample. or sufficient to measure the amount thereof, functional and / or physical and / or This includes, but is not limited to, chemical characterization.

[0088] If the sample has been physically characterized, the characterization can be selected from the following: (a) plant Quantification of physical chemical components (multiple); and / or measurement of the purity of component (b); and / or ( c) Molecular weight (or, in the case of a fraction containing multiple different phytochemical components, its molecular weight distribution or Determination of its various statistical functions; and / or (d) determination of the molecular formula (for example, (e) by nuclear magnetic resonance; and / or spectral analysis.

[0089] Spectral analysis is particularly preferred, and it is possible to generate any or all of the following spectra. There is a possibility: (a) Mass spectrum (e.g., mass-to-charge ratio (m / z) value versus abundance), and / or (b) Chromatography data (spectrum, column retention time, elution profile) etc.), and / or (c) Photodiode array (PDA) spectrum (e.g., UV range and visible range) Both), and / or (d) Electrochemical detection or evaporation light scattering detection, (e) Nuclear magnetic resonance (NMR) spectrum (for example, 1 H and / or 13 C NMR (Spectral dataset obtained via [method]).

[0090] When used in accordance with the present invention, spectral analysis is performed, for example, by GC-MS and / or It can be combined with fractionation and / or derivatization of the sample by using HPLC-PDA-MS. It can be combined.

[0091] Particularly preferred is the use of GC-MS to detect the presence or absence of 4-hydroxymethyl-N-methyl-proline in the sample or to measure its amount. Or to measure its amount.

[0092] When the sample is chemically characterized, the characterization can be selected from the measurement of the chemical reactivity of the phytochemical(s), the solubility of the phytochemical(s), the stability and melting point of the phytochemical(s), Or any combination thereof. Or any combination thereof.

[0093] When the sample is functionally characterized, the characterization can include, for example, in vivo or in vitro assays, enzyme inhibition assays (such as sialidase inhibition), receptor binding assays, cell Assays (such as cell replication, cell pathogens, cell-cell interactions, and cell secretion assays), immune Assays, antibacterial activity (such as cell binding and / or replication of bacteria and viruses) assays, Immune assays, antibacterial activity (such as cell binding and / or replication of bacteria and viruses) assays, Toxicity assays (such as LD 50 Assays) or any combination thereof Selected biological assays.

[0094] Solvent extraction Polar solvents suitable for use in the process of the present invention include, but are not limited to, organic solvents such as organic alcohols. Preferred are ethanol and methanol, as well as Mixtures of ethanol / water or methanol / water. Preferably, the polar solvent is 51 ~80% ethanol / water, 31~50% ethanol / water, and up to 30% ethanol / Water. It is selected from water. Particularly preferred is a polar solvent which is about 50% ethanol / water. Suitable non-polar solvents for use in removing unwanted components in the process of the present invention include organic solvents such as hexane and dichloromethane (DCM) or chloroform but are not limited thereto. Particularly preferred is dichloromethane. The conditions (time, temperature, degree of stirring, etc.) under which extraction(s) is / are carried out can be readily determined empirically and can vary depending on the nature of the sample, the nature of any pretreatment, and the solvent system selected.

[0095] Chromatographic fractions Chromatographic fractions can include gas-liquid chromatography. Gas-liquid chromatography is a process of separating a complex mixture of volatile substances into its components by partitioning the sample between a pressurized inert gas and a thin layer of a non-volatile liquid coated on an inert support within a heated column . To effect good separation of a particular compound in the mixture, it is important to use a column with the correct characteristics. The nature of the solid support, the type and amount of the liquid phase , the packing method, the overall length, and the column temperature are important factors. One of ordinary skill in the art can readily determine appropriate column characteristics by routine trial and error and by using general common knowledge , particularly in view of the nature of the extract under study and the nature of the solvents used in the extraction and the types of chemicals expected in those solvents.

[0096] One of ordinary skill in the art can readily determine appropriate column characteristics by routine trial and error and by using general common knowledge , particularly in view of the nature of the extract under study and the nature of the solvents used in the extraction and the types of chemicals expected in those solvents. One of ordinary skill in the art can readily determine appropriate column characteristics by routine trial and error and by using general common knowledge Particularly preferred and useful in many situations is a capillary column coated with a non-polar liquid phase (25m × 0.22mm id × 0.25μm BPX5 stationary phase, manufactured by SGE Ltd . or equivalent).

[0097] Many compounds are highly polar, low in volatility, and thermally unstable, making them difficult to analyze in gas chromatography. Not suitable for direct injection into graphs. Highly hydroxylated compounds contain intermolecular water. It is difficult to vaporize due to the elementary bond. However, if the hydroxyl hydrogen is replaced by another chemical group... By changing this, it can be made sufficiently volatile for GC analysis. Hydroxyl group is derived. The two most common methods of emulation are acetylation and silylation, and acetylate [CH4] 3CO-OR] or silyl ether, for example, trimethylsilyl (TMS) ether [ A (CH3)3Si-OR is formed. Therefore, the concentrated extract is chromatographic on an analytical scale. In embodiments fractionated by matrix, the phytochemical components of the concentrated extract are preferably, For example, derivatization is performed by acylation or silylation. Particularly preferred is trimeth This involves derivatization with lucilyl (TMS).

[0098] Chromatographic fractionation may also include ion exchange chromatography. Conversion chromatography partially purifies and concentrates ionic species and removes contaminants. The contractors, through daily trial and error and by using general knowledge, Depending on the amount to be separated and fractionated, the extract being studied, and the properties of the solvent used for extraction, The appropriate column packing material and mobile phase(s) can be easily identified. Method of the present invention Particularly preferred in this case is free acid or hydrogen (H + Form or ammonium (NH4 + ) This is a strongly acidic cation exchange resin that can be used in any of the following salt forms. It adsorbs cations from a solution and releases an equal number of counterions into the solution (depending on the form in which it is used). H+ or NH4 + (either of the ions). When used in hydroxide form (OH-), strong basic anion exchange resins are also preferred.

[0099] Characterization of fractions The form of characterization varies depending on the nature of the crude drug under study and the characterization method employed. Generally, any or all of the following approaches can be used:

[0100] (a) Functional characterization Functional characterization may include biological assays. Biological assays can 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 pathogens, and cell-cell interactions, and cell secretion assays), immunoassays, antibacterial activity (e.g., cell binding and / or replication of bacteria and viruses) assays, as well as toxicity assays (e.g., LD 50 assays).

[0101] Functional characterization can also be performed indirectly by forms of characterization that enable the identification of one or more indicators of biological activity.

[0102] (b) Physical characterization This can involve quantification of the phytochemical component(s) present in any given fraction or other stage of the process, measurement of the purity of the component, or determination of the molecular weight distribution (or various statistical functions thereof) in the case of fractions containing multiple different phytochemical components, determination of the molecular formula(s) (e.g., by nuclear magnetic resonance), and various forms of spectral analysis.

[0103] Particularly useful spectral characterization includes: ● Mass spectrum (e.g., mass-to-charge ratio (m / z) value versus abundance), and / or ● Chromatography data (spectrum, column retention time, elution profile, etc.) , and / or ● Photodiode array (PDA) spectrum (e.g., both UV and visible ranges) ), and / or ● Electrochemical detection or evaporative light scattering detection, and / or ● Nuclear magnetic resonance (NMR) spectrum ( 1 H and / or 13 Obtained via 13C NMR (Including spectral datasets).

[0104] Spectral characterization can be combined with a separation step. For example, GC- Using MS and HPLC-PDA-MS-ED-ELSD (as described herein) (to) the fractions, and their mass spectra, UV-visible spectra, electrochemical responses, or fractional mass spectra. This can be combined with the acquisition of data and chromatography spectral data.

[0105] Using any or all of the above characteristics, any sample (or its fraction or This allows us to define the "chemical fingerprint" of plant chemical components.

[0106] (c) Chemical characterization This is particularly related to the chemical reactivity of plant chemical components (multiple), their solubility, stability, and It can be done in the form of measuring the melting point.

[0107] Medical Use of the Compounds of the Present Invention neoplasm The compounds of the present invention are sialidase inhibitors, and therefore sialidase activity and / or It will find uses in the treatment or prevention of diseases and disorders mediated by sialic acid.

[0108] Sialidase is used in various pathological conditions, including bacterial and viral infections and neoplasms. Because they are involved in the process, these enzymes have become attractive therapeutic targets. - The expression of enzymes Neu1 and Neu3 appears to be altered in diabetes (for example, Ne u1 activity discussed by Natori, Y., et al, 2013, Biol.Pharm., Bull., 36, 1027). Sialidase is involved in atheroma formation (Sukhorukov, VN, et al., (2017, Curr. Pharm. Des., 23, 4696) and bones and joints Flame (Katoh, S., et al., 1999, J Immunol., 1 It is also thought to be involved in 62, 5058).

[0109] Therefore, the compounds of the present invention are neoplastic / proliferative, as described in more detail below. To find applications in the treatment or prevention of disorders.

[0110] As used herein, the term “neoplasm” refers to a condition involving the abnormal proliferation of tumor cells. It is used strictly to define diseases. This term includes benign, precancerous, and malignant It includes neoplasms (as defined above) and is used as a synonym for the term "proliferative disorder."

[0111] Neoplasms are genetically or epigenetically freed from normal physiological control. Inappropriate behavior in tumor cells that have acquired changes (i.e., cells that have been "transformed"). A neoplasm arising from high levels of cell division and / or low levels of apoptosis or senescence. It typically produces the following structures known as neoplasms: abnormal clumps of tissue and their proliferation. It outpaces the proliferation of normal tissue and is unregulated; its proliferation is in response to the stimulus that caused the change. The same excessive persistence continues even after cessation. Most neoplasms form large tissue masses (solid tumors). However, some neoplasms do not form such individual tissue masses. These include the cervix. This includes carcinoma in situ, anal carcinoma in situ, and leukemia.

[0112] Neoplasms can be benign, potentially malignant, or malignant. Benign tumors include uterine fibroids and pigment tumors. This includes cellular nevi (skin moles), but these are not invasive and do not transform into malignant neoplasms. It does not progress or worsen. Potentially malignant (precancerous) neoplasms do not metastasize. This includes carcinoma in situ, which is not invasive but can transform into a malignant neoplasm over time. .

[0113] Malignant neoplasms can invade and destroy surrounding tissues, potentially forming metastases, and ultimately It causes the development of new organisms (tumors) that can potentially kill or injure the host. ("Malignant neoplasms" and "cancer") The terms are used synonymously in this specification.

[0114] The terms "proliferative disorder" and "neoplasm" are used herein to refer to cellular pathology in vivo. It can be used synonymously to define a class of diseases involving plaque growth.

[0115] Therefore, proliferative disorders include cancer, cancer metastasis, smooth muscle cell proliferation, systemic sclerosis, and liver cirrhosis. Adult dyspnea syndrome, idiopathic cardiomyopathy, erythematous lupus, retinopathy (e.g., diabetic retinopathy) (Membranous disease), cardiac hyperplasia, benign prostatic hyperplasia, ovarian cysts, pulmonary fibrosis, endometriosis, fibromatosis, This includes lumatomas, lymphangiomatosis, sarcoidosis, and desmoid tumors. Smooth muscle Neoplasms involving cell proliferation include excessive cell proliferation in the vascular system (e.g., intimal smooth muscle cells). Certain conditions after biological or mechanically mediated vascular injury such as hyperplasia, restenosis, and angioplasty. This includes vascular occlusion, including stenosis. Furthermore, intimal smooth muscle cell hyperplasia is a condition that affects smooth muscles outside the vascular system. Muscle hyperplasia (e.g., bile ducts, bronchial airways, and kidney obstruction in patients with renal interstitial fibrosis) This may include (blockage). Non-cancerous proliferative diseases include psoriasis and its various clinical forms, and Reiter's syndrome. Group, actinic keratosis, and hyperproliferative variants of keratinization disorders (actinic keratosis, senile keratosis, This also includes excessive cell proliferation in the skin, such as in scleroderma.

[0116] The term “neoplasm” also, as used herein, refers to abnormal cell proliferation and / or other abnormalities in vivo. It is used in a broad sense to define diseases involving differentiation, and therefore hyperplasia, metaplasia, and It includes dysplasia.

[0117] Hyperplasia is the condition in which normal (untransformed) cells in an organ or tissue become abnormally advanced. It defines a proliferative state, thus leading to the overall enlargement of an organ and the formation of a benign tumor. It is possible, or it may only be visible under a microscope. Hyperplasia is a characteristic of certain stinging This is a physiological response to stress, and hyperplastic cells continue to be affected by normal regulatory mechanisms. (This differs from neoplastic growth, in which cells proliferate in an abnormal way that does not respond to normal physiological control.) Examples include congenital adrenal hyperplasia, endometrial hyperplasia, and benign prostatic hyperplasia (BPH). , breast hyperplasia (including ductal hyperplasia), focal epithelial hyperplasia (Heck's disease), sebaceous gland hyperplasia, liver Hyperplasia is one example.

[0118] Metaplasia is defined as the process in which one mature differentiated cell type is replaced by another mature differentiated cell type. To mean. For example, squamous metaplasia of columnar epithelial cells in salivary gland ducts (when calculi are present). Squamous metaplasia of the transitional epithelium of the bladder (similarly, in the presence of stones or in areas associated with infection) (Integrated), esophageal glandular metaplasia and bone in connective tissue in patients with gastric reflux disease (Barrett's esophagus) One example is chemical transformation.

[0119] Dysplasia is defined as a condition characterized by the abnormal maturation of cells within a tissue. This is generally premature. It consists of an expansion of mature cells and a corresponding decrease in the number and arrangement 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 cell dysplasia, is characterized by an increase in the number of immature cells in the bone marrow and blood cells. It shows a decrease in mature, functional cells. Other examples include neurofibromatosis.

