CN114835662B - Gallate novel compound in myrobalan and preparation method and application thereof - Google Patents
Gallate novel compound in myrobalan and preparation method and application thereof Download PDFInfo
- Publication number
- CN114835662B CN114835662B CN202210304343.1A CN202210304343A CN114835662B CN 114835662 B CN114835662 B CN 114835662B CN 202210304343 A CN202210304343 A CN 202210304343A CN 114835662 B CN114835662 B CN 114835662B
- Authority
- CN
- China
- Prior art keywords
- myrobalan
- novel
- silica gel
- methanol
- chloroform
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 235000011517 Terminalia chebula Nutrition 0.000 title claims abstract description 63
- 235000015489 Emblica officinalis Nutrition 0.000 title claims abstract description 62
- 244000277583 Terminalia catappa Species 0.000 title claims abstract 11
- LNTHITQWFMADLM-UHFFFAOYSA-N gallic acid Chemical compound OC(=O)C1=CC(O)=C(O)C(O)=C1 LNTHITQWFMADLM-UHFFFAOYSA-N 0.000 title abstract description 14
- 238000002360 preparation method Methods 0.000 title abstract description 13
- 150000001875 compounds Chemical class 0.000 title description 6
- -1 gallate compound Chemical class 0.000 claims abstract description 39
- 238000000926 separation method Methods 0.000 claims abstract description 17
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 15
- 230000000694 effects Effects 0.000 claims abstract description 15
- 239000000741 silica gel Substances 0.000 claims abstract description 15
- 229910002027 silica gel Inorganic materials 0.000 claims abstract description 15
- 101710184309 Probable sucrose-6-phosphate hydrolase Proteins 0.000 claims abstract description 13
- 102400000472 Sucrase Human genes 0.000 claims abstract description 13
- 101710112652 Sucrose-6-phosphate hydrolase Proteins 0.000 claims abstract description 13
- 102000016679 alpha-Glucosidases Human genes 0.000 claims abstract description 13
- 108010028144 alpha-Glucosidases Proteins 0.000 claims abstract description 13
- 235000011073 invertase Nutrition 0.000 claims abstract description 13
- 239000003814 drug Substances 0.000 claims abstract description 9
- 239000011347 resin Substances 0.000 claims abstract description 8
- 229920005989 resin Polymers 0.000 claims abstract description 8
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 69
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 36
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 32
- 238000010898 silica gel chromatography Methods 0.000 claims description 25
- 239000000243 solution Substances 0.000 claims description 20
- 239000012141 concentrate Substances 0.000 claims description 14
- 239000011259 mixed solution Substances 0.000 claims description 14
- 238000010828 elution Methods 0.000 claims description 13
- 239000003480 eluent Substances 0.000 claims description 11
- 238000010829 isocratic elution Methods 0.000 claims description 10
- 239000002904 solvent Substances 0.000 claims description 10
- WORJEOGGNQDSOE-UHFFFAOYSA-N chloroform;methanol Chemical compound OC.ClC(Cl)Cl WORJEOGGNQDSOE-UHFFFAOYSA-N 0.000 claims description 8
- 230000002401 inhibitory effect Effects 0.000 claims description 7
- GBMDVOWEEQVZKZ-UHFFFAOYSA-N methanol;hydrate Chemical compound O.OC GBMDVOWEEQVZKZ-UHFFFAOYSA-N 0.000 claims description 7
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 claims description 5
- 238000000034 method Methods 0.000 claims description 5
- 238000010992 reflux Methods 0.000 claims description 5
- 238000010298 pulverizing process Methods 0.000 claims description 4
- 238000005406 washing Methods 0.000 claims description 3
- 238000004519 manufacturing process Methods 0.000 claims 1
- 238000000605 extraction Methods 0.000 abstract description 8
- 239000003472 antidiabetic agent Substances 0.000 abstract description 6
- 230000005764 inhibitory process Effects 0.000 abstract description 6
- 230000002441 reversible effect Effects 0.000 abstract description 5
- 238000000746 purification Methods 0.000 abstract description 2
- 229940079593 drug Drugs 0.000 abstract 1
- 241000001522 Terminalia chebula Species 0.000 description 54
- 102000004190 Enzymes Human genes 0.000 description 7
- 108090000790 Enzymes Proteins 0.000 description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 7
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 6
- 235000004515 gallic acid Nutrition 0.000 description 6
- 229940074391 gallic acid Drugs 0.000 description 6
- 239000008103 glucose Substances 0.000 description 6
- 230000002829 reductive effect Effects 0.000 description 6
- 235000013399 edible fruits Nutrition 0.000 description 5
- 238000011160 research Methods 0.000 description 4
- 238000001228 spectrum Methods 0.000 description 4
- 239000007864 aqueous solution Substances 0.000 description 3
- 239000005457 ice water Substances 0.000 description 3
- 238000012544 monitoring process Methods 0.000 description 3
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 2
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 2
- 241000700159 Rattus Species 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 2
- 239000003405 delayed action preparation Substances 0.000 description 2
- 239000002552 dosage form Substances 0.000 description 2
- 238000001052 heteronuclear multiple bond coherence spectrum Methods 0.000 description 2
- 238000000990 heteronuclear single quantum coherence spectrum Methods 0.000 description 2
