CN107468760B - Application of guava leaf extract in preparing anti-rotavirus drugs - Google Patents
Application of guava leaf extract in preparing anti-rotavirus drugs Download PDFInfo
- Publication number
- CN107468760B CN107468760B CN201710769928.XA CN201710769928A CN107468760B CN 107468760 B CN107468760 B CN 107468760B CN 201710769928 A CN201710769928 A CN 201710769928A CN 107468760 B CN107468760 B CN 107468760B
- Authority
- CN
- China
- Prior art keywords
- acid
- guava leaf
- alpha
- leaf extract
- hydroxy
- 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
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/61—Myrtaceae (Myrtle family), e.g. teatree or eucalyptus
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
Landscapes
- Health & Medical Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Alternative & Traditional Medicine (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Medical Informatics (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Steroid Compounds (AREA)
- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention discloses an application of guava leaf extract in preparing rotavirus resisting drugs, wherein the guava leaf extract contains 7-15wt% of 2 alpha-hydroxy oleanolic acid, 20-40wt% of 2 alpha-hydroxy ursolic acid and 6-14 wt% of asiatic acid, and is prepared by the following steps: extracting folium Psidii Guajavae with ethanol or methanol under reflux or percolation, concentrating under reduced pressure to obtain extract, adding water into the extract, stirring, centrifuging, and collecting precipitate to obtain folium Psidii Guajavae extract. The invention discovers for the first time that 2 alpha-hydroxy oleanolic acid, 2 alpha-hydroxy ursolic acid and asiatic acid in the guava leaf extract have obvious rotavirus resisting effect, thereby widening the medicinal application of the guava leaf extract.
Description
Technical Field
The invention belongs to the field of medicines, and particularly relates to an application of a guava leaf extract in preparation of an anti-rotavirus medicine.
Background
Rotavirus (RV for short) is a double-stranded ribonucleic acid virus, belonging to the reoviridae family. It is the single leading cause of diarrhea in infants and young children, and almost every child about five years of age in the world has been infected with rotavirus at least once. It mainly infects small intestine epithelial cells, causing cell damage, causing diarrhea. Rotavirus is epidemic in summer, autumn and winter every year, the infection route is a feces-oral route, and the clinical manifestation is acute gastroenteritis which is permeable diarrhea disease.
Guava leaf, which is the leaf of Psidium guajava L. belonging to Myrtaceae, has the effects of eliminating dampness, strengthening spleen, clearing away heat and toxic materials, and relieving diarrhea with astringents. The inventor of the application discovers that the guava leaves have the effect of reducing blood sugar in the early work, and reports that the guava leaves have the effect of resisting rotavirus are not found at present.
Disclosure of Invention
The invention aims to provide application of guava leaf extract in preparing anti-rotavirus medicaments.
The purpose of the invention is realized by the following technical scheme:
the application of guava leaf extract in preparing anti-rotavirus medicaments;
the guava leaf extract contains 7-15wt% of 2 alpha-hydroxy oleanolic acid, 20-40wt% of 2 alpha-hydroxy ursolic acid and 6-14 wt% of asiatic acid;
the guava leaf extract is prepared by the following steps: extracting folium Psidii Guajava (Psidium guajava L.) with ethanol or methanol under reflux or percolation, concentrating under reduced pressure to obtain extract, adding water into the extract, stirring, centrifuging, and collecting precipitate to obtain folium Psidii Guajava extract;
the rotavirus resisting medicine also comprises acceptable auxiliary materials and other effective components which play a role in compatibility and synergy;
the rotavirus resisting medicine may be in various forms, such as capsule, tablet, powder, wine, suspension, emulsion, syrup, aerosol, injection, etc.
Compared with the prior art, the invention has the following advantages and effects:
1. the invention discovers for the first time that the 2 alpha-hydroxy oleanolic acid, the 2 alpha-hydroxy ursolic acid and the asiatic acid in the guava leaf extract have obvious rotavirus resisting effect, and widens the medicinal application of the guava leaf extract.
