CN110156693B - Methylated allantoin alkaloid and preparation method and application thereof - Google Patents
Methylated allantoin alkaloid and preparation method and application thereof Download PDFInfo
- Publication number
- CN110156693B CN110156693B CN201910508889.7A CN201910508889A CN110156693B CN 110156693 B CN110156693 B CN 110156693B CN 201910508889 A CN201910508889 A CN 201910508889A CN 110156693 B CN110156693 B CN 110156693B
- Authority
- CN
- China
- Prior art keywords
- allantoin
- dichloromethane
- methylated
- methanol
- alkaloid
- 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
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/16—Amides, e.g. hydroxamic acids
- A61K31/17—Amides, e.g. hydroxamic acids having the group >N—C(O)—N< or >N—C(S)—N<, e.g. urea, thiourea, carmustine
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
- A61K31/4164—1,3-Diazoles
- A61K31/4166—1,3-Diazoles having oxo groups directly attached to the heterocyclic ring, e.g. phenytoin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
- A61K31/4164—1,3-Diazoles
- A61K31/4178—1,3-Diazoles not condensed 1,3-diazoles and containing further heterocyclic rings, e.g. pilocarpine, nitrofurantoin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C273/00—Preparation of urea or its derivatives, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups
- C07C273/18—Preparation of urea or its derivatives, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups of substituted ureas
- C07C273/1872—Preparation of compounds comprising a -N-C(O)-N-C(O)-N- moiety
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C273/00—Preparation of urea or its derivatives, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups
- C07C273/18—Preparation of urea or its derivatives, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups of substituted ureas
- C07C273/189—Purification, separation, stabilisation, use of additives
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C275/00—Derivatives of urea, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups
- C07C275/46—Derivatives of urea, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups containing any of the groups, X being a hetero atom, Y being any atom, e.g. acylureas
- C07C275/58—Y being a hetero atom
- C07C275/62—Y being a nitrogen atom, e.g. biuret
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D233/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings
- C07D233/54—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members
- C07D233/66—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
- C07D233/88—Nitrogen atoms, e.g. allantoin
-
- 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
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Epidemiology (AREA)
- Toxicology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The methylated allantoin alkaloid has medical activity, has a certain protection effect in a cell aging model induced by high sugar, can be used for preparing cell aging protection medicines, and has important significance in the fields of agriculture and medicine; and provides wider prospect for effectively developing and utilizing the Qimen black tea.
Description
Technical Field
The invention relates to the technical field of natural medicines, in particular to a methylated allantoin alkaloid and a preparation method and application thereof.
Background
The tea contains abundant alkaloids, which account for about 3% -5% of the dry matter of the tea. Wherein, most of the alkaloids are purine alkaloids, and the little amount of the pyrimidine alkaloids. Wherein, the caffeine accounts for 3 to 4 percent, the theobromine accounts for 0.15 to 0.2 percent, and the theophylline accounts for 0.02 to 0.04 percent. Research has shown that caffeine is an important flavor substance in tea. In addition, caffeine has pharmacological functions of tonifying heart, promoting urination, removing toxic substances, relieving asthma and the like.
In addition to the three purine bases, reports on chemical bases of other alkaloids with biological activity in tea leaves are relatively few, and if the alkaloid derivative components with biological activity can be successfully researched and developed from the tea leaves, the alkaloid derivative components can make an important contribution to the fields of agriculture, medicine and the like.
Disclosure of Invention
The invention aims to solve the technical problem of providing a methylated allantoin alkaloid with a cell aging protection effect, and a preparation method and application thereof.
In order to solve the technical problems, the invention adopts the following technical scheme: methylated allantoin alkaloids are represented by the structure shown in formula I or formula II or formula III or formula IV or formula V or formula VI:
the invention also provides application of the methylated allantoin alkaloid in preparation of cell aging protection medicines.
In particular to a cell aging protective medicament which is prepared from the methylated allantoin alkaloid and pharmaceutical excipients.
