A lipid-lowering traditional Chinese medicine composition for inhibiting pancreatic lipase and cholesterol esterase and a preparation method thereof

Abstract

The present application relates to the technical field of traditional Chinese medicine composition, and specifically puts forward a kind of lipid-lowering traditional Chinese medicine composition for inhibiting pancreatic lipase and cholesterol esterase and a preparation method thereof, the raw materials of the traditional Chinese medicine composition are composed of cinnamon 30%-35%, safflower 50%-55% and dogwood 10%-20% in proportion. It uses the medicinal and edible medicinal materials, has good pancreatic lipase, cholesterol esterase inhibitory effect, and significantly reduces the enzyme reaction efficiency.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine composition technology, and in particular to a lipid-lowering traditional Chinese medicine composition that inhibits pancreatic lipase and cholesterol esterase and its preparation method. Background Technology

[0002] Obesity is widespread globally, having surpassed malnutrition and infectious diseases as a leading cause of death worldwide. In 1999, the WHO officially declared obesity a disease. Obesity is closely related to hyperlipidemia, which can lead to cardiovascular disease, insulin resistance, type 2 diabetes, non-alcoholic fatty liver disease, and other serious threats to human health.

[0003] Long-term excessive intake of high-calorie foods is one of the main factors leading to lipid metabolism disorders in the human body, thus causing hyperlipidemia. The activity of related enzymes affects lipid absorption in lipid metabolism, and reducing lipid absorption plays a positive role in regulating lipid metabolism. Pancreatic lipase is an important enzyme in the hydrolysis of dietary fat, ultimately hydrolyzing triglycerides into monoglycerides and fatty acids. The latter are absorbed in the intestine and then resynthesized into fat in the body. Therefore, inhibiting the activity of pancreatic lipase can effectively reduce the absorption of fat in the small intestine, thereby lowering lipids. Cholesterol esterase plays an important role in the hydrolysis of cholesterol esters. Cholesterol esters are hydrolyzed into free cholesterol before absorption; therefore, inhibiting the activity of cholesterol esterase can reduce cholesterol absorption to a certain extent. Summary of the Invention

[0004] In view of this, the present invention proposes a lipid-lowering traditional Chinese medicine composition that inhibits pancreatic lipase and cholesterol esterase and its preparation method. In response to the obesity problem in Chinese people caused by excessive absorption of triglycerides and cholesterol, natural ingredients are screened to obtain a traditional Chinese medicine composition with inhibitory effects on pancreatic lipase and cholesterol esterase.

[0005] The technical solution of the present invention is achieved as follows: The present invention provides a lipid-lowering traditional Chinese medicine composition that inhibits pancreatic lipase and cholesterol esterase. The raw materials of the traditional Chinese medicine composition are composed of 30%-35% cinnamon, 50%-55% saffron, and 10%-20% cornus officinalis by weight percentage.

[0006] In some embodiments, the raw materials of the traditional Chinese medicine composition are composed of 30% cinnamon, 50% saffron, and 20% cornus officinalis by weight percentage.

[0007] In some embodiments, the raw materials of the traditional Chinese medicine composition are composed of 32% cinnamon, 53% saffron, and 15% cornus officinalis by weight percentage.

[0008] In some embodiments, the raw materials of the traditional Chinese medicine composition are composed of 35% cinnamon, 55% saffron, and 10% cornus officinalis by weight percentage.

[0009] Cinnamon (Cinnamomum cassia) is the dried bark of the cinnamon tree, a plant belonging to the genus Cinnamomum in the Lauraceae family. It is used both medicinally and as a food flavoring and seasoning, making it a food and medicine homology product. In my country, it is widely distributed in tropical and subtropical regions such as Guangxi. First recorded in the *Shennong Bencao Jing* (Shennong's Classic of Materia Medica), it is described as having the effects of "tonifying fire and assisting yang, guiding fire back to its source, dispelling cold and relieving pain, and warming and unblocking the meridians." Modern pharmacological studies have shown that cinnamon possesses various pharmacological effects, including lowering blood sugar and lipids, anti-tumor activity, antioxidant properties, antibacterial activity, antiviral activity, reducing stomach damage, and improving learning and memory abilities.

[0010] Saffron is a traditional Chinese medicine. It is the dried stigma of saffron (Crocus sativus L.), a perennial bulbous herbaceous plant belonging to the genus Crocus in the Iridaceae family. It is not only a world-renowned natural fragrance and food dye, but also a valuable medicinal herb, listed in the 2020 edition of the *Chinese Pharmacopoeia (Part I)*. It has the effects of promoting blood circulation, removing blood stasis, cooling blood and detoxifying, relieving depression and calming the mind, and is used to treat amenorrhea, abdominal masses, febrile diseases with rashes, and depression. Currently, many active components of saffron have been isolated, mainly including crocin, crocinaldehyde, crocinoside, and crocinin, which have various effects such as anti-oxidation, anti-inflammation, and anti-tumor activity.