2. The preparation process is simple and easy for large-scale production, and the total triterpene acid content in the obtained extract is up to more than 50%. And the effective part has definite components, low toxicity and strong pharmacological action of resisting rotavirus, thereby having good medicinal prospect.
Detailed Description
The present invention will be described in further detail with reference to examples, but the embodiments of the present invention are not limited thereto.
Example 1
The guava leaf extract separating and extracting method comprises the following steps:
pulverizing dried folium Psidii Guajavae (produced in Guangdong Convolvulaceae), percolating with 95% (volume concentration) ethanol, filtering to obtain extractive solution, concentrating under reduced pressure, and drying to obtain extract; the dosage ratio of the guava leaves to the ethanol is 1g (8-15) mL; adding water into the extract, stirring, centrifuging, and collecting precipitate to obtain guava leaf extract.
Subjecting the obtained guava leaf extract to a macroporous adsorption resin column, and eluting by using ethanol with the volume concentration of 0-10% to remove impurities; eluting with ethanol with the volume concentration of 60-80%, and collecting the eluent;
determining the contents of 2 alpha-hydroxy oleanolic acid, 2 alpha-hydroxy ursolic acid and asiatic acid in the eluate by liquid chromatography; measuring by using 2 alpha-hydroxy oleanolic acid, 2 alpha-hydroxy ursolic acid and asiatic acid as standard substances by adopting an external standard method; wherein the liquid chromatogram comprises the following determination conditions: the column was an Agilent Cosmosil-C18 column (4.6X 250mm,5 μm) with a mobile phase of methanol-0.1% formic acid in water (86:214), a flow rate of 1.0ml/min, a column temperature of 30 ℃ using an Agilent1200Series HPLC pump, an Agilent1200Series autosampler, and an ultraviolet detector. Wherein the retention time of 2 alpha-hydroxy oleanolic acid, 2 alpha-hydroxy ursolic acid and asiatic acid is 12.9 min, 14.1 min and 6.0min respectively.
The guava leaf extract contains 9.1 wt% of 2 alpha-hydroxy oleanolic acid, 29.6 wt% of 2 alpha-hydroxy ursolic acid and 7.0 wt% of asiatic acid through calculation and conversion.
Example 2
The guava leaf extract was isolated and extracted from different batches of guava leaves (produced in Guangdong Convolvulus Cantor.) in the same manner as in example 1, and the contents of 2 α -hydroxy oleanolic acid, 2 α -hydroxy ursolic acid, and asiatic acid were measured in the same manner as in example 1.
The guava leaf extract obtained by measurement, calculation and conversion contains 8.7 wt% of 2 alpha-hydroxy oleanolic acid, 28.9 wt% of 2 alpha-hydroxy ursolic acid and 6.9 wt% of asiatic acid.
Example 3
The guava leaf extract was isolated and extracted from different batches of guava leaves (produced in Guangdong Convolvulus Cantor.) in the same manner as in example 1, and the contents of 2 α -hydroxy oleanolic acid, 2 α -hydroxy ursolic acid, and asiatic acid were measured in the same manner as in example 1.
The guava leaf extract obtained by measurement, calculation and conversion contains 9.3 wt% of 2 alpha-hydroxy oleanolic acid, 31.2 wt% of 2 alpha-hydroxy ursolic acid and 7.3 wt% of asiatic acid.
Example 4
The guava leaf extract was isolated and extracted from different batches of guava leaves (produced in Guangdong Convolvulus Cantor.) in the same manner as in example 1, and the contents of 2 α -hydroxy oleanolic acid, 2 α -hydroxy ursolic acid, and asiatic acid were measured in the same manner as in example 1.