The pharmaceutical dosage forms of the medicament include oral administration type and injection type. The oral preparation comprises tablets, capsules, granules, dripping pills and the like; the injection type comprises injection, mixed rotation liquid and the like. The specific preparation method can refer to the conventional method in the pharmaceutical field, and the used pharmaceutical auxiliary materials are selected from the auxiliary materials which are common in the pharmaceutical field according to different dosage forms.
The invention also provides a preparation method of the methylated allantoin alkaloid, which is characterized by comprising the following steps of:
(1) Crushing of raw materials
Pulverizing QIMEN black tea to obtain QIMEN black tea powder;
(2) Leaching
Leaching the Qimen black tea powder with ethanol aqueous solution, separating out an extracting solution, and drying the extracting solution to obtain a Qimen black tea extract;
(3) Separation and purification
Dissolving the Keemun black tea extract with methanol aqueous solution, concentrating to remove most of methanol, extracting with petroleum ether to remove small polar substances and pigment, and extracting with dichloromethane to obtain dichloromethane extract A;
dissolving the dichloromethane part extract A with water, adding an aqueous solution of tartaric acid, removing insoluble substances, extracting with dichloromethane, removing the aqueous layer, adding an equal volume of aqueous solution of sodium hydroxide, extracting to obtain a dichloromethane layer and an alkaline water layer, adding ammonium chloride into the alkaline water layer, and extracting with dichloromethane to obtain a dichloromethane part extract B;
and (3) separating and purifying the dichloromethane part extract B to obtain the methylated allantoin alkaloid.
Further, in the step (2), the concentration of the aqueous ethanol solution used was 95%.
Further, in the step (3), the concentration of the aqueous tartaric acid solution used is 2%, and the concentration of the aqueous sodium hydroxide solution is 1-2%.
Further, in the step (3), the separation and purification treatment comprises silica gel column chromatography, sephadex LH-20 gel column chromatography and Toyopearl column chromatography.
Further, the specific steps of the separation and purification treatment are as follows: dissolving the dichloromethane part extract B, performing first silica gel column chromatography by taking a dichloromethane-methanol volume ratio of 500:1 to 1:1 as gradient elution, performing second silica gel column chromatography by taking a dichloromethane-methanol volume ratio of 30:1 to 20:1 elution component obtained by the first silica gel column chromatography, performing second silica gel column chromatography by taking a dichloromethane-methanol volume ratio of 1000:1 to 1:1 as gradient elution, performing methanol recrystallization on a dichloromethane-methanol volume ratio of 20:1 elution component obtained by the second silica gel column chromatography, and obtaining a racemic trimethyl allantoin dimer, resolving the racemic trimethyl allantoin dimer by using a chiral column by using HPLC, and eluting by acetonitrile to obtain methylated allantoin alkaloids shown in a formula IV and methylated allantoin alkaloids shown in a formula V;
eluting the residual component of methanol recrystallization with methanol by using Toyopearl column chromatography, collecting the component eluted at the first time, purifying by using HPLC to obtain methylated allantoin alkaloids shown in a formula III, eluting the collected component eluted at the second time by using methanol by using Sephadex LH-20 gel column chromatography to obtain racemic 1,3,8-trimethylallantoin, resolving the racemic 1,3,8-trimethylallantoin by using chiral column by using HPLC, eluting by using acetonitrile to obtain methylated allantoin alkaloids shown in a formula I and methylated allantoin alkaloids shown in a formula II;
performing a third silica gel column chromatography on the dichloromethane-methanol eluting component obtained by the first silica gel column chromatography with the volume ratio of 50:1 to 30:1, performing a fourth silica gel column chromatography on the ethyl acetate-acetone 20:1 eluting component obtained by the third silica gel column chromatography with the ethyl acetate-acetone 20:1 as gradient elution, performing an HPLC purification on the ethyl acetate-acetone 70:1 eluting component obtained by the fourth silica gel column chromatography to obtain the methylated allantoin alkaloid shown in the formula VI.
"first..second..times" herein is not intended to limit the order of sequence, but is merely intended to distinguish, and as long as the corresponding components are separated, the next chromatographic separation can be performed.
The beneficial effects of the invention are as follows:
the methylated allantoin alkaloid provided by the invention has medical activity, has a certain protection effect in a cell aging model induced by high sugar, can be used for preparing cell aging protection medicines, and has important significance in the fields of agriculture and medicine; and provides wider prospect for effectively developing and utilizing the Qimen black tea.
