Finestem vanilla saponin compositions for inhibiting lipase and xanthine oxidase and methods of making the saponin compositions

Abstract

A method of preparing a trilodax saponin composition is provided. Also provided are a trilodax saponin composition obtained by the method, an oral composition comprising the trilodax saponin composition, use of the trilodax saponin composition in the manufacture of a medicament for treating hyperuricemia and / or hyperlipidemia in a subject, and a method of inhibiting one or both of lipase and xanthine oxidase in a subject. The trilodax saponin composition comprises, consists essentially of, or is one or more isolated saponins having a particular structure described herein.

Description

Technical Field

[0001] The present invention generally relates to compositions and methods for inhibiting lipases and / or xanthine oxidases, and more specifically to methods for preparing Lysimachia capillipes saponin compositions for inhibiting lipases and / or xanthine oxidases in a target. Background Technology

[0002] Pancreatic lipase (PL) is a key enzyme responsible for the hydrolysis of 50%-70% of dietary fats in the human gastrointestinal system and is widely recognized as one of the key enzymes for fat digestion and absorption. PL inhibitors can reduce the breakdown (metabolism) and absorption of dietary fats in the human digestive organs by lowering the hydrolytic activity of this key enzyme, thereby alleviating and preventing symptoms of metabolic diseases such as hyperlipidemia.

[0003] Xanthine oxidase (XO) is an enzyme that catalyzes the oxidation of hypoxanthine to xanthine, which is then oxidized to uric acid. Hydrogen peroxide (H₂O₂) is produced as a byproduct, and sometimes superoxide ions (O₂·4H₂O) are also generated. - The production of byproducts of uric acid and / or xanthine oxidation can contribute to pathological disorders such as hyperuricemia, gout, ulcers, cancer, ischemia, hypertension, cardiovascular disease, and oxidative stress.

[0004] In light of the foregoing, there remains an opportunity to provide novel and useful pancreatic lipase inhibitors. Such inhibitors could offer potential preventative and therapeutic pathways to reduce the risk of hyperlipidemia or other pathogenic complications caused by pancreatic lipase activity. Furthermore, there remains an opportunity to provide novel and useful xanthine oxidase inhibitors. Such inhibitors could offer potential preventative and therapeutic pathways to reduce the risk of hyperuricemia, gout, ulcers, cancer, ischemia, hypertension, cardiovascular disease, oxidative stress, oxidative damage, or other pathogenic complications caused by xanthine oxidation. Therefore, there remains an opportunity to provide novel and useful compositions and methods for inhibiting one or more of pancreatic lipases and xanthine oxidases. Summary of the Invention

[0005] Invention Summary

[0006] A method for preparing a saponin composition of *Hemiberlesia serratifolia* is disclosed. The *Hemiberlesia serratifolia* saponin composition may also be referred to as a composition comprising, optionally substantially comprising, or optionally composed of, saponins obtained or isolated from *Hemiberlesia serratifolia*.

[0007] The method includes a first extraction of the herb *Heliotropium indicum* material with a first solvent composition to obtain a crude extract, gradient elution of the crude extract with a microporous adsorption resin and an eluent containing an alcohol to obtain a second extract, separation of the second extract by normal-phase silica gel chromatography to obtain crude saponins, and separation of the crude saponins by medium-pressure liquid chromatography and high-performance liquid chromatography to obtain one or more isolated saponins of *Heliotropium indicum*.

[0008] The isolated saponins are also disclosed. These isolated saponins are obtained from *Vigna arvense* material. For example, the isolated saponins can be obtained by the preparation method described above. The isolated saponins comprise one or more of the chemical formulas (Ia) to (Ih) described herein.

[0009] In various embodiments, the herbaceous stalk saponin composition and the isolated saponin are the same, or they are substantially the same. Therefore, references herein to the herbaceous stalk saponin composition may be used interchangeably with references herein to the isolated saponin, and vice versa.

[0010] Also provided are a herbal saponin composition obtained by the method, an oral composition (including the herbal saponin composition) for oral administration to a subject, the use of the herbal saponin composition in the preparation of a medicament (or pharmaceutical agent) for treating hyperuricemia and / or hyperlipidemia in a subject, and a method for inhibiting one or both of pancreatic lipase and xanthine oxidase in a subject. The composition can be used to inhibit one or both of pancreatic lipase and xanthine oxidase in a subject. The method includes administering an effective amount of the composition to the subject, the composition being a herbal saponin composition or an oral care composition. In various embodiments, the composition is administered orally to the subject. Brief description of the attached diagram

[0011] Figure 1 The flowchart illustrates a method for preparing a schizocarpine saponin composition according to an embodiment of the present disclosure;

[0012] Figure 2 The figure shows the inhibition rate of pancreatic lipase (PL) using vanilla saponin DT05;

[0013] Figure 3 The figure shows the inhibition rate of pancreatic lipase (PL) using vanilla saponin DT06;

[0014] Figure 4 The figure shows the inhibition rate of pancreatic lipase (PL) using vanilla saponin DT09;

[0015] Figure 5The figure shows the inhibition rate of pancreatic lipase (PL) using vanilla saponin DT11;

[0016] Figure 6 The figure shows the inhibition rate of pancreatic lipase (PL) using vanilla saponin DT15;

[0017] Figure 7 The figure shows the inhibition rate of xanthine oxidase (XO) using vanilla saponin DT01;

[0018] Figure 8 The figure shows the inhibition rate of xanthine oxidase (XO) using vanilla saponin DT02;

[0019] Figure 9 The figure shows the inhibition rate of xanthine oxidase (XO) using vanilla saponin DT19; and

[0020] Figure 10 The figure shows the inhibition rate of xanthine oxidase (XO) using vanilla saponin DT11.

[0021] Detailed description of the current implementation plan

[0022] A method for preparing a saponin composition of *Heliotropium indicum* (which may be simply referred to herein as "the composition") is disclosed. Isolated saponins from *Heliotropium indicum* are also disclosed. The composition contains at least one phytoactive component. The at least one phytoactive component may be one or a combination of the isolated saponins. The composition and its preparation method are described below, followed by a description of the isolated saponins, and related uses and methods. The *Heliotropium indicum* saponin composition contains one or a combination of isolated saponins, is substantially composed of, or is composed of.

[0023] As will be understood from this disclosure, the composition is not particularly limited except for the plant-active components (e.g., isolated vanillin saponins), and in particular their extracts, and the methods for obtaining said plant-active components. Therefore, the composition can be formulated, for example, into an oral composition or contained therein, or into a nutritional product, medicine, or supplement or contained therein, and can be used as a unique and independent therapeutic agent or in combination with other compatible therapeutic agents.

[0024] The composition can be used to treat, prevent, and / or improve various conditions, such as those related to pancreatic lipase activity. Specifically, it will be understood from the following description and examples that the composition of this embodiment is believed to be able to inhibit pancreatic lipase, and more specifically, inhibit pancreatic lipase in the target.

[0025] The composition can also be used to treat, prevent, and / or improve various conditions, such as those related to xanthine oxidase activity. Specifically, it will be understood from the following description and examples that the composition of this embodiment is believed to be able to inhibit xanthine oxidase, and more specifically, inhibit xanthine oxidase in the target.

