CN110452110B - Phloroglucinol natural medicine and preparation method and application thereof - Google Patents
Phloroglucinol natural medicine and preparation method and application thereof Download PDFInfo
- Publication number
- CN110452110B CN110452110B CN201810425488.0A CN201810425488A CN110452110B CN 110452110 B CN110452110 B CN 110452110B CN 201810425488 A CN201810425488 A CN 201810425488A CN 110452110 B CN110452110 B CN 110452110B
- Authority
- CN
- China
- Prior art keywords
- petroleum ether
- ethyl acetate
- phloroglucinol
- silica gel
- volume ratio
- 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
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C49/00—Ketones; Ketenes; Dimeric ketenes; Ketonic chelates
- C07C49/587—Unsaturated compounds containing a keto groups being part of a ring
- C07C49/657—Unsaturated compounds containing a keto groups being part of a ring containing six-membered aromatic rings
- C07C49/683—Unsaturated compounds containing a keto groups being part of a ring containing six-membered aromatic rings having unsaturation outside the aromatic rings
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C2603/00—Systems containing at least three condensed rings
- C07C2603/93—Spiro compounds
- C07C2603/95—Spiro compounds containing "not free" spiro atoms
- C07C2603/96—Spiro compounds containing "not free" spiro atoms containing at least one ring with less than six members
- C07C2603/97—Spiro compounds containing "not free" spiro atoms containing at least one ring with less than six members containing five-membered rings
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Gastroenterology & Hepatology (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pharmacology & Pharmacy (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
Abstract
The invention relates to a phloroglucinol compound, namely, acyonone C, which is extracted and separated from Hypericum ascyron L for the first time, a preparation method and application thereof. The biological activity test shows that: the compound has protective activity on hepatocyte damage caused by APAP, so that it can be used for preparing hepatoprotective drugs.
Description
Technical Field
The invention relates to the technical field of medicines. Relates to a novel phloroglucinol natural drug, namely asyconone C with liver protection activity extracted and separated from the overground part of Hypericum ascyron L, and a preparation method and application thereof in liver protection treatment
Background
Hypericum ascyron is a plant of Hypericum of Guttiferae, and is distributed throughout the country except Qinghai and Xinjiang. Herba Hyperici Japonici is bitter and cold in nature; has effects in cooling blood, stopping bleeding, purging pathogenic fire, and removing toxic materials; it can be used for treating hematemesis, hemoptysis, hemorrhage, rheumatism, dysentery, hepatitis, headache due to liver fire, jaundice, skin ulcer, and furuncle.
Disclosure of Invention
The invention aims to provide a phloroglucinol compound asconone C extracted and separated from herba Hyperici Erecti for the first time, a preparation method and application thereof.
In order to solve the technical problem, the invention provides the following technical scheme:
the first aspect of the technical proposal of the invention provides a phloroglucinol compound, namely acyanone C,
the second aspect of the technical scheme of the invention provides a preparation method of phloroglucinol compound acyanone C, which is obtained by separating from the overground part of herba ecliptae and comprises the following specific steps:
extraction: extracting the ground overground part of the herba ecliptae with ethanol, and concentrating the obtained extracting solution to obtain a crude extract; dissolving the crude extract in water, suspending uniformly, extracting with petroleum ether, and concentrating the obtained extract to obtain petroleum ether extract.
Separation: performing silica gel column chromatography on the petroleum ether extract, performing gradient elution by using petroleum ether/ethyl acetate, and performing color development according to TLC (thin layer chromatography) and combining similar fractions to obtain 12 components Fr.1-12; wherein the component Fr.1 is petroleum ether/ethyl acetate volume ratio of 1: the 0 eluted part was further subjected to silica gel column chromatography in a petroleum ether/chloroform volume ratio of 4: elution was performed at 1 isocratic and similar fractions were combined according to TLC visualization to give 5 fractions A, B, C, D, E, F. And the component F is subjected to silica gel column chromatography and eluted by petroleum ether/ethyl acetate gradient, wherein the volume ratio of the petroleum ether to the ethyl acetate is 9: 1 elution fraction F5 was subjected to preparative HPLC with a methanol/water volume ratio of 95: and 5, eluting to obtain the compound of the invention, namely the acyonone C.