The preparation method of the methylated allantoin alkaloid has the advantages of simple process, easy implementation, low cost and good application prospect.
Drawings
FIG. 1 is a diagram of the x-ray crystal structure of (+ -) 1,3, 8-trimethylllantoin.
FIG. 2 is a diagram of the x-ray crystal structure of 1,3, 8-trimethyllantoin 4R,4' S-dimer.
FIG. 3 is a diagram of the x-ray crystal structure of 1,3, 8-trimethyllantoin 4,4' -dimer.
FIG. 4 is a diagram of the structure of the x-ray crystal of 1,3, 7-trimethylrilet.
FIG. 5 is a schematic diagram showing the experiments of (. + -.) 1,3,8-Trimethylallantoin, S- (+) -1,3,8-Trimethylallantoin, R- (-) -1,3, 8-trimethyllantoin, 1,3, 8-trimethyllantoin 4R,4'S-dimer, 1,3, 8-trimethyllantoin 4,4' -dimer, 1,3, 8-trimethyllantoin 4S,4'S-dimer, 1,3, 8-trimethyllantoin 4R,4' R-dimer and 1,3,7-Trimethyltr iuret inhibiting high sugar induced human umbilical vein endothelial cell senescence.
Detailed Description
The invention is further described below with reference to examples:
the various materials used in the examples below, unless otherwise specified, are commercially available products known in the art.
Example 1
Preparation of methylated allantoin alkaloids
1.1 description of methylated allantoin alkaloids
The methylated allantoin alkaloid is represented by a structure shown in a formula I or a formula II or a formula III or a formula IV or a formula V or a formula VI:
1.2 preparation method and results
(1) Crushing of raw materials
Taking 200 kg of Qimen black tea, and crushing the powder to obtain the Qimen black tea powder through a screen with the aperture of 10.5 mm;
(2) Leaching
Adding the Qimen black tea powder into 1000 kg of 95% ethanol water solution, stirring and extracting for 10 hours, filtering to obtain an extracting solution, concentrating the extracting solution to 400L, passing through a 200-mesh screen, continuously concentrating, passing through a 200-mesh screen, finally performing spray drying on the obtained concentrated solution, wherein the air inlet temperature is 190 ℃, the air outlet temperature is 90 ℃, and collecting the dried powder, and passing through a 40-mesh screen to obtain the Qimen black tea extract;
(3) Separation and purification
Dissolving the extract of Keemun black tea with 70% methanol water solution, concentrating to remove most of methanol (as far as possible, without affecting subsequent extraction layering), extracting with petroleum ether, removing petroleum ether layer to remove small polar substances and pigment, and extracting with dichloromethane to obtain dichloromethane extract A;
dissolving the dichloromethane part extract A with water, adding 2% tartaric acid aqueous solution, removing insoluble substances, extracting with dichloromethane, removing the aqueous layer, adding 1% -2% sodium hydroxide aqueous solution with equal volume, extracting to obtain a dichloromethane layer and an alkaline water layer, adding ammonium chloride into the alkaline water layer, and extracting with dichloromethane to obtain a dichloromethane part extract B;
dissolving and sampling 500 g of dichloromethane part extract B with chloroform, performing first silica gel column chromatography, eluting with dichloromethane-methanol volume ratio of 500:1 to 1:1 as gradient, performing second silica gel column chromatography with dichloromethane-methanol volume ratio of 30:1 to 20:1 as gradient, eluting with dichloromethane-methanol volume ratio of 1000:1 to 1:1, recrystallizing with methanol to obtain 10.0mg (named as 1,3,8-Trimethylallantoin 4,4' -dimer in fig. 5), and performing chiral column chromatography
OX-3R) was resolved by HPLC and eluted with acetonitrile to give 2.5mg (designated as 1,3,8-Trimethylallantoin 4S,4'S-dimer, 3a in FIG. 5) of the methylated allantoin alkaloid of formula IV and 2.6mg (designated as 1,3,8-Trimethylallantoin 4R,4' R-dimer, 3b in FIG. 5) of the methylated allantoin alkaloid of formula V;
