CN110218208A - A kind of diels-Alder type compound and its preparation method and application - Google Patents
A kind of diels-Alder type compound and its preparation method and application Download PDFInfo
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- CN110218208A CN110218208A CN201810177450.6A CN201810177450A CN110218208A CN 110218208 A CN110218208 A CN 110218208A CN 201810177450 A CN201810177450 A CN 201810177450A CN 110218208 A CN110218208 A CN 110218208A
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- C07—ORGANIC CHEMISTRY
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- C07D407/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having oxygen atoms as the only ring hetero atoms, not provided for by group C07D405/00
- C07D407/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having oxygen atoms as the only ring hetero atoms, not provided for by group C07D405/00 containing three or more hetero rings
Abstract
The invention discloses a kind of diels-Alder type compounds or its pharmaceutically acceptable salt or ester and preparation method and application shown in formula I.Diels shown in formula I-Alder type compound is alpha-glucosidase restrainer, suitable for the pharmaceutical composition with hypoglycemic curative effect.
Description
Technical field
The present invention relates to drug fields, and in particular to a kind of diels-Alder type compound and preparation method thereof and answers
With.
Background technique
Diels-Alder (Diels-Alder) type compound, is the distinctive chemical component of Mulberry plant, in structure
It is the α by chalcone or chalcone derivative, after with the compound containing prenyl [4+2] addition reaction occurs for β double bond
The methyl cyclohexane ene derivative of formation, some compounds also occur further to react, such as double-bond shift, oxidation, cyclisation, condensation
Deng, thus form the more complicated diels-Alder type compound of structure.This kind of structure-activity report seldom, is concentrated mainly on
It is anti-oxidant, anti-inflammatory activity.
Alpha-glucosidase (α-glucosidase) is called alpha-D-glucose glycosides hydrolase.It can be from oligosaccharides bottom
The non-reducing end of object cuts α-Isosorbide-5-Nitrae glycosidic bond, releases glucose, or free glucose residue out is transferred to another sugar
Class substrate forms α -1,6 glycosidic bonds, to obtain oligoisomaltose or sugar ester, glycopeptide of non-fermented etc..Detailed process are as follows:
Polysaccharide in food, as starch direct oral cavity saliva, amylopsin are digested to the oligosaccharide containing a small number of glucose molecules, phlorose
Glycosides enzyme just cuts α-Isosorbide-5-Nitrae glycosidic bond in the non-reducing end of these oligosaccharide, releases glucose, glucose is inhaled by small intestine epithelium
Enter blood circulation after receipts, just becomes blood glucose.It plays an important role during food absorption, it is necessary to after in combination,
Food could be digested and be absorbed.
Currently, alpha-glucosidase restrainer be widely used for reduce postprandial hyperglycemia, clinically apply such
Drug mainly has acarbose, voglibose and Miglitol, by clinic application, achieves preferable curative effect, is considered
Be diabetes B drug of first choice and type 1 diabetes insulin therapy ancillary drug, have broad application prospects.It is this kind of
Contain multiple sugar unit bodies in alpha-glucosidase restrainer molecule, synthesis step is more complex, and after taking these three types of medicines, warp
It often will appear the adverse reactions such as abdominal discomfort, flatulence, exhaust.
Therefore, there is an urgent need in the art to develop a kind of Small side effects, efficiently novel alpha-glucosidase restrainer.
Summary of the invention
The present invention provides a kind of diels different from the prior art-Alder type compound and preparation method thereof and answer
With.Diels of the invention-Alder type compound is to extract to obtain from a variety of Mulberry plants, and show to α-Portugal
Polyglycoside enzyme inhibition activity is high, source is diversified and obtains the advantages that facilitating.
The present invention provides a kind of diels-Alder type compounds or its pharmaceutically acceptable salt shown in formula I
Or ester:
Wherein, R R1Or R2；
When R is R1When be compound I-1；When R is R1When be compound I-2.