In the above preparation method, in the extraction step, the ethanol used is 95% ethanol.
In the above preparation method, in the extraction step, the extraction is performed by heating reflux extraction.
In the preparation method, in the separation step, the concentration of the first petroleum ether/ethyl acetate gradient elution is sequentially 1: 0,50: 1,20: 1,9: 1,4: 1,1: 1.
in the preparation method, in the separation step, the concentration of the second petroleum ether/ethyl acetate gradient elution is 1: 0,9: 1,4: 1,2: 1,0: 1.
in the above preparation method, in the separation step, the chromatographic column packing of the preparative HPLC is reverse-phase octadecyl bonded silica gel.
In a third aspect of the present invention, there is provided a pharmaceutical composition, wherein the pharmaceutical composition comprises the phloroglucinol compound according to claim 1, and a pharmaceutically acceptable salt or an additive.
The pharmaceutical composition may be prepared according to methods well known in the art. The compounds of the invention may be formulated into any dosage form suitable for human or animal use by combining them with one or more pharmaceutically acceptable solid or liquid excipients and/or adjuvants. The compounds of the present invention are generally present in the pharmaceutical compositions in an amount of from 0.1 to 95% by weight.
The compounds of the present invention or pharmaceutical compositions containing them may be administered in unit dosage form by enteral or parenteral routes, such as oral, intravenous, intramuscular, subcutaneous, nasal, oromucosal, ophthalmic, pulmonary and respiratory, dermal, vaginal, rectal, and the like.
The dosage form for administration may be a liquid dosage form, a solid dosage form, or a semi-solid dosage form. The liquid dosage forms can be solution (including true solution and colloidal solution), emulsion (including o/w type, w/o type and multiple emulsion), suspension, injection (including water injection, powder injection and infusion), eye drop, nose drop, lotion, liniment, etc.; the solid dosage form can be tablet (including common tablet, enteric coated tablet, buccal tablet, dispersible tablet, chewable tablet, effervescent tablet, orally disintegrating tablet), capsule (including hard capsule, soft capsule, and enteric coated capsule), granule, powder, pellet, dripping pill, suppository, pellicle, patch, aerosol (powder), spray, etc.; semisolid dosage forms can be ointments, gels, pastes, and the like.
The compound can be prepared into common preparations, sustained release preparations, controlled release preparations, targeting preparations and various particle drug delivery systems.
For tableting the compound of the present invention, a wide variety of excipients known in the art may be used, including diluents, binders, wetting agents, disintegrants, lubricants, and solubilizers. The diluent can be starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, calcium hydrogen phosphate, calcium carbonate, etc.; the humectant can be water, ethanol, isopropanol, etc.; the binder can be starch slurry, dextrin, syrup, Mel, glucose solution, microcrystalline cellulose, acacia slurry, gelatin slurry, sodium carboxymethylcellulose, methylcellulose, hydroxypropyl methylcellulose, ethyl cellulose, acrylic resin, carbomer, polyvinylpyrrolidone, polyethylene glycol, etc.; the disintegrant may be dry starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, crosslinked polyvinylpyrrolidone, crosslinked sodium carboxymethylcellulose, sodium carboxymethyl starch, sodium bicarbonate and citric acid, polyoxyethylene sorbitol fatty acid ester, sodium dodecyl sulfate, etc.; the lubricant and cosolvent may be talc, silica, stearate, tartaric acid, liquid paraffin, polyethylene glycol, etc.
The tablets may be further formulated into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or double-layer and multi-layer tablets.
To encapsulate the administration unit, the active ingredient of the compound of the present invention may be mixed with a diluent and a cosolvent, and the mixture may be directly placed in a hard capsule or soft capsule. Or the effective component of the compound of the invention can be prepared into granules or pellets with diluent, adhesive and disintegrating agent, and then placed into hard capsules or soft capsules. The diluents, binders, wetting agents, disintegrants, and cosolvents used to prepare the compound tablets of the present invention can also be used to prepare capsules of the compounds of the present invention.