subjecting the residual component of methanol recrystallization to Toyopearl column chromatography, eluting with methanol, collecting the first time eluted component, purifying by HPLC to obtain 6.0mg (named 1,3,8-Trimethylallantoin 4R,4' S-dimer, shown in FIG. 5) of methylated allantoin alkaloid shown in formula III, collecting the second time eluted component, eluting with methanol by SephadexLH-20 gel column chromatography to obtain 57.5mg (named (+ -) 1,3,8-Trimethylallantoin, shown in FIG. 5) of racemic 1,3,8-Trimethylallantoin, and subjecting to chiral column
OX-3R) was resolved by HPLC and eluted with acetonitrile to give7.4mg (designated as S- (+) -1,3,8-Trimethylallantoin, designated as 1a in FIG. 5) of the methylated allantoins of formula I and 7.5mg (designated as R- (-) -1,3,8-Trimethylallantoin, designated as 1b in FIG. 5) of the methylated allantoins of formula II;
performing a third silica gel main procedure on the dichloromethane-methanol volume ratio 50:1 to 30:1 elution component obtained by the first silica gel column chromatography, performing a fourth silica gel column chromatography on the ethyl acetate-acetone 20:1 elution component obtained by the third silica gel column chromatography by taking ethyl acetate-acetone 20:1 to 1:1 as gradient, performing HPLC purification on the ethyl acetate-acetone 70:1 elution component obtained by the fourth silica gel column chromatography by taking ethyl acetate-acetone 90:1 to 70:1 as gradient, and obtaining 6.5mg (named as 1,3, 7-trimethyllauret in fig. 5) of methylated allantoin alkaloids shown in a formula VI.
1.3 verification of the Properties of methylated allantoin alkaloids
1.3.1: the characteristics of (+ -) 1,3, 8-trimethylllantoin are as follows:
1) White powder, soluble in methanol and water;
2) Melting point 158-159 ℃; HR-ESI-MS: M/z 223.0808 ([ M+Na)] + ,calcd for C 7 H 12 N 4 O 3 Na + ,223.0807). Nuclear magnetic resonance spectral data are shown in table 1; crystal data and structure refinement are shown in table 2; the X-ray crystal structure is shown in figure 1.
TABLE 1 Nuclear magnetic resonance Spectrum data of (+ -) 1,3, 8-Trimethyllantoin 1 The H NMR was at 600MHz, 13 c NMR was measured at 125MHz with delta units in ppm, coupling constant J units in Hz and solvent in deuterated methanol).
| Position of | δ H (J in Hz) | δ C |
| 2 | 158.3 | |
| 4 | 5.18s | 67.5 |
| 5 | 173.2 | |
| 7 | 160.3 | |
| 9 | 2.98s | 25.0 |
| 10 | 2.87s | 26.8 |
| 11 | 2.69s | 26.9 |
TABLE 2 Crystal data of (+ -) 1,3, 8-Trimethyllantoin
All spectral data were passed through 1 H- 1 The two-dimensional nuclear magnetic resonance spectra of H COSY, HMQC, HMBC and the like belong to, and the structure of the obtained compound is proved.
1.3.2: the S- (+) -1,3, 8-Trimethyllantoin of formula I has the following characteristics:
1) White powder, soluble in methanol and water;
2)、
+60.8°(c 0.7MeOH)；CD(MeOH)λ(Δε)218(+16.1),242(-2.3)
1.3.3: the characteristics of R- (-) -1,3, 8-Trimethyllantoin of formula II are as follows:
1) White powder, soluble in methanol and water;
2)、
-61.3°(c 0.7MeOH)；CD(MeOH)λ(Δε)218(-17.1),242(+2.3).
1.3.4: the characteristics of the 1,3, 8-Trimethyllantoin 4R,4' S-dimer shown in formula III are as follows
1) White powder, soluble in pyridine, slightly soluble in methanol;
2) Melting point 214-215 ℃; HR-ESI-MS: M/z 421.1560 ([ M+Na)] + ,calcd for C 14 H 22 N 8 O 6 Na + ,421.1560),819.3201([2M+Na] + ,calcd for C 28 H 44 N 16 O 12 Na + ,819.3222). Nuclear magnetic resonance spectral data are shown in table 3; crystal data and structure refinement are shown in table 4; the X-ray crystal structure is shown in figure 2.