[0011] Cornus officinalis (Sieb. et Zucc.) is a deciduous tree or shrub belonging to the Cornaceae family. Its fruit, known as "yurou," is a famous traditional Chinese medicine, and its dried, ripe pulp is used in traditional Chinese medicine. It has a sour and astringent taste, is slightly warm in nature, and enters the liver and kidney meridians. It is believed to tonify the liver and kidneys, astringe essence and reduce urination, consolidate essence and stop bleeding, and astringe sweat and prevent prolapse. Chemical analysis shows that Cornus officinalis mainly contains iridoids and their glycosides, triterpenes, organic acids, flavonoids, and tannins. Modern medical research indicates that Cornus officinalis has immunomodulatory, anti-AIDS, anti-tumor, antioxidant, hypoglycemic, and hypolipidemic effects.

[0012] Secondly, the present invention also provides a method for preparing the above-mentioned traditional Chinese medicine composition, comprising the following steps:

[0013] Step 1: Weigh out cinnamon, saffron and cornelian cherry according to their mass percentages, pulverize them, pass them through a 40-80 mesh sieve, add 10-15 times the amount of 75% ethanol, reflux in a water bath at 60-80℃ for 1-2 hours, repeat the extraction 2-3 times, combine the extracts, and filter.

[0014] Step 2: The filtered extract is concentrated and dried under negative pressure vacuum to obtain the crude extract;

[0015] Step 3: Suspend the crude extract in water until the crude extract concentration is 1g / ml, and extract it sequentially with petroleum ether, ethyl acetate and n-butanol;

[0016] Step 4: Concentrate and dry the ethyl acetate extract from Step 3 under negative pressure to obtain the extract;

[0017] Step 5: Dissolve the extract obtained in Step 4 in ethanol solution, then adsorb it with HPD-600 resin, and elute with ultrapure water and 50% ethanol in sequence, and collect the ethanol eluent.

[0018] Step 6: Concentrate and dry the ethanol eluent collected in Step 5 under negative pressure vacuum to obtain the traditional Chinese medicine composition.

[0019] In some implementations, the drying temperature for steps two, four, and six is ​​40-60°C.

[0020] In some embodiments, step five includes: dissolving the extract obtained in step four in 75% ethanol solution to a concentration of 1 g / ml, loading the sample onto HPD-600 resin for adsorption for 5 h, eluting with ultrapure water for 5 PV, then eluting with 5 PV of 50% ethanol, and collecting the ethanol eluent.

[0021] The present invention has the following advantages over the prior art:

[0022] (1) The preparation process of this product is conventional and does not involve special or complicated process steps. Compared with other lipid-lowering products, the Chinese medicines used in this composition are all from medicinal and edible materials, which are relatively safe and have a wide range of applications.

[0023] (2) No synthesis or structural modification of natural products is required during the preparation of this product, which reduces the occurrence of side effects on the human body; no additional auxiliary drugs are required, which maintains the pure naturalness of the ingredient source;

[0024] (3) The traditional Chinese medicine composition of the present invention is based on the results of high-throughput screening of pancreatic lipase and cholesterol lipase of all the original medicinal materials. Under the same experimental conditions, it was found that it has a good inhibitory effect on pancreatic lipase and cholesterol lipase, and significantly reduces the enzyme reaction efficiency. Detailed Implementation

[0025] The technical solutions of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0026] It should be noted that when a component is referred to as being "fixed to" or "set on" another component, it can be directly on or indirectly on that other component. When a component is referred to as being "connected to" another component, it can be directly connected to or indirectly connected to that other component.

[0027] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the embodiments of this invention pertain. If any definition stated in this section is contrary to or otherwise inconsistent with a definition stated in a patent, patent application, published patent application, or other publication incorporated herein by reference, the definitions listed here shall prevail over those incorporated herein by reference.

[0028] Unless otherwise specified, the methods used in the following embodiments are conventional methods. Unless otherwise specified, the materials, reagents, and instruments used are conventional materials, reagents, and instruments in the art, and can be obtained commercially by those skilled in the art.

[0029] When a quantity, concentration, or other value or parameter is expressed as a range, a preferred range, or a range defined by a series of upper and lower preferred values, this should be understood as specifically disclosing all ranges formed by any pair of any upper or preferred value with any lower or preferred value, regardless of whether the range is disclosed individually. For example, when the range “1 to 5” is disclosed, the described range should be interpreted as including ranges “1 to 4”, “1 to 3”, “1 to 2”, “1 to 2 and 4 to 5”, “1 to 3 and 5”, etc. When numerical ranges are described herein, unless otherwise stated, the range is intended to include its endpoints and all integers and fractions within that range. In this specification and claims, range definitions may be combined and / or interchanged, unless otherwise stated, these ranges include all subranges contained therein.

[0030] Example 1

[0031] This embodiment proposes a method for preparing a lipid-lowering traditional Chinese medicine composition that inhibits lipase and cholesterol esterase activity. The specific preparation steps are as follows:

[0032] S1. Extraction of medicinal materials: Weigh out 30% cinnamon, 50% saffron and 20% cornus officinalis according to the mass percentage. Crush the medicinal materials and pass them through a 60-mesh sieve. Then add 10 times the weight of 75% ethanol to the crushed medicinal materials and reflux in a water bath at 70°C for 1.5 hours. Repeat the extraction 3 times. Combine the extracts and then filter them to collect the filtrate.