[0120] Hyperplasia, metaplasia, and dysplasia are generally reversible conditions, triggered by irritation (e.g., seizures or This is the result of injury. In contrast, neoplasms are generally irreversible and related to cell transformation. They are doing it.

[0121] The compounds of the present invention are used to treat proliferative disorders, benign, precancerous, and malignant neoplasms, hyperplasia, metaplasia, and other conditions. Furthermore, it finds common use in the treatment of any neoplasm, including dysplasia.

[0122] Therefore, the present invention relates to cancer, cancer metastasis, smooth muscle cell proliferation, systemic sclerosis, liver cirrhosis, and Human respiratory dyspnea syndrome, idiopathic cardiomyopathy, erythematous lupus, retinopathy (e.g., diabetic retinopathy) Cardiac hyperplasia, benign prostatic hyperplasia, ovarian cysts, pulmonary fibrosis, endometriosis, fibromatosis, Harmat This includes, but is not limited to, desmoid tumors, lymphangiomatosis, sarcoidosis, and desmoid tumors. It finds applications in treating proliferative disorders that do not involve the proliferation of smooth muscle cells. Excessive proliferation of cells (e.g., hyperplasia of intima smooth muscle cells, restenosis, and angioplasty) This includes vascular occlusion (including certain stenosis following biological or mechanically mediated vascular injury). Intima-mediated smooth muscle cell hyperplasia is hyperplasia in smooth muscle outside the vascular system (e.g., bile ducts, bronchi). This may include obstruction of the airways and within the kidneys in patients with renal interstitial fibrosis. Non-cancerous proliferative disorders include Psoriasis and its various clinical forms, Reiter's syndrome, actinic keratosis, and hyperkeratotic disorders. Skin microparticles such as sex variants (including actinic keratosis, senile keratosis, and scleroderma) This also includes excessive proliferation of cells.

[0123] Particularly preferred is the treatment of malignant neoplasms (cancer). The present invention relates to the following major classifications: Finding applications in the treatment of all cancers, including those selected from: (a) carcinoma; ( b) blastoma; (c) leukemia; (d) lymphoma; (e) myeloma; (f) sarcoma; and (g) Mixed-type cancer.

[0124] Carcinoma refers to a malignant neoplasm of epithelial origin, or cancer of the inner or outer layers of the body. Carcinomas, which are malignant tumors of the epithelium, account for 80-90 percent of all cancer cases. The weave is found throughout the body. Not only in the skin, but also in the covering and inner layers of organs and internal passages such as the gastrointestinal tract. There are also. In a preferred embodiment, the cancers treated according to the present invention are the following cancers Select from: colon; rectum; appendix; lungs; thymus; breasts; cervix; bladder; and eyes.

[0125] This invention relates to hepatoblastoma (e.g., nephroblastoma, non-epithelial renal tumor, rhabdoid renal tumor, kidney Sarcoma, and pPNETs of the kidney, medulloblastoma, pancreatic blastoma, pulmonary blastoma, pleuropulmonary blastoma, Neuroblastoma (common peripheral nerve cell tumors, as well as ganglion neuroblastoma and retinoblastoma) It is found to have applications in the treatment of all blastomas, including (including tumors).

[0126] The present invention relates to the treatment of all leukemias, myeloproliferative disorders, and myelodysplastic disorders, including the following: Finding applications in: lymphocytic leukemia (e.g., progenitor cell leukemia, mature B-cell leukemia, adult Mature T-cell leukemia and NK-cell leukemia; acute myeloid leukemia; chronic myeloproliferative disorders; bone marrow Dysplastic syndromes and other myeloproliferative disorders. Therefore, the present invention relates to lymphocytic leukemia, Lymphocytic leukemia, or lymphoblastic leukemia (malignant lymphoid and lymphocytic hematopoietic cells) Tumors), as well as polycythemia vera or erythrocytemia (various blood cell products, but red blood cells) It has found applications in the treatment of various types of leukemia, including those with predominantly malignant tumors.

[0127] Lymphoma affects the glands or lymph nodes of the lymphatic system, blood vessels, lymph nodes, and organs (especially the spleen and tonsils). White blood cells or lymphocytes that originate in the network of the tonsils and thymus, and fight infection by purifying bodily fluids. Unlike leukemia, which is often called "liquid cancer," lymphoma is a "solid cancer." Lymphoma can also occur in certain organs such as the stomach, breasts, or brain. These lymphomas are called extranodal lymphomas. Lymphomas are divided into two categories. Classified as: Hodgkin lymphoma and non-Hodgkin lymphoma. Lead in Hodgkin lymphoma. The presence of Sternberg cells is used to diagnostically differentiate Hodgkin lymphoma from non-Hodgkin lymphoma. The present invention relates to the treatment of all such lymphomas and reticuloendothelial neoplasms, including the following. Finding applications for: (a) Hodgkin lymphoma; (b) Non-Hodgkin lymphoma (e.g., progenitor lymphoma) Bolar lymphoma, mature B-cell lymphoma, mature T-cell lymphoma and NK-cell lymphoma; (c) - Kitt lymphoma, and (d) other lymphotrichal neoplasms, including mantle cell lymphoma.

[0128] Therefore, the present invention relates, for example, to lymphatic glands or nodules (spleen, tonsils, and thymus) Tumors of the stomach, breast, and brain, as well as a wide range of lymphomas. To find applications in treatment.

[0129] Myeloma is a cancer that develops in plasma cells of the bone marrow. Therefore, the present invention relates to the formation of the lymphatic system. Hematological malignancies and blood-related malignancies (e.g., leukemia, acute lymphoblastic leukemia, chronic lymphoblastic leukemia) Blood diseases, B-cell lymphoma (diffuse large B-cell lymphoma, etc.), T-cell lymphoma, Hodg Hodgkin lymphoma, non-Hodgkin lymphoma (Reedsternberg in Hodgkin lymphoma) The presence of cells distinguishes Hodgkin lymphoma from non-Hodgkin lymphoma), pilose cell lymphoma and Burkitt lymphoma), as well as myeloid hematopoietic malignancies (e.g., acute myeloid leukemia, chronic bone marrow malignancy). Myelin leukemia, myeloid leukemia, and imatinib-sensitive and refractory chronic myeloid leukemia, myelodysplasia Multiple myeloma syndrome, bortezomib-sensitive and refractory multiple myeloma, myeloproliferative disorders or promyelocytes It is expected to find applications in the treatment of hematopoietic malignancies and blood-related malignancies, including leukemia and follicular thyroid carcinoma.

[0130] This invention finds applications in the treatment of all sarcomas. Sarcomas are those of bone, tendon, cartilage, muscle, This refers to cancer that develops in supporting tissues such as fat and connective tissue. It generally occurs in young adults. The most common type of sarcoma presents as a bone mass, often accompanied by pain. Sarcoma tumors are usually... These are similar to the tissues in which they proliferate. Exemplary sarcomas for treatment according to the present invention include osteosarcoma ( or osteogenic sarcoma); chondrosarcoma; leiomyosarcoma (smooth muscle); rhabdomyosarcoma (skeletal muscle); mesomyosarcoma Tumor or mesothelioma (membranous inner layer of the body cavity); fibrosarcoma (fibrous tissue); angiosarcoma or intravascular If you have a skin tumor (vascular); liposarcoma; glioma; astrocytoma; myxosarcoma (primary embryonic connective tissue) This includes mesenchymal or mixed mesodermal tumors (mixed connective tissue type). Fibrosarcomas include, Peripheral nerve sheath tumors and other fibrous neoplasms, such as fibroblastic and myofibroblastic tumors, This includes nerve sheath tumors and other fibromatous neoplasms. Kaposi's sarcoma is also included. Soft tissue sarcomas, For example, Ewing tumors and Askin tumors of soft tissue, pPNETs of soft tissue, and extrarenal rhabdomyoplasia. Idoid tumors, fibrohistiocytic tumors; synovial sarcomas; bone and chondromatous soft tissue and alveolar soft tissue sarcomas. This also includes sexual neoplasms. Osteosarcoma (malignant bone tumor) includes malignant fibrous neoplasms of bone; malignant chordoma and This includes odontogenic malignancies. Gliomas include oligodendrogliomas, mixed and unspecified gliomas. This includes neuroepithelial glial tumors.

[0131] The present invention includes, for example, adenosquamous carcinoma, mixed mesodermal tumor, carcinosarcoma, and teratoma. To find applications in the treatment of certain types of cancer. Therefore, the present invention relates to astrocytoma, neuroblastoma, and other cancers. Tumors, gliomas, schwannomas, ependymomas and choroidal nerve plexus tumors (e.g., ependymomas and choroidal nerve plexus tumors) Plexus tumors; intracranial and spinal cord endodermal tumors (e.g., medulloblastoma, primitive neuroectodermal tumor (PNET)). ), medullary tumors, atypical teratomas / rhabdoid tumors and other intracranial and intraspinal tumors (for example) Generally, this includes pituitary adenomas and carcinomas, tumors of the cellar region (craniopharyngiomas), nerves, and mixed tumors. Various CNS, PN (including neuroglial tumors, medulloblastomas, and intracranial and intraspinal neoplasms) To find applications in the treatment of S and other intracranial and intraspinal neoplasms.

[0132] Therefore, the present invention relates to intracranial and intraspinal germ cell tumors; intracranial and intraspinal germ cell tumors; head Intracranial and intraspinal teratomas; intracranial and intraspinal embryonic carcinomas; intracranial and intraspinal yolk sac tumors; intracranial and specific uses in the treatment of intraspinal choriocarcinoma, as well as mixed-type intracranial and intraspinal tumors. To find.

[0133] The present invention also has applications in the treatment of various germ cell tumors, trophoblast tumors, and gonadal neoplasms. Therefore, the present invention generally identifies malignant germ cells in the extracranial and extragonadal regions. Tumors, extracranial and extragonadal malignant teratomas, extracranial and extragonadal embryonic carcinomas, extracranial and Extragonadal yolk sac tumors; extracranial and extragonadal choriocarcinomas, and extracranial and extragonadal It has found applications in the treatment of malignant extracranial and extragonadal germ cell tumors, including malignant mixed germ cell tumors. The present invention also applies, for example, to malignant adenoma, seminomas, malignant gonadal teratomas, gonadal embryonic carcinomas, This includes gonadal yolk sac tumors, gonadal cholangiocarcinoma, mixed-form malignant gonadal tumors, and malignant gonadal adenoblastoma. , it will find applications in the treatment of malignant gonadal germ cell tumors.

[0134] infectious disease The compound of the present invention is a sialidase inhibitor, and therefore sialidase activity and / or Alternatively, find uses in the treatment or prevention of diseases and disorders mediated by sialic acid. These diseases and disorders include infectious diseases (including bacterial and viral infections).

[0135] The compounds of the present invention exhibit antiinfective properties against any infectious agent (e.g., pathogen inhibition or pathogen killing). ) may have activity. Therefore, the compounds of the present invention can target a wide range of different infectious agents. It is possible (i.e., it has activity against it). Therefore, the present invention relates to viruses, particles Infections involving fungal, protozoan, prion, or metazoan factors, It will find a wide range of applications in the treatment or prevention of mild infectious diseases or infectious illnesses.

[0136] Therefore, the present invention relates to the treatment or prevention of viral infections; the treatment or prevention of bacterial infections. Prevention; treatment or prevention of protozoal infections; treatment or prevention of fungal infections; treatment of prion infections or prevention; and / or treatment or prevention of infection or invasion of metazoans (e.g., helminths). The compounds of this invention also find a wide range of applications in the treatment of chronic, dormant, or latent viruses. Infection or invasion by bacteria, protozoa, fungi, prions, or metazoans (e.g., helminths) It can be found to have applications in the treatment or prevention of [the disease].

[0137] ● The virus's targets include the following viruses (or virus classes), but These are not limited to: Retroviridae (e.g., human immunodeficiency virus including HIV-1) S); Picornaviridae (e.g., poliovirus, hepatitis A virus, enterovirus) Russus, dromedary rhinovirus, echovirus; calicivirus Family (e.g., strains that cause gastroenteritis); Togaviridae (e.g., equine encephalitis virus, Rubella virus; Flaviviridae (e.g., dengue virus, encephalitis virus, yellow fever virus) Viruses; Coronavirus family (e.g., coronavirus); Rhabdoviridae family (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) Enzavirus; Bungaviridae (e.g., Hantan virus, Bungavirus, F Levovirus and nairovirus; Arenaviridae (hemorrhagic fever viruses); Leovirus Rusidae (e.g., reovirus, orbivirus, and rotavirus); Birnavirus Family; Hepadnaviridae (Hepatitis B virus); Parvoviridae (Parvovirus); Papillomaviridae (papillomaviruses, polyomaviruses); Adenoviridae (Most adenoviruses); Herpesviridae (Herpes simplex virus (HSV)) 1 and 2, Varicella herpes zoster virus, Cytomegalovirus (CMV), Herpes virus Rus; Poxviridae family (smallpox virus, vaccinia virus, poxvirus); and Iridoviridae (e.g., African swine fever virus); and unclassified viruses (e.g., For example, the causative agent of spongiform encephalopathy, the pathogen of hepatitis delta (defective satellite of the hepatitis B virus) It is believed that the HCV virus (which causes non-A and non-B hepatitis) is the cause of the disease. (HIV and related viruses, and astroviruses). Of these, HIV is particularly preferred. Hepatitis A, Hepatitis B, Hepatitis C, Rabies virus, Poliovirus, Influenza virus Russus, meningitis virus, measles virus, mumps virus, rubella, pertussis, encephalitis virus S, papillomavirus, yellow fever virus, respiratory syncytial virus, parvovirus, Kungunya virus, hemorrhagic fever virus and herpesvirus, especially varicella and cytomega. These are loviruses and Epstein-Barr viruses.