Table 3.1,3,8 Nuclear magnetic resonance Spectrum data of Trimethyllantoin 4R,4' S-dimer [ (] 1 The H NMR was at 600MHz, 13 c NMR was measured at 125MHz with delta units in ppm, coupling constant JIn Hz, the solvent is deuterated methanol).
| Position of | δH(J in Hz) | δC |
| 2 | 159.2 | |
| 2' | 159.2 | |
| 4 | 77.7 | |
| 4' | 77.7 | |
| 5 | 172.0 | |
| 5' | 172.0 | |
| 7 | 158.9 | |
| 7' | 158.9 | |
| 9 | 2.97s | 25.2 |
| 9' | 2.97s | 25.2 |
| 10 | 2.88s | 26.6 |
| 10' | 2.88s | 26.6 |
| 11 | 2.65s | 27.1 |
| 11' | 2.65s | 27.1 |
Table 4.1,3,8 Crystal data of Trimethyllantoin 4R,4' S-dimer
All spectral data were passed through 1 H- 1 Two-dimensional nuclear magnetic resonance spectra such as H COSY, HSQC, HMBC and the like belong to the field, and the structure of the obtained compound is proved by analysis of X-ray crystal data.
1.3.5: the characteristics of the 1,3, 8-trimethyllantoin 4,4' -dimer are as follows:
1) White powder, soluble in pyridine;
2)、HR-ESI-MS:m/z 421.1560([M+Na] + ,calcd for C 14 H 22 N 8 O 6 Na + ,421.1560),819.3233([2M+Na] + ,calcd for C 28 H 44 N 16 O 12 Na + ,819.3222). The nuclear magnetic resonance spectrum data are shown in Table 5. Crystal data and structure refinement are shown in table 6; the X-ray crystal structure is shown in figure 3.
Table 5.1,3,8-Nuclear magnetic resonance Spectrum data of Trimethyllantoin 4,4' -dimer [ (] 1 The H NMR was at 600MHz, 13 CNMR was tested at 125MHz with delta units in ppm, coupling constant J units in Hz, and solvent in deuterated pyridine).
Table 6.1,3,8 Crystal data of Trimethyllantoin 4,4' -dimer
All spectral data were passed through 1 H- 1 The two-dimensional nuclear magnetic resonance spectra of H COSY, HSQC, HMBC and the like belong to, and the structure of the obtained compound is proved.
1.3.6: the characteristics of the 1,3, 8-Trimethyllantoin 4S,4' S-dimer shown in formula IV are as follows:
1) White powder, soluble in water;
2) Melting point 196-197 ℃;
+32°(c 0.3MeOH)；CD(MeOH)λ(Δε)220(+23.6),245(-11.3).
1.3.7: the characteristics of the 1,3, 8-trimethyllantoin 4R,4' R-dimer shown in formula V are as follows:
1) White powder, soluble in water;
2) Melting point is 192-193 ℃;
-28°(c 0.3MeOH)；CD(MeOH)λ(Δε)220(-21.8),245(+10.6).
1.3.8: the characteristics of 1,3, 7-Trimethylriluret of formula VI are as follows
1) White powder, soluble in pyridine;
2) Melting point 185-186 ℃; HR-ESI-MS: M/z 211.0806 ([ M+Na)] + ,calcd for C 6 H 12 N 4 O 3 Na + ,211.0807),399.1706([2M+Na] + ,calcd for C 12 H 24 N 8 O 6 Na + ,399.1717). Nuclear magnetic resonance spectral data are shown in table 7; crystal data and structure refinement are shown in table 8; the X-ray crystal structure is shown in figure 4.