[0033] S2. Concentration and drying: The filtrate after S1 is filtered is concentrated and dried under reduced pressure at a temperature of 50°C to obtain the crude extract.

[0034] S3. Extraction and separation: The crude extract obtained in S2 is suspended in water to a concentration of 1 g / ml, and then extracted sequentially with petroleum ether, ethyl acetate and n-butanol, and the ethyl acetate extract phase is collected.

[0035] S4. Concentration and drying: The ethyl acetate extract phase obtained in S3 is concentrated and dried under reduced pressure at a temperature of 50°C.

[0036] S5. Purification preparation: Add 75% ethanol solution to the extract obtained by drying in S4 to dissolve it to a concentration of 1g / ml, adsorb it with HPD-600 resin for 5h, then elute with 5 column volumes of ultrapure water, and then elute with 5 column volumes of 50% ethanol solution. Collect the ethanol eluent.

[0037] S6. Concentration and Drying: The 50% ethanol eluent collected in S5 is concentrated and dried under reduced pressure at a temperature of 50°C to obtain the traditional Chinese medicine composition.

[0038] High-throughput screening of the prepared traditional Chinese medicine composition for pancreatic lipase inhibition was conducted: based on the defatting of 4-methylumbelliferone oleate (4-MUO) to 4-methylumbelliferone (4-MU) under the action of porcine pancreatic lipase (PPL), the dynamic fluorescence value of 4-MU at an excitation wavelength of 320 nm (λem = 450 nm) was measured using an enzyme-linked immunosorbent assay (ELISA) reader, and the inhibitory ability of the medicinal material on the enzyme was determined according to the net area value of its fluorescence emission intensity.

[0039] High-throughput screening of cholesterol esterase inhibition was performed on the prepared traditional Chinese medicine composition: based on the cholesterol esterase catalyzing the hydrolysis of 4-nitrophenylbutyrate (pNPB), the catalytic activity of cholesterol esterase was determined by measuring the absorption of the product pNP at 405 nm using an enzyme-linked immunosorbent assay (ELISA) reader.

[0040] Calculations show that the combined effect of 30% cinnamon, 50% saffron, and 20% cornus officinalis is as follows:

[0041] Inhibition rate 62.1％ 10.8％

[0042] Example 2

[0043] This embodiment proposes a method for preparing a lipid-lowering traditional Chinese medicine composition that inhibits lipase and cholesterol esterase activity. The specific preparation steps are as follows:

[0044] S1. Extraction of medicinal materials: Weigh out 32% cinnamon, 53% saffron and 15% cornus officinalis according to the mass percentage. Crush the medicinal materials and pass them through a 60-mesh sieve. Then add 12 times the weight of 75% ethanol to the crushed medicinal materials and reflux in a water bath at 70°C for 1.5 hours. Repeat the extraction 3 times. Combine the extracts and then filter them to collect the filtrate.

[0045] S2. Concentration and drying: The filtrate after S1 is filtered is concentrated and dried under reduced pressure at a temperature of 50°C to obtain the crude extract.

[0046] S3. Extraction and separation: The crude extract obtained in S2 is suspended in water to a concentration of 1 g / ml, and then extracted sequentially with petroleum ether, ethyl acetate and n-butanol, and the ethyl acetate extract phase is collected.

[0047] S4. Concentration and drying: The ethyl acetate extract phase obtained in S3 is concentrated and dried under reduced pressure at a temperature of 50°C.

[0048] S5. Purification preparation: Add 75% ethanol solution to the extract obtained by drying in S4 to dissolve it to a concentration of 1g / ml, adsorb it with HPD-600 resin for 5h, then elute with 5 column volumes of ultrapure water, and then elute with 5 column volumes of 50% ethanol solution. Collect the ethanol eluent.

[0049] S6. Concentration and Drying: The 50% ethanol eluent collected in S5 is concentrated and dried under reduced pressure at a temperature of 50°C to obtain the traditional Chinese medicine composition.

[0050] High-throughput screening of the prepared traditional Chinese medicine composition for pancreatic lipase inhibition was conducted: based on the defatting of 4-methylumbelliferone oleate (4-MUO) to 4-methylumbelliferone (4-MU) under the action of porcine pancreatic lipase (PPL), the dynamic fluorescence value of 4-MU at an excitation wavelength of 320 nm (λem = 450 nm) was measured using an enzyme-linked immunosorbent assay (ELISA) reader, and the inhibitory ability of the medicinal material on the enzyme was determined according to the net area value of its fluorescence emission intensity.

[0051] High-throughput screening of cholesterol esterase inhibition was performed on the prepared traditional Chinese medicine composition: based on the cholesterol esterase catalyzing the hydrolysis of 4-nitrophenylbutyrate (pNPB), the catalytic activity of cholesterol esterase was determined by measuring the absorption of the product pNP at 405 nm using an enzyme-linked immunosorbent assay (ELISA) reader.