- 229940126904 hypoglycaemic agent Drugs 0.000 description 2
- 210000004347 intestinal mucosa Anatomy 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 230000000144 pharmacologic effect Effects 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 210000000813 small intestine Anatomy 0.000 description 2
- 238000012916 structural analysis Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 238000004809 thin layer chromatography Methods 0.000 description 2
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 1
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- 241000221032 Combretaceae Species 0.000 description 1
- 206010068319 Oropharyngeal pain Diseases 0.000 description 1
- 201000007100 Pharyngitis Diseases 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 230000002155 anti-virotic effect Effects 0.000 description 1
- 238000003287 bathing Methods 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 229930003935 flavonoid Natural products 0.000 description 1
- 150000002215 flavonoids Chemical class 0.000 description 1
- 235000017173 flavonoids Nutrition 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 125000002791 glucosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)* 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 210000000936 intestine Anatomy 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000004112 neuroprotection Effects 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 150000007965 phenolic acids Chemical class 0.000 description 1
- 235000009048 phenolic acids Nutrition 0.000 description 1
- 239000006187 pill Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000008213 purified water Substances 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000007873 sieving Methods 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 235000018553 tannin Nutrition 0.000 description 1
- 229920001864 tannin Polymers 0.000 description 1
- 239000001648 tannin Substances 0.000 description 1
- 150000003648 triterpenes Chemical class 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D307/00—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom
- C07D307/02—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom not condensed with other rings
- C07D307/34—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members
- C07D307/38—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members with substituted hydrocarbon radicals attached to ring carbon atoms
- C07D307/40—Radicals substituted by oxygen atoms
- C07D307/46—Doubly bound oxygen atoms, or two oxygen atoms singly bound to the same carbon atom
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Diabetes (AREA)
- Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Hematology (AREA)
- Obesity (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Endocrinology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pharmacology & Pharmacy (AREA)
- Emergency Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Botany (AREA)
- Mycology (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Saccharide Compounds (AREA)
Abstract
The invention provides a novel gallate compound in myrobalan and a preparation method and application thereof, belonging to the technical field of extraction and separation of traditional Chinese and Tibetan medicines. The invention provides a novel gallate compound in myrobalan, which has better sucrase and maltase inhibition activity and can be used for preparing hypoglycemic drugs and health care products. The invention also provides a preparation method of the novel gallate compound in the myrobalan, which comprises the steps of extraction, macroporous resin separation and silica gel forward and reverse column purification in sequence, and the novel gallate compound in the myrobalan is successfully separated. The preparation method is simple, and can realize the rapid and high-purity extraction and separation of the novel gallate compounds in the myrobalan.
Description
Technical Field
The invention relates to the technical field of extraction and separation of traditional Chinese medicine and Tibetan medicine, in particular to a novel gallate compound in myrobalan and a preparation method and application thereof.
Background
The Tibetan medicine myrobalan is a dry mature fruit of the plant myrobalan (Terminalia chebula Retz.) of the family Combretaceae, which is called chebula, and fructus Aristolochiae, is native to India, myanmar, etc., and is distributed in Tibet, yunnan, guangdong, guangxi, etc. places in China. The myrobalan has the effects of astringing intestines and astringing lung, reducing pathogenic fire and relieving sore throat, and modern pharmacological researches prove that the myrobalan has various pharmacological effects of antioxidation, neuroprotection, anti-tumor, antivirus, antibiosis and the like, and the main chemical components of the myrobalan comprise tannins, phenolic acids, triterpenes, flavonoids and the like. In Tibetan medicine and Mongolian medicine, chebulae fructus is most commonly used, and its frequency of use is almost equal to that of Glycyrrhrizae radix in Chinese medicinal prescription, which is regarded as "Tibetan medicine king". The myrobalan has various effects, wide clinical application and high medicinal value, but the research on the myrobalan is less and scattered at present, and the medicinal components of the myrobalan are not clear.