For preparing the compound of the present invention into injection, water, ethanol, isopropanol, propylene glycol or their mixture can be used as solvent, and appropriate amount of solubilizer, cosolvent, pH regulator, and osmotic pressure regulator commonly used in the art can be added. The solubilizer or cosolvent can be poloxamer, lecithin, hydroxypropyl-beta-cyclodextrin, etc.; the pH regulator can be phosphate, acetate, hydrochloric acid, sodium hydroxide, etc.; the osmotic pressure regulator can be sodium chloride, mannitol, glucose, phosphate, acetate, etc. For example, mannitol and glucose can be added as proppant for preparing lyophilized powder for injection.
In addition, colorants, preservatives, flavors, or other additives may also be added to the pharmaceutical preparation, if desired.
For the purpose of administration and enhancing the therapeutic effect, the drug or pharmaceutical composition of the present invention can be administered by any known administration method.
The dosage of the pharmaceutical composition of the compound of the present invention to be administered may vary widely depending on the nature and severity of the disease to be prevented or treated, the individual condition of the patient or animal, the route and dosage form of administration, and the like. Generally, a suitable daily dosage range for a compound of the invention is 0.001-5mg/Kg body weight. The above-described dosage may be administered in one dosage unit or divided into several dosage units, depending on the clinical experience of the physician and the dosage regimen including the use of other therapeutic means.
The compounds or compositions of the present invention may be administered alone or in combination with other therapeutic or symptomatic agents. When the compound of the present invention is used in a synergistic manner with other therapeutic agents, the dosage thereof should be adjusted according to the actual circumstances.
The fourth aspect of the technical scheme of the invention provides application of phloroglucinol compound acyanone C in preparing liver-protecting medicaments.
Advantageous technical effects
The invention adopts an MTT method to test the protective activity of the acyonone C on HepG2 cells damaged by paracetamol. Experiments prove that the acyonone C has obvious protective activity on the damaged liver cells caused by the APAP. Therefore, the acyonone C can be used for preparing liver-protecting medicaments.
Detailed Description
The chemical structure of acyconone C referred to in the examples below (the arabic numerals in the structure are the index positions of carbon atoms in the chemical structure):
preparation of acyconone C as described in example 1
Extraction: pulverizing 75.0kg of dried aerial parts of Hypericum ascyron L, extracting with 95% ethanol under reflux for 3 times, and concentrating the extractive solution under reduced pressure to obtain crude extract; dissolving the crude extract in water, suspending uniformly, extracting with petroleum ether for three times, and concentrating the obtained extract under reduced pressure to obtain petroleum ether extract.
Separation: performing silica gel column chromatography on the petroleum ether extract, performing gradient elution by using petroleum ether/ethyl acetate, and performing color development and combination of similar fractions according to TLC to obtain 12 components Fr.1-12; wherein the component Fr.1 is petroleum ether/ethyl acetate volume ratio of 1: the 0-eluted fraction was further subjected to silica gel column chromatography using petroleum ether/chloroform 4: elution was performed at 1 isocratic and similar fractions were combined according to TLC visualization to give 5 fractions A, B, C, D, E, F. And the component F is subjected to silica gel column chromatography and eluted by petroleum ether/ethyl acetate gradient, wherein the volume ratio of the petroleum ether to the ethyl acetate is 9: fraction 1, fraction F5, was purified by preparative HPLC [ C18 column (10 μm, 250 × 20mm) methanol/water 95: 5 volume ratio elution, flow rate 6mL/min, detection at 254nm wavelength ] to give compound acyonone C (5mg) of the present invention.
And (3) structural identification: the chemical structure of the compound acyonone C is determined by various modern spectral techniques such as NMR, HRESIMS, UV, IR, optical rotation and the like according to the conventional method, and the physicochemical properties are as follows:
colorless oil, molecular formula C26H34O2,[α]20D+76.9(c 0.05,MeOH)；
Ultraviolet spectrum UV (MeOH) lambdamax(logε)258(4.12)，292(3.31)nm；
Infrared Spectrum IR (KBr) vmax：1704、1666、1598、1450；
High resolution mass spectrum HRESIMS M/z 379.2637[ M + H ]]+(calculated for C26H35O2,379.2637)；
Hydrogen spectrum of nuclear magnetic resonance1H NMR (500MHz) and NMR carbon Spectroscopy13C NMR (125MHz) data are shown in Table I,
TABLE 1 of said acyconone C1H 13C and NMR (ppm in CDCl)3)
Example 2 inhibition of the protective activity of acyonone C against APAP-induced hepatocyte injury in vitro.