The present invention also provides a kind of diels-Alder type compound preparation method shown in formula I, including it is following
Step:
Step 1: Mulberry plant is raw material, extracts to obtain medicinal extract with alcoholic solution, medicinal extract pure water is dispersed, is extracted with organic solvent
To get crude extract after taking；
Step 2: crude extract obtained by step (1) being separated through column chromatography, is further purified up to Di Er shown in Formulas I
This-Alder type compound；
Wherein, in compound I, R R1When compound (compound I-1) and compound I in, R R2When change
The detection parameters for closing object (i.e. compound I-2) are as follows:
High-efficient liquid phase chromatogram condition: C18 bonded silica gel column, Detection wavelength 254nm, flow velocity 2.5mL/min, mobile phase are body
The acetonitrile solution that product content is 55%, compound I-1 retention time are tR=20.3min；Mobile phase volume is that content is
56% acetonitrile solution, compound I-2 retention time are tR=18.5min.
Those skilled in the art can be obtained according to above-mentioned detection parameters, the condition of corresponding selection and adjustment column chromatography
When such as preparing using high-efficient liquid phase technique, the efficient liquid phase point in above-mentioned detection parameters is can be used in compound I-1 and compound I-2
From condition, other separation conditions can also be used, as long as the detection parameters of the product obtained are consistent with above-mentioned detection parameters.
Wherein, in step (1), described raw material this field is known, can be in the root of Mulberry plant, stem, leaf, skin and fruit
One or more, the preferably stem skin of milk mulberry；The alcoholic solution be ethanol water, the ethanol water it is dense
Degree preferably 95%；The operation of the extraction and condition are the operation and condition of the extraction of this field routine, and described mentions
Number is taken preferably to carry out 3 times；Each extraction time is preferably 1.5 hours；The ethanol water that the extraction uses
The volume of solution is preferably 10 with the ratio for being the quality that Mulberry plant is raw material；The process for adding pure water to disperse medicinal extract
In, the mass ratio of the pure water and the medicinal extract is the pure water of this field routine and the mass ratio of medicinal extract, and the present invention is preferably
4；The operation of the extraction and condition are the operation and condition of this field routine；The organic solvent be alkane solvents,
One of chlorinated hydrocarbon solvent, esters solvent and alcohol reagent are a variety of；The alkane solvents are preferably petroleum ether；
The chlorinated hydrocarbon solvent is preferably methylene chloride；The esters solvent is preferably ethyl acetate；The alcohols
It is preferred solvents n-butanol；The operation of the filtering and condition are the operation and condition of the filtering of this field routine；It is described to subtract
Press operation and the condition of concentration for the operation and condition of the concentration of this field routine, the concentration is preferably concentrated under reduced pressure.
Wherein, in step (2), freeze-drying is still further comprised, the freeze-drying is preferably vacuum freeze-drying technology, described
Vacuum freeze-drying technology is the vacuum freeze-drying technology of this field routine；It is described that preferably high performance liquid chromatography is further purified is pure
Change；The high-efficient liquid phase chromatogram purification is isolated and purified under the conditions of Detection wavelength 254nm, flow velocity 2.5mL/min, flowing
It is mutually the acetonitrile solution that volume content is 55%, collection retention time is tR=20.3min component, obtains compound I-1；Stream
Dynamic phase volume is the acetonitrile solution that content is 56%, and collection retention time is tRThe component of=18.5min obtains compound I-
2；The column chromatography for separation is followed successively by by positive silica gel column chromatography, gel filtration chromatography and reversed-phase silica gel column chromatography separation；Institute
The eluent for the positive silica gel column chromatography stated is methylene chloride-methanol, the volume ratio of the eluent dichloromethane and methanol
It is followed successively by 95:5,80:20,70:30,60:40 and 50:50；The eluent of the gel filtration chromatography is methylene chloride-methanol,
The volume ratio of the eluent dichloromethane and methanol is 1:1；The eluent of the reversed-phase silica gel column chromatography is methanol-
The volume ratio of water, the eluant methanol and water is followed successively by 55:45,65:35,75:25 and 85:15；The silica gel column chromatography
In column, the routine that the specification of chromatographic column is referred to this field is selected, and column internal diameter × column length of the present invention is preferably 10cm
×100cm；In the silica gel column chromatography, the routine that the specification of silica gel is referred to this field is selected, and preferably 200
~300 mesh；In the silica gel column chromatography, the quality of silica gel and the mass ratio of extract crude product can refer to this field it is conventional into
Row selection, preferably 10；The dosage of each gradient eluent can refer to this field with column volume ratio and routinely be selected
It selects, preferably 3；The routine that the gel column can refer to this field is selected；The reverse phase silica gel column can refer to this
The routine in field is selected.