[0138] ● Bacterial targets include both Gram-negative and Gram-positive bacteria, but are not limited to these. Not applicable. Examples of bacteria that the compounds of the present invention may target include, but are not limited to, the following. Undetermined: Helicobacter pylori, Borelia burgdo rferi, Legionella pneumophilia, Mycobacterium genus Species (e.g., M. tuberculosis, M. leprae, M. avium) , M. intracellulare, M. kansaii, and M. gordo nae), Staphylococcus aureus, Neisseria gon orrhoeae, Neisseria meningitidis, Listeria monocytogenes, Streptococcus pyogenes (group A Streptococcus agalactiae (Group B Streptococcus) , Streptococcus viridans, Streptococcus fa ecalis, Streptococcus bovis, an anaerobic species of the Streptococcus genus, S treptococcus pneumoniae, Campylobacter genus, Enterococcus Cass species, Haemophilus influenzae, Bacillus an thracis, Corynebacterium species (including C. diphtheriae), E rysipelothrix rhusiopathiae, Clostridium perfringens, Clostridium tetani, Enterobac ter aerogenes, Klebsiella species (including K. pneumoniae) , Pasturella multocida, Bacteroides sp., Fusobact erium nucleatum, Streptobacillus monilijo rmis, Treponema pallidium, Treponema perte nue, Leptospira species, Rickettsia species, and Actinomyces species (A. isra) (including elii). Bacteria that form biofilms in vivo are identified as compounds of the present invention. These are the targets of Tannerella forsythia, Tanner ella denticola, Porphyromonas gingivalis, This includes Gardnerella vaginalis.

[0139] ● Fungal targets include Cryptococcus neoformans and Histo plasma capsulatum, Coccidioides immitis, B lastomyces dermatitidis, Chlamydia tracho This includes, but is not limited to, matis and Candida albicans. do not have.

[0140] ● Protozoa target species include the malaria parasite genus (Plasmodium falcipa) rum, Plasmodium malariae, Plasmodium ovale , and including Plasmodium vivax), species of the genus Toxoplasma (T. gon (Including dii and T. cruzii), species of the genus Leishmania, and the genus Cryptosporidium. Species (including C. parvum), Cyclospora species (C. cayetanensis) (including), Entomoeba species (including E. histolytica), and Dia This includes, but is not limited to, species of the genus Luzia (including G. lamblia).

[0141] ● Targets of metazoans include parasites or pathogens such as helminths (e.g., Cystosoma species). The body is included.

[0142] Inhibition of bacterial growth in vivo The sialidase inhibitory properties of the compounds of the present invention also affect symbiotic and / or pathogenicity in vivo. Inhibition of bacterial growth, particularly inhibition of bacterial biofilms in mammalian (e.g., human) hosts. Alternatively, find applications in disrupting host-bacterial cell interactions, including elimination.

[0143] Therefore, these compounds are found in bacterial biofilms (e.g., subgingival plaque biofilms). Diseases and disorders mediated or characterized by the presence of films and fascial biofilms To find uses in the treatment or prevention of harm.

[0144] These diseases include periodontal disease, bacterial vaginosis, and Tannerella forsythia. hia, Tannerella denticola, Porphyromonas g ingivalis and Gardnerella vaginalis (the latter species is thin) This includes diseases caused by infections (associated with bacterial vaginosis and premature birth).

[0145] Atheroma formation The compound of the present invention is a sialidase inhibitor, and sialidase is involved in this process. Therefore, atheroma formation (Sukhorukov, VN, et al., 2 017, Curr. Pharm. Des., 23, 4696) and osteoarthritis ( Katoh, S., et al., 1999, J Immunol., 162 To find use in the treatment or prevention of , 5058). Therefore, the compounds of the present invention find use in ate To find applications in the treatment and prevention of Roam's arteriosclerosis.

[0146] Regulation of symbiotic bacterial growth The sialidase inhibitory properties of the compounds of the present invention also affect the microbial community (and especially symbiotic) in the host. Regulation of the composition of bacteria, for example, the composition of symbiotic bacteria in mammalian (e.g., human) hosts. Find applications in regulation. Particularly preferred is the regulation of the gut microbiota.

[0147] inflammation The compounds of this invention are thought to be involved in the TNF-α-induced inflammatory process in osteoarthritis. It inhibits sialidase (Gee, K. et al., 2003, J Biol Chem., 37275). Furthermore, the compounds of the present invention exhibit TNF-α activity. They can suppress or inhibit inflammation. Thus, they can suppress inflammation and inhibit the function of physiological functions. It finds use in any disorder that plays a role in the impairment and / or symptoms and / or pain. Example For example, the compounds of the present invention reduce acute, chronic, localized, or systemic inflammation. It can be used as an anti-inflammatory agent to eliminate it.

[0148] Inflammation can be caused by microorganisms, trauma, chemicals, heat, cold, sunburn, or other harmful events. It occurs when tissue is damaged by endogenous chemicals (e.g., bradykinin, hi). Stamina and serotonin are released in the event of injury or illness, and such chemicals It activates and attracts macrophages and other white blood cells in the tissue. In this process, TNF-α Chemical mediators such as these are released, causing inflammation.

[0149] Inflammatory disorders are characterized by persistent or chronic inflammation. In such situations, long-term Inflammation causes tissue destruction, leading to widespread damage and / or final damage to the affected tissues and / or organs. It causes a certain kind of dysfunction.

[0150] Therefore, the compounds of the present invention affect delocalized inflammatory disorders, for example, multiple organs. To find applications in the treatment of such disorders. Such disorders include immune dysfunction (therefore This includes disorders resulting from (possibly containing autoimmune components). Such conditions include systemic estrogen This includes conditions such as systemic lupus erythematosus (SLE), scleroderma, and hypersensitivity reactions.

[0151] The compounds of the present invention also have effects on skin inflammation and chronic prostatitis, glomerulonephritis, inflammatory bowel disease, and pelvic Inflammatory diseases, reperfusion injury, rheumatoid arthritis, graft rejection, vasculitis, asthma, acne, degenerative joints Joint inflammation, inflammation of the oral mucosa, gastrointestinal tract, inflammation of the eyes, nose, and ears, and other steroid reactions. To find applications in the treatment of localized inflammatory disorders, including inflammatory diseases.

[0152] In particular, the compounds of the present invention find applications in the treatment of inflammatory skin diseases. For example, actinic keratosis, acne (acne vulgaris, comedones, rosacea, and nodular cystic acne) (including acne), allergic contact dermatitis, angioedema, bullous pemifigoids, cutaneous drug reactions Erythema multiforme, erythematous scleroderma, photodermatitis, psoriatic arthritis, scleroderma and urticaria, psoriasis, skin Inflammation (e.g., atopic dermatitis), scleroderma, steroid-responsive inflammatory skin disorders (e.g., Urinary tract pruritus, as well as conditions related to exposure to sunlight, radiation, chemotherapy, and environmental irritants. This includes skin condition.

[0153] The compounds of the present invention also find applications in the treatment of inflammatory autoimmune diseases. Diseases affect specific tissues or organs (such as musculoskeletal arthritis and ankylosing spondylitis). (various tissues, etc.), gastrointestinal tract (for example, Crohn's disease and ulcerative colitis), CNS (for example) For example, Alzheimer's disease, multiple sclerosis, motor neuron disorders, Parkinson's disease, and chronic illnesses. Sexual fatigue syndrome), pancreatic beta cells (for example, insulin-dependent diabetes mellitus), adrenal glands (for example) (For example, Addison's disease), kidney (e.g., Goodpasture syndrome, IgA nephropathy and interstitial kidney) Inflammation), exocrine glands (e.g., Sjögren's syndrome and autoimmune pancreatitis), and skin (e.g.) This may include psoriasis and atopic dermatitis.

[0154] Other inflammatory disorders treatable according to the present invention include osteoarthritis, periodontal disease, diabetic nephropathy, and chronic diseases. Obstructive pulmonary disease, arthrsclerosis, graft-versus-host disease, chronic pelvic inflammatory disease, endometriosis, chronic This includes conditions such as hepatitis and tuberculosis.

[0155] Energy-related diseases The compounds of the present invention have blood glucose regulating activity. Therefore, the present invention is useful for various energy To find applications in the treatment of diseases. These diseases include a wide range of illnesses and disorders. For example, this includes homeostatic disorders, metabolic disorders, glucose metabolism dysfunction, and appetite disorders.

[0156] Therefore, examples of energy utilization disorders include insulin resistance and various forms of diabetes. Diseases (including type 1 and type 2 diabetes), metabolic syndrome, obesity, wasting syndrome (for example) (and cancer-related cachexia), muscle disorders, gastrointestinal disorders, growth retardation, hypercholesterolemia, atherosclerosis This includes arteriosclerosis and age-related metabolic disorders.

[0157] Energy-utilization diseases also include metabolic syndrome, obesity, and / or diabetes. This includes conditions related to hyperglycemia, impaired glucose tolerance, hyperinsulinemia, diabetes, metabolic disorders, etc. Acidosis, cataracts, diabetic neuropathy, diabetic nephropathy, diabetic retinopathy, macula Degenerative diseases, glomerulosclerosis, diabetic cardiomyopathy, insulin resistance, glucose metabolism disorders, joints inflammation, hypertension, hyperlipidemia, osteoporosis, osteopenia, bone loss, brittle bone syndrome, acute coronary syndrome, This includes infertility, short bowel syndrome, chronic fatigue, eating disorders, and bowel motility disorders.

[0158] Insulin resistance, metabolic syndrome, diabetes In healthy individuals, blood glucose levels are controlled by insulin (produced by the beta cells of the pancreas) and glucagon. It is maintained within a narrow range by two pancreatic hormones (produced by the alpha cells of the pancreas). The pancreatic beta cells sense an increase in blood glucose levels and respond by secreting insulin. Insulin promotes glucose uptake by the body's tissues, thereby increasing blood glucose concentration. It returns to a physiological range. Glucagon interacts with other substances, primarily stimulating glucose production in the liver. This raises blood sugar levels in a fasting state.

[0159] Insulin resistance is a decrease in the action of insulin in skeletal muscle, adipocytes, and hepatocytes. This is characterized by the fact that, as a result, normal amounts of insulin are released from the cells of these tissues. It becomes insufficient to produce a phosphorus response. In adipocytes, due to insulin resistance, storage The triglycerides that are present are hydrolyzed, and the amount of free fatty acids in the plasma increases. In muscle, Nsulin resistance reduces glucose uptake and, in hepatocytes, reduces glucose storage. In both of the latter cases, an increase in blood glucose concentration occurs.

[0160] High plasma levels of insulin and glucose due to insulin resistance are often associated with metabolic syndrome. It progresses to Rick's syndrome and type 2 diabetes.

[0161] Metabolic syndrome is a set of abnormalities and disorders that increase the risk of cardiovascular disease and diabetes. It is a collection. The incidence rate is very high in many developed countries: some studies suggest it affects up to 25% of the population. This shows the prevalence of this condition in the United States. This disorder is (metabolic) syndrome X, insulin Also known as phosphorus resistance syndrome, Lieben syndrome, and CHAOS. Obesity syndrome is diagnosed by the presence of three or more of the following symptoms: central obesity (in men) For men, the minimum height is 101.6 centimeters (40 inches), and for women, it is 88.9 centimeters. Waist measurement of 35 inches or more); high levels of triglycerides (150 mg / dL or higher); Low HDL levels (less than 40 mg / dL in men, 50 mg / dL in women) (Below 130 / 85 mmHg), and hypertension (130 / 85 mmHg or higher). Related diseases and signs are as follows: Present: Fatty liver (often progressing to non-alcoholic fatty liver disease), polycystic ovary syndrome Symptoms include hemochromatosis (iron overload) and acanthosis nigricans (acanthosis nigricans).

[0162] The primary treatment for metabolic syndrome involves lifestyle changes (calorie restriction and physical changes). It is an active condition. However, drug treatment is often necessary. Generally, metabolic syndrome The individual diseases that make up the disease are treated separately (for example, diuretics and hypertension) ACE inhibitors). Cholesterol drugs affect LDL cholesterol and triglyceride levels. If levels are rising, lower them; if HDL levels are falling, raise them. It can be used for the use of drugs that reduce insulin resistance (e.g., metformin and thi). Azolidinedione is controversial. Aerobic exercise treats less than 31% of cases. It is effective and generally does not cause a decrease in fasting plasma glucose or insulin resistance. .

[0163] Therefore, new and / or alternative treatments for metabolic syndrome, particularly obesity and / or effective treatment is needed for elevated triglyceride levels.

[0164] Type 2 diabetes is a chronic disease characterized by persistently elevated blood glucose levels (hyperglycemia). Phosphate resistance, along with impaired insulin secretion from pancreatic beta cells, is a characteristic feature of this disease. The progression of insulin resistance to type 2 diabetes is due to the pancreatic beta cells being unable to maintain normal blood glucose levels (euglycemia). When the body can no longer produce enough insulin to maintain blood sugar levels, postprandial hyperglycemia occurs. Characterized.