[0026] Therefore, the composition can be used to treat (i.e., alleviate, prevent, reverse, etc.) conditions commonly associated with pancreatic lipase activity, such as complications associated with hyperlipidemia and other health disorders. The composition can also be used to treat (i.e., alleviate, prevent, reverse, etc.) conditions commonly associated with xanthine oxidase activity, such as complications associated with hyperuricemia, gout, and other health disorders such as ulcers, cancer, ischemia, hypertension, cardiovascular disease, oxidative stress, and oxidative damage.

[0027] As described above, the composition comprises a plant-active component. More specifically, the plant-active component comprises at least one extract of *Heliotropium indicum*, optionally consisting substantially of or optionally consisting of therefrom. In various embodiments, the plant-active component consists of a single isolated extract of *Heliotropium indicum* or a combination of single isolated extracts of *Heliotropium indicum* as described herein. As described above, the plant-active component may be an isolated saponin or a combination of isolated saponins as described herein.

[0028] Thin-stemmed vanilla

[0029] The plant-active component and therefore the composition comprises an extract of *Hemiberlesia lataniae*, i.e., an extract comprising material from the plant species *Hemiberlesia lataniae*, optionally consisting substantially of it, or optionally composed of it. There are no particular limitations on the *Hemiberlesia lataniae* extract, and it may comprise or be obtained by the methods disclosed herein and suitable for use in the embodiments herein, any extract or combination of extracts from the *Hemiberlesia lataniae* plant. More specifically, exemplary *Hemiberlesia lataniae* extracts include those that are capable of inhibiting pancreatic lipases and / or xanthine oxidases and / or causing / exhibiting any other such activities, as described herein as part of the plant-active component.

[0030] It has been reported that *Lysimachia capillipes* contains various bioactive components, such as triterpenoids, saponins, phenolic acids, flavonoids, flavonols, flavones, organic acids, and other compounds. *Lysimachia capillipes* can be simply called *L. capillipes*, or by various other names such as *Lysimachia capillipes* Hemsl., *Hairystalk Loosestrife*, "Xi Geng Xiang Cao", *Lysimachia capillipes*, "Pai Cao Xiang", or *Pai Cao Xiang*.

[0031] Those skilled in the art will understand that *Heliotropium indicum* is cultivated primarily for its plant material, which includes, but is not limited to, its roots, stems, rhizomes, leaves, buds, flowers, seeds, and / or fruits. Therefore, in various embodiments, the extract of *Heliotropium indicum* is an extract of the whole plant. The plant material can be derived from one or more plants and can be fresh or dried, typically dried to prevent decay. The dried plant material can then be formed into a powder, which is further processed to form the extract, as described below.

[0032] In various embodiments, the plant-active component comprises an extract from the whole *Heliotropium indicum* plant. In these embodiments, the plant material of the whole *Heliotropium indicum* plant can be, for example, the roots, stems, rhizomes, leaves, buds, flowers, seeds, and / or fruits of the *Heliotropium indicum* plant. Without being bound by any particular theory, it is believed that the whole plant of *Heliotropium indicum* is most useful for inhibiting pancreatic lipase and / or xanthine oxidase; while individual parts of *Heliotropium indicum* are not (as explained in the Examples section below).

[0033] In other embodiments, the *Heliotropium indicum* extract may comprise material from any part or combination of parts of the plant, and is not limited to extracts from the whole plant. For example, the *Heliotropium indicum* extract may comprise material extracted from one or more parts of the *Heliotropium indicum* plant, including its roots, stems, rhizomes, leaves, buds, flowers, seeds, and / or fruits. Furthermore, the extract may be further processed (e.g., defatting, partial defatting, grinding, drying, precipitation, washing, filtering, sieving, extraction, distillation, concentration, etc.) to obtain the *Heliotropium indicum* extract. Similarly, the *Heliotropium indicum* plant may be extracted in an unprocessed form or processed prior to the extraction of the *Heliotropium indicum* extract (e.g., used in an unprocessed form, suspension form, dehydrated form, concentrated form, etc.). In some embodiments, the plant active component comprises the *Heliotropium indicum* extract, which comprises material obtained (i.e., extracted) from the whole plant of *Heliotropium indicum*.

[0034] The amount of *Heliotropium indicum* extract used in the plant active ingredient can vary and will be selected based on the number and type of components used in the plant active ingredient. In some embodiments, the plant active ingredient comprises 0.1 to 5000 mg of *Heliotropium indicum* extract, such as 0.1 to 4000 mg, optionally 1 to 3500 mg, optionally 10 to 3000 mg, optionally 20 to 2500 mg, or optionally 50 to 2000 mg. However, amounts outside these ranges may also be used. For example, in some embodiments, the plant active ingredient comprises an amount of *Heliotropium indicum* extract of at least 0.1 mg, optionally at least about 1 mg, optionally at least about 20 mg, optionally at least about 50 mg, optionally at least 100 mg, optionally at least 250 mg, optionally at least 500 mg, optionally at least 1000 mg, optionally at least 1500 mg, optionally at least 2000 mg, optionally at least 2500 mg, optionally at least 3000 mg, optionally at least 3500 mg, optionally at least 4000 mg, or optionally at least 4500 mg. In these or other embodiments, the upper limit may be selected such that the plant active ingredient comprises an amount of *Heliotropium indicum* extract of ≤100, ≤250, ≤500, ≤750, ≤1000, ≤2000, or ≤5000 mg. In various embodiments, the plant-active component may include an amount of *Heliotropium indicum* extract, optionally greater than 1% by weight, optionally greater than 5% by weight, optionally greater than 10% by weight, optionally greater than 25% by weight, optionally greater than 50% by weight, optionally greater than 75% by weight, optionally greater than 80% by weight, or optionally greater than 95% by weight, based on the total weight of the plant-active component. In such embodiments, the upper limits can be selected typically as ≤10% by weight, ≤20% by weight, ≤30% by weight, ≤40% by weight, ≤50% by weight, ≤60% by weight, ≤70% by weight, ≤80% by weight, ≤90% by weight, and ≤99% by weight, based on the total weight of the plant-active component.

[0035] In some embodiments, the plant active ingredient comprises more than one herb extract, such as two, three, four or more herb extracts. In such embodiments, each herb extract is selected independently and may be the same as or different from any other herb extract, and each is used in the amount described above.

[0036] The herb extract can be used in any form, such as pure (i.e., without solvents, carrier media, diluents, etc.) or in a carrier media such as a solvent or dispersant. If present, the carrier media may comprise an aqueous solvent (e.g., water), an organic solvent, a fluid, or an oil, or a combination thereof. When used, the carrier media will be selected based on the specific components of the plant active ingredient and / or composition (such as the specific herb extract used). It should be understood that the herb extract can be combined with a carrier media (if used) before, during, or after combination with the plant active ingredient and / or any other components of the composition.

[0037] Method for preparing herb saponin compositions

[0038] refer to Figure 1 Methods for preparing vanilla saponin compositions typically involve extracting vanilla material, subsequently separating the crude extract, and then purifying it. In specific embodiments of the method, the vanilla material is the whole vanilla plant. Optionally, the vanilla plant material may be steamed before the extraction step. For example, fresh vanilla plants may be steamed or blanched in hot water for a period of several minutes, then dried, and subsequently cut into pieces or alternatively left intact. However, in other embodiments, the vanilla plant material is not steamed or blanched before extraction begins.