The present invention also provides a kind of diels shown in Formulas I-Alder type compound or its pharmaceutically acceptable salts
Or ester is preparing the application in alpha-glucosidase restrainer.
The present invention also provides a kind of diels-Alder type compound shown in formula I or its is pharmaceutically acceptable
Salt or ester are preparing the application in hypoglycemic drug.
The present invention also provides a kind of pharmaceutical composition, including diels shown in formula I containing treatment effective dose-Ah
That moral type compound or its pharmaceutically acceptable salt or ester and pharmaceutically acceptable carrier.
Preferably, the pharmaceutical composition further comprises other hypoglycemic active constituents and can pharmaceutically connect
The carrier received；Wherein, the preferred acarbose of other hypoglycemic active constituents, voglibose or combinations thereof.
In a preferred example, diels shown in formula I-Alder type compound activity ingredient and other hypoglycemic work
The mass ratio of property ingredient is preferably 1:100~100:1, is more preferably 1:10~10:1.
In a preferred example, the diels shown in formula I-Alder type compound or its pharmaceutically acceptable salt
Or content of the ester in pharmaceutical composition can be mass percent 0.1%~99.5%, preferably mass percent 10%~
99.9%, more preferably 70%~99.9%.
In another preferred example, the diels shown in formula I-Alder type compound or its is pharmaceutically acceptable
The content of salt or ester in pharmaceutical composition can be mass percent 60.0%~99.5%, preferably mass percent 70%
~99.5%, more preferably 80%~99.5%.
" pharmaceutically acceptable carrier " refers to: one or more biocompatible solids or liquid filler or gel
Substance, they are suitable for people's use and it is necessary to have enough purity and sufficiently low toxicity." compatibility " referred to herein as group
Close in object each component energy and the compound of the present invention and they between mutually admix, and significantly reduce the drug effect of compound.
Pharmaceutically acceptable carrier part example have cellulose and its derivates (such as sodium carboxymethylcellulose, ethyl cellulose sodium,
Cellulose ethanoate etc.), gelatin, talcum, solid lubricant (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oil (such as soya-bean oil,
Sesame oil, peanut oil, olive oil etc.), polyalcohol (such as propylene glycol, glycerol, mannitol, sorbierite), emulsifier (such as), wetting agent (such as lauryl sodium sulfate), colorant, flavoring agent, stabilizer, antioxidant, preservative, apyrogeneity
Water etc..
Pharmaceutical composition of the invention includes diels-Alder type compound or its pharmacology within the scope of safe and effective amount
Upper acceptable salt or ester and pharmacologically acceptable excipient or carrier.Wherein " safe and effective amount " refers to: compound
Amount be enough to be obviously improved the state of an illness, and be unlikely to generate serious side effect.In general, pharmaceutical composition contains diels-A Er
Moral type compound or its pharmacologically acceptable salt or ester 1-2000mg/ agent, preferably, more preferably containing containing 5-200mg/ agent
There is 10-100mg/ agent." one " is preferably a capsule or tablet.
The method of application of pharmaceutical composition of the invention is not particularly limited, and representative method of application includes (but not
Be limited to): oral, rectum, parenteral (intravenous, intramuscular or subcutaneous) and local administration, preferably administration mode be take orally to
Medicine.
The solid dosage forms for oral administration includes capsule, tablet, pill, powder and granule.At these
In solid dosage forms, reactive compound is mixed at least one conventional inert excipients or carrier, such as sodium citrate or Dicalcium Phosphate,
Or it is mixed with following compositions: (a) filler or expanding material, for example, starch, lactose, sucrose, glucose, mannitol and silicic acid；(b)
Adhesive, for example, hydroxymethyl cellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and Arabic gum；(c) moisturizing
Agent, for example, glycerol；(d) disintegrating agent, for example, agar, calcium carbonate, potato starch or tapioca, alginic acid, certain comprehensive silicons
Hydrochlorate and sodium carbonate；(e) retarding solvent, such as paraffin；(f) absorbsion accelerator, for example, quaternary ammonium compound；(g) wetting agent, such as
Cetanol and glycerin monostearate；(h) adsorbent, for example, kaolin；(i) lubricant, for example, talcum, calcium stearate,
Or mixtures thereof magnesium stearate, solid polyethylene glycol, lauryl sodium sulfate,.In capsule, tablet and pill, dosage form can also
Include buffer.