[0165] The most important drug currently used to treat type 2 diabetes is metformin (Gluco) phage, Diabex, Diaformin, Fortamet, Riomet, G These include lumetza and cidophage. Metformin is a biguanide class drug. It is an oral antihyperglycemic agent. Other biguanides include phenformin and buformin (now discontinued). It contains (stops). Metformin primarily suppresses the hepatic release of glucose from glycogen stores. It works by reducing it, but has some effect in increasing glucose uptake. It also has other widely used drug classes, such as the sulfonylurea group of drugs. These drugs include (glibenclamide and gliclazide). These drugs are produced by the pancreas. It increases glucose-stimulated insulin secretion. A new class of drugs includes receptors in the cell nucleus. By binding to PPAR (peroxisome proliferator-activated receptor), a group of molecules Thiazolidinediones that act by (e.g., rosiglitazone, pioglitazone, and This includes β-troglitazone. Other classes include α-glucosidase inhibitors (acarbol). (L-), meglitinide (stimulates insulin release), nateglinide, repaglinide, and so (including their analogues), peptide analogues (for example, those that act as insulin secretagogues) Incretin mimetic drugs, glucagon-like peptide analogs (e.g., exenatide), dipeptide Tidyl peptidase-4 (DPP-4) inhibitors (increase incretin levels (for example) For example, sitagliptin, as well as amylin agonist analogs (which slow gastric emptying), This includes treatments that inhibit lucagon (e.g., pramlintide).

[0166] However, existing treatments for various forms of type 2 diabetes primarily target β-cells. It does not appear to improve the function of endogenous factors, and all existing treatments cannot stop the progression of the disease. The wound, over time, may fail to normalize blood sugar levels and / or prevent subsequent complications. It is also not possible to do so. Existing treatments are also associated with undesirable side effects. For example, Insulin secretagogues and insulin injections can cause hypoglycemia and weight gain. Patients may also become unresponsive to insulin secretagogues over time. Metformin and α-glucosidase inhibitors often cause gastrointestinal problems, PPA R-agonists tend to cause weight gain and increased edema. Exenatide causes nausea. It has also been reported to cause vomiting.

[0167] Glycosylation plays a crucial role in regulating protein properties and is involved in many diseases. Itoh, N. et al., 2007 (Am J Physio Endocrinol Metab 293: E1069-E1077) is type 2. We report on serum N-glycan profiles in humans with diabetes, and the bifurcated N-acetylglucan Along with lucosamine, the amount of branched N-glycans containing α1,6-fucose is increasing. They discovered that. Copeland, RJ et al., 2008 (Am J Physiol Endocrinol Metab 295: E17-E28) This article reviews the importance of O-linked N-acetylglucosamine in diabetes, and GlcNA We concluded that there is a strong positive correlation between c-formation and the development of insulin resistance. O-bound type- β-N-acetylglucosamine (O-GlcNAc) undergoes phosphorylation, as well as serine and / or it is a dynamic post-translational modification that cycles between the on and off of the threonine hydroxyl group. -The circulation of GlcNAc involves O-GlcNAc transferase and O-GlcNAc agent. It is regulated by the synergistic action of ze. GlcNAcization is a major characteristic of type 2 diabetes. Hexosamine is involved in the pathogenesis of glucose toxicity and chronic hyperglycemia-induced insulin resistance. Nidase activity has been shown to be elevated in the serum of diabetic patients (e.g., Agard h, CD et al., 1982, Acta Med Scand.212 :39-41).

[0168] Type 1 diabetes (or insulin-dependent diabetes) is a condition in which insulin is released from the islets of Langerhans in the pancreas. This condition is characterized by the loss of insulin-producing beta cells, leading to insulin deficiency. The main cause of the disease is an autoimmune attack mediated by T cells. In diabetic cases in North America and Europe... There are no known preventive measures that can be taken for type 1 diabetes, which accounts for up to 10% of all diabetes cases. Those affected are otherwise healthy and at a healthy weight at the time of onset. Sensitivity and responsiveness to the virus are usually normal, especially in the initial stages.

[0169] The main treatment for type 1 diabetes is to monitor blood glucose levels using blood test monitors, even in the early stages. This involves insulin supplementation combined with careful monitoring. This can lead to ketosis and diabetic ketoacidosis, potentially resulting in coma or death. In addition to general subcutaneous injections, insulin can also be delivered using a pump. This allows for continuous insulin delivery 24 hours a day at a pre-set level, and at meal times. The insulin dose (bolus) can be programmed as needed. With inhaled insulin... One particular product was recently approved by the FDA.

[0170] Treatment for type 1 diabetes needs to be continued indefinitely. Treatment should not impair normal activities. However, sufficient awareness, appropriate care, and adherence to regulations regarding testing and medications are necessary.

[0171] Therefore, novel and / or alternative antidiabetic drug therapies, particularly those that restore β-cell function, A treatment that can do this is needed. In particular, a treatment with fewer side effects than existing drug therapies for type 2 diabetes. For effective drugs that can treat both type 1 diabetes and related conditions, There are clinical needs that are not being met in a practical and substantial sense.

[0172] This invention has broad applications in the treatment of all energy-dependent diseases.

[0173] Therefore, diseases that can be treated by the present invention include, for example, disorders of homeostasis and metabolic disorders. This includes sexually transmitted diseases, impaired glucose metabolism, and appetite disorders.

[0174] In preferred embodiments, the present invention addresses insulin resistance, various forms of diabetes, and metastasis. Bolic syndrome, obesity, wasting syndrome (e.g., cancer-related cachexia), muscle disorders, gastrointestinal diseases Diseases, growth retardation, hypercholesterolemia, atherosclerosis, and age-related metabolic disorders To find applications in treating harm.

[0175] The present invention also relates to conditions associated with metabolic syndrome, obesity, and / or diabetes. For example, hyperglycemia, impaired glucose tolerance, hyperinsulinemia, diabetes, metabolic acidosis, cataracts Diabetic neuropathy, diabetic nephropathy, diabetic retinopathy, macular degeneration, glomerulosclerosis Diabetic cardiomyopathy, insulin resistance, glucose metabolism disorders, arthritis, hypertension, hyperlipidemia , osteoporosis, osteopenia, bone loss, brittle bone syndrome, acute coronary syndrome, infertility, short bowel syndrome, It can be used to treat chronic fatigue, eating disorders, bowel motility disorders, and glucose metabolism disorders.

[0176] This invention can also be used to suppress appetite. We do not wish to be bound by theory. However, this can even affect blood sugar levels, leading to fatigue / lethargy and a craving for sweets. Eliminate the major dips that cause the desire to eat (i.e., "improved (By promoting "energy balance").

[0177] Particularly preferred are insulin resistance, metabolic syndrome, obesity, and diabetes. This is a treatment for (especially type 2 diabetes).

[0178] This invention finds applications in the treatment of insulin resistance. Insulin resistance is related to the skeletal structure. It is characterized by a decrease in the action of insulin in muscle, adipocytes, and hepatocytes, resulting in a normal state. Insufficient insulin is needed to elicit a normal insulin response from the cells of these tissues. This takes several minutes. In fat cells, due to insulin resistance, stored triglycerides are added. Hydrolysis leads to an increase in free fatty acids in the plasma. In muscles, insulin resistance is related to glucose It reduces uptake and, in hepatocytes, reduces glucose storage. In both of the latter cases... This results in an increase in blood glucose concentration. High levels of insulin and glucose in the plasma due to insulin resistance. The condition often progresses to metabolic syndrome and type 2 diabetes.

[0179] The present invention has applications in the treatment of metabolic syndrome (as defined herein). Discover. This disorder is (metabolic) syndrome X, insulin resistance syndrome, Lee It is also known as Benn syndrome and CHAOS.

[0180] The present invention relates to the treatment of diseases associated with metabolic syndrome, including, for example, the following. Uses include: fatty liver (often progressing to non-alcoholic fatty liver disease), polycystic venereformin. Ovarian syndrome, hemochromatosis (iron overload), and acanthosis nigricans (acanthosis nigricans).

[0181] This invention finds applications in the treatment of diabetes, including type 1 and type 2 diabetes. It is a chronic disease characterized by a persistent rise in blood glucose levels (hyperglycemia). Insulin resistance is This, along with impaired insulin secretion from pancreatic β-cells, is a characteristic feature of this disease. (Type 2 diabetes) The progression of insulin resistance is due to the pancreatic beta cells' inability to maintain normal blood glucose levels (euglycemia). When the body is unable to produce enough insulin, it is characterized by the development of postprandial hyperglycemia. ru.

[0182] type 1 diabetes This invention seeks to find applications in the treatment of type 1 diabetes (or insulin-dependent diabetes). Type 1 diabetes is characterized by the loss of insulin-producing beta cells in the islets of Langerhans in the pancreas. This causes insulin deficiency. The main cause of this beta cell loss is self-mediated by T cells. It is an immune attack. There are no known preventive measures that can be taken. Most people who are affected will otherwise The patient is healthy and at a healthy weight at the time of onset. Sensitivity and responsiveness to insulin are usually not particularly... In the initial stages, this is normal.

[0183] As described below, the compounds of the present invention may have anti-diabetic viral activity. Since diabetes-inducing viruses are the causative factors of type 1 diabetes, anti-diabetic-inducing viral activity is important. Compounds having are particularly preferred for use in the treatment or prevention of type 1 diabetes, or in the following: (a) Treatment of virus-induced type 1 diabetes; (b) Delay in the onset of viral diabetes. (c) Prevention; (d) Treatment of type 1 diabetes; (e.g., non-immune cells) Treatment or prevention of (by cytolysis or immune-mediated cytolysis); (e) virus-mediated infection (f) Prevention, reduction, or elimination of intrinsic interferon production; (f) Autoreactions targeting β-cells (g) Prevention, reduction, or elimination of virus-mediated bystander activation of sex T cells; (g) β cells Prevention, reduction, or elimination of viral activation and / or spread of targeted autoreactive T cells. ; and / or (h)β cells exposed to immune-mediated cytolysis by regulatory T cells as a viral medium Prevention or reduction of loss of ability.

[0184] Such patients are at high risk of progressing to type 1 diabetes, and as mentioned above, diabetes-induced viruses Because there is a possibility of infection with Russ, such use is not suitable for subjects with pancreatic islet autoantibodies. It is particularly useful in treatment.

[0185] The present invention also relates to acquired mitochondrial diseases, particularly those associated with the aforementioned energy utilization disorders. To find applications in the treatment and management of tochondria.

[0186] Pharmacology The compounds of this invention are administered intravenously, intramuscularly, intraperitoneally, subcutaneously, transdermally, through the respiratory tract (aerosol), and rectally. , including vaginal and topical (including buccal and sublingual) administration, via topical, oral or parenteral routes. Therefore, it can be administered.

[0187] The amount administered depends on the specific dosage unit used, the duration of treatment, and the age and sex of the patient being treated. Furthermore, it varies greatly depending on the nature and severity of the disorder being treated, and the specific compound selected. obtain.

[0188] Generally, the effective dose of the administered compound is typically about 0.01 mg / kg per day. The range is 500 mg / kg. The unit dose may contain 0.05 to 500 mg of the compound. It can be taken at least once a day. The compound is taken orally as described below. Using conventional dosage unit forms, either topically, parenterally, or topically, the pharmaceutical carrier It can be administered together with [another drug].

[0189] The preferred route of administration is oral administration. Generally, the appropriate dose is per day of the recipient. The recommended daily dose is in the range of 0.01 to 500 mg per kilogram of body weight, preferably per kilogram of body weight. The range is 0.1 to 50 mg per program, most preferably per kilogram of body weight per day. The dosage ranges from 1 to 5 mg.

[0190] The desired dose is preferably provided as a single dose for daily administration. However, However, 2, 3, 4, 5, or 6 or more doses are administered at appropriate intervals throughout the day. The above subdoses can also be used. These subdoses are, for example, 0 per unit dosage form. 0.001 to 100 mg, preferably 0.01 to 10 mg, most preferably 0.5 to 1.0 It can be administered in a single dosage form containing mg of the active ingredient.

[0191] formulation The compounds for use in accordance with the present invention can take any form. From cultivated or natural sources (for example, plants of the Rhamnaceae and Sapindaceae families (for example, Na Plants of the genus Zizyphus or Guarana, for example, Zizyphus jujuba or Pauli It may be a plant of the species *Nia cupana*. Particularly preferred as a plant source is These include the fruits, parts of the fruits, fruit extracts, and fruit juices of the aforementioned plants.

[0192] When isolated from natural sources, the compound can be purified. However, the composition of the present invention As mentioned above, the substance can take the form of a crude drug. Such a crude drug is preferably, They are analyzed before use to determine whether they meet standard specifications.

[0193] The crude drugs to be used in accordance with the present invention may be dried plant materials. Alternatively, the crude drugs may be , physical or chemical pretreatment, e.g., powdering, grinding, freezing, evaporation, filtration, pressing, spraying Processed plant material, including misting, extrusion, supercritical solvent extraction, and tincture production. To obtain. When herbal medicines are managed or sold in the form of the whole plant (or a part thereof), the plant material is It can be dried before use in any convenient form, including freeze-drying, spray-drying, or air-drying. The drying process can be used.

[0194] The compounds of the present invention are found in polar solvents (ethanol / water mixture, for example, 50% v / v or higher (e.g.) For example, extraction with an ethanol / water mixture (up to approximately 70% v / v) can extract proteins and It can be separated from high molecular weight components such as polysaccharides. Other suitable techniques include various This includes membrane technologies. These include microfiltration, ultrafiltration, and nanofiltration. Alternatively, Alternatively, charged compounds can be concentrated using electrodialysis. The method either allows only molecules below a certain size to pass through, or depends on the charge of the molecules. Using a membrane with pore sizes that allow molecules to pass through or prevent them from passing through. Compounds can also be concentrated using anion and cation exchange resins.