[0039] Extraction of saponins from *Heliotropium indicum* plant material comprises a series of extraction steps to obtain an extract. The term "extract" is used herein in its conventional sense to refer to a composition obtained from a source material by fluid extraction. Therefore, the term "extract" or "plant extract" should be understood as a composition obtained from a plant source (i.e., plant material) by fluid extraction (e.g., solvent extraction, gas extraction, CO2 extraction, etc.). Extraction methods include water extraction, steam extraction, solvent extraction, etc., known in the art. Exemplary extraction techniques include solvent extraction, for example, using polar solvents such as alcohols (e.g., methanol, ethanol, butanediol, etc.), ethers (e.g., diethyl ether, methyl tert-butyl ether, etc.), ketones (e.g., acetone), esters (e.g., ethyl acetate), phenol, water, etc.), nonpolar solvents such as benzene, xylene, toluene, etc., and their derivatives, modifications, and combinations thereof (e.g., solvent-water blends, including alcohol-water, acetone-water, etc.). Other and alternative extraction techniques include sequential fractionation, total hydro-ethanolic extraction, single-stage extraction, supercritical fluid extraction (e.g., with CO2), and those or other processing techniques (such as filtration, purification, distillation, dehydration, evaporation, concentration, drying, etc.) utilizing sequential or secondary extraction from a first extract (e.g., a non-polar solvent extract of a plant extract obtained from a polar solvent extract). Specific examples of suitable extraction methods are described in U.S. Patent No. 7,897,184, which is incorporated herein by reference.

[0040] In one embodiment, organic solvent extraction techniques can be used to obtain plant extracts. In another embodiment, sequential fractionation of the solvent can be used to obtain plant extracts. Water-ethanol extraction techniques can also be used to obtain plant extracts. This is typically referred to as a single-pass extraction. The plant extract produced in this process will contain a variety of phytochemicals present in the extracted material, including fats and water-soluble phytochemicals. After collecting the plant extract solution, the solvent is evaporated to obtain the plant extract.

[0041] Extraction with pure ethanol can also be used. This technique uses ethanol as a solvent. The plant extract produced by this extraction technique may contain lipid-soluble and / or lipophilic compounds in addition to water-soluble compounds. Extraction with pure methanol can also be used in a similar manner and similar results can be obtained.

[0042] Another example of an extraction technique that can be used to obtain plant extracts is supercritical fluid carbon dioxide extraction (SFE). In this extraction procedure, the material to be extracted is not exposed to any organic solvent. Instead, under supercritical conditions (e.g., >31.3 °C and >73.8 bar), the extraction solvent is carbon dioxide (CO2), with or without a modifier. Those skilled in the art will understand that temperature and pressure conditions can be varied to obtain the optimal yield of the plant extract. This technique produces plant extracts of lipophilic and / or fat-soluble compounds, similar to the all-hexane and ethyl acetate extraction techniques that can also be used.

[0043] Each of the above extraction methods may also include one or more additional processing steps and / or combinations thereof as understood in the art. For example, the plant material may be pulverized, crushed, ground, etc. One or more filtration steps may also be included to remove, for example, cellulose / fiber or other solid materials. One or more purification steps may also be included to remove, for example, certain components and / or contaminants. Such purification may be accomplished, for example, by distillation, evaporation, centrifugation, etc. One or more concentration and / or drying steps may also be included to remove water and / or other volatiles, such as alcohols, lighter compounds, VOCs, etc. Furthermore, acids and / or bases may be added to adjust the pH or neutralize it. Depending on the desired form of the final / final plant extract, various additional steps as understood in the art may also be utilized, such as screening, pressing, grinding, milling, mixing, dispersing, etc. It should be understood that combinations of these additional processing steps, repeated and / or in different orders, are also contemplated.

[0044] In various embodiments, the solvent used to obtain a suitable plant extract of this disclosure is a solvent in which the obtained plant extract and / or its subsequent forms (e.g., plant extract powder) are suitable for ingestion. For example, the solvent is water or ethanol.

[0045] Further reference Figure 1In this method, a first extraction of steamed or uncooked *Heliotropium indicum* plant material is performed using a first solvent composition to obtain a crude extract of *Heliotropium indicum*. In specific embodiments, the first solvent composition comprises an alcohol, preferably ethanol (EtOH). In some embodiments, the first solvent composition is preferably 75% ethanol. Alternatively, the first solvent composition is 70% to 80% ethanol, 71% to 79% ethanol, 72% to 78% ethanol, 73% to 77% ethanol, or 74% to 76% ethanol. However, it should be understood that the extraction can be carried out with 0.1% to 100% ethanol. In specific embodiments, the extraction is performed with 10 times the concentration of 75% ethanol for at least 0.5 hours, alternatively at least 1.0 hour, at least 1.5 hours, at least 2.0 hours, at least 2.5 hours, at least 3.0 hours, at least 3.5 hours, or at least 4.0 hours. Optionally, the first extraction is performed over a period of 3.5 to 4.5 hours, alternatively 3.6 to 4.4 hours, 3.7 to 4.3 hours, 3.8 to 4.2 hours, or 3.9 to 4.1 hours. Preferably, the first extraction lasts at least 4 hours. In some embodiments, the first extraction is repeated at least once more, such that the first extraction comprises two or more similar or nearly identical sub-extraction steps. For example, the first extraction may be performed twice consecutively.

[0046] After the first extraction, the crude extract is eluted with a gradient using a macroporous adsorption resin and an eluent containing an alcohol to obtain a second extract. In a specific embodiment, the alcohol is ethanol, and the eluent comprises a gradient of ethanol solutions, including 20% ​​ethanol, 40% ethanol, 70% ethanol, and 95% ethanol. The crude extract is added to a bed of microporous adsorption resin using water as the washing solvent. The eluent is then gradually introduced into the macroporous adsorption resin, starting with 20% ethanol, then 40% ethanol, then 70% ethanol, and then 95% ethanol. The macroporous adsorption resin can be, for example, a styrene-type nonpolar copolymer, such as D101 macroporous adsorption resin. In a specific embodiment, the second extract is obtained from a sub-step of elution with 70% ethanol.

[0047] Following gradient elution, the second extract is subjected to a separation process. In a specific embodiment, the second extract is first separated using normal-phase silica gel chromatography to obtain crude vanilla saponins. Subsequently, the crude saponins are further separated using medium-pressure liquid chromatography (MPLC) and subsequent high-performance liquid chromatography (HPLC) to obtain each of the isolated saponins from vanilla. For example, a C18 column can be used for medium-pressure liquid chromatography. The C18 column comprises an adsorbent having an octadecylalkyl substituent. In various embodiments, the adsorbent is functionalized to silica gel, namely octadecylsilyl (ODS). The silica gel can be modified with additional functional groups. A pump can be used to force the solvent through the C18 column at pressures from 50 to 3500 psi. The pump can also be used for high-performance liquid chromatography and can operate at pressures up to 6000 psi or higher. As described above, the vanilla saponin composition can be one or more of the saponins isolated in this disclosure. The isolated saponins are described in more detail below.