Coating and shell material preparation can be used in solid dosage forms such as tablet, sugar-pill, capsule, pill and granule, such as casing and
Other materials well known in the art.They may include opacifying agent, also, reactive compound or compound in such composition
Release can discharge in certain a part in the digestive tract in a delayed fashion.The example of adoptable embedding component is polymeric material
And wax material.When necessary, reactive compound can also be with one of above-mentioned excipient or a variety of formation microencapsulation forms.
The liquid dosage form of the oral administration includes pharmaceutically acceptable lotion, solution, suspension, syrup or tincture
Agent.In addition to active compounds, liquid dosage form may include the inert diluent routinely used in this field, such as water or other molten
Agent, solubilizer and emulsifier, example know, ethyl alcohol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-BDO, dimethyl
Formamide and oil, especially cottonseed oil, peanut oil, maize germ, olive oil, castor oil and sesame oil or these substances it is mixed
Close object etc..
Other than these inert diluents, composition also may include auxiliary agent, such as wetting agent, emulsifier and suspending agent, sweet taste
Agent, corrigent and fragrance.
In addition to active compounds, suspension may include suspending agent, for example, ethoxylation isooctadecane alcohol, polyoxyethylene
Or mixtures thereof sorbierite, Isosorbide Dinitrate, microcrystalline cellulose, aluminium methoxide and agar etc..
Composition for parenteral injection may include physiologically acceptable sterile, aqueous or anhydrous solution, dispersion liquid,
Suspension or lotion, and the aseptic powdery for re-dissolving into sterile Injectable solution or dispersion liquid.It is suitable aqueous and
Nonaqueous carrier, diluent, solvent or excipient include water, ethyl alcohol, polyalcohol and its suitable mixture.
Dosage form for local administration includes ointment, powder, patch, stock solution and inhalant.Active constituent is sterile
Under the conditions of with physiologically acceptable carrier and any preservative, buffer, or when necessary may need propellant mix together
It closes.
The pharmaceutical composition can be administered alone, or be administered in combination with other pharmaceutically acceptable compounds,
Such as acarbose.
It is the mammal that the compounds of this invention of safe and effective amount is applicable to treatment when using pharmaceutical composition
(such as people), wherein dosage is the effective dosage pharmaceutically thought when application, for the people of 60kg weight, day is to medicament
Amount is usually 1~2000mg, preferably 5~500mg, more preferably 10~100mg.Certainly, specific dosage is also contemplated that administration way
The factors such as diameter, patient health situation, within the scope of these are all skilled practitioners technical ability.
A method of screening hypoglycemic drug candidate, comprising the following steps:
Step 1: providing untested compound and positive reference compound, the positive reference compound is shown in Formulas I
Diels-Alder type compound or its pharmaceutically acceptable salt or ester；
Step 2: in test group, detecting influence of the untested compound to alpha-glucosidase, and and positive control
Corresponding experimental result is compared in group and negative control group, wherein in positive controls, detects positive control chemical combination
Influence of the object to alpha-glucosidase；
Wherein, if the untested compound is significantly higher than negative control group to the inhibition level of alpha-glucosidase,
Prompting the untested compound is hypoglycemic drug candidate.
In another preferred example, in step (2), by test group compared with positive controls, and compare the ratio of I1 and I2
Value, wherein I1 is inhibition level of the untested compound to alpha-glucosidase, and I2 is positive reference compound to α-Portugal
The inhibition level of polyglycoside enzyme, if I1/I2 >=80%, prompting the untested compound is hypoglycemic drug candidate.
On the basis of common knowledge of the art, above-mentioned each optimum condition, can any combination to get each preferable reality of the present invention
Example.
The reagents and materials used in the present invention are commercially available.
The positive effect of the present invention is that: diels of the invention-Alder type compound is new construction type
Alpha-glucosidase restrainer can be used to prevent or treat diabetes.Diels of the invention-Alder type compound has height
The alpha-glucosaccharase enzyme inhibition activity of effect is 840 times of positive compound acarbose or so.Diels-A Er of the invention
Moral type compound can be extracted from a variety of Mulberry plants and be obtained, and source diversification, it is convenient to obtain.