[0195] When isolated from natural sources, purify the compound for use according to the present invention. This is possible. In embodiments in which the compound is formulated with pharmaceutically acceptable excipients, for example, This includes inert diluents, disintegrants, binders, lubricants, sweeteners, flavorings, colorings, and preservatives. Any suitable excipient can be used. Suitable inert diluents include sodium carbonate. It also contains calcium carbonate, sodium phosphate and calcium, and lactose. On the other hand, corn starch and alginic acid are suitable disintegrants. The binder is starch and It may contain gelatin, while the lubricant, if present, is generally magnesium stearate. It is stearic acid or talc.

[0196] The pharmaceutical composition can take any suitable form, for example, tablets, elixirs, etc. It may include capsules, solutions, suspensions, powders, granules, and aerosols.

[0197] The pharmaceutical composition may take the form of a kit of components, the kit of which includes instructions for use and / or the composition of the present invention may comprise multiple different components in unit dosage forms.

[0198] Tablets for oral use contain inert diluents, disintegrants, binders, lubricants, sweeteners, flavorings, Mixed with pharmaceutically acceptable excipients such as colorants and preservatives, for use according to the present invention It may contain compounds for this purpose. Suitable inert diluents include sodium carbonate and calcium carbonate. It contains sodium phosphate and calcium, as well as lactose, while corn starch Ti and alginic acid are suitable disintegrants. The binder may include starch and gelatin. On the other hand, lubricants, if present, are generally magnesium stearate, stearic acid, and It is talc. If necessary, the tablets are monostearate to slow down absorption in the gastrointestinal tract. Coating with materials such as glyceryl distearate or glyceryl distearate. This can be done. Oral capsules contain a hard compound mixed with a solid diluent. Gelatin capsules and active ingredients are dissolved in water or peanut oil, liquid paraffin, or oil. It contains soft gelatin capsules mixed with oils such as olive oil.

[0199] Formulations for rectal administration have a suitable base, such as cocoa butter or salicylate. It may be provided as a suppository. Formulations suitable for vaginal administration may be provided in addition to the active ingredient in the art. Pessaries, tampons, and creams containing carriers known to be suitable in the context of They may be provided as gels, pastes, foams, or spray formulations.

[0200] For intramuscular, intraperitoneal, subcutaneous, and intravenous use, the compounds of the present invention are generally suitable. Provided in a sterile aqueous solution or suspension buffered to a suitable pH and isotonicity. The solution contains Ringer's solution and isotonic sodium chloride. The aqueous suspension according to the present invention is Lulurose derivatives, sodium alginate, polyvinylpyrrolidone, and tragacanth gum It may contain suspending agents such as lecithin, as well as humectants such as lecithin. It contains ethyl and n-propyl p-hydroxybenzoate.

[0201] The compounds of the present invention may also be provided as liposome formulations.

[0202] When administered orally, the compound or multiple compounds may be in the form of capsules, pills, tablets, lozenges, or other forms. Zengi, melts, powders, granules, solutions, suspensions, dispersions, or emulsions (these dissolves Liquids, suspensions, dispersions, or emulsions (which may be aqueous or non-aqueous) are solids. It can also be formulated as a liquid formulation. Solid unit formulations include, for example, surfactants and lubricants. , and inactive additives such as lactose, sucrose, calcium phosphate, and corn starch. Capsules that can be of the usual hard-shell or soft-shell gelatin type, containing a filler. could be.

[0203] In another embodiment, the compound of the present invention is acacia, corn starch, or gelatin Binders such as corn, and potato starch intended to aid in the breakdown and dissolution of tablets after administration. Disintegrants such as alginate, corn starch, and guar gum improve the flow of tablet granules, and tablets A lubricant intended to prevent tablet material from adhering to the surface of the die and punch of the agent, for example, Luc, stearic acid, or magnesium stearate, calcium stearate, if Zinc stearate enhances the aesthetic quality of tablets, making them more palatable to patients. In combination with dyes, colorants, and flavorings intended for this purpose, lactose, sucrose, They are also formulated into tablets using conventional tablet bases such as cornstarch.

[0204] Suitable excipients for use in oral liquid dosage forms include water and alcohol, such as ethanol. It contains diluents such as benzyl alcohol and polyethylene alcohol, and is pharmaceutically acceptable. With or without the addition of a surfactant, suspension agent, or emulsifier. That is the case.

[0205] The compounds of the present invention can also be administered parenterally, i.e., subcutaneously, intravenously, intramuscularly, or intraperitoneally. It can be administered to [the patient].

[0206] In such embodiments, the compound is a pharmaceutical carrier (a sterile liquid or a mixture of liquids). It is provided as an injectable dose in a physiologically acceptable diluent. The liquids include water, physiological saline, dextrose aqueous solution and related sugar solutions, and alcohol. Ethanol, isopropanol, or hexadecyl alcohol, etc., glycol ( (such as polypropylene glycol or polyethylene glycol), glycerol ketal (2, 2-dimethyl-1,3-dioxolane-4-methanol), ether (poly(ethylene) (Recall) 400 etc.), oil, fatty acid, fatty acid ester or glyceride, or acetyl It contains glyceridized fatty acid glycerides and is a pharmaceutically acceptable surfactant (soap). (e.g., detergents), suspending agents (pectin, caromer, methylcellulose) (such as hydroxypropyl methylcellulose or carboxymethylcellulose), This may or may not involve the addition of emulsifiers and other pharmaceutical adjuvants. Suitable oils that can be used in the parenteral formulation of the present invention are petroleum, animal, plant, or synthetic. For example, peanut oil, soybean oil, sesame oil, cottonseed oil, corn oil, olive oil, Petroleum and mineral oil. Suitable fatty acids include oleic acid, stearic acid, and isostearate. It contains phosphoric acid. Suitable fatty acid esters include, for example, ethyl oleate and myristic acid. It is isopropyl acid.

[0207] Suitable soaps contain fatty alkali metals, ammonium, and triethanolamine salts. In rare cases, suitable detergents include cationic detergents, such as dimethyldialkylammonium halogen. Iodides, alkylpyridinium halides, and alkylamine acetates; ionic washes Agents, for example, alkyl, aryl, and olefin sulfonates, alkyl, olefin ethers, and monoglyceride sulfates, as well as sulfosuccinates; nonionic detergents, For example, fatty amine oxides, fatty acid alkanolamides, and polyoxyethylene poly Propylene copolymers; and amphoteric detergents, for example, alkyl-beta-aminopropion The compounds include nates, 2-alkylimidazoline quaternary ammonium salts, and mixtures. ru.

[0208] The parenteral compositions of the present invention are typically compounds for use in solution according to the present invention. Contains approximately 0.5 to 25% by weight. Preservatives and buffer solutions can also be used. Avoid irritation at the injection site. To minimize or eliminate them, such compositions have about 12 to about 17 hydrophilic-lipophilic components. The formulation may contain a nonionic surfactant having a lance (HLB). The amount of the herbicide is in the range of approximately 5 to approximately 15% by weight. The surfactant is a single compound having the HLB described above. It may be a single component, or a mixture of two or more components having the desired HLB. Examples of surfactants used in the parent formulation include polyethylene sorbitan fatty acid esters. Classes such as sorbitan monooleate and propylene oxide and propylene High molecular weight adducts of ethylene oxide with hydrophobic bases formed by condensation with lichol That is the case.

[0209] The compounds or a plurality of compounds for use in accordance with the present invention may also be administered topically. When so, the carrier may appropriately contain a solution, ointment, or gel base. The base is For example, petrolatum, lanolin, polyethylene glycol, beeswax, mineral oil, water and alcohol The topical formulation may contain diluents such as ethanol, as well as one or more emulsifiers and stabilizers. It may contain a compound concentration of approximately 0.1 to 10% w / v (weight per unit volume).

[0210] When used auxiliaryly, the compounds or multiple compounds for use in accordance with the present invention It can be formulated for use with one or more other drugs. Therefore, it can be supplemented. Adjunctive use is designed to be compatible with (or synergistically compatible with) other drugs(s). A specific unit dose, or a compound or multiple compounds mixed with one or more enzymes. This can be reflected in the formulation. Auxiliary use is also possible when the compound of the present invention is packaged together with an enzyme (for example, (As part of a series of unit doses) This may be reflected in the composition of the pharmaceutical kit of the present invention. (Auxiliary use) Furthermore, information regarding the co-administration of a compound or multiple compounds and / or enzymes and / or This may be reflected in the instruction manual.

[0211] Cosmetic formulation The cosmetic compositions of the present invention include, for example, moisturizing compositions, cleansing compositions, or compositions that are beneficial to the skin. Any composition that may provide benefits can be selected. The cosmetic compositions of the present invention include, for example, If, then, select from the following, an excipient or carrier that is acceptable as a cosmetic, It may include.

[0212] In one embodiment, the cosmetic composition is a cleansing composition. Suitable cleansing composition The substance is solid or semi-solid at room temperature. An example of a useful cleansing composition is glycerin. Fatty acids, including soap, detergent, and mixtures thereof. Solid cleansing compositions include, but are not limited to, soap. This is widely taught in "Chemology for the 1990s," and its contents are as follows: As incorporated herein by reference, the cleansing composition is preferably fluid. .

[0213] In one embodiment of the present invention, the cleansing composition comprises glycerin soap. Examples of useful glycerin soaps include the U.S. 4, incorporated herein by reference. This includes what is disclosed in Patent Nos. 405,492 and Nos. 4,879,063. , but not limited to these.

[0214] Examples of suitable fatty acid soaps include those with a hydrocarbon chain length of approximately 10-22 (including carboxyl carbons). It contains soap derived from, for example, sodium salts, which may be saturated or unsaturated. potassium salts, ammonium salts, triethanolammonium salts, and mixtures thereof. It is possible.

[0215] Suitable synthetic detergents include those known in the art for the desired purpose. Examples of cleansing agents useful for personal cleansing include isethionate, sarcosinate, and It contains glyceryl ether sulfonates, which are pure chain-length variants or co- It may also be derived from commercially available oils such as coconut oil. Other suitable surfactants include: Anionic acyl sarcosinates, methyl acyl taurates, N-acyl glutamic acid, Alkyl sulfosuccinates, alkyl phosphate esters, ethoxylated alkyl phosphates phosphate esters, trideceth sulfates, protein condensates, ethoxylated alkyl groups Sulfate and alkylamine oxide mixtures, betaine, sultene and mixtures thereof It contains substances. Specifically, alkyl ether sulfates having 1 to 12 ethoxy groups. In particular, ammonium lauryl ether sulfate and sodium lauryl ether sulfate.

[0216] The cosmetic composition may be a moisturizing composition.

[0217] Other optional components of the cosmetic composition of the present invention include fragrances, scents, preservatives, and colorants. This product includes, but is not limited to, agents, dyes, anticaking agents, and skincare and haircare ingredients. It contains, but is not limited to, personal care ingredients.

[0218] Examples of suitable personal care ingredients useful in the present invention include a safe and effective amount of humidifier. Agents, sunscreen active ingredients, skin soothing agents, anti-irritants, anti-inflammatory agents, skin emollients, conditioning agents Skincare products, moisturizers, deodorants, antiperspirants, artificial sunscreens, antibacterial agents, anti-acne agents, anti-wrinkle agents, Anti-cutaneous atrophy agents, cutaneous sclerosing agents, antipruritics, antifungal agents, local anesthetics, evening skin tone treatments Agents, active natural ingredients, agents to minimize the appearance of unwanted hair or slow down regrowth, This includes, but is not limited to, skin texture modifiers and additional cleansing agents. do not have.

[0219] In one embodiment, the compound is used, for example, in the treatment of dry skin and as a skin emollient. By using a water-in-oil (w / o) emulsion, water or alcohol extract It can be used from the time of purchase.

[0220] Skin emollients remain on the skin surface or stratum corneum, acting as lubricants and reducing peeling. They work by their ability to improve the appearance of the skin. Typical emollients include fatty acids. It contains sterols, fatty alcohols, mineral oil, polyethersiloxane copolymers, etc. Examples of effective emollients include polypropylene glycol ("PPG")-15 stearyl ester. Cetyl ether, PPG-10 cetyl ether, steareth-10, oleth-8, PP G-4 Lauryl Ether, Vitamin E Acetate, PEG-7 Glyceryl Cocoate, Lanolin This includes, but is not limited to, combinations thereof, of vitamin E acetate. PEG-7 glyceryl cocoate and combinations thereof are preferred.

[0221] Examples of suitable humectants include polyhydric alcohols. Suitable polyhydric alcohols include glycan Cerol (also known as glycerin), polyalkylene glycol, alkylene poly All and their derivatives (propylene glycol, dipropylene glycol, polypropylene Pyrene glycol, polyethylene glycol and their derivatives, sorbitol Hydroxypropyl sorbitol, hexylene glycol, 1,3-dibutylene glyco 1,2,6-Hexanetriol, Ethoxylated Glycerin, Propoxylated Glycerin This includes, but is not limited to, serine and mixtures thereof.

[0222] Appropriate skin soothing agents include panthenol, bisabolol, allantoin, aloe, and so These combinations are included, but are not limited to them.

[0223] Suitable conditioning agents include dimethiconepropyl PG-betaine and dimethicone. Polyols, polyquaternium-10, guar, guar derivatives, and combinations thereof This includes, but is not limited to, salicylic acid and sulfur. Suitable anti-acne active ingredients include salicylic acid and sulfur. Lactic acid, glycolic acid, pyruvate, urea, resorcinol, N-acetylcysteine, Retinoic acid, benzoyl peroxide, octopirox, triclosan, azelaic acid, f Phenoxyethanol, phenoxypropanol, flavonoids, their derivatives, and so These combinations include, but are not limited to, salicylic acid and benzoyl peroxide. It is preferable. [Examples]

[0224] Here, the present invention will be described with reference to specific embodiments. These are merely examples and not explanatory. For the sole purpose: They are not limited to the claimed exclusive scope or the described invention. It is by no means intended to do so. These embodiments are currently intended to carry out the present invention. Configure the best possible mode.