[0048] Isolated saponins

[0049] The plant-active components and therefore the *Heliotropium indicum* saponin compositions comprise isolated saponins or combinations of isolated saponins. The compositions may also consist substantially of one or more of the isolated saponins, or of one or more of the isolated saponins. Each isolated saponin is obtained from or extracted from the *Heliotropium indicum* plant. For example, isolated saponins can be obtained by the methods described above.

[0050] The isolated schizocarpine saponins contain the structural formulas shown below (Ia) to (Ih) and have the corresponding chemical names.

[0051] Formula (Ia)-Saponin "DT01"; (Z)-2-Methylbut-2-enoic acid (2R,3S,4R,5R,6R)-6-(((3S,4aR,6aR,6bS,8R,8aS,9S,12aR,12bS,13S,14aR,14bR)-8-acetoxy-3-(((2R,3R,4S,5S)-4,5-dihydroxy-3-(((2S,3R,4S,5S,6R)-3,4,5-tri- Hydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)-13-hydroxy-4,4,6a,6b,11,11,14b-heptamethyl-16-oxooctadecylhydro-1H,9H-12b,8a-(epoxymethano)picen-9-yl)oxy)-4,5-dihydroxy-2-(hydroxymethyl)tetrahydro-2H-pyran-3-yl ester:

[0052]

[0053] Formula (Ib)-Saponin “DT02”; (Z)-2-Methylbut-2-enoic acid (2R,3S,4R,5R,6R)-6-(((3S,4aR,6aR,6bS,8R,8aS,9S,12aR,12bS,13S,14aR,14bR)-8-acetoxy-3-(((2S,3R,4S,5S)-3-(((2S,3R,4S,5S,6R)-6-(acetoxy) Methyl)-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)oxy)-4,5-dihydroxytetrahydro-2H-pyran-2-yl)oxy)-13-hydroxy-4,4,6a,6b,11,11,14b-heptamethyl-16-oxooctadecylhydro-1H,9H-12b,8a-(epoxybridged methylene)) succinyl-9-yl)oxy)-4,5-dihydroxy-2-(hydroxymethyl)tetrahydro-2H-pyran-3-yl ester:

[0054]

[0055] Formula (Ic)-Saponin "DT05"); Capilliposide C:

[0056]

[0057] Formula (Id)-Saponin "DT06"; Herba styracifolium saponin B:

[0058]

[0059] Formula (Ie)-Saponin "DT09"; (Z)-2-Methylbut-2-enoic acid (3S,4aR,6aR,6bS,8R,8aR,9S,12aR,12bS,14aR,14bR)-3-(((2S,3R,4S,5S)-5-(((2S,3R,4S,5S,6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-(((2S,3R,4S,5R)-3,4,5-trihydroxytetrahydro-2H-pyran- 2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)-4-hydroxy-3-(((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)-8-hydroxy-4,4,6a,6b,11,11,14b-heptamethyl-16-oxooctadecylhydro-1H,9H-12b,8a-(epoxybridged methylene)) succinyl-9-yl ester:

[0060]

[0061] Formula (If)-Saponin “DT19”; (Z)-2-Methylbut-2-enoic acid (2R,3S,4R,5R,6R)-6-(((3S,4aR,6aR,6bS,8R,9S,12aR,12bS,14aR,14bR)-8-acetoxy-3-(((2S,3R,4S,5S)-5-(((2S,3R,4S,5S,6R)-6-(acetoxymethyl)-4,5-dihydroxy-3-(((2S,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H -pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)-4-hydroxy-3-(((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)-4,4,6a,6b,11,11,14b-heptamethyl-16-oxooctadecyl-1H,9H-12b,8a-(epoxybridged methylene)) pyran-9-yl)oxy)-4,5-dihydroxy-2-(hydroxymethyl)tetrahydro-2H-pyran-3-yl ester:

[0062]

[0063] Formula (Ig)-Saponin “DT11”; 4-Methylpentanoic acid (3S,4aR,6aR,6bS,8R,8aR,9S,12aR,12bS,14aR,14bR)-3-(((2S,3R,4S,5R)-5-(((2R,3R,4S,5S,6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-(((2S,3R,4S,5R)-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl) )oxy)tetrahydro-2H-pyran-2-yl)oxy)-4-methyl-3-(((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)-8,16-dihydroxy-4,4,6a,6b,11,11,14b-heptamethyloctadecyl-1H,9H-12b,8a-(epoxybridged methylene)) succinyl-9-yl ester:

[0064]

[0065] Formula (Ih)-Saponin "DT15"; (Z)-2-Methylbut-2-enoic acid (3S,4aR,6aR,6bS,8R,8aR,9S,12aR,12bS,13S,14aR,14bR)-3-

[0066] (((2S,3R,4S,5S)-5-(((2S,3R,4S,5S,6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-(((2S,3R,4S,5R)-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)-4-hydroxy-3-(((2S,3R,4S,5S)-5 ... S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)-8,13-dihydroxy-4,4,6a,6b,11,11,14b-heptamethyl-16-oxooctadecylhydro-1H,9H-12b,8a-(epoxybridged methylene)) succinyl-9-yl ester:

[0067]

[0068] These isolated schizocarpine saponins are also summarized in Table 1 below.

[0069] Table 1:

[0070]

[0071]

[0072] definition

[0073] To provide a clear and consistent understanding of the specification and claims, the following definitions are provided.

[0074] The terms "composition" or "formulation" refer to products that treat, improve, promote, increase, manage, control, maintain, optimize, modify, reduce, inhibit, or prevent a specific condition associated with a natural state, biological process, or disease or disorder. For example, a composition or formulation improves, minimizes, inhibits, or prevents at least one of hyperglycemia, diabetes, hyperlipidemia, hyperuricemia, gout, ulcers, cancer, ischemia, and hypertension. The terms composition and formulation include, but are not limited to, pharmaceutical (i.e., drug), over-the-counter (OTC) drugs, cosmetics, food, food ingredients, or dietary additive compositions that include an effective amount of a plant-based active ingredient, and particularly saponins from vanilla. Exemplary compositions and / or formulations include creams, cosmetic washes, packets, or powders, or as emulsions, lotions, liniments, foams, tablets, plasters, granules, or ointments. Preferred compositions are formulated for topical application / application and for oral application / ingestion.

[0075] As used herein, the terms "effective amount" or "therapeutic effective amount" for pure compounds, compositions, extracts, mixtures of extracts, extract components and / or active agents or ingredients or combinations thereof refer to an effective amount whose dosage and duration are sufficient to achieve the desired result. For example, an "effective amount" or "therapeutic effective amount" refers to the amount of such pure compounds, compositions, extracts, plant extracts, mixtures of extracts, mixtures of plant extracts, extract components and / or active agents or ingredients or combinations thereof of the present invention that, when applied to a subject (e.g., a mammal, such as a human), are sufficient to achieve a treatment, such as improving, minimizing, inhibiting or preventing at least one of hyperlipidemia, hyperuricemia, gout, ulcers, cancer, ischemia, and hypertension. The amounts of compositions, extracts, plant extracts, mixtures of extracts, mixtures of plant extracts, extract components, plant active ingredients and / or active agents or ingredients constituting an "effective amount" or "therapeutically effective treatment" of this disclosure will vary depending on the active agent or compound, the condition being treated and its severity, the method of application, the duration of treatment, or the age of the subject to be treated, but can be conventionally determined by one of ordinary skill in the art in consideration of his own knowledge and this disclosure.