Detailed description of the invention
Fig. 1 is the ECD map of compound I-1 made from embodiment 1
Fig. 2 is the ECD map of compound I-2 made from embodiment 1
Specific embodiment
The present invention is further illustrated below by the mode of embodiment, but does not therefore limit the present invention to the reality
It applies among a range.In the following examples, the experimental methods for specific conditions are not specified, according to conventional methods and conditions, or according to quotient
The selection of product specification.
Embodiment 1
Following embodiment is statistically considered as significant difference with P < 0.05 *.
Instrument and reagent
Nuclear Magnetic Resonance: Bruker DRX 400 (Germany), TMS internal standard；High-resolution mass spectrometer: Waters Xevo G2-
XS (U.S.)；Medium and low pressure column chromatograph (Japanese YMC company)；Microplate reader: Bio-TekSynergy2 (U.S.).
Silica gel (200-300 mesh), thin-layer chromatographic analysis prefabricated board are Haiyang Chemical Plant, Qingdao's production with prefabricated board is prepared；
C18 filler (Japanese YMC company).Agents useful for same is to analyze pure (Sinopharm Chemical Reagent Co., Ltd.).
1) it extracts: taking milk mulberry stem skin (10kg), crush, 95% alcohol reflux is added to extract, alcohol dosage is 10 (v/ of medicinal material amount
M) again, extract 3 times, 1.5 hours every time, merge after extracting solution is filtered, it is dry, obtain milk mulberry dry extract.Medicinal extract is suspended in 4 times of amounts
In water, petroleum ether, methylene chloride, ethyl acetate and extracting n-butyl alcohol are used respectively, obtains different positions.To ethyl acetate extract
It is concentrated under reduced pressure, calculating dry weight is 70g.
2) column chromatographs: the ethyl acetate extract that will be obtained, and carrying out silica gel column chromatography, (10 times of volumes are in the silicon of sample quality
Glue, the glass column of 100cm × 10cm), respectively with dichloro methane-methanol (95:5 → 80:20 → 70:30 → 60:40 →
50:50) gradient elution, mixed solvent amount used in each gradient are 3 times of column volumes, successively collect, merge, are obtained 10 big group
Point.Component Fr.D (6.0g) the 4th big is taken to pass sequentially through gel filtration chromatography (methylene chloride: methanol=1:1) and reverse phase silica gel column
Chromatography (methanol: water=55:45 → 65:35 → 75:25 → 85:15) obtains secondary component D-g (58mg) and D-e (50mg).
3) prepared by HPLC: above-mentioned secondary component D-g (58mg) is prepared (55% acetonitrile solution, Detection wavelength with HPLC
254nm, flow velocity 2.5mL/min), obtain compound I-1:macrourin I (tR=20.3min)；By above-mentioned secondary component D-e
(50mg) prepares (56% acetonitrile solution, Detection wavelength 254nm, flow velocity 2.5mL/min) through HPLC, obtains compound I-2:
macrourin J(tR=18.5min).
Compound I-1 (macrourin I), Yellow amorphous powder are slightly soluble in water, dissolve in acetone.HRESI-MS m/
z 647.2288[M-H]-, determine that compound molecular weight is 648；[α]2 D 5+305.6°(c0.4,MeOH)；ECD(MeOH,nm)λ
max(Δε)229(+1.41),282(-5.35),323(+15.28),334(+16.48)；1H-NMR(acetone-d6,
400MHz) δ: 13.21 (1H, s, 10 "-OH), 7.99 (1H, d, J=8.8Hz, H-14 "), 7.40 (1H, d, J=8.4Hz, H-
4), 7.02 (1H, brs, H-3), 6.98 (1H, d, J=2.0Hz, H-7), 6.89 (1H, brs, H-20 "), 6.84 (1H, d, J=
2.0Hz, H-6 '), 6.81 (1H, dd, J=8.4,2.0Hz, H-5), 6.80 (1H, d, J=2.0Hz, H-2 '), 6.32 (1H, d, J
=8.8Hz, H-13 "), 6.24 (1H, d, J=2.4Hz, H-17 "), 6.14 (1H, dd, J=8.4,2.4Hz, H-19 "), 4.60
(1H, brs, H-4 "), 3.83 (1H, brs, H-3 "), 3.58 (1H, brs, H-5 "), 2.55 (2H, brt, J=7.2Hz, H-
), 2.43 21 " (1H, brd, J=12.4Hz, H-2 " a), 2.19 (1H, brs, H-6 " a), 1.88 (1H, brd, J=12.4Hz, H-
2 " b), 1.77 (2H, brt, J=7.2Hz, H-22 "), 1.43 (3H, s, H3- 7 "), 1.33 (3H, s, H3- 24 "), 1.30 (3H s,
H3-25″)。13C-NMR data are shown in Table 1.