[0225] Example 1: Isolation and identification of compounds 1-3 All three 4-hydroxymethyl-N-methyl-L-proline compounds (1-3) are Z. Observed in the fruits of jujuba, the seeds of guarana, and the leaves of Z. spina-christi. It was isolated, and its structure was elucidated.

[0226] Guarana seeds mainly contain compound 1 (approximately 0.1% w / w), with small amounts of compounds 2 and 3. In the genus Ziziphus, the ratio of compounds 1 to 3 was typically about 2:1. (Approximately 0.1% w / w). Compounds 1 and 2 are from the fruit of Z. jujuba, and compound 3 is from Z. s It was isolated from pina-christi.

[0227] None of these compounds have been isolated from other sources.

[0228] Preparation of extracts from plants of the genus Ziziphus. Dried Z. spina-christi leaf sample (BEZ01) (200g) The substance was extracted overnight in 4 liters of 50% ethanol aqueous solution. The filtered extract was then cationized. Replacement column (H + The captured fraction is passed through an Amberlite IR120 (of the same type) and then converted to an Amberlite IR120 (of the same type). It was eluted with Monia. Then, it was subjected to anion exchange column chromatography (OH). - type The mixture was subjected to Amberlite CG400, and the bound fraction was eluted with acetic acid. It was applied to a acetate column.

[0229] Analysis of the fractions revealed that compounds 1 (HS0810 / 146 / 36-50 mg) and 3 (HS The compound 1 (0810 / 146 / 41-44mg) was identified using the same method. Are JH0806 / 42 / 4) and 2 (LJ746 / 18 / 7) also fruits of Z.jujuba? It was isolated from guarana (JH0420 / 8 / 20).

[0230] Guarana extract (Blue Sky Botanics batch 150701A) Under the GC conditions described below, the trimethylsilyl derivative (main ion 1) was cleaved for a retention time of 9.9 minutes. It has a major compound that matches caffeine as determined by GCMS of 94 amu, and is novel N-methylated pipecolic acid (N-methyl-4R,5S-dihydroxy-2S-pipecoli) The extract of the genus Ziziphus has phenicol (JH0806 / 90 / 5; retention time 8.4 minutes). It was different.

[0231] GC analysis of extracts and pure compounds All samples were lyophilized before derivatization. Trimethylsilyl (TMS) derivatization The body contains hexamethyldisilazane (HMDS) and trimethylchlorosilane (T) in pyridine. A mixture of MCS (Pierce "Tri-Sil" silylation reagents in a 2:1:10 ratio). The sample was prepared using HMDS:TMCS:pyridine. The sample was incubated at 60°C for 15 minutes. After heating, the mixture was left at room temperature for at least 60 minutes. The insoluble reaction product was removed by centrifugation. The solution was allowed to settle, and the supernatant was transferred to a new vial using a syringe.

[0232] The analysis was performed using a highly polar fused silica column (Varian 'Factor Four'). Equipped with a VF-5ms column (25m x 0.25mm ID, 0.25mm layer thickness). G using Perkin Elmer Autosystem XL gas chromatograph The test was performed using C-MS. The carrier gas (helium) flow rate was 1 ml / min. Trimethylsilyl (TMS) derivatives are incubated at 160°C for 5 minutes, then at 10°C / min. Separation was performed using a temperature program that linearly increased the temperature up to 300°C at a rapid rate. The solution was held at 0°C for an additional 10 minutes. The total analysis time was 29 minutes. Electron shock of the column eluate. The mass spectrometry system features a quadrupole ion filter system that operates at 250°C throughout the analysis. The measurements were taken using a Perkin Elmer TurboMassGold mass spectrometer. The detector's mass range was set to 100-650 amu. Transfer line The temperature (GC to MS) was maintained at 250°C. The sample was inactivated quartz wool. Through a fused silica narrow bore injection liner filled with this material, the split vent (min It was injected into the column via a 50:1 ratio. The temperature at the injection port was maintained at 200°C. The input volume was 1 μl. System control, data acquisition, and mass spectral analysis were performed by Per kin Elmer TurboMass software (TurboMass v. 4) It was executed using .4).

[0233] Compounds 1-3 are described below as trimethylsilyl derivatives prepared as described above. Under these conditions, GC-MS exhibits characteristic mass spectra at different retention times. Compound 1, characteristic ions 186 (100%), 260 (10%), and 288 (10%) %)amu(retention time 4.8 minutes). Compound 2, characteristic ions 170 (10%) and 186 (100%) amu (retention time) 5.6 minutes). Compound 3, characteristic ions: 170 (10%), 274 (100%), 288 (15%) , and 376 (10%) amu (holding time 7 minutes).

[0234] Determination of structure The compounds purified by ion exchange were then used with heavy water as the solvent in Bruker A The analysis was performed using a Vance 500MHz instrument. The NMR findings are as follows:

[0235] Structural elucidation of compound 1 [ka] [Table 1] 1 H NMR (500MHz, D2O) δ / ppm 3.89 (1H, t, J = 9.14 Hz, 2), 3.48-3.58 (2H, m, 5), 3. 46-3.50 (1H, m, 6), 3.29 (1H, dd, J = 12 .0, 8.8 Hz, 5), 2.87 (3H, s, 7), 2.65-2. 77 (1H, m, 4), 2.60 (1H, dt, J = 13.6, 8 .4 Hz, 3), 1.71-1.82 (1H, m, 3) 13 C NMR (126MHz, D2O) δ / ppm 173.6 (8), 7 1.2 (2), 62.8 (6), 58.2 (5), 41.4 (7), 3 8.6 (4), 32.2 (3)

[0236] The attachment of protons to individual carbon atoms is determined from the pendant and HMQC spectra. The COZY correlation was determined to be the correlation from 2 to 3 to 4, and the correlation from 4 to both 5 and 6. This shows the relationship. CH(2) at δ3.89 ppm, methyl(7) at δ2.87 ppm, Furthermore, the hydrogen chemical shifts of methylene(5) at δ3.29 ppm and δ3.52 ppm are all This indicates that it binds to nitrogen, forming an N-methylpyrrolidine ring structure.

[0237] The hydrogenochemical shift of methylene at δ3.50 ppm (6) shows that methylene is converted to hydroxyl. This indicates that they bond and thus form a hydroxylmethyl bonded to 4. The quaternary HMBC correlation from 3 to δ173.6 ppm indicates that the acid binds to 2. They are doing it.

[0238] The NOESY correlation between protons with δ3.89 ppm (2) and δ2.70 ppm (4) is: This shows that they are on the same side of the ring and form an S,S configuration. Based on these interpretations... The structure is N-methyl-4S-(hydroxymethyl)-2S-pyrrolidinecarboxylic acid ( It is assumed that it is a compound 1).

[0239] The chemical shift of the proton is in good agreement with what is expected from the standard table of substituent effects. The carbon shift is in strict agreement with the predicted value (ACD CNMR predictor). This type of construction had not been reported before.

[0240] Structural elucidation of compound 2 [ka] [Table 2] 1 H NMR (500MHz, D2O) δ / ppm 3.39-3.53 (2H, m, 6), 2.96-3.05 (1H, m, 2), 2.91 (1H, d, J = 10.7 Hz, 5), 2.42-2.57 (1H, m, 5) , 2.29-2.42 (1H, m, 4), 2.22-2.29 (1H, m , 3), 2.22 (3H, s, 7), 1.46(1H, dt, J = 8.8, 4.7Hz, 3). 13 C NMR (126MHz, D2O) δ / ppm 173.6 (8), 6 8.6 (2), 64.5 (6), 57.8 (5), 41.4 (7), 3 8.6 (4), 33.2 (3)

[0241] The attachment of protons to individual carbon atoms is determined from the pendant and HMQC spectra. The COZY correlation was determined to be the correlation from 2 to 3 to 4, and the correlation from 4 to both 5 and 6. This shows the relationship. CH(2) at δ3.01 ppm, methyl(7) at δ2.22 ppm, Furthermore, the hydrogen chemical shifts of methylene(5) at δ2.91 ppm and δ2.51 ppm are all This indicates that it binds to nitrogen, forming an N-methylpyrrolidine ring structure. δ3.47 The hydrogen chemical shift of methylene in ppm (6) shows that methylene is bonded to hydroxyl. This indicates that it forms a hydroxylmethyl bonded to 4. From 7 to 2 and 5 The N-methylpyrrolidine ring structure is confirmed by HMBC correlation. From 2 and 3, δ1 The HMBC correlation up to quaternary at 73.6 ppm indicates that the acid is bound to δ. There is no NOESY correlation between the protons at 3.01 ppm (2) and δ 2.34 ppm (4). It is on the opposite side of the ring and indicates that it will be an S,R configuration. Based on these interpretations, the structure This is N-methyl-4R-(hydroxymethyl)-2S-pyrrolidinecarboxylic acid (compound 2) It is assumed that this is the case.

[0242] The chemical shift of the proton is in good agreement with what is expected from the standard table of substituent effects. The carbon shift is in strict agreement with the predicted value (ACD CNMR predictor).

[0243] The structure that was elucidated had not been reported before.

[0244] Structural analysis of compound 3 [ka] [Table 3] 1 H NMR (500MHz, D2O) δ / ppm 4.21 (1H, dd, J = 11.3, 7.3 Hz, 2), 3.81 (1H, d, J = 1 2.9 Hz, 5), 3.58 (2H, s, 6), 3.14 (1H, d , J = 12.9 Hz, 5), 2.99 (3H, s, 7), 2.41 (1H, dd, J = 13.9, 7.3 Hz, 3), 2.13 (1H dd, J = 13.9, 11.3 Hz, 3) 13 C NMR (126MHz, D2O) δ / ppm 172.8 (8), 79 .9 (4), 70.6 (2), 64.9 (6), 63.3 (5), 43 .1(7), 39.4 (3)

[0245] The attachment of protons to individual carbon atoms is determined from the pendant and HMQC spectra. The COZY correlation shows a linkage from 2 to 3. C at δ 4.21 ppm H(2), methyl(7) at δ2.99 ppm, and δ3.14 ppm and δ3.81 The hydrogen chemical shift of methylene(5) at ppm is such that all are bonded to nitrogen, forming an N-methyl ring structure. This indicates the formation of HMBC from methyl δ2.99pm(7) to 2 and 5. The correlation confirms this configuration. The singlet δ at 3.58 ppm is likely due to the chemical shifts between carbon and hydrogen. These are integrated as two protons that represent hydroxymethyl. This results in HMB The correlations of C are 3, 4, and 5. The fact that it is a singlet means that it is a fourth-order correlation. Therefore, it means that it is attached. The carbon chemical shift of 4 is hydroxyl uni It means that the parts are joined together.

[0246] Based on these interpretations, the structure is N-methyl-4-hydroxy-4-(hydroxymethyl It is assumed to be (L)-2-pyrrolidinecarboxylic acid. There is a NOESY correlation between 2 and 6. Yes, this means that the hydroxymethyl group is on the same side as the proton of the ring, and therefore the structure N-methyl-4-hydroxy-4S-(hydroxymethyl)-2S-pyrrolidinecarb It is assumed to be acid (compound 3).

[0247] The chemical shift of the proton is in good agreement with what is expected from the standard table of substituent effects. The carbon shift is in strict agreement with the predicted value (ACD CNMR predictor). This type of construction had not been reported before.

[0248] Proton spectra of compound 1 and compound 3 and extraction of Z. spina-christi By comparing it with the proton NMR spectrum of the extracted material (NMR file K101613), A Along with amino acids, it contains a mixture of two compounds as the main N-methylproline component. This was confirmed.

[0249] Example 2: Using compounds 1 and 3 and Z. spina-christi extract Diglycerid load test in S introduction Purified compounds 1 and 3 contain both compounds from Z. spina-christi. The blood glucose regulating activity of the extract in mice was studied.

[0250] method The animal experiment protocol for this study was approved by the Animal Experiment Committee of Toyama University (S -2010UH-2). Male ddy mice (29-33g) that had been fasted overnight were subjected to acute biopsy. Used in glucose tolerance tests. Maltose (2.5g / kg body weight) or sucrose (2.5g ( / kg body weight), and the test sample was dissolved in a 0.9% NaCl solution and administered via a gastric tube. The drug was administered in a vial. The control group received only physiological saline.

[0251] Blood samples for glucose measurement were prepared at 0, 15, 30, 60, and 120 minutes after disaccharide loading. Blood glucose levels were collected from the tail vein. Blood glucose levels were measured using the Antsence II (trademark) portable kit. Measured by Sankyo Co. Ltd., Tokyo, Japan. Extracts from jujube plants HS0810 / 142 / 41 and purified compounds were tested by this method. It was done.

[0252] [Table 4]

[0253] [Table 5]

[0254] [Table 6]

[0255] [Table 7]

[0256] conclusion A mixture of jujube plants, like compound 1, shows blood after a disaccharide challenge with maltose. It had the effect of lowering blood sugar levels. Compound 3 lowered blood sugar more quickly in 30 minutes. Although there appeared to be complex effects, blood sugar levels were clearly revealed early in the sucrose challenge. Although it increased, by 2 hours it was below the higher starting point than other groups, likely due to DMDP It shows better regulation than controls with lower blood glucose levels.

[0257] Example 3: Absence of glucosidase inhibition by compounds 1 and 3 introduction The glycosidase assay was performed using compounds 1 and 3, and the na used in the study of disaccharide-loaded mice. The study was conducted using extracts from plants of the genus Cinnamomum, and investigated whether glucosidase inhibition could explain the effect on blood glucose. We determined whether the activity was nonspecific to the enzyme. Using a glycosidase panel, we determined that the activity found was nonspecific to the enzyme. We confirmed that it not only binds but also selectively inhibits the reaction.