[0052] Calculations showed that the combined effect of 32% cinnamon, 53% saffron, and 15% cornus officinalis was as follows:

[0053] Inhibition rate 64.3％ 11.5％

[0054] Example 3

[0055] This embodiment proposes a method for preparing a lipid-lowering traditional Chinese medicine composition that inhibits lipase and cholesterol esterase activity. The specific preparation steps are as follows:

[0056] S1. Extraction of medicinal materials: Weigh out 35% cinnamon, 55% saffron and 10% cornus officinalis according to the mass percentage. Crush the medicinal materials and pass them through a 60-mesh sieve. Then add 15 times the weight of 75% ethanol to the crushed medicinal materials and reflux in a water bath at 70°C for 1 hour. Repeat the extraction twice, combine the extracts, filter and take the filtrate.

[0057] S2. Concentration and drying: The filtrate after S1 is filtered is concentrated and dried under reduced pressure at a temperature of 50°C to obtain the crude extract.

[0058] S3. Extraction and separation: The crude extract obtained in S2 is suspended in water to a concentration of 1 g / ml, and then extracted sequentially with petroleum ether, ethyl acetate and n-butanol, and the ethyl acetate extract phase is collected.

[0059] S4. Concentration and drying: The ethyl acetate extract phase obtained in S3 is concentrated and dried under reduced pressure at a temperature of 50°C.

[0060] S5. Purification preparation: Add 75% ethanol solution to the extract obtained by drying in S4 to dissolve it to a concentration of 1g / ml, adsorb it with HPD-600 resin for 5h, then elute with 5 column volumes of ultrapure water, and then elute with 5 column volumes of 50% ethanol solution. Collect the ethanol eluent.

[0061] S6. Concentration and Drying: The 50% ethanol eluent collected in S5 is concentrated and dried under reduced pressure at a temperature of 50°C to obtain the traditional Chinese medicine composition.

[0062] High-throughput screening of the prepared traditional Chinese medicine composition for pancreatic lipase inhibition was conducted: based on the defatting of 4-methylumbelliferone oleate (4-MUO) to 4-methylumbelliferone (4-MU) under the action of porcine pancreatic lipase (PPL), the dynamic fluorescence value of 4-MU at an excitation wavelength of 320 nm (λem = 450 nm) was measured using an enzyme-linked immunosorbent assay (ELISA) reader, and the inhibitory ability of the medicinal material on the enzyme was determined according to the net area value of its fluorescence emission intensity.

[0063] High-throughput screening of cholesterol esterase inhibition was performed on the prepared traditional Chinese medicine composition: based on the cholesterol esterase catalyzing the hydrolysis of 4-nitrophenylbutyrate (pNPB), the catalytic activity of cholesterol esterase was determined by measuring the absorption of the product pNP at 405 nm using an enzyme-linked immunosorbent assay (ELISA) reader.

[0064] Calculations show that the combined effect of 35% cinnamon, 55% saffron, and 10% cornus officinalis is as follows:

[0065] Inhibition rate 65.9％ 11.9％

[0066] Comparative Example 1

[0067] This comparative example presents a method for preparing a lipid-lowering traditional Chinese medicine composition that inhibits lipase and cholesterol esterase activity. The specific steps are as follows:

[0068] This embodiment proposes a method for preparing a lipid-lowering traditional Chinese medicine composition that inhibits lipase and cholesterol esterase activity. The specific preparation steps are as follows:

[0069] S1. Extraction of medicinal materials: Weigh out 35% cinnamon, 10% saffron and 55% cornus officinalis according to the mass percentage. Crush the medicinal materials and pass them through a 60-mesh sieve. Then add 10 times the weight of 75% ethanol to the crushed medicinal materials and reflux in a water bath at 70°C for 1.5 hours. Repeat the extraction 3 times. Combine the extracts and then filter them to collect the filtrate.

[0070] S2. Concentration and drying: The filtrate after S1 is filtered is concentrated and dried under reduced pressure at a temperature of 50°C to obtain the crude extract.

[0071] S3. Extraction and separation: The crude extract obtained in S2 is suspended in water to a concentration of 1 g / ml, and then extracted sequentially with petroleum ether, ethyl acetate and n-butanol, and the ethyl acetate extract phase is collected.

[0072] S4. Concentration and drying: The ethyl acetate extract phase obtained in S3 is concentrated and dried under reduced pressure at a temperature of 50°C.

[0073] S5. Purification preparation: Add 75% ethanol solution to the extract obtained by drying in S4 to dissolve it to a concentration of 1g / ml, adsorb it with HPD-600 resin for 5h, then elute with 5 column volumes of ultrapure water, and then elute with 5 column volumes of 50% ethanol solution. Collect the ethanol eluent.

[0074] S6. Concentration and Drying: The 50% ethanol eluent collected in S5 is concentrated and dried under reduced pressure at a temperature of 50°C to obtain the traditional Chinese medicine composition.

[0075] High-throughput screening of the prepared traditional Chinese medicine composition for pancreatic lipase inhibition was conducted: based on the defatting of 4-methylumbelliferone oleate (4-MUO) to 4-methylumbelliferone (4-MU) under the action of porcine pancreatic lipase (PPL), the dynamic fluorescence value of 4-MU at an excitation wavelength of 320 nm (λem = 450 nm) was measured using an enzyme-linked immunosorbent assay (ELISA) reader, and the inhibitory ability of the medicinal material on the enzyme was determined according to the net area value of its fluorescence emission intensity.