[0076] The term "pharmaceutically acceptable" refers to a drug, preparation, extract, or inert ingredient that is suitable for use in contact with the tissues of humans and lower animals, without undue toxicity, incompatibility, instability, irritation, etc., and is commensurate with a reasonable benefit / risk ratio.

[0077] The terms “application” and “administration” are defined as providing the composition to a subject via routes known in the art, including but not limited to local, intravenous, intra-arterial, oral, parenteral, sublingual, transdermal, rectal, intramuscular, subcutaneous, intraosseous, transmucosal, or intraperitoneal administration. In a preferred embodiment, local and / or oral administration of the described composition is suitable.

[0078] The terms “minimize,” “reduce,” “inhibit,” “lower,” and / or “inhibit” indicate that, in the presence of the plant (or plant) components or plant extracts described herein, pancreatic lipase activity and / or xanthine oxidase activity are reduced or decreased by inhibiting pancreatic lipase, and / or xanthine oxidation is reduced or decreased by inhibiting xanthine oxidase, and / or their downstream effects, compared to pancreatic lipase activity in the absence of the plant components or plant extracts described herein (such as in control samples). The extent to which pancreatic lipase activity is reduced and fat metabolism and absorption is decreased, and / or the extent to which xanthine oxidation is reduced, and / or their downstream effects, are influenced by the nature and quantity of the plant ingredient or plant extract present, but will be significant, for example, as a detectable reduction in pancreatic lipase activity and / or xanthine oxidase activity; ideally, the reduction in pancreatic lipase activity and / or xanthine oxidase activity is greater than about 5%, about 10%, about 15%, about 20%, about 25%, about 50%, about 75%, about 90%, about 95%, or about 99% (or any reduction in the range of about 5% to about 99%) compared to the absence of said plant ingredient or plant extract. For example, compositions containing plant ingredients or plant extracts of *Hemiberlesia oleracea*, particularly saponins of *Hemiberlesia oleracea*, can minimize or reduce pancreatic lipase activity and / or xanthine oxidase activity.

[0079] As used herein, the term "object" or "individual" includes mammals to which the composition may be administered. Non-limiting examples of mammals include humans, non-human primates, rodents (including transgenic and non-transgenic mice), etc. In some embodiments, the object is a mammal, and in some embodiments, the object is a human.

[0080] Oral composition (or oral preparation)

[0081] The oral composition may include any amount of plant-active components (e.g., isolated saponins or combinations of isolated saponins), selected based on the number and type of components used as a whole in the oral composition. Generally, the plant-active components are present in the oral composition in an amount that effectively inhibits one or both of pancreatic lipase and xanthine oxidase (XO) in the target.

[0082] In some embodiments, the oral composition comprises an amount of the plant active ingredient of 0.1 to 5000 mg, optionally 1 to 3000 mg, optionally 2 to 2000 mg, optionally 5 to 1750 mg, optionally 10 to 1500 mg, optionally 15 to 1250 mg, optionally 20 to 1000 mg, optionally 25 to 750 mg, optionally 30 to 500 mg, optionally 35 to 500 mg, optionally 40 to 500 mg, optionally 45 to 450 mg, optionally 50 to 450 mg, or optionally 50 to 400 mg. However, amounts outside and / or overlapping with these ranges may also be used. For example, it should be understood that the ranges described above regarding the amount of each plant extract in the plant active ingredient can be equally applied to the amount of each plant extract in the oral composition as a whole, such as when the plant active ingredient consists of only one plant extract.

[0083] In some embodiments of the oral composition, the formulation comprises saponins or combinations of saponins from *Hedyotis diffusa*, which can be obtained as an extract of *Hedyotis diffusa*. However, saponins from *Hedyotis diffusa* can be obtained from other suitable sources. Without being bound by any particular theory, the stated formulation is believed to be particularly useful, and even more useful, for the treatment, relief, and / or prevention of gout, due to one or both of lipase inhibition and xanthine oxidase inhibition.

[0084] Flavorings and / or sugar substitutes may be included in the oral composition and may be any type of conventional component as understood in the art, such as flavoring agents. Plant (or plant) extracts may each be as described above. Examples of suitable flavoring agents are further described below.

[0085] Generally, oral compositions are not limited in terms of formulation, peripheral ingredients, form, or number of functions, except that they contain plant-based active ingredients and their extracts. Conversely, the oral compositions may be different and can be formulated in any manner consistent with this disclosure.

[0086] Typically, the oral composition is formulated or otherwise modified for administration to a mammalian subject (e.g., a human). For example, in various embodiments, the oral composition is modified for consumption and / or oral administration to a human subject. Thus, the oral composition is formulated for oral administration to a subject. In such embodiments, the oral composition may also be referred to as an ingestible composition and typically contains at least one pharmaceutically acceptable additive in addition to a bioactive agent composition. In specific embodiments, the pharmaceutically acceptable additive is not naturally occurring. In other words, in these specific embodiments, the pharmaceutically acceptable additive is not a natural product. In other embodiments, the pharmaceutically acceptable additive is selected from conventional additives understood in the art and can be used in conventional amounts.

[0087] Therefore, it should be understood that the specific additives, carriers, excipients, fillers, etc., present in or in combination with the oral composition can vary. Furthermore, the physical form of the oral composition is not limited and will be selected based on the specific components of the oral composition, the intended use of the oral composition, etc. Thus, as will be understood in consideration of the description herein, the oral composition can be formulated as a liquid, dry powder, suspension, emulsion, gel, paste, etc., and combinations thereof. In some embodiments, the oral composition is formulated as a sterile, pyrogen-free liquid solution or suspension, coated capsule, suppository, lyophilized powder, transdermal patch, soft capsule, or other known form. Other examples of suitable forms include solids, gels, liquids, creams, lotions, hair oils, mousses, powders, foams, sprays, ointments, or other such articles, wherein the plant-active ingredient is placed in a suitable carrier medium (such as any of those described herein).

[0088] Oral compositions can be prepared using various methods. For example, the active ingredient (such as a plant extract) and optionally one or more inactive ingredients (such as one or more conventional components, additives, excipients, etc.) of the oral composition can be mixed or blended and compressed or co-blended using various techniques understood in the art. The oral compositions of this disclosure are not limited to a specific order of manufacturing steps or methods.

[0089] In various embodiments, the oral composition is administered orally to a subject by ingestion. The subject is typically a person and may include men and women of various ages. The methods / compositions of this disclosure are not limited to a specific subject.

[0090] Excipients can be further classified into other components. Specifically, excipients used in oral solid dosage forms have been classified into several groups based on their functionality, such as diluents, disintegrants, binders, compression aids, granulation agents, flow aids, lubricants, release-controlling polymers, stabilizers (such as antioxidants, chelating agents, and pH adjusters), film-coating polymers, coating agents, mediators, plasticizers, surfactants, colorants, sweeteners, and flavorings.