Experimental sample
Preparation of a tested sample solution: the test sample was the pure compound prepared in example 1 (acyconone C). An appropriate amount of sample is accurately weighed and prepared into a 10mM/mL solution by using DMSO for a pharmacological activity test.
Cell lines: human liver cancer HepG2 cells. Growth was carried out in DMEM medium (100U/ml penicillin, 100. mu.g/ml streptomycin) containing 10% fetal bovine serum at 37 ℃ under 5% CO2 saturated humidity. The passage was digested with a solution containing 0.25% trypsin and 0.02% EDTA.
Experimental methods
The MTT method is used. HepG2 cells were inoculated in 96-well cell culture plates, and after 24 hours of culture, the non-toxic concentration of the test compound and paracetamol (APAP, final concentration 8mM) were added, while positive drug control (bicyclol), solvent blank control and model were set. The action on the cells was continued for 48 h. Discarding the culture solution, adding 100 μ l of MTT (0.5mg/ml) solution into each well, culturing for 4h, discarding the MTT solution, adding 150 μ l of DMSO into each well, mixing with oscillator, and measuring absorbance at 570nm wavelength of microplate reader.
Cell survival (%) × (mean OD of administration group/mean OD of solvent control group) × 100%.
Results of the experiment
The APAP 8mM acts on HepG2 cells for 48h, obvious damage is caused to HepG2 cells, and the cell survival rate is 20.96%. Under the current experimental scheme, the combination of acyonone C and APAP at the concentration of 10 μ M has obvious protective effect on the damage of HepG2 cells caused by the APAP (the cell survival rate is 31.58%), and has statistical difference compared with the model group.
Conclusion of the experiment
The compound asyconone C has a better protective effect on liver cell injury caused by in vitro APAP, so the asyconone C can be used for preparing a liver-protecting medicament.
Claims (4)
1. A phloroglucinol compound with the following chemical structural formula:
2. the method of claim 1, comprising the steps of:
extraction: extracting the ground overground part of the herba ecliptae with ethanol, and concentrating the obtained extracting solution to obtain a crude extract; dissolving the crude extract in water, suspending uniformly, extracting with petroleum ether, and concentrating the obtained extract to obtain petroleum ether extract;
separation: performing silica gel column chromatography on the petroleum ether extract, and performing gradient elution by using petroleum ether/ethyl acetate, wherein the volume ratio of the petroleum ether to the ethyl acetate is 1: the eluted fraction of 0 was further subjected to silica gel column chromatography in a petroleum ether/chloroform volume ratio of 4: 1, and further performing silica gel column chromatography on the eluent, and performing gradient elution by using petroleum ether/ethyl acetate, wherein the volume ratio of the petroleum ether to the ethyl acetate is 9: 1 by preparative HPLC, in a methanol/water volume ratio of 95: 5, eluting to obtain phloroglucinol compounds;
in the extraction step, the adopted ethanol is 95 percent ethanol;
the extraction adopts a heating reflux extraction method;
in the separation step, the concentration of the first petroleum ether/ethyl acetate gradient elution is sequentially 1: 0,50: 1,20: 1,9: 1,4: 1,1: 1; the concentration of the second petroleum ether/ethyl acetate gradient elution is 1: 0,9: 1,4: 1,2: 1,0: 1;
the chromatographic column packing of the preparative HPLC is reverse-phase octadecyl bonded silica gel.
3. A pharmaceutical composition comprising the phloroglucinol compound of claim 1, and a pharmaceutically acceptable salt or additive.