Table 7.1,3,7-Nuclear magnetic resonance Spectrum data of Trimethylriluret [ ] 1 The H NMR was at 600MHz, 13 c NMR was measured at 125MHz with delta units in ppm, coupling constant J units in Hz and solvent in deuterated pyridine).
| Position of | δ H (J in Hz) | δ C |
| 2 | 157.7 | |
| 4 | 154.7 | |
| 6 | 154.2 | |
| 8 | 2.78d(4.2) | 27.4 |
| 9 | 3.23s | 30.2 |
| 10 | 2.86d(4.8) | 26.4 |
Table 8.1,3,7 Crystal data of Trimethylriluret
All spectral data were passed through 1 H- 1 The two-dimensional nuclear magnetic resonance spectra of H COSY, HSQC, HMBC and the like belong to, and the structure of the obtained compound is proved.
Example 2
In vitro high sugar induced cell senescence protection test of methylated allantoin alkaloids of the invention
Cell culture: human Umbilical Vein Endothelial Cells (HUVECs) were cultured in a carbon dioxide incubator under certain conditions (37 ℃ C., carbon dioxide concentration 5%, humidity 95%) using Dulbecco's Modified Eagle's (DMEM) medium containing 10% fetal bovine serum. The culture solution is changed for 1 time every 1-2 days, and cells grow densely in a single layer after 3-5 days after inoculation, so that fusion and island connection occur. When the cells were grown to confluence, they were passaged with 0.25% trypsin digestion at 1: and 4, subculturing. HUVEC cells in logarithmic growth phase were digested into single cell suspension, resuspended in DMEM medium containing 10% FBS, and plated in 6-well plates. The test is divided into a Control group (Control), a Model group (Model), a metformin group (methyl) and each drug group, wherein the Control group is a blank Control without adding any substances, the Model group is formed by co-culturing HUVECs with 33mM glucose, the metformin group is formed by co-culturing metformin with HUVECs and high sugar, each drug group is respectively (. + -.) 1,3,8-Trimethylallantoin, S- (+) -1,3,8-Trimethylallantoin, R- (-) -1,3, 8-trimethylolantin, 1,3, 8-trimethylolantin 4R,4'S-dimer, 1,3, 8-trimethylolantin 4, 4S, 1,3, 8-trimethylolantin 4S,4' R-dimer, 1,3, 7-trimethylolantin and dimethyldiguanide prepared by the invention are respectively cultured with the same amount as the metformin group and high sugar. After 48h of co-culture, the culture supernatants were discarded, each group was operated according to the instructions of the beta-galactosidase staining kit, and finally the cell senescence was observed by means of an inverted fluorescence microscope.
As shown in fig. 5, compared with the control group, the glucose can successfully induce nerve cell injury after being added, the aging model is successfully established, and the metformin group has obvious cell aging protection effect; the drug groups to which (+ -) -1,3,8-Trimethylallantoin, S- (+) -1,3,8-Trimethylallantoin, R- (-) -1,3, 8-trimethyllaxoiin, 1,3, 8-trimethyllaxoiin 4R,4'S-dimer, 1,3, 8-trimethyllaxoiin 4,4' -dimer, 1,3, 8-trimethyllaxoiin 4S,4'S-dimer, 1,3, 8-trimethyllaxoiin 4R,4' R-dimer and 1,3, 7-trimethylritlet were added respectively also showed a superior protective effect against cell senescence, thereby demonstrating that they have a protective effect against cell senescence induced by excessive sugar.
The experiments prove that the products (+/-) -1,3,8-Trimethylallantoin, S- (+) -1,3,8-Trimethylallantoin, R- (-) -1,3, 8-trimethyllaxoid, 1,3, 8-trimethyllaxoid 4R,4' S-dimer, 1,3, 8-trimethyllaxoid 4S,4' S-dimer, 1,3, 8-trimethyllaxoid 4R,4' R-dimer and 1,3, 7-trimethyllaxoid prepared by the invention have stronger protection effect on cell aging.
Therefore, the methylated allantoin alkaloid compound can be applied to the preparation of medicaments for inhibiting cell aging.
In the specific implementation, the methylated allantoin alkaloid compound is prepared into a cell aging protection medicament according to the pharmaceutically acceptable dosage and pharmaceutically common auxiliary materials. The pharmaceutical dosage forms include oral forms, injection forms, and the like. The oral preparation comprises tablets, capsules, granules, dripping pills and the like; the injection type comprises injection, mixed rotation liquid and the like. The specific preparation method refers to the conventional method in the pharmaceutical field.