[0091] In various embodiments, the oral composition comprises at least one component selected from binders, lubricants, flow aids, and combinations thereof. In some embodiments, the oral composition comprises one or more compounds, including but not limited to methylcellulose, hydroxypropyl methylcellulose, ethylcellulose, cellulose acetate, gum arabic, gums, waxes, glyceryl monostearate, acrylic polymers and copolymers, methacrylic acid, methyl acrylate, ethyl acrylate, methyl methacrylate, ethyl methacrylate, lactose, calcium sulfate, calcium hydrogen phosphate, sugars, microcrystalline cellulose (MCC), starch, sodium starch glycolate, polyvinylpyrrolidone, polyethylene glycol, and magnesium stearate. Combinations of such components can be used, and such components and other components used in conventional tablets are understood in the art.

[0092] The term "diluent" as used herein can refer to an inert substance added to increase the volume of the oral composition, giving the tablet a practically compressible size. Therefore, they may also be referred to as fillers. Commonly used diluents include, but are not limited to, microcrystalline cellulose (MCC), wood cellulose, corn starch, modified corn starch, tricalcium phosphate, calcium sulfate, lactose, kaolin, mannitol, sodium chloride, dried starch, (powdered) sugar, dextrose, mannitol, sorbitol, etc. These diluents / fillers can be used alone or in various mixtures and in any amount known in the art for use in oral compositions.

[0093] As used herein, "flavoring agent" refers to a compound intended to impart a more palatable taste to an oral composition. Flavoring agents vary widely in their chemical structure, ranging from simple esters, alcohols, and aldehydes to carbohydrates and complex volatile oils. Almost any desired type of synthetic flavoring is now available and is well known in the art. If a hard, sour, or bitter taste originating from the starting material can be suppressed by flavoring or flavoring, an acidifier (e.g., citric acid, tartaric acid, malic acid, ascorbic acid, etc.), a sweetener (e.g., sodium saccharin, dipotassium glycyrrhizate, aspartame, stevia, thaumatin, etc.), or a flavoring (e.g., various fruit flavorings containing lemon oil, orange oil, or strawberry, as well as yogurt, mint, menthol, etc.) may be used alone or in various mixtures, and in any amount known in the art to be used in an oral composition.

[0094] As used herein, "lubricant" refers to a material that performs many functions associated with oral compositions. In some embodiments, such as tablet manufacturing, the lubricant performs one or more functions, such as increasing the flow rate of tablet granulation, preventing tablet material from adhering to the surfaces of the die and punch, reducing intergranular friction, and facilitating tablet ejection from the die cavity. Examples of suitable lubricants include, but are not limited to, zinc stearate, gum arabic powder, cocoa butter, carnauba wax, calcium carboxymethyl cellulose, sodium carboxymethyl cellulose, caropeptide, aqueous silica solution, dried aluminum hydroxide gel, glycerin, magnesium silicate, light anhydrous silicate, light liquid paraffin, crystalline cellulose, hardened oil, synthetic aluminum silicate, sesame oil, flour starch, beeswax, magnesium oxide, dimethyl polysiloxane, potassium sodium tartrate, sucrose fatty acid esters, glyceryl fatty acid esters, silicone resins, and aluminum hydroxide. The lubricant comprises gel, stearyl alcohol, stearic acid, aluminum stearate, calcium stearate, polyoxyethylene stearate, magnesium stearate, cetyl alcohol, gelatin, talc, magnesium carbonate, precipitated calcium carbonate, corn starch, lactose, stearate, sucrose, potato starch, hydroxypropyl cellulose, fumaric acid, sodium stearate fumarate, polyethylene glycol, polyoxyethylene polyoxypropylene glycol, polysorbate, beeswax, magnesium aluminum metasilicate, methylcellulose, Japanese wax, glyceryl monostearate, sodium lauryl sulfate, calcium sulfate, magnesium sulfate, liquid paraffin, phosphoric acid, palmitic acid, and hydrogenated vegetable oils and fats. The lubricant can be used alone or in various mixtures and in any amount known in the art for oral compositions.

[0095] As used herein, "binder" is an agent used to impart binding properties to powdered materials. Binders, or sometimes referred to as "granulators," impart binding properties to tablet formulations, ensuring that the tablets remain intact after compression and improving free-flow quality by formulating granules of desired hardness and size. Materials commonly used as binders include starches, such as corn starch and pregelatinized starch; gelatin; sugars, such as sucrose, glucose, dextrose, molasses, and lactose; natural and synthetic gums, such as gum arabic, sodium alginate, Irish moss extract, panval gum, gum indica, mucilage of isapol husks, carboxymethyl cellulose, methyl cellulose, polyvinylpyrrolidone (PVP), magnesium aluminum silicate, microcrystalline cellulose, microcrystalline dextrose, amylose, larch arabinogalactan, ethyl cellulose, cellulose acetate, etc. These binders can be used alone or in various mixtures and in any amount known in the art for oral compositions.

[0096] As used herein, "coloring agent" is an agent that imparts a more aesthetically pleasing appearance to oral compositions and, in addition, helps manufacturers control the product during its preparation and assists users in identifying it. Any approved, certified water-soluble FD&C dye, mixtures thereof, or their corresponding lakes can be used to color tablets. A lake is a combination formed by adsorbing a water-soluble dye onto a hydrated oxide of a heavy metal, thereby producing an insoluble form of the dye. The coloring agent can be used alone or in various mixtures and in any amount known in the art for use in oral compositions.

[0097] Other conventional ingredients that may optionally be present in oral compositions include preservatives, stabilizers, anti-adhesion agents, or silica flow modifiers or flow aids, such as silica. Such ingredients may be used alone or in various mixtures and in any amount known in the art in oral compositions.

[0098] It should be understood that certain components or additives may be classified under different technical terms, and the fact that a component or additive is classified under such terms does not imply that it is limited to that function. One or more additives may be present in oral compositions in various amounts, if used. Additional ingredients for optional use in oral compositions, such as when suitable for oral administration, are described in the following documents: U.S. Patent Nos. 5,747,006, 5,980,904, 6,994,874, 7,060,304, 7,247,321, 7,348,034, 7,364,759, 7,700,110, 7,722,904, 8,202,556, 8,916,212, 9,445,975, 9,801,809, 10,307,366, 10,532,024, and 10,537,516. The publications of these publications, including U.S. Publications Nos. 2006 / 0257509, 2007 / 0224154, 2008 / 0081082, 2008 / 0124409, 2013 / 0302265, 2017 / 0252293, 2017 / 0281666, 2018 / 0200285, 2019 / 0083566, 2019 / 0160117, 2020 / 0171117, 2020 / 0383898, 2021 / 0017240 and 2021 / 0212926, are hereby incorporated in their entirety by reference.

[0099] Application method

[0100] The oral composition can be administered or applied as needed, once daily, several times daily, or in any suitable regimen to achieve the desired results. In the methods of this disclosure, the frequency of administration (e.g., oral ingestion) can depend on several factors, including the desired level of pancreatic lipase inhibition and / or xanthine oxidase (XO) inhibition. The oral composition may comprise isolated saponins or combinations of isolated saponins as described herein. Typically, a regimen involves taking the oral composition once or twice daily, including administration in the morning and / or evening. The amount and / or frequency of application of the oral composition can depend on several factors, including the level of desired results and the specific oral composition.