The guava leaf extract obtained by measurement, calculation and conversion contains 8.5 wt% of 2 alpha-hydroxy oleanolic acid, 28.1 wt% of 2 alpha-hydroxy ursolic acid and 6.8 wt% of asiatic acid.
Example 5
The guava leaf extract was isolated and extracted from different batches of guava leaves (produced in Guangdong Convolvulus Cantor.) in the same manner as in example 1, and the contents of 2 α -hydroxy oleanolic acid, 2 α -hydroxy ursolic acid, and asiatic acid were measured in the same manner as in example 1.
The guava leaf extract obtained by measurement, calculation and conversion contains 9.6 wt% of 2 alpha-hydroxy oleanolic acid, 33.2 wt% of 2 alpha-hydroxy ursolic acid and 7.4 wt% of asiatic acid.
Example 6
The guava leaf extract obtained in the examples 1 to 5 is tested for rotavirus resisting effect by adopting a plaque reduction experiment method, and the specific method is as follows:
(1) HEp-2 cells were seeded in 24-well cell culture plates at a cell density of 2X 105cell/mL, 1 mL/well, at 37 ℃ with 5% CO2Culturing in an incubator for about 20h to grow into a cell monolayer.
(2) About 60 to about100 mu L of 80PFU/mL rotavirus suspension and 100 mu L of different concentration drugs, and a cell control group and a virus control group are arranged at the same time; standing at 37 deg.C for 5% CO2Culturing in an incubator for 2 h. The crosses were gently shaken every 15min to allow the virus to be evenly and well adsorbed and penetrate into the cells.
(3) And after the virus is adsorbed for 2 hours, sucking away virus liquid, washing with PBS for 2 times, adding 0.5mL of agarose gel covering liquid into each hole, adding 0.5mL of medicine with corresponding concentration after the covering liquid is solidified, and adding equal-volume maintenance liquid into a cell control group and a virus control group.
(4) And (3) placing the 24-hole plate in a cell culture box for culturing for 4-5 days, adding 10% formaldehyde for fixing overnight, removing agarose gel, dyeing with 1% crystal violet for 30min, slightly washing the 24-hole plate with tap water, air-drying and counting the number of plaques. The plaque inhibition rate of the sample was calculated, and the median inhibitory concentration IC50 was calculated. Plaque inhibition rate ═ [ (number of plaques in virogroup-number of plaques in drug group)/number of plaques in virogroup ] × 100%
The following drug groups were tested for anti-rotavirus activity according to the method described above:
drug group 1: 2 α -hydroxy oleanolic acid alone;
drug group 2: 2 alpha-hydroxy ursolic acid is used alone;
drug group 3: asiatic acid is used alone;
experimental group 4: a composition consisting of 2 alpha-hydroxy oleanolic acid, 2 alpha-hydroxy ursolic acid and asiatic acid in a weight ratio of 3:5:1 is used;
experimental group 5: a composition consisting of 2 alpha-hydroxy oleanolic acid, 2 alpha-hydroxy ursolic acid and asiatic acid in a weight ratio of 1:3:3 is used;
experimental group 6: the composition is prepared from 2 alpha-hydroxy oleanolic acid, 2 alpha-hydroxy ursolic acid and asiatic acid according to the weight ratio of 1:3: 1;
experimental group 7: the effective part of the guava leaf prepared in example 1 was used;
experimental group 8: the effective part of the guava leaf prepared in example 2 is used;
experimental group 9: the effective part of the guava leaf prepared in example 3 was used;
experimental group 10: the effective part of the guava leaf prepared in example 4 was used;
experimental group 11: the effective part of the guava leaf prepared in example 5 was used;
experimental group 12: ribavirin.
The results of the anti-rotavirus test of each experimental group are shown in table 1.