It should be understood that the examples and embodiments described herein are for illustrative purposes only and are not intended to limit the present invention, and that various modifications or changes in light thereof will be suggested to persons skilled in the art and are to be included within the spirit and purview of this application.
Claims (7)
1. A methylated allantoin alkaloid characterized by being represented by a structure represented by formula iii or formula iv or formula v:
2. use of a methylated allantoin alkaloid according to claim 1 in the preparation of a cell senescence protective medicament.
3. A cell senescence protective medicament, characterized by being prepared from the methylated allantoin alkaloid according to claim 1 and a pharmaceutical excipient.
4. A cytoprotective drug according to claim 3, wherein the drug is in a pharmaceutical dosage form selected from the group consisting of oral and injectable.
5. The method for preparing the methylated allantoin alkaloids according to claim 1, comprising the following steps:
(1) Crushing of raw materials
Pulverizing QIMEN black tea to obtain QIMEN black tea powder;
(2) Leaching
Leaching the Qimen black tea powder with ethanol aqueous solution, separating out an extracting solution, and drying the extracting solution to obtain a Qimen black tea extract;
(3) Separation and purification
Dissolving the Keemun black tea extract with methanol aqueous solution, concentrating to remove most of methanol, extracting with petroleum ether to remove small polar substances and pigment, and extracting with dichloromethane to obtain dichloromethane extract A;
dissolving the dichloromethane part extract A with water, adding an aqueous solution of tartaric acid, removing insoluble substances, extracting with dichloromethane, removing the aqueous layer, adding an equal volume of aqueous solution of sodium hydroxide, extracting to obtain a dichloromethane layer and an alkaline water layer, adding ammonium chloride into the alkaline water layer, and extracting with dichloromethane to obtain a dichloromethane part extract B;
separating and purifying the dichloromethane part extract B to obtain the methylated allantoin alkaloid;
the specific steps of separation and purification treatment are as follows: dissolving the dichloromethane part extract B, performing first silica gel column chromatography by taking a dichloromethane-methanol volume ratio of 500:1 to 1:1 as gradient elution, performing second silica gel column chromatography by taking a dichloromethane-methanol volume ratio of 30:1 to 20:1 elution component obtained by the first silica gel column chromatography, performing second silica gel column chromatography by taking a dichloromethane-methanol volume ratio of 1000:1 to 1:1 as gradient elution, performing methanol recrystallization on a dichloromethane-methanol volume ratio of 20:1 elution component obtained by the second silica gel column chromatography, and obtaining a racemic trimethyl allantoin dimer, resolving the racemic trimethyl allantoin dimer by using a chiral column by using HPLC, and eluting by acetonitrile to obtain methylated allantoin alkaloids shown in a formula IV and methylated allantoin alkaloids shown in a formula V;
eluting the residual component of methanol recrystallization with methanol by using a Toyopearl column chromatography, collecting the component eluted at the first time, and purifying by using HPLC to obtain the methylated allantoin alkaloid shown in the formula III.
6. The method for producing a methylated allantoin-based alkaloid according to claim 5, wherein the concentration of the aqueous ethanol solution used in the step (2) is 95%.