[0101] Improved inhibition of lipase and / or xanthine oxidase can be achieved by applying the formulations of the present invention externally, internally, or in some combination thereof. Preferably, the formulations of the present invention are applied together with an acceptable carrier. For example, the formulations of the present invention can be administered internally in the form of pills, tablets, powders, bars, beverages, etc., together with an acceptable carrier. Therefore, the formulations described herein can be used in a variety of finished products, including pharmaceutical products, food products, and beverage compositions. Preferably, the products can be used to provide lipase and / or xanthine oxidase inhibition in mammals.

[0102] When the formulations of the present invention are administered orally in liquid form, the liquid may be water-based, milk-based, tea-based, fruit juice-based, or some combination thereof. Solid and liquid formulations for internal administration according to the present invention may further comprise thickeners, including xanthan gum, carboxymethyl cellulose, carboxyethyl cellulose, hydroxypropyl cellulose, methyl cellulose, microcrystalline cellulose, starch, dextrin, fermented whey, tofu, maltodextrin, polyols including sugar alcohols (e.g., sorbitol and mannitol), carbohydrates (e.g., lactose), propylene glycol alginate, gellan gum, guar gum, pectin, tragacanth gum, gum arabic, locust bean gum, gum arabic, gelatin, and mixtures of these thickeners. These thickeners are typically included in the formulations of the present invention at a level of up to about 0.1%, depending on the specific thickener involved and the desired viscosity effect.

[0103] The solid and liquid (food and beverage) formulations of the present invention may and typically contain effective amounts of one or more sweeteners, including carbohydrate sweeteners and natural and / or artificial zero / low-calorie sweeteners. The amount of sweetener used in the formulations of the present invention varies, but generally depends on the type of sweetener used and the desired sweetness intensity.

[0104] Typically, the formulation of the present invention is administered orally at least daily for a period of time sufficient to inhibit lipase and / or xanthine oxidase to the desired level. Oral administration of the formulation of the present invention can be continued for any suitable period of time. More specifically, within hours to days of initial intake, the user may notice an improvement in symptoms of hyperlipidemia, hyperuricemia, gout, ulcers, cancer, ischemia, and / or hypertension. It should be understood that the frequency of administration of the formulation of the present invention will vary depending on the desired level of improvement. Specifically, the degree of symptom relief will vary directly with the total amount of the oral composition used.

[0105] Useful dosage forms can be prepared using methods and techniques known to those skilled in the art, and additional ingredients can be included in the production of tablets, capsules, or liquid dosage forms.

[0106] Industrial applicability

[0107] This disclosure provides novel and useful inhibitors of advanced pancreatic lipase (PL) and / or xanthine oxidase (XO), including the plant-active components described herein. Such PL inhibitors can provide potential preventative and therapeutic pathways to reduce the risk of hyperlipidemia or other pathogenic complications caused by PL activity. Such XO inhibitors can provide potential preventative and therapeutic pathways to reduce the risk of hyperuricemia, gout, ulcers, cancer, ischemia, hypertension, oxidative stress, oxidative damage, or other pathogenic complications caused by XO activity. Therefore, the compositions and methods described herein can be used to inhibit PL and / or XO.

[0108] The general compositions and product lines provided by this disclosure relate to personal care, nutritional, skin care, and nutritional beauty products utilizing such PL and / or XO inhibitors, and specific examples include pills and liquids utilizing such PL and / or XO inhibitors.

[0109] Lipids (PL) are a metabolic pathway involved in the breakdown and absorption of dietary fats. Elevated lipid levels in the body can lead to serious diseases, potentially resulting in death. For example, elevated lipid levels can contribute to the development of atherosclerosis and thus may affect cardiovascular health. The compositions and methods of this disclosure provide potential preventative and therapeutic pathways for reducing the risk of hyperlipidemia or other pathogenic complications caused by PL.

[0110] XO is a metabolic pathway involved in uric acid formation. Elevated uric acid levels in the blood can lead to gout when high levels of uric acid crystallize and deposit in joints, tendons, and surrounding tissues. Elevated uric acid in the blood can also affect cardiovascular health. Furthermore, byproducts of XO activity, such as hydrogen peroxide, can cause oxidative stress. The compositions and methods of this disclosure provide potential preventative and therapeutic pathways for reducing the risk of hyperuricemia, gout, ulcers, cancer, ischemia, hypertension, oxidative stress, oxidative damage, or other pathogenic complications caused by XO. Detailed Implementation

[0111] The following examples illustrating the compositions and methods of this disclosure are intended to be illustrative and not limiting of this disclosure.

[0112] Example

[0113] The inhibitory effects of saponins from *Heliotropium indicum* on pancreatic lipase (PL) and xanthine oxidase (XO) were evaluated. The saponins from *Heliotropium indicum* exhibited inhibitory effects on pancreatic lipase (PL) and xanthine oxidase (XO), as shown in the table and figure below.

[0114] Inhibition of pancreatic lipase (PL)

[0115] Add 25 μL of the test sample to 25 μL of lipase solution (1 mg / mL) and mix thoroughly. Then, add 50 μL of 4-methylumbelliferyl oleate (0.1 mmol / L) as the lipase substrate to the mixture and incubate at 23 °C for 20 min. The reaction is then terminated with 100 μL of sodium citrate (0.1 mol / L, pH 4.2). To measure absorbance, excitation and emission wavelengths were set at 320 nm and 450 nm, respectively. Tris buffer (pH 8.0) containing the sample (without enzyme) represents the background group, Tris buffer containing the enzyme (without sample) represents the control group, and Tris buffer alone represents the blank group. All experiments were repeated three times. The absorbance of the solutions was measured. The inhibition rate of PL was calculated using the following equation:

[0116]

[0117] Where A1 is the absorbance of the mixture of sample and PL, A2 is the absorbance of the mixture of sample and buffer solution, A3 is the absorbance of the mixture of buffer solution and PL, and A4 is the absorbance of the buffer solution.

[0118] The results are shown in Table 2-6 below. The average inhibition rate can also be referenced. Figure 2-6 To understand.

[0119] Table 2:

[0120]

[0121]

[0122] *See also Figure 2 (Where the X-axis represents concentration and the Y-axis represents % inhibition)

[0123] Table 3:

[0124]

[0125] *See also Figure 3 (Where the X-axis represents concentration and the Y-axis represents % inhibition)

[0126] Table 4:

[0127]

[0128]

[0129] *See also Figure 4 (Where the X-axis represents concentration and the Y-axis represents % inhibition)

[0130] Table 5:

[0131]

[0132] *See also Figure 5 (Where the X-axis represents concentration and the Y-axis represents % inhibition)

[0133] Table 6:

[0134]

[0135] *See also Figure 6 (Where the X-axis represents concentration and the Y-axis represents % inhibition)

[0136] Inhibition of xanthine oxidase (XO)

[0137] The inhibitory effect of xanthine oxidase activity on saponins from *Vigna scabra* was determined by spectrophotometry using a 96-well plate. The reaction mixture consisted of 50 μL of the sample and 50 μL of 0.02 U / mL xanthine oxidase solution, which was shaken appropriately for 30 seconds and incubated at 25 °C for 5 minutes. Then, 150 μL of 0.48 mM xanthine solution was added, and the mixture was again carefully shaken for 30 seconds and incubated at 37 °C for 30 minutes. The absorbance of the incubated solution was measured at 290 nm. The inhibition rate of XO was calculated using the following equation:

[0138]

[0139] Where A1 is the absorbance of the mixture of sample and XO, A2 is the absorbance of the mixture of sample and buffer solution, A3 is the absorbance of the mixture of buffer solution and XO, and A4 is the absorbance of the buffer solution.