TABLE 1 anti-rotavirus Activity IC50 test results
As can be seen from the experimental data of experimental groups 1-3 in Table 1, 2 alpha-hydroxy oleanolic acid, 2 alpha-hydroxy ursolic acid and asiatic acid show certain anti-rotavirus activity, and the anti-rotavirus activity of the compounds is equivalent to that of ribavirin; this indicates that the composition contains 2 alpha-hydroxy oleanolic acid, 2 alpha-hydroxy ursolic acid and asiatic acid which all have anti-rotavirus activity.
As can be seen from the experimental data of experimental groups 4-6 in Table 1, after 2 alpha-hydroxy oleanolic acid, 2 alpha-hydroxy ursolic acid and asiatic acid are combined, the activity of resisting rotavirus is further improved and is better than that of any monomer, which shows that 2 alpha-hydroxy oleanolic acid, 2 alpha-hydroxy ursolic acid and asiatic acid can generate synergistic anti-rotavirus effect after being combined. This also indicates that the composition contains a combination of 2 α -hydroxyoleanolic acid, 2 α -hydroxyursolic acid and asiatic acid as an active ingredient, and the anti-rotavirus activity thereof is further enhanced.
As can be seen from the experimental data of experimental groups 7-11 in Table 1, the effective part of the guava leaf contains 2 alpha-hydroxy oleanolic acid, 2 alpha-hydroxy ursolic acid and asiatic acid as active ingredients, so that the effective part of the guava leaf has anti-rotavirus activity, and the antiviral activity of the effective part of the guava leaf is in direct proportion to the contents of the active ingredients, namely 2 alpha-hydroxy oleanolic acid, 2 alpha-hydroxy ursolic acid and asiatic acid.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (4)
1. The application of the guava leaf extract in preparing the rotavirus resisting medicine is characterized in that: the guava leaf extract contains 7-15wt% of 2 alpha-hydroxy oleanolic acid, 20-40wt% of 2 alpha-hydroxy ursolic acid and 6-14 wt% of asiatic acid.
2. The use of guava leaf extract according to claim 1 for the preparation of an anti-rotavirus agent, wherein: the guava leaf extract is prepared by the following steps: extracting folium Psidii Guajavae with ethanol or methanol under reflux or percolation, concentrating under reduced pressure to obtain extract, adding water into the extract, stirring, centrifuging, and collecting precipitate to obtain folium Psidii Guajavae extract.
3. The use of guava leaf extract according to claim 1 for the preparation of an anti-rotavirus agent, wherein: the rotavirus resisting medicine contains acceptable supplementary material and other effective components with synergistic effect.
4. The use of guava leaf extract according to claim 1 for the preparation of an anti-rotavirus agent, wherein: the rotavirus resisting medicine is capsule, tablet, powder, wine, suspension, emulsion, syrup, aerosol or injection.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710769928.XA CN107468760B (en) | 2017-08-31 | 2017-08-31 | Application of guava leaf extract in preparing anti-rotavirus drugs |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710769928.XA CN107468760B (en) | 2017-08-31 | 2017-08-31 | Application of guava leaf extract in preparing anti-rotavirus drugs |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107468760A CN107468760A (en) | 2017-12-15 |
| CN107468760B true CN107468760B (en) | 2020-10-27 |
Family
ID=60604366
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710769928.XA Active CN107468760B (en) | 2017-08-31 | 2017-08-31 | Application of guava leaf extract in preparing anti-rotavirus drugs |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107468760B (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102178741A (en) * | 2011-04-11 | 2011-09-14 | 暨南大学 | Guava leaf extract with function of reducing blood sugar as well as preparation method and application thereof |
| CN102475731A (en) * | 2010-11-29 | 2012-05-30 | 上海奇羚芊生物科技发展有限公司 | Miao ethnic medicine raw material for treating children's (rotavirus, dysentery and malaria) disease molecules |