[0076] High-throughput screening of cholesterol esterase inhibition was performed on the prepared traditional Chinese medicine composition: based on the cholesterol esterase catalyzing the hydrolysis of 4-nitrophenylbutyrate (pNPB), the catalytic activity of cholesterol esterase was determined by measuring the absorption of the product pNP at 405 nm using an enzyme-linked immunosorbent assay (ELISA) reader.

[0077] Calculations show that the combined effect of 35% cinnamon, 10% saffron, and 55% cornus officinalis is as follows:

[0078] Inhibition rate 35.8％ 5.3％

[0079] Compared with the previous example, changing the mass percentages of cinnamon, saffron, and cornus officinalis reduced pancreatic lipase inhibitory activity from 62.1% to 35.8% and cholesterol esterase inhibitory activity from 10.8% to 5.3%.

[0080] Comparative Example 2

[0081] This comparative example presents a method for preparing a lipid-lowering traditional Chinese medicine composition that inhibits lipase and cholesterol esterase activity. The specific steps are as follows:

[0082] This embodiment proposes a method for preparing a lipid-lowering traditional Chinese medicine composition that inhibits lipase and cholesterol esterase activity. The specific preparation steps are as follows:

[0083] S1. Extraction of medicinal materials: Weigh out 10% cinnamon, 55% saffron and 35% cornus officinalis according to the mass percentage. Crush the medicinal materials and pass them through a 60-mesh sieve. Then add 10 times the weight of 75% ethanol to the crushed medicinal materials and reflux in a water bath at 70°C for 1.5 hours. Repeat the extraction 3 times. Combine the extracts and then filter them to collect the filtrate.

[0084] S2. Concentration and drying: The filtrate after S1 is filtered is concentrated and dried under reduced pressure at a temperature of 50°C to obtain the crude extract.

[0085] S3. Extraction and separation: The crude extract obtained in S2 is suspended in water to a concentration of 1 g / ml, and then extracted sequentially with petroleum ether, ethyl acetate and n-butanol, and the ethyl acetate extract phase is collected.

[0086] S4. Concentration and drying: The ethyl acetate extract phase obtained in S3 is concentrated and dried under reduced pressure at a temperature of 50°C.

[0087] S5. Purification preparation: Add 75% ethanol solution to the extract obtained by drying in S4 to dissolve it to a concentration of 1g / ml, adsorb it with HPD-600 resin for 5h, then elute with 5 column volumes of ultrapure water, and then elute with 5 column volumes of 50% ethanol solution. Collect the ethanol eluent.

[0088] S6. Concentration and Drying: The 50% ethanol eluent collected in S5 is concentrated and dried under reduced pressure at a temperature of 50°C to obtain the traditional Chinese medicine composition.

[0089] High-throughput screening of the prepared traditional Chinese medicine composition for pancreatic lipase inhibition was conducted: based on the defatting of 4-methylumbelliferone oleate (4-MUO) to 4-methylumbelliferone (4-MU) under the action of porcine pancreatic lipase (PPL), the dynamic fluorescence value of 4-MU at an excitation wavelength of 320 nm (λem = 450 nm) was measured using an enzyme-linked immunosorbent assay (ELISA) reader, and the inhibitory ability of the medicinal material on the enzyme was determined according to the net area value of its fluorescence emission intensity.

[0090] High-throughput screening of cholesterol esterase inhibition was performed on the prepared traditional Chinese medicine composition: based on the cholesterol esterase catalyzing the hydrolysis of 4-nitrophenylbutyrate (pNPB), the catalytic activity of cholesterol esterase was determined by measuring the absorption of the product pNP at 405 nm using an enzyme-linked immunosorbent assay (ELISA) reader.

[0091] Calculations show that the combined effect of 10% cinnamon, 55% saffron, and 35% cornus officinalis is as follows:

[0092] Inhibition rate 60.9％ 9.6％

[0093] Compared with Example 1, by changing the mass percentages of cinnamon, saffron, and cornus officinalis, the pancreatic lipase inhibitory activity decreased from 62.1% to 60.9%, and the cholesterol esterase inhibitory activity decreased from 10.8% to 9.6%.

[0094] Comparative Example 3

[0095] This comparative example presents a method for preparing a lipid-lowering traditional Chinese medicine composition that inhibits lipase and cholesterol esterase activity. The specific steps are as follows:

[0096] This embodiment proposes a method for preparing a lipid-lowering traditional Chinese medicine composition that inhibits lipase and cholesterol esterase activity. The specific preparation steps are as follows:

[0097] S1. Extraction of medicinal materials: Weigh out 55% cinnamon, 35% saffron and 10% cornus officinalis according to the mass percentage. Crush the medicinal materials and pass them through a 60-mesh sieve. Then add 10 times the weight of 75% ethanol to the crushed medicinal materials and reflux in a water bath at 70°C for 1.5 hours. Repeat the extraction 3 times. Combine the extracts and then filter them to collect the filtrate.