4. Use of the phloroglucinol compounds according to claim 1 for the preparation of a liver-protecting medicament.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810425488.0A CN110452110B (en) | 2018-05-07 | 2018-05-07 | Phloroglucinol natural medicine and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810425488.0A CN110452110B (en) | 2018-05-07 | 2018-05-07 | Phloroglucinol natural medicine and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110452110A CN110452110A (en) | 2019-11-15 |
| CN110452110B true CN110452110B (en) | 2022-01-11 |
Family
ID=68471736
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810425488.0A Active CN110452110B (en) | 2018-05-07 | 2018-05-07 | Phloroglucinol natural medicine and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110452110B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113444062A (en) * | 2020-03-27 | 2021-09-28 | 中国医学科学院药物研究所 | Phlebendrin F as phloroglucinol compound and application thereof in resisting liver injury |
| CN115181024A (en) * | 2021-04-07 | 2022-10-14 | 中国医学科学院药物研究所 | Phloroglucinol compound and application thereof in resisting liver injury |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101879195A (en) * | 2009-05-06 | 2010-11-10 | 凌沛学 | Process for preparing general flavone extract of effective part of hypericum ascyron linne and new medicament appliaciton |
| CN107488162B (en) * | 2015-10-19 | 2020-09-18 | 中国医学科学院药物研究所 | Bicyclic alcohol derivatives, and preparation and application thereof |
| CN107569527A (en) * | 2017-10-31 | 2018-01-12 | 桂林纽泰生物科技有限公司 | The method that general flavone is extracted from hyperici,herba |
-
2018
- 2018-05-07 CN CN201810425488.0A patent/CN110452110B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN110452110A (en) | 2019-11-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JPH0543469A (en) | Beta-glucuronidase inhibitor | |
| CN108191616B (en) | Monomer component with selective butyrylcholine esterase inhibition effect in bletilla striata and application thereof | |
| CN110452110B (en) | Phloroglucinol natural medicine and preparation method and application thereof | |
| CN111647003B (en) | Triepoxyhexahydrochromone A, and pharmaceutical composition and application thereof | |
| CN111943919B (en) | Phloroglucinol compound Hyperacmosin C and preparation method and application thereof | |
| CN108659089B (en) | Sterol compound with antioxidant effect and application thereof in preparation of medicines | |
| CN106928299B (en) | Compound from cortex lycii radicis, preparation method and application thereof in aspect of reducing blood sugar | |
| CN105534971B (en) | Dibenzo oxepin class compound screens or prepares in vitro the application in drug | |
| US20150010664A1 (en) | Application of albizzia chinensis extract in preparation of medicine for treatment of gastric ulcer | |
| CN111718359B (en) | Hyperterpnoid A compound and application thereof in neuroprotection | |
| CN111714492B (en) | Application of natural heteroterpenoid drug hypemone A in preparation of antidiabetic drugs | |
| CN111943920B (en) | Phloroglucinol compound Hyperacmosin D and application thereof in preparation of antidiabetic drugs | |
| CN111718360B (en) | Mixed-source terpenoid and preparation method and application thereof | |
| CN112939759B (en) | Application of phloroglucinol compound in preparation of liver protection drugs | |
| CN117551160A (en) | Application of pentacyclic triterpene compounds in treatment of neurodegenerative diseases | |
| CN112939914A (en) | Pheracimin H as phloroglucinol compound and preparation method and application thereof | |
| CN111529515A (en) | Application of 12, 15-dioxo-alpha-cnidiene in pharmacy | |
| CN101407456A (en) | Effective ingredient having beta-secretase inhibition effect in Homalonema occulta | |
| CN115181024A (en) | Phloroglucinol compound and application thereof in resisting liver injury | |
| CN100387234C (en) | Medicine containing forrest silk vine root orherb aglycone, and its preparing method and use | |
| CN110204477B (en) | Diterpene alkaloid with antioxidant effect and application thereof in preparation of medicines | |
| CN113444062A (en) | Phlebendrin F as phloroglucinol compound and application thereof in resisting liver injury | |
| CN103450011B (en) | The diterpenoid acid of anti-Coxsackie virus and preparation method thereof and its pharmaceutical composition and purposes | |
| CN112457284B (en) | Oligo-lignans compound, preparation method thereof, pharmaceutical composition thereof and application thereof | |
| CN110680826A (en) | Application of triterpenoid saponin compound and salt thereof |
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 |