Disclosure of Invention
Therefore, the invention aims to provide a novel gallate compound in myrobalan, and a preparation method and application thereof. The novel gallate compound in the myrobalan has better sucrase and maltase inhibition activity, and can be used for preparing medicaments and health-care products for reducing blood sugar.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a novel gallic acid ester compound in myrobalan, which has a structure shown in a formula I:
the invention also provides a preparation method of the novel gallic acid ester compound in the myrobalan, which comprises the following steps:
pulverizing fructus Chebulae, reflux extracting with ethanol water solution, and concentrating to obtain fructus Chebulae extract;
washing the myrobalan extract with AB-8 macroporous resin, eluting with ethanol, and concentrating to obtain concentrate;
after the concentrate is mixed with silica gel, performing silica gel column chromatography rough separation, wherein the silica gel column chromatography rough separation is segmented by using a chloroform/methanol solvent system to respectively obtain Fr.1, fr.2, fr.3, fr.4, fr.5 and Fr.6, and the volume ratio of chloroform to methanol in the chloroform/methanol solvent is 9: 1. 8: 2. 7: 3. 6: 4. 1:1 and 0:1, a step of;
and sequentially performing normal phase silica gel column chromatography gradient elution and reversed phase silica gel column chromatography isocratic elution on the Fr.6 to obtain the novel gallate compound in the myrobalan.
Preferably, the volume fraction of the ethanol aqueous solution is 75-95%.
Preferably, the mass ratio of the concentrate to the silica gel is 1:1.5.
Preferably, the silica gel column chromatography is performed by using 10 times of 100-mesh silica gel by weight.
Preferably, the eluent of the gradient elution is chloroform-methanol mixed solution, and the volume ratio of chloroform to methanol in the chloroform-methanol mixed solution is 20:1 to 0:1.
Preferably, the mass of the gradient elution eluent is 500 times fr.6.
Preferably, the eluent for isocratic elution is a methanol-water mixed solution, and the volume percentage of methanol in the methanol-water mixed solution is 20%.
The invention also provides application of the novel gallate compound in the myrobalan in inhibiting activity of sucrase and maltase.
The invention also provides application of the novel gallate compound in the myrobalan in preparing hypoglycemic drugs.
The invention provides a novel gallate compound in myrobalan, which has better sucrase and maltase inhibition activity and can be used for preparing hypoglycemic drugs and health care products.
The invention also provides a preparation method of the novel gallate compound in the myrobalan, which comprises the steps of extraction, macroporous resin separation and silica gel forward and reverse column purification in sequence, and the novel gallate compound in the myrobalan is successfully separated. The preparation method is simple, and can realize the rapid and high-purity extraction and separation of the novel gallate compounds in the myrobalan.
Drawings
FIG. 1 is a DEPT135 spectrum of a novel gallate compound in myrobalan;
FIG. 2 is a DEPT90 spectrum of a novel gallate compound in myrobalan;
FIG. 3 is a HSQC spectrum of the novel gallate compounds in myrobalan;
FIG. 4 is a HMBC spectrum of a novel gallate compound in myrobalan;
FIG. 5 sucrase IC of novel gallate compounds in myrobalan 50 A value;
FIG. 6 is a schematic diagram of maltase IC of novel gallate compounds in myrobalan 50 Values.
Detailed Description
The invention provides a novel gallic acid ester compound in myrobalan, which has a structure shown in a formula I:
the molecular formula of the novel gallate compound in the myrobalan is C 13 H 10 O 7 The chemical name is gallic acid-5-hydroxymethyl furfural ester.