Compound I-2 (macrourin J), Yellow amorphous powder are slightly soluble in water, dissolve in acetone.HRESI-MS m/
z 647.2285[M-H]-, determine that compound molecular weight is 648；[α]2 D 5+302.3°(c 0.2,MeOH)；ECD(MeOH,nm)
λmax(Δε)225(+0.23),236(-4.94),303(-5.90),324(+15.71,sh),334(+19.91)；1H-NMR
(acetone-d6, 400MHz) and δ: 7.43 (1H, d, J=8.8Hz, H-14 "), 7.41 (1H, d, J=8.4Hz, H-4), 7.00
(1H, brs, H-3), 6.98 (1H, d, J=2.0Hz, H-7), 6.97 (1H, brs, H-20 "), 6.83 (1H, d, J=2.0Hz, H-
3), 6.81 (1H, dd, J=8.4,2.0Hz, H-5), 6.75 (1H, d, J=1.6Hz, H-2 '), 6.34 (1H, d, J=8.4Hz,
H-13 "), 6.20 (1H, d, J=2.4Hz, H-17 "), 6.19 (1H, dd, J=8.4,2.4Hz, H-19 "), 4.52 (1H, brs,
H-4 "), 4.13 (1H, brs, H-3 "), 3.54 (1H, brs, H-5 "), 2.71 (2H, brt, J=7.2Hz, H-21 "), 2.31
A), 2.19 ((a), 1.93 (1H, brd, J=12.8Hz, H-2 are " b), by 1H, brs, H-6 " by 1H, dd, J=12.8,2.4Hz, H-2 "
1.90 (2H, brt, J=7.2Hz, H-22 "), 1.58 (3H, s, H3- 24 "), 1.53 (3H, s, H3- 25 "), 1.45 (3H, s, H3-
7″)；13C-NMR data are shown in Table 1.
1. compound I-1~2 of table13C-NMR tables of data
aSignal is not observed
bSignal interchangeable
Embodiment 2
1, instrument and material
Microplate reader (Bio-Tek Synergy2), centrifuge KA-1000 (Shanghai flying pigeon), double superclean bench (Suzhou
Purification), adjustable pipette (Finland, 2-20uL, 20-200uL, 100-1000uL), 96 orifice plates (Corning, USA), import
Pipette tips (Axygen, USA), positive compound (acarbose).
2, method
1) successively mixed phosphate salt buffer (pH=7.6), p-nitrophenyl α-D- pyrans in the reaction system of 250 μ L
Glucoside, alpha-glucosidase and untested compound, 30 DEG C of temperature are incubated 30 minutes.
2) terminator sodium carbonate liquor is added stops reaction.
3) it is detected at 405nm wavelength using microplate reader.
3, sample to be tested, positive drug processing method
Sample to be tested and acarbose accurately claim sample, are diluted to series of tasks concentration using DMSO before testing.Substrate is to nitre
Base phenyl α-D- glucopyranoside weighs 30.125mg addition 50mL volumetric flask to specifications and is diluted to detection buffer
Working solution.
AC: the light absorption value of control group, AB: blank group light absorption value；AS: sample sets light absorption value；ASB: sample blank group light absorption value
4, experimental result
4.1, primary dcreening operation
2. inhibiting rate primary dcreening operation table of table
4.1、IC50Value measurement
The 3. alpha-glucosaccharase enzyme inhibition activity of compound I-1~2 IC of table50Table
Remarks: experiment is measured in parallel 3 times
As a result as shown in table 2 and table 3.The result shows that diels of the invention-Alder type compound shown in formula I
It is 840 times of positive compound acarbose or so with efficient alpha-glucosaccharase enzyme inhibition activity.Therefore, of the invention
Diels-Alder compounds of formula I can prevent or treat diabetes, while can develop as new and effective alpha-glucosaccharase
Enzyme inhibitor.