[0140] The results are shown in Table 7-10 below. The average inhibition rate can also be referenced. Figure 7-10 To understand.

[0141] Table 7:

[0142]

[0143] *See also Figure 7 (Where the X-axis represents concentration and the Y-axis represents % inhibition)

[0144] Table 8:

[0145]

[0146]

[0147] *See also Figure 8 (Where the X-axis represents concentration and the Y-axis represents % inhibition)

[0148] Table 9:

[0149]

[0150] *See also Figure 9 (Where the X-axis represents concentration and the Y-axis represents % inhibition)

[0151] Table 10:

[0152]

[0153] *See also Figure 10 (Where the X-axis represents concentration and the Y-axis represents % inhibition)

[0154] The terms “comprising” or “including” are used in their broadest sense to refer to and encompass the concepts of “including,” “containing,” “substantially consisting of,” and “consisting of.” The use of “for example,” “such as,” and “including” to list exemplary examples is not limited to the examples listed. Thus, “for example” or “such as” means “for example, but not limited to” or “such as, but not limited to,” and covers other similar or equivalent examples. The term “about” as used herein is used to appropriately cover or describe minute variations in numerical values ​​measured by instrumental analysis or as a result of sample processing. Such minute variations can be on the order of ±0-10%, ±0-5%, or ±0-2.5% of the numerical value. Furthermore, the term “about” is applied to two numerical values ​​when relating to a range of values. Additionally, the term “about” can be applied to numerical values ​​even when not explicitly stated.

[0155] Generally, as used herein, a hyphen “-” or dash “—” in a range of values ​​means “to” or “up to”; ">” means “above” or “greater than”; “≥” means “at least” or “greater than or equal to”; “<” means “below” or “less than”; and “≤” means “at most” or “less than or equal to”. On an individual basis, each of the patent applications, patents and / or patent application publications mentioned above is expressly incorporated herein in its entirety by reference in one or more non-limiting embodiments.

[0156] It should be understood that the appended claims are not limited to the specific compounds, compositions, or methods described in the detailed description, and may vary among specific embodiments falling within the scope of the appended claims. Regarding any Markush group herein relied upon for describing specific features or aspects of different embodiments, it should be understood that different, particular, and / or unpredictable results can be obtained from each member of the respective Markush group, independently of all other Markush members. Reliance on each member of the Markush group may be individual and / or combined, and may provide sufficient support for specific embodiments within the scope of the appended claims.

[0157] It should also be understood that any scopes and subscopes relied upon in describing the different embodiments of the invention fall independently and collectively within the scope of the appended claims, and are to be understood as describing and taking into account all scopes, including integer and / or fractional values ​​therein, even if such values ​​are not explicitly stated herein. Those skilled in the art will readily recognize that the enumerated scopes and subscopes are sufficient to describe and implement the different embodiments of the invention, and that such scopes and subscopes can be further described as relating to 1 / 2, 1 / 3, 1 / 4, 1 / 5, and so on. As merely an example, the range “0.1-0.9” can be further described as a smaller 1 / 3 (i.e., 0.1-0.3), a middle 1 / 3 (i.e., 0.4-0.6), and a larger 1 / 3 (i.e., 0.7-0.9), which individually and collectively fall within the scope of the appended claims and can be individually and / or collectively relied upon, providing sufficient support for specific embodiments within the scope of the appended claims. Furthermore, regarding the wording that limits or modifies the scope, such as "at least," "greater than," "less than," "not exceeding," etc., it should be understood that such language includes sub-ranges and / or upper or lower limits. As another example, the scope of "at least 10" inherently includes sub-ranges of at least 10-35, at least 10-25, 25-35, and so on, and each sub-range can be relied upon individually and / or collectively to adequately support specific embodiments within the scope of the appended claims. Finally, a single number within the disclosed scope can be relied upon to adequately support specific embodiments within the scope of the appended claims. For example, the scope of "1-9" includes various individual integers (such as 3) and individual numbers, including decimal points (or fractions), such as 4.1, which can be relied upon to adequately support specific embodiments within the scope of the appended claims.

[0158] The invention has been described herein by way of example, and it should be understood that the terminology used is intended to be descriptive rather than limiting. Many modifications and variations of the invention are possible in light of the above teachings. The invention may be practiced in ways other than those specifically described within the scope of the appended claims. The subject matter of all combinations of independent and dependent claims (including single and multiple dependent claims) is expressly considered herein.

Claims

1. A method for preparing a composition of vanilla saponins, the method comprising: The first extraction of the fine-stemmed vanilla material was carried out using the first solvent composition to obtain a crude extract; The crude extract was subjected to gradient elution with a macroporous adsorption resin and an eluent containing alcohol to obtain a second extract. The second extract was separated by normal-phase silica gel chromatography to obtain crude saponins; The crude saponins were separated by medium-pressure liquid chromatography and high-performance liquid chromatography to obtain one or more isolated saponins from *Vigna arvense*.

2. The method of claim 1, wherein the herbaceous stalk material is further defined as the whole plant of herbaceous stalk.

3. The method according to claim 1 or 2, wherein the fine-stemmed vanilla material is steamed prior to the first extraction.

4. The method according to any one of claims 1 to 3, wherein the first solvent composition comprises an alcohol.

5. The method according to claim 4, wherein the alcohol is ethanol.

6. The method according to claim 4 or 5, wherein the first solvent composition is 75% ethanol.

7. The method according to any one of claims 1 to 6, wherein the eluent comprises 20% ethanol, 40% ethanol, 70% ethanol and 95% ethanol.

8. The method according to any one of claims 1 to 7, wherein the first extraction is performed for at least 3.5 hours, optionally for a period of time ranging from 3.5 to 4.5 hours.

9. The method according to any one of claims 1 to 8, wherein the first extraction is repeated.

10. A vanilla saponin composition obtained by the method according to any one of claims 1 to 9, wherein the vanilla saponin composition optionally comprises isolated saponins, consists substantially of isolated saponins, or is composed of isolated saponins.

11. An isolated saponin obtained from *Heliotropium indicum* material, said isolated saponin comprising one of the following structures (Ia) to (Ih):

12. An oral composition for oral administration to a subject, the oral care composition comprising: i) the herbaceous saponin composition according to claim 10; or ii) The saponins isolated according to claim 11.

13. The use of the following substances in the preparation of medicaments for the treatment of hyperuricemia and / or hyperlipidemia: i) the herbaceous saponin composition according to claim 10; or ii) The saponins isolated according to claim 11.

14. A method for inhibiting one or both of lipase and xanthine oxidase in a subject, said method comprising administering an effective amount of a composition to said subject, wherein said composition is: i) the herbaceous saponin composition according to claim 10; or ii) The saponins isolated according to claim 11.

15. The method of claim 14, wherein the composition is administered orally to the subject.

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