[0098] S2. Concentration and drying: The filtrate after S1 is filtered is concentrated and dried under reduced pressure at a temperature of 50°C to obtain the crude extract.

[0099] S3. Extraction and separation: The crude extract obtained in S2 is suspended in water to a concentration of 1 g / ml, and then extracted sequentially with petroleum ether, ethyl acetate and n-butanol, and the ethyl acetate extract phase is collected.

[0100] S4. Concentration and drying: The ethyl acetate extract phase obtained in S3 is concentrated and dried under reduced pressure at a temperature of 50°C.

[0101] S5. Purification preparation: Add 75% ethanol solution to the extract obtained by drying in S4 to dissolve it to a concentration of 1g / ml, adsorb it with HPD-600 resin for 5h, then elute with 5 column volumes of ultrapure water, and then elute with 5 column volumes of 50% ethanol solution. Collect the ethanol eluent.

[0102] S6. Concentration and Drying: The 50% ethanol eluent collected in S5 is concentrated and dried under reduced pressure at a temperature of 50°C to obtain the traditional Chinese medicine composition.

[0103] High-throughput screening of the prepared traditional Chinese medicine composition for pancreatic lipase inhibition was conducted: based on the defatting of 4-methylumbelliferone oleate (4-MUO) to 4-methylumbelliferone (4-MU) under the action of porcine pancreatic lipase (PPL), the dynamic fluorescence value of 4-MU at an excitation wavelength of 320 nm (λem = 450 nm) was measured using an enzyme-linked immunosorbent assay (ELISA) reader, and the inhibitory ability of the medicinal material on the enzyme was determined according to the net area value of its fluorescence emission intensity.

[0104] High-throughput screening of cholesterol esterase inhibition was performed on the prepared traditional Chinese medicine composition: based on the cholesterol esterase catalyzing the hydrolysis of 4-nitrophenylbutyrate (pNPB), the catalytic activity of cholesterol esterase was determined by measuring the absorption of the product pNP at 405 nm using an enzyme-linked immunosorbent assay (ELISA) reader.

[0105] Calculations show that the combined effect of 55% cinnamon, 35% saffron, and 10% cornus officinalis is as follows:

[0106] Inhibition rate 56.9％ 10.4％

[0107] Compared with Example 1, after changing the mass percentages of cinnamon, saffron, and cornus officinalis, the pancreatic lipase inhibitory activity decreased from 62.1% to 56.9%, and the cholesterol esterase inhibitory activity decreased from 10.8% to 10.4%.

[0108] Comparative Example 4

[0109] This comparative example presents a method for preparing a lipid-lowering traditional Chinese medicine composition that inhibits lipase and cholesterol esterase activity. The specific preparation steps are as follows:

[0110] S1. Extraction of medicinal materials: Weigh out 60% cinnamon, 0% saffron and 40% cornus officinalis according to the mass percentage. Crush the medicinal materials and pass them through a 60-mesh sieve. Then add 10 times the weight of 75% ethanol to the crushed medicinal materials and reflux in a water bath at 70°C for 1.5 hours. Repeat the extraction 3 times. Combine the extracts and then filter them. Take the filtrate.

[0111] S2. Concentration and drying: The filtrate after S1 is filtered is concentrated and dried under reduced pressure at a temperature of 50°C to obtain the crude extract.

[0112] S3. Extraction and separation: The crude extract obtained in S2 is suspended in water to a concentration of 1 g / ml, and then extracted sequentially with petroleum ether, ethyl acetate and n-butanol, and the ethyl acetate extract phase is collected.

[0113] S4. Concentration and drying: The ethyl acetate extract phase obtained in S3 is concentrated and dried under reduced pressure at a temperature of 50°C.

[0114] S5. Purification preparation: Add 75% ethanol solution to the extract obtained by drying in S4 to dissolve it to a concentration of 1g / ml, adsorb it with HPD-600 resin for 5h, then elute with 5 column volumes of ultrapure water, and then elute with 5 column volumes of 50% ethanol solution. Collect the ethanol eluent.

[0115] S6. Concentration and Drying: The 50% ethanol eluent collected in S5 is concentrated and dried under reduced pressure at a temperature of 50°C to obtain the traditional Chinese medicine composition.

[0116] High-throughput screening of the prepared traditional Chinese medicine composition for pancreatic lipase inhibition was conducted: based on the defatting of 4-methylumbelliferone oleate (4-MUO) to 4-methylumbelliferone (4-MU) under the action of porcine pancreatic lipase (PPL), the dynamic fluorescence value of 4-MU at an excitation wavelength of 320 nm (λem = 450 nm) was measured using an enzyme-linked immunosorbent assay (ELISA) reader, and the inhibitory ability of the medicinal material on the enzyme was determined according to the net area value of its fluorescence emission intensity.

[0117] High-throughput screening of cholesterol esterase inhibition was performed on the prepared traditional Chinese medicine composition: based on the cholesterol esterase catalyzing the hydrolysis of 4-nitrophenylbutyrate (pNPB), the catalytic activity of cholesterol esterase was determined by measuring the absorption of the product pNP at 405 nm using an enzyme-linked immunosorbent assay (ELISA) reader.