The invention also provides a preparation method of the novel gallic acid ester compound in the myrobalan, which comprises the following steps:
pulverizing fructus Chebulae, reflux extracting with ethanol water solution, and concentrating to obtain fructus Chebulae extract;
washing the myrobalan extract with AB-8 macroporous resin, eluting with ethanol, and concentrating to obtain concentrate;
after the concentrate is mixed with silica gel, performing silica gel column chromatography rough separation, wherein the silica gel column chromatography rough separation is segmented by using a chloroform/methanol solvent system to respectively obtain Fr.1, fr.2, fr.3, fr.4, fr.5 and Fr.6, and the volume ratio of chloroform to methanol in the chloroform/methanol solvent is 9: 1. 8: 2. 7: 3. 6: 4. 1:1 and 0:1, a step of;
and sequentially performing normal phase silica gel column chromatography gradient elution and reversed phase silica gel column chromatography isocratic elution on the Fr.6 to obtain the novel gallate compound in the myrobalan.
The invention is characterized in that the dry myrobalan fruit is crushed and then is extracted by ethanol water solution in a reflux way, and the myrobalan fruit extract is obtained by concentration.
The specific method for crushing the dry myrobalan fruits is not particularly limited, and the method is well known to those skilled in the art.
In the present invention, the volume fraction of the aqueous ethanol solution is preferably 75 to 95%. The amount of the ethanol aqueous solution is not particularly limited, and the ethanol aqueous solution can be completely extracted.
In the present invention, the number of times of the reflux extraction is preferably 3, and the time of each time is preferably 2 hours.
In the present invention, the concentration is preferably reduced pressure concentration, and the time and temperature of the reduced pressure concentration are not particularly limited, and the present invention may be carried out so as to completely remove ethanol and water.
After the myrobalan extract is obtained, the myrobalan extract is washed by AB-8 macroporous resin, eluted by ethanol and then concentrated to obtain a concentrate.
In the invention, the myrobalan extract is preferably dissolved with water and then put on an AB-8 macroporous resin column, and the sugar and protein are removed by water elution.
In the present invention, the ethanol is preferably an aqueous ethanol solution having a volume percentage of 95% used for the ethanol elution.
In the present invention, the concentration is preferably reduced pressure concentration, and the time and temperature of the reduced pressure concentration are not particularly limited, and the present invention may be carried out so as to completely remove ethanol and water.
After the concentrate is obtained, the concentrate is mixed with silica gel, and then the silica gel column chromatography rough separation is carried out, wherein the silica gel column chromatography rough separation is segmented by using a chloroform/methanol solvent system to respectively obtain Fr.1, fr.2, fr.3, fr.4, fr.5 and Fr.6, and the volume ratio of chloroform to methanol in the chloroform/methanol solvent is 9: 1. 8: 2. 7: 3. 6: 4. 1:1 and 0:1.
in the present invention, the mass ratio of the concentrate to the silica gel is preferably 1:1.5. In the present invention, the particle size of the silica gel is preferably 100 mesh.
In the present invention, the sample is preferably further dried after being mixed.
In the present invention, the silica gel column chromatography is preferably performed by using 10 times by weight of 100 mesh silica gel.
The amount of the chloroform/methanol solvent used in the present invention is not particularly limited, and it is sufficient to completely separate each part.
In the invention, the silica gel column chromatography is preferably carried out in the course of rough separation, and the Fr.1, fr.2, fr.3, fr.4, fr.5 and Fr.6 are obtained by combining the same parts of the monitoring of the thin layer plate.
After Fr.6 is obtained, the Fr.6 is sequentially subjected to normal phase silica gel column chromatography gradient elution and reversed phase silica gel column chromatography isocratic elution to obtain the novel gallate compound in the myrobalan.
In the invention, the eluent of the gradient elution is preferably chloroform-methanol mixed solution, and the volume ratio of chloroform to methanol in the chloroform-methanol mixed solution is preferably 20:1 to 0:1.
In the invention, the chloroform-methanol mixed solution is preferably prepared from the following components in percentage by volume: 1. 10: 1. 5: 1. 2:1 and 0: the elution was performed with 1 gradient, each gradient using 100 times the mass of fr.6, each gradient eluting for 1 hour.
In the present invention, the mass of the gradient elution eluent is preferably 500 times that of fr.6.
In the present invention, the reverse phase silica gel column chromatography is preferably reverse phase silica gel column chromatography RP-18.
In the present invention, the eluent for isocratic elution is preferably a methanol-water mixed solution, and the volume percentage of methanol in the methanol-water mixed solution is preferably 20%.
In the present invention, the isocratic elution is preferably performed at an atmospheric flow rate.
In the present invention, thin layer chromatography monitoring is preferably performed during the isocratic elution.