[0258] Enzyme assay The glycosidase assay was developed by Watson et al. (Phytochemist As described in try 1997, 46, 255), p-nitrophenyl substrate (substrate) Sialidase using methylumbelliferyl-N-acetylneuraminic acid as an example The assay was performed using (k). The assay was performed in a microtiter plate. The enzyme was, Enzyme in 0.1M citrate / 0.2M disodium hydrogen phosphate (makilben) buffer The assay was performed at the optimal pH. All assays were carried out at 20°C. All enzymes and substrates were tested. It was purchased from Sigma Aldrich Chemicals Limited. The incubation mixture consists of 10 μl of enzyme solution, 10 μl of iminosaccharide solution, and enzyme 50 μl of appropriate 5 mM p-nitrophenate in a makilben buffer with optimal pH It consisted of a substrate. During the exponential steps of the reaction, the reaction was controlled by 0.4M glycine (pH The process was stopped at 10.4). Absorbance was read at 405 nm. Blank alkaloid Water was used instead. The reaction was performed three times.

[0259] Enzyme inhibitor solution - 1 mg mL of compound -1 Dissolve in distilled water at the following concentration, then add the substrate 1 mg mL per serving -1 After dilution, 0.14 mg mL -1 Screenin at this concentration I did it.

[0260] In the sialidase assay, a 1 mM inhibitor (or water without the inhibitor) and a 2.5 nM inhibitor were used. Sialidase (containing NanH from T. forsythia) is used in 20 mM sodium phosphate solution. 0.1 mM methylumbelliferyl-N-acetylneuraminic acid in um-buffer, pH 7 Incubated in the presence of 0.2. Added to pH 10.5 60 mM sodium carbonate buffer. The reaction was stopped at 30 and 60 seconds. The emission of fluorescent MU was observed at 450 nm. Sialidase activity was quantified by measuring fluorescence emission at excitation at 350 nm. The percentage represents the change in fluorescence between 30 and 60 seconds, compared to the reaction without the inhibitor. It was expressed as a chemical reaction. The reaction was carried out three times.

[0261] result The results are expressed as inhibition percentage. Extracts from jujube plants, or jujube plants and guarana. No glucosidase inhibition was observed with compounds 1 and 3. Glucosidase inhibitors DMDP and DNJ (1-deoxynojirimycin) are precursors of various glucosidases. It showed the expected potent inhibition. Of the other glycosidases tested, the isolated compound showed... The only inhibition observed was from the shearing of bacteria T. forsythia by compound 1. This was due to Ze (compound 3 and extracts from jujube plants could not be used in the assay). (It was.) [Table 8]

[0262] The numbers in bold indicate strong inhibition at a concentration of 0.4 mM.

[0263] Negative values ​​likely indicate increased enzyme activity due to improved stabilization or folding.

[0264] DMDP and DNJ are the most commonly existing plant iminosaccharides, and both are α-glucosyl A potent inhibitor of γ-gase (Watson, AA, Fleet, G. W) J., Asano, N., Molyneux, R.J.;Nash, R. J. Phytochemistry 2001, 56, 265-295). D NJ (1-deoxynojirimycin) was first isolated from mulberry and used in traditional Chinese medicine for diabetes. It was used to treat illness.

[0265] Example 4: Decrease in TNF-α by Compound 1 introduction Measuring TNF-α production in human blood requires systemic administration to control TNF-α levels. Predict the effects of either drug, i.e., shadows in the blood ex vivo or in vitro. Hibiki precisely mimics the effects in vivo. This raises ethical concerns or, more importantly, safety concerns. If it is difficult to conduct research directly in vivo in humans due to any of the considerations It is particularly useful. Whole blood incubation is an in vitro pro-inflammatory drug in humans. It has been shown to be useful in demonstrating the stimulation of the dermatologist (Finch-Arie). tta, MB& Cochran, FR, 1991, Agents a (and Actions 34, 49). This approach is for any of the systemically administered drugs. It is also useful for monitoring the bioavailability of drugs (Song, HJ) et al., 2005, European Journal of Clini Cal Nutrition 59, 508).

[0266] There are several approaches to suppressing inflammation, and currently the most commonly used therapeutically is... The two are a) steroids (glucocorticoid analogs) and b) etanercept ( (Soluble TNF-α receptor fusion protein) and infliximab (mono-TNFα) These are drugs that neutralize the effects of TNF-α, such as clonal antibodies. Both involve suppression of pro-inflammatory cytokine production by steroids, and etanercept. It targets the prevention of TNF-α action by infliximab. This also addresses inflammation. This demonstrates the importance of targeting TNF-α to suppress the response (Feldma nn, M. & Maini, RN, 2001, Annual Review (of Immunology 19, 163). Similarly, glucocorticoid analogs It also suppresses the production of cytokines and dexamethasone, specifically LPS from human monocytes. It can reduce the production of TNF-α in response to stimulation (Waage, A. & B. Akke, O., 1988, Immunology 63: 299).

[0267] The purpose of this study is to investigate the ability of compounds to regulate TNF-α levels in human whole blood. The objective was to evaluate the anti-inflammatory activity of substance 1.

[0268] material Blood and buffy coat fractions are collected from the National Blood Service of Scotland in Glasgow, UK. Provided by SNBTS. Ficoll Histopaque (1.077g / l) ), lipopolysaccharide (from Salmonella abortus equi) is Sigma- Purchased from Aldrich Co. Ltd. (UK). PGE is Cayman C Obtained from chemical Co. (Ann Arbor, MI). TNFα-EL Human TNFα antibody pairs for ISA assays are available from Invitrogen / Life Science. Obtained from iences Europe. All drugs are from Gibco BRL in the UK. It was dissolved in RPMI1640 medium.

[0269] method In experiments using whole blood incubation, the blood was incubated without any further processing after donation. Used. The blood was from the Scottish National Blood Service, and was tested for HIV, hepatitis B, and hepatitis C. All tests for parasitic diseases such as inflammation, CMV, and malaria (conducted by the National Blood Transfusion Service) are negative. When defined as confirming that it was provided by a healthy donor, it was also always 50 By measuring the baseline level of TNF-α, which was less than pg / ml, acute inflammation at the time of blood collection was detected. Our laboratory also confirmed that there are no symptomatic diseases.

[0270] Cellular stimulation and measurement of TNF-α A whole blood aliquot (800 μl) was taken, as shown in the results, for RPMI 1640. Incubate the dissolved compound with the appropriate pre-incubation period, and then LPS was added and the atmosphere was maintained at 37°C for 20 hours in a humidified (100%) air with 5% CO2. The incubation was continued. At the end of the incubation period, plasma or The supernatant, consisting of one of the culture media, is centrifuged at 10,000 g for 30 seconds at room temperature. The data was collected and TNF-α levels were measured using the human TNF-α ELISA system (BioSourc (supplied from Invitrogen, Europe SA, Belgium) It was measured using the method described previously in accordance with the manufacturer's instructions for use (Brown e (t al., 2013).

[0271] Results and Conclusions Compound 1 reduces LPS-stimulated TNFα- production in a concentration-dependent manner in human blood. The trend in values ​​was shown.

[0272] Figure 1 shows the effect of compound 1 (μM) on LPS-stimulated TNF-α production in human blood. Whole blood was collected in the absence of LPS (10 μg / ml) with varying concentrations of the compound (white space). Incubated for 24 hours in the presence of (black circle) or (symbol). 37°C (5% CO2, 10 After incubation at 0% humidity, plasma is collected from the blood by centrifugation, and plasma samples are taken. The level of TNF-α in the sample was measured by ELISA. The value is the mean ± standard value for n=3. This represents the semi-deviation.

[0273] Although not important at the low μM concentrations used here, the concentration of the jujube plant extract is important. Depending on how they are produced, this and other 4-hydroxymethyl-N -Methyl-L-proline (compounds 1, 2, and 3) is claimed to have anti-inflammatory properties in plants. This may help explain the symptomatic effects.

[0274] Example 5: 4-hydroxymethyl-N-methyl sialidase and the proliferation of periodontal pathogens The effects of thyl-L-proline Periodontal disease affects an estimated 700 million people worldwide, and approximately 10-20% of the UK population. This is a significant health burden. Recent studies have shown that, in addition to being a cause of tooth loss, The increasing incidence of periodontal disease is linked to various important factors such as cardiovascular disease, diabetes, and rheumatoid arthritis. It has been shown to be associated with various systemic diseases. The incidence increases with age.

[0275] As part of the progression of the disease, the ability of periodontal bacteria to form subgingival plaque biofilms The ability of periodontal bacteria to stimulate innate immunity through force and interaction with oral epithelial cells is a cause of disease. It is considered a key. Periodontal pathogen Tannerella forsythia, T. Diseases from Denticola and Porphyromonas gingivalis It has been shown that the original sialidase is involved in this process. Therefore, shea Ridase inhibition represents a promising candidate for the treatment or prevention of periodontitis. The enzyme involved is It is present as a major pathogenic factor of the influenza virus (i.e., the N in H1N1 is ta (This represents neuraminidase, the target of miflu.)

[0276] The purpose of this study is to determine how 4-hydroxymethyl-N-methyl-L-proline interacts with periodontal bacteria and The ability to inhibit the sialidase activity of purified sialidase, and extracts containing compounds (e.g.) The objective was to investigate the effects of the fruits of plants in the genus Ziziphus. Furthermore, biofilm formation and We conducted preliminary studies on in vitro models of host-bacterial binding to investigate the toxicity of periodontal pathogens. The effects of compounds and extracts on the survival and proliferation of pathogens in the oral cavity were evaluated. This is because it is considered important.

[0277] The effects in these assays can also predict other medical conditions involving sialidase activity. Human microbiomes that have a more common potential relevance to prevention or cure This may show the effects of the compounds and extracts containing them.

[0278] Sialidase activity assay In these assays, the fluorescence-generating substrate is methylumbelliferyl-N-acetyl Iramic acid (MUNANA) was used. This is cleaved by the action of sialic acid, N - Releases acetylneuraminic acid (Neu5Ac) and methylumbelliferone (MU), The latter exhibits maximum fluorescence at 450 nm when excited at 350 nm. MU emission is due to sialidation. It can be used to quantify enzyme activity.

[0279] Whole bacteria or purified sialidase, compound 1 (Example 3) or 1% N-methyl- In or without the presence of a 50% aqueous ethanol extract of the fruit of a jujube plant containing lorin. And it was revealed to MUNANA.

[0280] 1 mM inhibitor (or water without the inhibitor) and 2.5 nM sialidase (T. fo (Containing NanH derived from rsythia), 0.1 mM MUNANA, pH 7.2, Incubated in the presence of 20 mM sodium phosphate buffer. pH 10.5 60m The reaction was stopped at 30 and 60 seconds by adding M sodium carbonate buffer. Fluorescence generation. MU emission was quantified by measuring fluorescence emission at 450 nm and excitation at 350 nm. The percentage of sialidase activity was compared to that of the response without the inhibitor. The change in fluorescence was observed between 0 and 60 seconds. The reaction was performed three times.

[0281] Various compounds and commercially available sialidase (neuraminidase) inhibitors zanamivir (in The drug used is the pharmaceutical product "Relenza (trademark)" which is sold as a treatment for influenza. Initial screening for N1Neu inhibition was performed.

[0282] Initial screening of compounds and plant extracts for Neu1 activity A sialidase assay was performed, and the reaction was observed using 0.1 mM MUNANA, pH 7.4, 5 0 mM Tris, 5 mM CaCl2, 200 mM NaCl, plant extract or control It either contained the sialidase inhibitor zanamivir at the indicated concentration or did not contain the putative inhibitor. 1 Neu was present at 0.025 μg / ml during the reaction. 50 μl of these reactions yielded 10 Using 0 μl of 100 mM sodium carbonate buffer, pH 10.5, at 5 minutes and 10 minutes. Quenched. Sialidase activity was assessed using the 10-minute time point. The data shown is... The value of 'T' represents the average of one experiment in which each condition was repeated three times. The values ​​present in the quenched reaction are... The MU is excited at 340nm and emits light at 430nm, using the TECANM200 Infinite Microwave. Sialidase activity was quantified using a cross-plate reader and compared to conditions without an inhibitor. It was expressed as a percentage of sex (MU fluorescence).

[0283] host cell binding In addition to forming biofilms and being present on the mucosal surface, bacteria also bind to host cells. It can adhere and penetrate. This includes antimicrobial therapy, the immune system, or mechanical removal of the biofilm. For bacteria, persistence when faced with challenges, and access to nutrients within host cells, There are several advantages to this. The binding of bacteria to host cells also affects immunomodulation. Increased cell infiltration can lead to upcontrol of inflammation and may be a contributing factor to periodontal disease. Sialidase (and other products) are the host plants of P. gingivalis and T. forsythia. It is important for binding with the chief cell.

[0284] Adhesion of host cells by P. gingivalis and T. forsythia Antibiotic protection assays were used to test for inhibition of infiltration. These were performed using two sets of assays. This involves growing a monolayer of host cells, and then dividing them into a host cell:bacterial ratio of 1:100. The bacteria were infected at a rate (multiple infections). After 1.5 hours of incubation, one set was used. The bacteria were dissolved and seeded onto agar to obtain the number of viable bacteria. This is the number of bound cells. This brings up the total number of bacteria. The second set of infected monolayers is treated with the antibiotic metronidazole for 1 hour. Incubated. Metronidazole killed external (attached) bacteria and infiltrated ( Only the internalized bacteria remain. Then, the cells are lysed and seeded onto agar as described above. The number of invasive bacteria was subtracted from the total number of bacteria bound to obtain attached (external) bacteria.