[0118] Calculations show that the combined effect of 60% cinnamon, 0% saffron, and 40% cornus officinalis is as follows:

[0119] Inhibition rate 31.6％ 6.0％

[0120] Compared with Example 1, in Comparative Example 4, the absence of saffron reduced pancreatic lipase inhibitory activity from 62.1% to 31.6% and cholesterol esterase inhibitory activity from 10.8% to 6.0%.

[0121] Comparative Example 5

[0122] This comparative example presents a method for preparing a lipid-lowering traditional Chinese medicine composition that inhibits lipase and cholesterol esterase activity. The specific preparation steps are as follows:

[0123] S1. Extraction of medicinal materials: Weigh out 0% cinnamon, 40% saffron and 60% cornus officinalis according to the mass percentage. Crush the medicinal materials and pass them through a 60-mesh sieve. Then add 10 times the weight of 75% ethanol to the crushed medicinal materials and reflux in a water bath at 70°C for 1.5 hours. Repeat the extraction 3 times. Combine the extracts and then filter them to collect the filtrate.

[0124] S2. Concentration and drying: The filtrate after S1 is filtered is concentrated and dried under reduced pressure at a temperature of 50°C to obtain the crude extract.

[0125] S3. Extraction and separation: The crude extract obtained in S2 is suspended in water to a concentration of 1 g / ml, and then extracted sequentially with petroleum ether, ethyl acetate and n-butanol, and the ethyl acetate extract phase is collected.

[0126] S4. Concentration and drying: The ethyl acetate extract phase obtained in S3 is concentrated and dried under reduced pressure at a temperature of 50°C.

[0127] S5. Purification preparation: Add 75% ethanol solution to the extract obtained by drying in S4 to dissolve it to a concentration of 1g / ml, adsorb it with HPD-600 resin for 5h, then elute with 5 column volumes of ultrapure water, and then elute with 5 column volumes of 50% ethanol solution. Collect the ethanol eluent.

[0128] S6. Concentration and Drying: The 50% ethanol eluent collected in S5 is concentrated and dried under reduced pressure at a temperature of 50°C to obtain the traditional Chinese medicine composition.

[0129] High-throughput screening of the prepared traditional Chinese medicine composition for pancreatic lipase inhibition was conducted: based on the defatting of 4-methylumbelliferone oleate (4-MUO) to 4-methylumbelliferone (4-MU) under the action of porcine pancreatic lipase (PPL), the dynamic fluorescence value of 4-MU at an excitation wavelength of 320 nm (λem = 450 nm) was measured using an enzyme-linked immunosorbent assay (ELISA) reader, and the inhibitory ability of the medicinal material on the enzyme was determined according to the net area value of its fluorescence emission intensity.

[0130] High-throughput screening of cholesterol esterase inhibition was performed on the prepared traditional Chinese medicine composition: based on the cholesterol esterase catalyzing the hydrolysis of 4-nitrophenylbutyrate (pNPB), the catalytic activity of cholesterol esterase was determined by measuring the absorption of the product pNP at 405 nm using an enzyme-linked immunosorbent assay (ELISA) reader.

[0131] Calculations show that the combined effect of 0% cinnamon, 40% saffron, and 60% cornus officinalis is as follows:

[0132] Inhibition rate 51.2％ 6.4％

[0133] Compared with Example 1, in Comparative Example 5, the absence of cinnamon reduced pancreatic lipase inhibitory activity from 62.1% to 51.2% and cholesterol esterase inhibitory activity from 10.8% to 6.4%.

[0134] Comparative Example 6

[0135] This comparative example presents a method for preparing a lipid-lowering traditional Chinese medicine composition that inhibits lipase and cholesterol esterase activity. The specific preparation steps are as follows:

[0136] S1. Extraction of medicinal materials: Weigh out 60% cinnamon, 40% saffron and 0% cornus officinalis according to the mass percentage. Crush the medicinal materials and pass them through a 60-mesh sieve. Then add 10 times the weight of 75% ethanol to the crushed medicinal materials and reflux in a water bath at 70°C for 1.5 hours. Repeat the extraction 3 times. Combine the extracts and then filter them to collect the filtrate.

[0137] S2. Concentration and drying: The filtrate after S1 is filtered is concentrated and dried under reduced pressure at a temperature of 50°C to obtain the crude extract.

[0138] S3. Extraction and separation: The crude extract obtained in S2 is suspended in water to a concentration of 1 g / ml, and then extracted sequentially with petroleum ether, ethyl acetate and n-butanol, and the ethyl acetate extract phase is collected.

[0139] S4. Concentration and drying: The ethyl acetate extract phase obtained in S3 is concentrated and dried under reduced pressure at a temperature of 50°C.

[0140] S5. Purification preparation: Add 75% ethanol solution to the extract obtained by drying in S4 to dissolve it to a concentration of 1g / ml, adsorb it with HPD-600 resin for 5h, then elute with 5 column volumes of ultrapure water, and then elute with 5 column volumes of 50% ethanol solution. Collect the ethanol eluent.