7. The method for producing a methylated allantoin-based alkaloid according to claim 5 or 6, wherein in the step (3), the concentration of the aqueous tartaric acid solution used is 2% and the concentration of the aqueous sodium hydroxide solution is 1-2%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910508889.7A CN110156693B (en) | 2019-06-13 | 2019-06-13 | Methylated allantoin alkaloid and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910508889.7A CN110156693B (en) | 2019-06-13 | 2019-06-13 | Methylated allantoin alkaloid and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110156693A CN110156693A (en) | 2019-08-23 |
| CN110156693B true CN110156693B (en) | 2023-04-21 |
Family
ID=67628776
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910508889.7A Active CN110156693B (en) | 2019-06-13 | 2019-06-13 | Methylated allantoin alkaloid and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110156693B (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB889101A (en) * | 1959-12-01 | 1962-02-07 | Irwin Irville Lubowe | Allantoin compounds |
| DE19853842A1 (en) * | 1998-11-23 | 2000-05-25 | Erich Ruf | New water-soluble compound formed between allantoin and glyoxylic acid useful in cosmetic and/or pharmaceutical formulations |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104557958A (en) * | 2015-01-23 | 2015-04-29 | 广西壮族自治区药用植物园 | Preparation process of anthriscifolcone A and anthriscifolcone B and cytotoxic effect for tumor cells |
| CN108623524B (en) * | 2018-04-17 | 2021-02-26 | 安徽农业大学 | Imidazole dimer alkaloid, and preparation method and application thereof |
| CN108607000A (en) * | 2018-05-30 | 2018-10-02 | 新疆医科大学第附属医院 | The application of allantois extract and allantoin monomer in preparing blood lipid-lowering medicine |
| CN108530430B (en) * | 2018-06-08 | 2021-07-13 | 安徽农业大学 | Ester type catechin pyrrolidine alkaloid and preparation method and application thereof |
-
2019
- 2019-06-13 CN CN201910508889.7A patent/CN110156693B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB889101A (en) * | 1959-12-01 | 1962-02-07 | Irwin Irville Lubowe | Allantoin compounds |
| DE19853842A1 (en) * | 1998-11-23 | 2000-05-25 | Erich Ruf | New water-soluble compound formed between allantoin and glyoxylic acid useful in cosmetic and/or pharmaceutical formulations |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110156693A (en) | 2019-08-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102940687B (en) | A kind of Fructus Toosendan extract and uses thereof | |
| CN111961022B (en) | Biflavonoid compound with neuroprotective effect and extraction and separation method thereof | |
| CN107827864B (en) | Method for separating chemical components of methylene chloride extraction part of piper longum composition and application | |
| CN108530430A (en) | Ester catechin pyrrolidine alkaloid and its preparation method and application | |
| CN101259124A (en) | Pharmaceutical use of wedelolactone and its derivative | |
| CN104262465A (en) | Rubiaceae cyclopeptides used as TAK1 inhibitor and preparation method thereof | |
| CN110156693B (en) | Methylated allantoin alkaloid and preparation method and application thereof | |
| CN105198943B (en) | A kind of entitled tea hill how glycosides A acylated flavonoids glucosides and its preparation method and application | |
| CN115260148B (en) | Compound extracted and separated from long pepper and application of compound in preparation of anti-inflammatory drugs | |
| CN113754620B (en) | Lignan amide compound in fructus cannabis, and preparation method and application thereof | |
| CN101318937B (en) | Alkaloid compounds, preparation method and application thereof | |
| CN111153955A (en) | Gynostemma pentaphylla extract with lipid-lowering effect and preparation method and application thereof | |
| CN108623524B (en) | Imidazole dimer alkaloid, and preparation method and application thereof | |
| CN113214214B (en) | Preparation method and application of terpenoid in Atractylodes lancea | |
| CN112830936B (en) | Riboflavin compound, preparation method and application thereof | |
| CN111377933B (en) | Alkaloid compound extracted from orychophragmus violaceus seeds as well as extraction method and application thereof | |
| CN103275138B (en) | 16 carbon diacetyls are without double bond lactone type sophorolipid and application thereof | |
| CN109206392B (en) | Coumarin compound and preparation method and application thereof | |
| CN110638822A (en) | Albizzia julibrissin glycoside compound for promoting endothelial cell proliferation and application thereof | |
| CN110585221A (en) | Albizzia julibrissin new lignan compound for improving steatosis and application thereof | |
| CN112300185B (en) | Alkaloid compound with reduced hepatotoxicity, and preparation method and application thereof | |
| CN112402412B (en) | Application of inner ester compound of jingdao in preparing medicine for treating inflammation-caused diseases | |
| CN115073481B (en) | Furana quebracho base dimer or pharmaceutically acceptable salt thereof, and preparation method and application thereof, and pharmaceutical composition | |
| CN113149840B (en) | Wild pepper extract and preparation method and application thereof | |
| CN116693480B (en) | Dihydro-fraxinenone A and preparation method and application 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 |