-
2017
- 2017-08-31 CN CN201710769928.XA patent/CN107468760B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102475731A (en) * | 2010-11-29 | 2012-05-30 | 上海奇羚芊生物科技发展有限公司 | Miao ethnic medicine raw material for treating children's (rotavirus, dysentery and malaria) disease molecules |
| CN102178741A (en) * | 2011-04-11 | 2011-09-14 | 暨南大学 | Guava leaf extract with function of reducing blood sugar as well as preparation method and application thereof |
Non-Patent Citations (4)
| Title |
|---|
| Guava extract (Psidium guajava) alters the labelling of blood constituents with technetium-99m;ABREU P.R.C.等;《J Zhejiang Univ SCIENCE B》;20061231;第7卷(第6期);第429-435页 * |
| Immunomodulatory activities of flavonoids monoterpenoids, triterpenoids, iridoid glycosides and phenolic compounds of plantago species;Lien-Chai Chiang等;《Planta Med》;20031231;第69卷;第600-604页，尤其是第600页摘要右栏第2-4行 * |
| 张文举等.番石榴叶中提取的槲皮素和2α-羟基熊果酸体外抗人轮状病毒作用.《中草药》.2005,第36卷(第1期), * |
| 番石榴叶中提取的槲皮素和2α-羟基熊果酸体外抗人轮状病毒作用;张文举等;《中草药》;20050131;第36卷(第1期);第76-79页，尤其是第77页左栏第2段、第79页左栏第1段第7-11行 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107468760A (en) | 2017-12-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US9687516B2 (en) | Preparation method for extractive of jinxuan hemorrhoid washing powder botanicals | |
| CN101062069A (en) | Whole coumarins extract from root of dahuriae angelica and the preparing method thereof | |
| CN103948659A (en) | Preparation method of notoginseng leaf total saponins | |
| CN111018821A (en) | Biflavonoid compound and preparation method and application thereof | |
| US9453041B2 (en) | Method for preparing albiflorin and paeoniflorin | |
| CN104873570B (en) | A kind of method for extraction and purification of Prunella vulgaris general flavone and its application | |
| CN104958330A (en) | Oroxylum indicum general flavone extraction and purification method and application thereof | |
| CN101880269B (en) | Diterpene monomers and method for separating and preparing diterpene monomers from Clerodendron cyrtophyllum Turcz | |
| CN112159451A (en) | Gynostemma pentaphylla saponin extract and preparation method thereof | |
| CN107468760B (en) | Application of guava leaf extract in preparing anti-rotavirus drugs | |
| CN104027403B (en) | A kind of preparation method of Radix Polygalae sugar ester effective site | |
| CN101703669B (en) | Smilax china effective fractions and extraction as well as purification process thereof | |
| CN100536868C (en) | Powder injection contg high content tanshin polyphenolic acid salts, and its preparation method | |
| CN103923043A (en) | Method for effective preparation of salvianolic acid B extract | |
| CN1189176C (en) | Astragalus root methyl-glycoside composition and preparation method | |
| CN105646638B (en) | The preparation method of pedunculoside | |
| CN110025659B (en) | Traditional Chinese medicine active site with anti-HBV effect, preparation method and application | |
| CN101468997A (en) | Method for extracting and separating bilobalide from ginkgo leaf | |
| CN109966339B (en) | Sophora alopecuroides L effective part with anti-HBV virus effect, preparation method and application thereof | |
| CN103936812B (en) | Lupinane type triterpene compound and pharmaceutical composition thereof are applied with it | |
| CN102885985A (en) | Preparation method and application of rhizoma gastrodiae and rhizoma chuanxiong compound extract | |
| CN101863953B (en) | Method for separating and preparing Liriope graminifolia saponin A from Liriope graminifolia | |
| CN101926835B (en) | Method for extracting gingko total terpene lactones and injection containing same | |
| CN106188215B (en) | Withaphysalin type compounds and application thereof | |
| CN104547148A (en) | Hedyotis diffusa extract used for preventing and treating senile dementia and preparation method of hedyotis diffusa extract |
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 |