[0141] S6. Concentration and Drying: The 50% ethanol eluent collected in S5 is concentrated and dried under reduced pressure at a temperature of 50°C to obtain the traditional Chinese medicine composition.

[0142] High-throughput screening of the prepared traditional Chinese medicine composition for pancreatic lipase inhibition was conducted: based on the defatting of 4-methylumbelliferone oleate (4-MUO) to 4-methylumbelliferone (4-MU) under the action of porcine pancreatic lipase (PPL), the dynamic fluorescence value of 4-MU at an excitation wavelength of 320 nm (λem = 450 nm) was measured using an enzyme-linked immunosorbent assay (ELISA) reader, and the inhibitory ability of the medicinal material on the enzyme was determined according to the net area value of its fluorescence emission intensity.

[0143] High-throughput screening of cholesterol esterase inhibition was performed on the prepared traditional Chinese medicine composition: based on the cholesterol esterase catalyzing the hydrolysis of 4-nitrophenylbutyrate (pNPB), the catalytic activity of cholesterol esterase was determined by measuring the absorption of the product pNP at 405 nm using an enzyme-linked immunosorbent assay (ELISA) reader.

[0144] Calculations show that the combined effect of 60% cinnamon, 40% saffron, and 0% dogwood is as follows:

[0145] Inhibition rate 59.6％ 10.7％

[0146] Compared with Example 1, in Comparative Example 6, the absence of Cornus officinalis reduced pancreatic lipase inhibitory activity from 62.1% to 59.6% and cholesterol esterase inhibitory activity from 10.8% to 10.7%.

[0147] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A lipid-lowering traditional Chinese medicine composition that inhibits pancreatic lipase and cholesterol esterase, characterized in that, The ingredients consist of 32% cinnamon, 53% saffron, and 15% cornus officinalis by weight percentage. The preparation method of the lipid-lowering traditional Chinese medicine composition that inhibits pancreatic lipase and cholesterol esterase includes the following steps: Step 1: Weigh out cinnamon, saffron and cornelian cherry according to their mass percentages, pulverize them, pass them through a 40-80 mesh sieve, add 10-15 times the amount of 75% ethanol, reflux in a water bath at 60-80℃ for 1-2 hours, repeat the extraction 2-3 times, combine the extracts, and filter. Step 2: The filtered extract is concentrated and dried under negative pressure vacuum to obtain the crude extract; Step 3: Suspend the crude extract in water until the crude extract concentration is 1g / ml, and extract it sequentially with petroleum ether, ethyl acetate and n-butanol; Step 4: Concentrate and dry the ethyl acetate extract from Step 3 under negative pressure vacuum to obtain the extract; Step 5: Dissolve the extract obtained in Step 4 in ethanol solution, then adsorb it with HPD-600 resin, and elute with ultrapure water and 50% ethanol in sequence, and collect the ethanol eluent. Step 6: Concentrate and dry the ethanol eluent collected in Step 5 under negative pressure vacuum to obtain the traditional Chinese medicine composition.

2. A lipid-lowering traditional Chinese medicine composition that inhibits pancreatic lipase and cholesterol esterase, characterized in that, The ingredients consist of 35% cinnamon, 55% saffron, and 10% cornus officinalis by weight percentage. The preparation method of the lipid-lowering traditional Chinese medicine composition that inhibits pancreatic lipase and cholesterol esterase includes the following steps: Step 1: Weigh out cinnamon, saffron and cornelian cherry according to their mass percentages, pulverize them, pass them through a 40-80 mesh sieve, add 10-15 times the amount of 75% ethanol, reflux in a water bath at 60-80℃ for 1-2 hours, repeat the extraction 2-3 times, combine the extracts, and filter. Step 2: The filtered extract is concentrated and dried under negative pressure vacuum to obtain the crude extract; Step 3: Suspend the crude extract in water until the crude extract concentration is 1g / ml, and extract it sequentially with petroleum ether, ethyl acetate and n-butanol; Step 4: Concentrate and dry the ethyl acetate extract from Step 3 under negative pressure vacuum to obtain the extract; Step 5: Dissolve the extract obtained in Step 4 in ethanol solution, then adsorb it with HPD-600 resin, and elute with ultrapure water and 50% ethanol in sequence, and collect the ethanol eluent. Step 6: Concentrate and dry the ethanol eluent collected in Step 5 under negative pressure vacuum to obtain the traditional Chinese medicine composition.

3. The lipid-lowering traditional Chinese medicine composition for inhibiting pancreatic lipase and cholesterol esterase as described in claim 1 or 2, characterized in that, The drying temperature for steps two, four, and six is ​​40-60℃.

4. The lipid-lowering traditional Chinese medicine composition for inhibiting pancreatic lipase and cholesterol esterase as described in claim 1 or 2, characterized in that, Step 5 includes: dissolving the extract obtained in step 4 in 75% ethanol solution to a concentration of 1 g / ml, loading the sample onto HPD-600 resin for adsorption for 5 h, eluting with ultrapure water for 5 PV, and then eluting with 5 PV of 50% ethanol, and collecting the ethanol eluent.