Claims (10)
1. a kind of diels-Alder type compound or its pharmaceutically acceptable salt or ester shown in formula I:
Wherein, R R1Or R2。
2. a kind of diels-Alder type compound preparation method shown in formula I as described in claim 1, including with
Lower step:
Step 1: Mulberry plant is raw material, extracts to obtain medicinal extract with alcoholic solution, medicinal extract pure water is dispersed, is extracted with organic solvent
Afterwards to get crude extract；
Step 2: crude extract obtained by step (1) being separated through column chromatography, is further purified up to diels-shown in Formulas I
Alder type compound；
Wherein, in compound I, R R1When compound and compound I in, R R2When compound detection parameters such as
Under:
High-efficient liquid phase chromatogram condition: C18 bonded silica gel column, Detection wavelength 254nm, flow velocity 2.5mL/min, mobile phase contain for volume
The acetonitrile solution for being 55% is measured, R is R in compound I1The retention time of the compound be tR=20.3min；Mobile phase volume
The acetonitrile solution for being 56% for content, R is R in compound I2The retention time of the compound be tR=18.5min.
3. diels-Alder type compound preparation method shown in formula I as claimed in claim 2, which is characterized in that
In the step (2), the column chromatography for separation is followed successively by by positive silica gel column chromatography, gel filtration chromatography and reverse phase silica gel
Column chromatography for separation.
4. diels-Alder type compound preparation method shown in formula I as claimed in claim 3, which is characterized in that
The eluent of the positive silica gel column chromatography is methylene chloride-methanol, the volume of the eluent dichloromethane and methanol
Than being followed successively by 95:5,80:20,70:30,60:40 and 50:50；
And/or the eluent of the gel filtration chromatography is methylene chloride-methanol, the eluent dichloromethane and methanol
Volume ratio be 1:1；
And/or the eluent of the reversed-phase silica gel column chromatography is methanol-water, the volume ratio of the eluant methanol and water
It is followed successively by 55:45,65:35,75:25 and 85:15.
5. diels-Alder type compound preparation method shown in formula I as claimed in claim 2, which is characterized in that
In the step (2), described is further purified as high-efficient liquid phase chromatogram purification.
6. diels-Alder type compound preparation method shown in formula I as claimed in claim 5, which is characterized in that
The high-efficient liquid phase chromatogram purification isolates and purifies under the conditions of Detection wavelength 254nm, flow velocity 2.5mL/min, and mobile phase is body
The acetonitrile solution that product content is 55%, collection retention time are tR=20.3min component, obtaining R in compound I is R1Chemical combination
Object；Mobile phase volume is the acetonitrile solution that content is 56%, and collection retention time is tRThe component of=18.5min, obtains chemical combination
R is R in object I2Compound.
7. as described in claim 1 diels-Alder type compound or its pharmaceutically acceptable salt shown in formula I or
Ester is preparing the application in alpha-glucosidase restrainer.
8. as described in claim 1 diels-Alder type compound or its pharmaceutically acceptable salt shown in formula I or
Ester is preparing the application in hypoglycemic drug.
9. a kind of pharmaceutical composition, including controlling for diels shown in formula I-Alder type compound described in claim 1
Diels shown in formula I-Alder type the compound or its pharmaceutically acceptable salt or ester and pharmacy for the treatment of effective dose
Upper acceptable carrier.
10. the pharmaceutical composition as claimed in claim 9, which is characterized in that the pharmaceutical composition is further wrapped
Containing other hypoglycemic active constituents；Other hypoglycemic active constituents are acarbose, voglibose or its group
It closes.
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103476408A (en) * | 2011-03-28 | 2013-12-25 | 同和药品株式会社 | Use of compounds isolated from morus bark |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103476408A (en) * | 2011-03-28 | 2013-12-25 | 同和药品株式会社 | Use of compounds isolated from morus bark |
Non-Patent Citations (3)
| Title |
|---|
| MENG WANG,ET AL.: "Diels–Alder adducts with PTP1B inhibition from Morus notabilis", 《PHYTOCHEMISTRY》 * |
| 何雪梅 等: "桑属植物的化学成分及药理活性综述", 《蚕业科学》 * |
| 唐本钦 等: "广东桑叶化学成分及其α-葡萄糖苷酶活性研究", 《中草药》 * |
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