[0285] Biofilm assay Periodontal pathogens are present in the biofilm in the oral cavity (as part of plaque), and in normal oral cavity Both hygiene and periodontal disease treatment involve destroying the biofilm. Therefore, potential Such treatments or preventive measures should also be able to destroy the biofilm of periodontal pathogens. This can be modeled in vitro as a single or mixed species biofilm. The effects of plant extracts and compounds may not be directly antibacterial, but they affect host glycoproteins. It acts to prevent adhesion to the substance or to prevent the acquisition of nutrients from glycoproteins, Various different approaches were taken. These involved using serum (fetal bovine serum - FBS) P. g in a nutrient-rich culture medium (trypsin soy broth - TSB) on a coated surface. To culture ingivalis and T. forsythia, as well as carbon and nitrogen P. gingiv was found in a nutrient-deficient medium containing only serum (10% v / v FBS) as the source. This involved culturing alis, considering its apparent activity as a sialidase inhibitor. The ability of extracts from jujube plants to prevent biofilm formation was tested. .

[0286] To quantify biofilm formation, we use a high-throughput, low-sensitivity CrystalBio Olette staining and crystal violet staining have higher sensitivity but lower throughput. Two approaches were used for bacterial counting. Crystal violet staining was used. It can also be used to image biofilms for qualitative observation.

[0287] P. gingivalis and T. forsythia were found in 1500 μg / ml of na Microtie pre-coated with FBS in the presence or absence of extracts from the genus Clawberry. The cells were cultured in tar plates for 2 days and 5 days in supplemented TSB, respectively. In the experiment, it became possible to quantify biofilm formation by counting bacteria, and crystal Violet staining made it possible to visualize biofilms. Under these conditions, Biofilm formation of P. gingivalis was not significantly inhibited. T. f Orsythia biofilm formation was reduced by at least twofold under all conditions, and inhibitors 2.2x10 of the control without the control 7 Bacteria / ml to 1.0 x 10 7 It decreased to [a certain level].

[0288] Biofilm inhibition assay - P. gingivalis + F. n in rich medium ucleatum (25586) mixed species biofilm T. forsythia and P. gi cultured on TSB on serum-coated surfaces The effect of extracts from the genus Ziziphus on Ziziphus ngivalis P. gingivalis or T. forsythia, FBS (50 μl) After adding 00% FBS and incubating overnight at 4°C, wash once with PBS to remove unbound cells. Pre-coated 96-well polylysine coated tissue cultures (with serum removed) In the well of the oat, 0.1% w / v yeast extract, 5 μg / ml hemin, 1 μg / ml mena Seeds were seeded at an OD600 of 0.05 in a TSB containing dione and 50 μg / ml gentamicin. Seeds were planted. Both seeds were grown at 37°C for 5 days for T. forsythia and P. gi. Ngivalis was cultured anaerobically for 48 hours. The supernatant was removed, and the wells were PB (Primary Bacteria). Washed twice with S. The biofilm was vigorously rinsed in PBS before dilution (if appropriate). The samples were resuspended and counted using a Herber chamber and a phase-contrast microscope at 400x magnification.

[0289] The imaging wells were stained with crystal violet. Remove the supernatant and the wells Incubate the mixture with 0.1% (w / v) crystal violet at room temperature for 15 minutes. The dye was removed, and the wells were washed three times with PBS. The biofilm was inverted. The image was captured at 400x magnification using a microscope and imaging software. The image capture parameters (light intensity and exposure time) were constant for each image. The data represents the average of three experiments, with each condition repeated three times in each experiment.

[0290] Possible explanations for the different effects of jujube plant extracts on biofilm formation are as follows: The difference lies in the nutritional requirements of these two organisms; T. forsythia is different from P. gingi It has stricter growth requirements than Valis. For example, T. forsythia Normally, an exogenous source of N-acetylneuraminic acid (NAM) is required, but this is bio It can be replaced with sialic acid during film formation (in this experiment, this source is FBS). (provided by), this can be destroyed by sialidase, while P Gingivalis can be found in supplemented TSB medium even in the presence of sialidase inhibitors. It may be possible to obtain the necessary nutrients from it. Therefore, as a source of carbon and nitrogen Limited media containing only serum (FBS) also showed efficacy in extracting jujube plants for biofilm formation. It was used to evaluate the effects of objects.

[0291] Biofilm Inhibition Assay - P. gingivalis in Limited Medium T. forsythia is more hardy than P. gingivalis, so use a limited medium. Attempts to culture it as a single-species biofilm using this medium were unsuccessful. Therefore, limited medium was used. When extracts from the genus Ziziphus are applied to P. gingivalis itself that has been cultured, P. gingivalis was treated by adding 50 μl of 100% FBS. After incubation at 4°C overnight, the sample was washed once with PBS to remove unbound serum. In the wells of a recorded 96-well polylysine-coated tissue culture plate, 10 Further supplementation with %v / v FBS, 5 μg / ml hemin, and 50 μg / ml gentamicin. P. gingivalis was sown in a limited culture medium at 0.05 OD600. The cells were cultured anaerobically at °C for 5 days.

[0292] Remove the supernatant and immerse the wells in 0.1% (w / v) crystal violet at room temperature for 1 Incubated for 5 minutes. Removed the dye and rinsed the wells with a stream of water. A) Crystal Bar Quantification of violet staining - Crystal violet is stained with ethanol:acetone (80:2 0) Extracted from the biofilm into a new 96-well microtiter plate. This was quantified by measuring the absorbance at 590 nm.

[0293] result The data shown represents the average of two experiments, and each condition was repeated 10 times in each experiment. It was returned.

[0294] N1 neurialidase inhibition [Table 9] [Table 10] Compound 1 at NB 1 mM exhibits 50% inhibition of NanH from T. forsythia. This resulted in (Example 3)

[0295] [Table 11] [Table 12] [Table 13] In jujube plants, 1500 μg / ml of the inhibitor reduced biofilm formation compared to no inhibitor. This showed [Table 14] In NB-rich medium, P. gingiv was stained with crystal violet. There was no clear effect against ALS biofilms.

[0296] Consideration Compound 1 (Example 3) and 1% of 4-hydroxymethyl-N-methyl-L-proline 1~ The extract of the jujube plant containing 3 is from Tannerella forsythia. It inhibited dase. The extract from the jujube plant did not appear to have a direct antibacterial effect. Despite having a high sugar content, it prevents the acquisition of nutrients and serum coating By limiting adhesion to the surface, biofilm formation can be inhibited. Biofilm formation in P. gingivalis was significantly inhibited by plant extracts. Although it was not possible, it was cultured in a limited medium (serum-FBS is the sole source of carbon and nitrogen). This was inhibited.

[0297] This indicates that the extract containing compounds 1-3 inhibits its sialidase and possibly other enzymes. This suggests that it was interfering with nutrient acquisition by P. gingivalis. Therefore, this may limit the protein catabolism of P. gingivalis. This is particularly detrimental to the acquisition of nutrients from serum, and to protein sources (soybean meal). However, this is not so harmful to obtaining nutrients from TSB, which has already undergone enzymatic digestion. Presumably, the limited culture medium contains P. gingivalis, which is composed of gingival crevicular fluid (GCF), serum, and other components. (Similar to) host glycoproteins, and other glycoproteins in blood and saliva, through catabolism. This better illustrates the situation in vivo, where nutrients are acquired.

[0298] T. forsythia is more strict than P. gingivalis, so use a limited medium. Attempts to culture it as a single-species biofilm using this method have so far failed. However, biofilms can form on the serum-coated surface of TSB. In this series of experiments, extracts from the jujube plant were found to be biofiltrating T. forsythia. It appeared to inhibit cell formation.

[0299] These findings also indicate that the compounds of the present invention are related to species Ga, which are associated with bacterial vaginosis and premature birth. Based on inhibition of sialidases from other bacteria, including Rdnerella vaginalis. This demonstrates that it can be found to have applications in various therapeutic procedures.

[0300] Example 6: Mouse samples of Zizyphus jujuba fruit 50% aqueous ethanol extract test The fruit extract of Zizyphus jujuba was prepared with 50% aqueous ethanol and dried. Analysis of the extract by gas chromatography-mass spectrometry (GCMS) revealed that monosaccharides The study showed a high sugar content (>5%) and the presence of compounds 1-3 at 1%. Z. spi Previous studies on the extraction of Na-Christi extracts have shown that maltose and sucrose are negative. This was performed in male ddy mice during a load test (Example 2).

[0301] In this study, male C57BL / 6J mice were subjected to glucose, maltose, and sugar. It was used in a load test of loin.

[0302] method The animal experiment protocol was approved by the Animal Experiment Committee of Toyama University. The animals were kept asleep overnight. They were fed either glucose (2.5g / kg body weight) or sucrose (2.5g / kg body weight). Furthermore, the extract was dissolved in a 0.9% NaCl solution and administered to mice via a gastric tube. The control group received only physiological saline. Blood samples for glucose measurement were prepared at 0 after glucose loading. Blood glucose levels were collected from the tail vein at 15, 30, 60, and 120 minutes. Antsence II (trademark) (Sankyo Co. Ltd. Tokyo, J Measured by (apan). The tests were as follows:

[0303] ● Glucose tolerance test: 1000 mg / kg body weight (Figure 2, Table 1) ● Glucose tolerance test: 500 mg / kg body weight (Figure 3, Table 2) ● Sucrose loading test: 1000 mg / kg body weight (Figure 4, Table 3) ● Maltose loading test 500 mg / kg body weight (Figure 5, Table 4)

[0304] result Figure 2 shows the effect of extracts from jujube plants on blood glucose levels. Each of the following 2.5 g / kg body weight glucose samples contains an extract (white circle) containing 1000 mg / kg body weight. Blood glucose concentration in male C57BL / 6J mice after oral feeding. The control group was fed physiological saline. (Black circle). Each value represents the mean ± SEM (n=5).

[0305] Figure 3 shows the effect of extracts from jujube plants on blood glucose levels. 2.5 g / kg body weight glucose, containing extract (white circle) (500 mg / kg body weight each), is administered orally. Blood glucose levels in male C57BL / 6J mice after loading. The control group was loaded with physiological saline. (Black circle). Each value represents the mean ± SEM (n=5).

[0306] Figure 4 shows the effect of extracts from jujube plants on blood glucose levels. Each of the following is a sucrose extract containing 1000 mg / kg body weight (white circle) and 2.5 g / kg body weight. Blood glucose concentration in male C57BL / 6J mice after oral feeding. The control group was fed physiological saline. (Black circle). Each value represents the mean ± SEM (n=5).

[0307] Figure 5 shows the effect of extracts from jujube plants on blood glucose levels. 2.5 g / kg body weight of maltose, including extract (white circle) (500 mg / kg body weight each), is administered orally. Blood glucose levels in male C57BL / 6J mice after loading. The control group was loaded with physiological saline. (Black circle). Each value represents the mean ± SEM (n=5).

[0308] [Table 15] [Table 16] [Table 17] [Table 18]

[0309] conclusion The fruit extract of Zizyphus jujub, which contains additional monosaccharides, contains glucose and sugars. - Suppressed blood glucose in both loading tests. The effect of maltose or sucrose on loading conditions. The results showed that, except for the fact that the extract did not show significant glucosidase inhibition, maltase and This may have been caused by sucrase inhibition. The results of the glucose tolerance test were glucosy A different mechanism independent of γ-dosage inhibition, and possibly insulin release or other mechanisms. This suggests an effect on rheumatism.

[0310] Equal portions The above description details currently preferred embodiments of the present invention. Consider these descriptions. Those skilled in the art will anticipate that numerous modifications and variations will occur in practice. These modifications and variations are intended to be included within the scope of the claims appended herein. It is.

Claims

1. A method for producing a composition, The composition is a pharmaceutical composition, herbal medicine, or nutritional supplement, and the process includes the step of monitoring the quality of the pharmaceutical composition, herbal medicine, or nutritional supplement by detecting the presence or absence of 4-hydroxymethylproline in a sample of the pharmaceutical composition, herbal medicine, or nutritional supplement, or by measuring the amount thereof, wherein the 4-hydroxymethylproline is 【Chemistry 1】 Alternatively, the method comprising selecting from pharmaceutically acceptable salts thereof or their N-oxides.

2. A method for monitoring the quality of a pharmaceutical composition, herbal medicine, or nutritional supplement, (a) A step of providing a sample of a pharmaceutical composition, herbal medicine, or nutritional supplement, (b) The step of detecting the presence or absence of 4-hydroxymethylproline in the sample, or measuring its amount, The aforementioned 4-hydroxymethylproline, 【Chemistry 2】 Alternatively, the method comprising selecting from pharmaceutically acceptable salts thereof or their N-oxides.

3. A method for selecting plant breeding lines and / or varieties, Step (1): A step of providing a material derived from the plant of the plant source, wherein the plant source is selected from (a) plants of the genus Ziziphus and (b) plants of the genus Guarana, and the plant source may be selected from the plant species of (i) Z. jujuba, Z. spina-christi, Z. lotus, Z. mauritiana, Z. joazeiro, and Paulinia cupana, and / or (ii) fruits, fruit parts, fruit extracts, fruit juice, seeds, bark, roots, and / or leaves. Step (2): A step of determining the presence or absence of 4-hydroxymethylproline in the material from step (1), and / or measuring its amount. Includes, The aforementioned 4-hydroxymethylproline, 【Transformation 3】 Alternatively, the method comprising selecting from pharmaceutically acceptable salts thereof or their N-oxides.

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