After the isocratic elution is completed, the obtained eluent is preferably concentrated to obtain the novel gallate compound in the myrobalan. The specific mode of the concentration is not particularly limited, and may be any mode known to those skilled in the art.
The invention also provides application of the novel gallate compound in the myrobalan in inhibiting activity of sucrase and maltase.
The invention also provides application of the novel gallate compound in the myrobalan in preparing hypoglycemic drugs.
In the invention, the hypoglycemic agent preferably comprises an effective dose of the novel gallate compound in the myrobalan, the stereoisomer, the medicinal salt and the pharmaceutically acceptable carrier, auxiliary materials, excipient and diluent.
In the invention, the dosage form of the hypoglycemic agent preferably comprises a pharmaceutically acceptable dosage form of a tablet, an injection, a capsule, a granule, a pill, a powder, an oral liquid, a sustained-release preparation, a controlled-release preparation or a nano preparation.
In order to further illustrate the present invention, the novel compounds of gallic acid ester in myrobalan, and the preparation method and application thereof, provided by the present invention, are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.
Example 1
(1) Pulverizing 3kg of natural dry fructus Chebulae fruit, sieving, reflux-extracting with 95% ethanol solution for 3 times each for 2 hr, and concentrating under reduced pressure to recover ethanol to obtain total extract.
(2) Dissolving the extract obtained in the step (1) with purified water, loading on an AB-8 macroporous resin column, eluting with water to remove sugar and protein, eluting with 95% ethanol solution with volume percentage, and concentrating the eluate under reduced pressure to obtain 320g of concentrate;
(3) And (3) mixing the concentrate obtained in the step (2) with 1.5 times of 100-mesh silica gel, airing, performing silica gel column chromatography coarse separation by using 10 times of 100-mesh silica gel, dividing by using a chloroform/methanol solvent system (volume ratio of 9:1, 8:2, 7:3, 6:4, 1:1 and 0:1), and combining the same parts by monitoring a thin layer plate to obtain six parts of Fr.1, fr.2, fr.3, fr.4, fr.5 and Fr.6. Fr.6 is eluted by normal phase silica gel column chromatography with a gradient of 500 times weight chloroform-methanol mixed solution (the volume ratio of chloroform to methanol is 20:1, 10:1, 5:1, 2:1 and 0:1 respectively), each gradient is 100 times of Fr.6 in mass, each gradient is eluted for 1 hour), then is eluted by reverse phase silica gel column chromatography (RP-18) with a constant pressure flow rate of methanol-water (the volume percentage of methanol is 20%), the thin layer chromatography is monitored, and the eluent is concentrated to obtain 11mg of new compound.
Structural analysis is carried out on the novel gallate compound in the myrobalan:
the specific data are: gallic acid-5-hydroxymethyl furfural ester, yellow solid. UV (MeOH) amax (logepsilon) 225 (4.66), 293 (4.19) nm; IR (KBr) v max 3415,1669,1384,1212,1050,1025,766cm-1; 1 H-NMR(400MHz,DMSO-d 6 )δ:9.48(1H,s,-CHO),7.30(1H,d,J＝3.6Hz,H-3),6.96(2H,s),6.65(1H,d,J＝3.6Hz,H-4),5.22(2H,s,H-6)； 13 C-NMR(100MHz,DMSO-d 6 )δ:178.3,166.2,156.4,153.0,145.2,122.6,119.3,112.3,108.9,57.6；HRESI-MS m/z 277.0350[M-H] - (calcd.For C 13 H 10 O 7 ,277.0354).
fig. 1 is a DEPT135 spectrum of a novel gallate compound in myrobalan, fig. 2 is a DEPT90 spectrum of a novel gallate compound in myrobalan, fig. 3 is an HSQC spectrum of a novel gallate compound in myrobalan, and fig. 4 is an HMBC spectrum of a novel gallate compound in myrobalan.
According to the structural analysis, the novel gallate compound in the myrobalan with the structure shown in the formula I is prepared.
Evaluation example of Activity
The novel compounds of gallic acid ester in the myrobalan obtained by separation in the example are taken for carrying out the research on the inhibition activity of sucrase and maltase.
Specific experimental method
(1) Rat intestinal sucrase and maltase extraction and activity determination: rats were fasted for 12 hours, sacrificed by cervical spine removal, immediately removed from the small intestine and placed on an ice table, the small intestine dissected and turned overThe intestinal mucosa is exposed, washed by PBS precooled at 4 ℃ and then wiped dry, and the small intestinal mucosa is scraped by a glass slide according to the mass volume ratio of 1:5 adding pre-cooled PBS at 4deg.C, homogenizing, centrifuging at 4deg.C at 8000r/min for 20min, collecting supernatant, packaging, and storing at-20deg.C. Adding 0.35mL PBS into 0.1mL supernatant enzyme solution, water-bathing at 37deg.C for 10min, adding 50 μL of 0.25mol/L sucrose (maltose) solution, immediately adding into ice water bath for 5min after 20min, and continuously adding Na 2 CO 3 The reaction was stopped at 0.5mL of solution. 3-well replicates, 1. Mu. MoL glucose produced per minute in 1L solution at 37℃and pH6.8 was defined as 1 enzyme activity unit. Enzyme activity = glucose concentration x 2 x 1000/20.
(2) Screening for sucrase inhibitory Activity: mu.L of the enzyme solution containing 17.5U/mL and 50. Mu.L of the 5mg/mL sample were added to a 48-well plate, and pre-incubated at 37℃for 10min. Then 50. Mu.L of a 0.5mol/L sucrose solution was added and the mixture was incubated at 37℃for 20min. Immediately putting into ice water bath for 5min, reducing enzyme activity, and continuously adding 0.1mol/L Na 2 CO 3 The reaction was stopped at 50. Mu.L of solution and repeated three times. Glucose concentration was determined using a glucose kit and the inhibitory activity of the sample on sucrase was calculated.
(3) Maltase inhibitory activity screening: mu.L of the enzyme solution containing 11.56U/mL and 50. Mu.L of the 5mg/mL sample were added to a 48-well plate, and pre-incubated at 37℃for 10min. Then, 50. Mu.L of a 1.39mmol/mL maltose solution was added and the mixture was incubated at 37℃for 20min. Immediately putting into ice water bath for 5min, reducing enzyme activity, and continuously adding 0.1mol/L Na 2 CO 3 The reaction was stopped at 50. Mu.L of solution and repeated three times. Glucose concentration was determined using a glucose kit and the inhibitory activity of the sample on maltase was calculated.
The research on the sucrase and maltase inhibition activities shows that the novel compound with the gallate structure has sucrase inhibition activity IC 50 The value was 499.1. Mu.M, and IC was inhibited against maltase half-maximal ratio 50 The value was 482.7. Mu.M, see FIGS. 5 and 6, respectively.
The foregoing is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. It should be noted that modifications and adaptations to the present invention may occur to one skilled in the art without departing from the principles of the present invention and are intended to be comprehended within the scope of the present invention.
Claims (5)
1. The novel gallate compound in myrobalan is characterized by having a structure shown in a formula I:
2. the method for preparing the novel gallate compound in myrobalan according to claim 1, comprising the steps of:
pulverizing fructus Chebulae, reflux extracting with ethanol water solution, and concentrating to obtain fructus Chebulae extract; the volume fraction of the ethanol water solution is 75-95%;
washing the myrobalan extract with AB-8 macroporous resin, eluting with ethanol, and concentrating to obtain concentrate;
after the concentrate is mixed with silica gel, performing silica gel column chromatography rough separation, wherein the silica gel column chromatography rough separation is segmented by using a chloroform/methanol solvent system to respectively obtain Fr.1, fr.2, fr.3, fr.4, fr.5 and Fr.6, and the volume ratio of chloroform to methanol in the chloroform/methanol solvent is 9: 1. 8: 2. 7: 3. 6: 4. 1:1 and 0:1, a step of;
sequentially performing normal phase silica gel column chromatography gradient elution and reversed phase silica gel column chromatography isocratic elution on the Fr.6 to obtain a novel gallate compound in the myrobalan; the eluent of the gradient elution is chloroform-methanol mixed solution, and the volume ratio of chloroform to methanol in the chloroform-methanol mixed solution is 20:1-0:1; the mass of the eluent of the gradient elution is 500 times of that of Fr.6; the eluent for isocratic elution is a methanol-water mixed solution, and the volume percentage of methanol in the methanol-water mixed solution is 20%.
3. The method of claim 2, wherein the mass ratio of concentrate to silica gel is 1:1.5.
4. A method according to claim 2 or 3, wherein the silica gel column chromatography is performed using 10 times by weight of 100 mesh silica gel.
5. Use of a novel gallate compound in myrobalan as claimed in claim 1 in the manufacture of a medicament for inhibiting the activity of sucrase and maltase.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210304343.1A CN114835662B (en) | 2022-03-25 | 2022-03-25 | Gallate novel compound in myrobalan and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210304343.1A CN114835662B (en) | 2022-03-25 | 2022-03-25 | Gallate novel compound in myrobalan and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114835662A CN114835662A (en) | 2022-08-02 |
| CN114835662B true CN114835662B (en) | 2024-04-05 |
Family
ID=82561620
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210304343.1A Active CN114835662B (en) | 2022-03-25 | 2022-03-25 | Gallate novel compound in myrobalan and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114835662B (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110279732A (en) * | 2019-07-23 | 2019-09-27 | 中国科学院西北高原生物研究所 | Application of the Fructus Chebulae extract in the food, drug or health care product that preparation inhibits amylase and glucosidase activity |
-
2022
- 2022-03-25 CN CN202210304343.1A patent/CN114835662B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110279732A (en) * | 2019-07-23 | 2019-09-27 | 中国科学院西北高原生物研究所 | Application of the Fructus Chebulae extract in the food, drug or health care product that preparation inhibits amylase and glucosidase activity |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114835662A (en) | 2022-08-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US9770479B2 (en) | Extract of Rehmannia glutinasa Libosch for reducing blood sugar, reducing blood fat, treating leukemia, and preparation method and uses thereof | |
| CN101035548B (en) | Steroidal saponin pharmaceutical composition, the preparation method and use thereof | |
| CN112209979B (en) | Production process for jointly separating high-purity glycyrrhizin, debitterized glycyrrhizin and total flavonoids from monoammonium glycyrrhizinate mother liquor | |
| CN102134268B (en) | Method for preparing panax japonicus saponin IVa and application of panax japonicus saponin IVa in preparing a medicament for protecting liver and lowering transaminase | |
| CN114835662B (en) | Gallate novel compound in myrobalan and preparation method and application thereof | |
| CN112159451A (en) | Gynostemma pentaphylla saponin extract and preparation method thereof | |
| CN109820873B (en) | Defatted periplaneta americana and preparation method and application thereof | |
| CN101732302B (en) | Application of chlorogenic acid in preparing medicines for preventing and curing pulmonary fibrosis | |
| CN101342186B (en) | Use of pentacyclic triterpene saponin compound | |
| CN109053756B (en) | Phenylpropanoid ester type catechin and preparation method and application thereof | |
| CN102579530A (en) | Preparation method of aralia taibaiensis total saponin having diabetes mellitus resisting effect and medicament | |
| CN110960569A (en) | Phyllanthus emblica extract and preparation method and application thereof | |
| CN1312170C (en) | Technique of preparing extract product of Radde Anemone Rhizome extract, and its application in preparing medication of treating cancer | |
| CN114470101A (en) | Traditional Chinese medicine composition, preparation method thereof and oral agent containing traditional Chinese medicine composition | |
| CN109467581B (en) | Method for extracting flavonoid glycoside from lotus leaves and pharmaceutical preparation containing flavonoid glycoside from lotus leaves | |
| CN103923042A (en) | Preparation method for salvianolic acid B extract | |
| CN114605370B (en) | Chebula acid cracking compound in chebula fruit, and preparation method and application thereof | |
| CN111920799A (en) | Kulecuo effective component composition and preparation method and application thereof | |
| CN101269123A (en) | Secondary development novel technique for thirst eliminating capsule for lowering blood sugar | |
| CN116178331B (en) | Antidiabetic xanthone compound 2-hydroxyxanthone, preparation method, pharmaceutical composition and application thereof | |
| CN110903270A (en) | 2, 6-epoxy diphenyl heptane compound, preparation method and application thereof, pharmaceutical composition and application thereof | |
| CN115429790B (en) | Application of isoflavone compound in preparing medicament for preventing or treating alcoholic liver injury or dispelling alcohol effect and protecting liver | |
| CN115010689B (en) | Compound kurarinol A and preparation method and application thereof | |
| CN104116753A (en) | Application of aucubin to preparation of medicines for treating idiopathic pulmonary fibrosis | |
| CN114213480B (en) | Glycoside compound, pharmaceutical composition, preparation method and application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |