CN110218208B - Diels-Alder type compound and preparation method and application thereof - Google Patents
Diels-Alder type compound and preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a Diels-Alder compound shown as a formula I or a pharmaceutically acceptable salt or ester thereof, and a preparation method and application thereof. The diels-alder type compound shown as the formula I is an alpha-glucosidase inhibitor and is suitable for a pharmaceutical composition with the effect of reducing blood sugar.
Description
Technical Field
The invention relates to the field of medicines, and particularly relates to a Diels-Alder compound and a preparation method and application thereof.
Background
Diels-Alder (Diels-Alder) type compounds are peculiar chemical components of Morus plants, and structurally are methyl cyclohexene derivatives formed by [4+2] addition reaction of alpha, beta double bonds of chalcone or chalcone derivatives and compounds containing isoprenyl, and some compounds are further reacted, such as double bond shift, oxidation, cyclization, condensation and the like, so that Diels-Alder type compounds with more complex structures are formed. The structural activity is rarely reported, and the structural activity is mainly focused on antioxidant and anti-inflammatory activity.
Alpha-glucosidase is also called alpha-D-glucoside hydrolase. It is possible to obtain non-fermentable isomaltooligosaccharides, sugar esters, glycopeptides, and the like by cleaving the alpha-1, 4 glycosidic bond from the non-reducing end of the oligosaccharide substrate to release glucose, or transferring the liberated glucose residue to another saccharide substrate to form an alpha-1, 6 glycosidic bond. The specific process is as follows: the food polysaccharide, such as starch, is digested into oligosaccharide containing a few glucose molecules by oral saliva and pancreatic amylase, alpha-glucosidase cuts alpha-1, 4 glycosidic bond at non-reducing end of the oligosaccharide to release glucose, and the glucose is absorbed by small intestine epithelium and enters blood circulation to become blood sugar. It plays an important role in the absorption process of food, and must be combined therewith for the food to be digested and absorbed.
At present, the alpha-glucosidase inhibitor is widely used for reducing postprandial hyperglycemia, the clinically applied medicines mainly comprise acarbose, voglibose and miglitol, and the clinical application shows that the alpha-glucosidase inhibitor has better curative effect, is considered as a first choice medicine for type 2 diabetes and an auxiliary medicine for insulin treatment for type 1 diabetes, and has wide application prospect. The molecules of the alpha-glucosidase inhibitor all contain a plurality of sugar unit bodies, the synthesis steps are complex, and adverse reactions such as abdominal discomfort, flatulence, exhaust and the like often occur after the three drugs are taken.
Therefore, there is an urgent need in the art to develop a novel α -glucosidase inhibitor with less side effects and high efficiency.
Disclosure of Invention
The invention provides a Diels-Alder compound different from the prior art and a preparation method and application thereof. The Diels-Alder compound can be extracted from various Morus plants, and has the advantages of high alpha-glucosidase inhibitory activity, diversified sources, convenient acquisition and the like.
The invention provides a Diels-Alder compound shown as a formula I or a pharmaceutically acceptable salt or ester thereof:
wherein R is R1Or R2；
When R is R1When the compound is the compound I-1; when R is R1When the compound is the compound I-2.
The invention also provides a preparation method of the Diels-Alder compound shown in the formula I, which comprises the following steps:
step 1: extracting Morus plant with alcoholic solution to obtain extract, dispersing the extract with pure water, and extracting with organic solvent to obtain crude extract;
step 2: separating the crude extract obtained in the step (1) by column chromatography, and further purifying to obtain a Diels-Alder compound shown in a formula I;
wherein, in the compound I, R is R1A compound of (i.e. compound I-1), and compound I wherein R is R2The detection parameters of the compound (i.e., compound I-2) were as follows:
high performance liquid chromatography conditions: c18 bonded silica gel column, detection wavelength of 254nm, flow rate of 2.5mL/min, mobile phase of acetonitrile water solution with volume content of 55%, retention time of compound I-1 is tR20.3 min; the mobile phase volume is 56% acetonitrile water solution, and the retention time of the compound I-2 is tR＝18.5min。
The skilled person can select and adjust the column chromatography conditions according to the above detection parameters to obtain compound I-1 and compound I-2, for example, when the high performance liquid chromatography is used for preparation, the high performance liquid separation conditions in the above detection parameters can be used, or other separation conditions can be used, as long as the detection parameters of the obtained product are consistent with the above detection parameters.
Wherein, in the step (1), the raw material is one or more of roots, stems, leaves, skins and fruits of the plants in the genus of Morus, preferably stem skins of Morus alba; the alcohol solution is ethanol water solution, and the concentration of the ethanol water solution is preferably 95%; the extraction operation and conditions are conventional in the field, and the extraction times are preferably 3 times; the time for each extraction is preferably 1.5 hours; the ratio of the volume of the ethanol aqueous solution used for extraction to the mass of the mulberry plant serving as the raw material is preferably 10; in the process of dispersing the extract by adding pure water, the mass ratio of the pure water to the extract is the conventional mass ratio of the pure water to the extract in the field, and the mass ratio is preferably 4; the operation and conditions of the extraction are the operation and conditions conventional in the field; the organic solvent is one or more of alkane solvents, chlorohydrocarbon solvents, ester solvents and alcohol reagents; the alkane solvent is preferably petroleum ether; the chlorinated hydrocarbon solvent is preferably dichloromethane; the ester solvent is preferably ethyl acetate; the alcohol solvent is preferably n-butanol; the filtration operations and conditions are those conventional in the art; the operation and conditions of the concentration under reduced pressure are those of the concentration conventional in the art, and the concentration is preferably concentration under reduced pressure.
Wherein, in the step (2), the freeze-drying is further included, the freeze-drying is preferably a decompression freeze-drying technology, and the decompression freeze-drying technology is a decompression freeze-drying technology which is conventional in the art; the further purification is preferably high performance liquid chromatography purification; the high performance liquid chromatography purification is carried out under the conditions that the detection wavelength is 254nm and the flow rate is 2.5mL/min, the mobile phase is acetonitrile water solution with the volume content of 55 percent, and the collection retention time is tR20.3min to give compound I-1; the volume of the mobile phase is 56 percent of acetonitrile water solution, and the collection retention time is tR18.5min to give compound I-2; the column chromatography separation sequentially comprises forward silica gel column chromatography, gel column chromatography and reverse phase silica gel column chromatography separation; the eluent of the forward silica gel column chromatography is dichloromethane-methanol, and the volume ratios of the dichloromethane to the methanol of the eluent are 95:5, 80:20, 70:30, 60:40 and 50:50 in sequence; the eluent of the gel column chromatography is dichloromethane-methanol, and the eluent isThe volume ratio of the dichloromethane to the methanol of the eluent is 1: 1; the eluent of the reverse phase silica gel column chromatography is methanol-water, and the volume ratio of the methanol to the water of the eluent is 55:45, 65:35, 75:25 and 85:15 in sequence; in the silica gel chromatographic column, the specification of the chromatographic column can be selected according to the routine in the field, and the inner diameter of the column multiplied by the length of the column is preferably 10cm multiplied by 100 cm; in the silica gel chromatographic column, the specification of silica gel can be selected according to the routine in the field, and is preferably 200-300 meshes; in the silica gel chromatographic column, the mass ratio of the silica gel to the crude extract can be selected according to the routine in the field, and is preferably 10; the ratio of the amount of each gradient eluent to the column volume can be selected in accordance with conventional practice in the art, and is preferably 3; the gel column may be selected according to the routine in the art; the reverse phase silica gel column may be selected as is conventional in the art.
The invention also provides an application of the Diels-Alder compound shown in the formula I or pharmaceutically acceptable salt or ester thereof in preparing an alpha-glucosidase inhibitor.
The invention also provides an application of the Diels-Alder compound shown in the formula I or the pharmaceutically acceptable salt or ester thereof in preparing a hypoglycemic medicament.
The invention also provides a pharmaceutical composition which comprises a therapeutically effective dose of the Diels-Alder compound shown in the formula I or a pharmaceutically acceptable salt or ester thereof and a pharmaceutically acceptable carrier.
Preferably, the pharmaceutical composition further comprises other hypoglycemic active ingredients and a pharmaceutically acceptable carrier; wherein, the other hypoglycemic active ingredients are preferably acarbose, voglibose or the combination thereof.
In a preferred embodiment, the mass ratio of the diels-alder type compound active ingredient shown in formula I to the other hypoglycemic active ingredients is preferably 1:100 to 100: 1, more preferably 1:10 to 10: 1.
in a preferred embodiment, the content of the diels-alder type compound represented by formula I or the pharmaceutically acceptable salt or ester thereof in the pharmaceutical composition may be 0.1-99.5% by mass, preferably 10-99.9% by mass, and more preferably 70-99.9% by mass.
In another preferred embodiment, the content of the diels-alder type compound represented by formula I or the pharmaceutically acceptable salt or ester thereof in the pharmaceutical composition may be 60.0-99.5% by mass, preferably 70-99.5% by mass, and more preferably 80-99.5% by mass.
The "pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances which are suitable for human use and must be of sufficient purity and sufficiently low toxicity. By "compatible" is meant herein that the components of the composition are capable of intermixing with and with the compounds of the present invention without significantly diminishing the efficacy of the compounds. Examples of pharmaceutically acceptable carrier moieties are cellulose and its derivatives (e.g., sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (e.g., stearic acid, magnesium stearate), calcium sulfate, vegetable oils (e.g., soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (e.g., propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers (e.g., propylene glycol, glycerin, mannitol, sorbitol, etc.), and the like
) Wetting agents (e.g., sodium lauryl sulfate), coloring agents, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, and the like.
The pharmaceutical composition of the present invention comprises a diels-alder type compound or a pharmacologically acceptable salt or ester thereof in a safe and effective amount range and a pharmacologically acceptable excipient or carrier. Wherein "safe and effective amount" means: the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. Generally, the pharmaceutical composition contains 1 to 2000mg, preferably 5 to 200mg, more preferably 10 to 100mg of the diels-alder type compound or a pharmacologically acceptable salt or ester thereof per dose. Said "dose" is preferably a capsule or tablet.
The mode of administration of the pharmaceutical composition of the present invention is not particularly limited, and representative modes of administration include (but are not limited to): oral, rectal, parenteral (intravenous, intramuscular or subcutaneous) and topical administration, preferably oral.
The solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient or carrier, such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and acacia; (c) humectants, for example, glycerol; (d) disintegrating agents, for example, agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) slow solvents, such as paraffin; (f) absorption accelerators, e.g., quaternary ammonium compounds; (g) wetting agents, such as cetyl alcohol and glycerol monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets and pills, the dosage forms may also comprise buffering agents.
Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared using coatings and shells such as enteric coatings and other materials well known in the art. They may contain opacifying agents and the release of the active compound or compounds in such compositions may be delayed in release in a certain part of the digestive tract. Examples of embedding components which can be used are polymeric substances and wax-like substances. If desired, the active compound may also be in microencapsulated form with one or more of the above-mentioned excipients.
The liquid dosage form for oral administration comprises pharmaceutically acceptable emulsion, solution, suspension, syrup or tincture. In addition to the active compounds, the liquid dosage forms may contain inert diluents commonly employed in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1, 3-butylene glycol, dimethylformamide and oils, in particular, cottonseed, groundnut, corn germ, olive, castor and sesame oils or mixtures of such materials and the like.
In addition to these inert diluents, the compositions can also contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.
Suspensions, in addition to the active compounds, may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol, sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures thereof and the like.
Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and nonaqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols and suitable mixtures thereof.
Dosage forms for topical administration include ointments, powders, patches, sprays, and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants which may be required if necessary.
The pharmaceutical compositions may be administered alone or in combination with other pharmaceutically acceptable compounds, such as acarbose.
When the pharmaceutical composition is used, a safe and effective amount of the compound of the present invention is suitable for mammals (such as human beings) to be treated, wherein the administration dose is a pharmaceutically effective administration dose, and for a human body with a weight of 60kg, the daily administration dose is usually 1 to 2000mg, preferably 5 to 500mg, and more preferably 10 to 100 mg. Of course, the particular dosage will depend upon such factors as the route of administration, the health of the patient, and the like, and is within the skill of the skilled practitioner.
A method of screening for hypoglycemic drug candidates comprising the steps of:
step 1: providing a test compound and a positive control compound, wherein the positive control compound is a Diels-Alder compound shown in a formula I or a pharmaceutically acceptable salt or ester thereof;
step 2: detecting the influence of the compound to be detected on the alpha-glucosidase in a test group, and comparing the test result with corresponding experimental results in a positive control group and a negative control group, wherein the influence of the positive control compound on the alpha-glucosidase is detected in the positive control group;
wherein if the degree of inhibition of the test compound on alpha-glucosidase is obviously higher than that of a negative control group, the test compound is suggested to be a candidate drug for reducing blood sugar.
In another preferred example, in step (2), the test group is compared with the positive control group, and the ratio of I1 to I2 is compared, wherein I1 is the degree of inhibition of the test compound on alpha-glucosidase, I2 is the degree of inhibition of the positive control compound on alpha-glucosidase, and if I1/I2 is more than or equal to 80%, the test compound is suggested to be the candidate drug for reducing blood sugar.
On the basis of the common knowledge in the field, the above preferred conditions can be combined randomly to obtain the preferred embodiments of the invention.
The reagents and starting materials used in the present invention are commercially available.
The positive progress effects of the invention are as follows: the Diels-Alder compound is an alpha-glucosidase inhibitor with a novel structure type, and can be used for preventing or treating diabetes. The Diels-Alder compound has high-efficiency alpha-glucosidase inhibition activity, which is about 840 times of that of the positive compound acarbose. The Diels-Alder compound can be extracted from various Morus plants, and has diversified sources and convenient acquisition.
Drawings
FIG. 1 is an ECD spectrum of Compound I-1 obtained in example 1
FIG. 2 is an ECD spectrum of Compound I-2 obtained in example 1
Detailed Description
The invention is further illustrated by the following examples, which are not intended to limit the scope of the invention. The experimental methods without specifying specific conditions in the following examples were selected according to the conventional methods and conditions, or according to the commercial instructions.
Example 1
The following examples all were statistically considered to be significantly different by P < 0.05.
Instrument and reagent
Nuclear magnetic resonance apparatus: bruker DRX 400 (germany), TMS internal standard; high resolution mass spectrometer: waters Xevo G2-XS (USA); medium-low pressure column chromatography (YMC corporation, japan); an enzyme-labeling instrument: Bio-TekSynergy2 (USA).
Silica gel (200-300 meshes), and the thin-layer chromatography analysis precast slab and the precast slab preparation are both produced by Qingdao ocean factories; c18 filler (japan YMC). All the reagents used were analytical grade (national pharmaceutical group chemical reagents, Inc.).
1) Extraction: pulverizing stem bark of Morus alba Linne (10kg), extracting with 95% ethanol under reflux for 3 times (1.5 hr each time) with the amount of ethanol 10(v/m) times of the amount of the medicinal materials, filtering the extractive solutions, mixing, and drying to obtain Morus alba Linne dry extract. Suspending the extract in 4 times of water, and extracting with petroleum ether, dichloromethane, ethyl acetate and n-butanol respectively to obtain different parts. The ethyl acetate fraction was concentrated under reduced pressure, and the dry weight was calculated to be 70 g.
2) Column chromatography: the ethyl acetate fractions obtained were subjected to silica gel column chromatography (10 times the volume of silica gel based on the mass of the sample, 100 cm. times.10 cm glass column), gradient-eluted with dichloromethane-methanol solutions (95:5 → 80:20 → 70:30 → 60:40 → 50:50) respectively, using 3 times the amount of the mixed solvent for each gradient, and collected and combined in order to obtain 10 large fractions. The fourth major fraction fr.d (6.0g) was taken to give secondary fractions D-g (58mg) and D-e (50mg) by gel column chromatography (dichloromethane: methanol: 1) and reverse phase silica gel column chromatography (methanol: water: 55:45 → 65:35 → 75:25 → 85:15) in this order.
3) HPLC preparation: HPLC preparation of the above secondary fraction D-g (58mg) (55% acetonitrile in water, detection wavelength 254nm, flow rate 2.5mL/min) provided Compound I-1: macrourin I (t)R20.3 min); will be at the topThe following secondary fraction D-e (50mg) was prepared by HPLC (56% acetonitrile in water, detection wavelength 254nm, flow rate 2.5mL/min) to give compound I-2: macrourin J (t)R＝18.5min)。
Compound I-1(macrourin I), a yellow amorphous powder, slightly soluble in water, soluble in acetone. HRESI-MS M/z 647.2288[ M-H ]]-Determining the molecular weight of the compound to be 648; [ alpha ] to]2 D 5+305.6°(c0.4,MeOH)；ECD(MeOH,nm)λmax(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-21″)，2.43(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″)。13The C-NMR data are shown in Table 1.
Compound I-2(macrourin J), a yellow amorphous powder, slightly soluble in water, soluble in acetone. HRESI-MS M/z 647.2285[ M-H ]]-Determining the molecular weight of the compound to be 648; [ alpha ] to]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)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(1H，dd，J＝12.8，2.4Hz，H-2″a)，2.19(1H，brs，H-6″a)，1.93(1H，brd，J＝12.8Hz，H-2″b)，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″)；13The C-NMR data are shown in Table 1.
TABLE 1 preparation of compounds I-1 to 213C-NMR data sheet
aNo signal was observed
bSignal conversion
Example 2
1. Instruments and materials
Enzyme labeling instrument (Bio-Tek Synergy2), centrifuge KA-1000 (Shanghai flying pigeon), double-person super clean bench (Suzhou purification), adjustable pipettor (Finland, 2-20uL, 20-200uL, 100-1000uL), 96-well plate (Corning, USA), inlet tip (Axygen, USA), and positive compound (acarbose).
2. Method of producing a composite material
1) Phosphate buffer (pH 7.6), p-nitrophenyl α -D-glucopyranoside, α -glucosidase and the test compound were mixed in sequence in a 250 μ L reaction system and incubated at 30 ℃ for 30 minutes.
2) The reaction was stopped by adding a solution of the terminating agent sodium carbonate.
3) Detection was performed using a microplate reader at a wavelength of 405 nm.
3. Sample to be tested, positive drug treatment method
The sample to be tested and acarbose were weighed out accurately and diluted to the working concentration series with DMSO before the experiment. The substrate p-nitrophenyl alpha-D-glucopyranoside was weighed 30.125mg as per instructions and added to a 50mL volumetric flask and diluted with assay buffer to make the working solution.
ACLight absorption value of control group, ABBlank group light absorption value; a. theS: the absorbance value of the sample set; a. theSB: absorbance of sample blank set
4. Results of the experiment
4.1 Primary Screen
TABLE 2 inhibition ratio prescreening table
4.1、IC50Value determination
TABLE 3 inhibitory Activity of Compounds I-1-2. alpha. -glucosidase IC50Watch (A)
Remarking: experimental replicates 3 times
The results are shown in tables 2 and 3. The result shows that the Diels-Alder type compound shown as the formula I has high-efficiency alpha-glucosidase inhibition activity which is about 840 times of that of the positive compound acarbose. Therefore, the Diels-Alder type I compound can prevent or treat diabetes, and can be developed into a novel high-efficiency alpha-glucosidase inhibitor.
Claims (8)
1. A Diels-Alder type compound of formula I:
wherein R is R1Or R2，
Represents R1And R2The position of attachment of the group to the parent.
2. A process for the preparation of a diels-alder type compound of formula I according to claim 1, comprising the steps of:
step 1: extracting milk mulberry serving as a raw material with an alcohol solution to obtain an extract, dispersing the extract with pure water, and extracting with an organic solvent to obtain a crude extract;
step 2: separating the crude extract obtained in the step 1 by column chromatography, and further purifying to obtain a Diels-Alder compound shown in the formula I, wherein the column chromatography separation is sequentially performed by forward silica gel column chromatography, gel column chromatography and reverse phase silica gel column chromatography, and the further purification is high performance liquid chromatography purification;
wherein, in the compound I, R is R1A compound of (II) and compound I wherein R is R2The test parameters for the compounds were as follows:
high performance liquid chromatography conditions: c18 bonded silica gel column, detecting wavelength of 254nm, flow rate of 2.5mL/min, mobile phase is acetonitrile water solution with volume content of 55%, and R in compound I is R1Has a retention time tR20.3 min; the mobile phase volume is acetonitrile water solution with the content of 56 percent, and R in the compound I is R2Has a retention time tR＝18.5min。
3. The process for the preparation of the diels-alder form of compound of formula I according to claim 2, wherein the forward silica gel column chromatography eluent is dichloromethane-methanol, the volume ratio of dichloromethane to methanol of said eluent is 95:5, 80:20, 70:30, 60:40 and 50:50 in sequence;
and/or the eluent of the gel column chromatography is dichloromethane-methanol, and the volume ratio of the dichloromethane to the methanol of the eluent is 1: 1;
and/or the eluent of the reverse phase silica gel column chromatography is methanol-water, and the volume ratio of the methanol to the water of the eluent is 55:45, 65:35, 75:25 and 85:15 in sequence.
4. The process for the preparation of the diels-alder form of compound of formula I according to claim 2, wherein the purification by hplc is carried out at a detection wavelength of 254nm and a flow rate of 2.5mL/min, the mobile phase is an aqueous acetonitrile solution with a volume content of 55%, the collection is carried out for a retention time tRGetting the compound I R as R in 20.3min1A compound of (1); the volume of the mobile phase is 56 percent of acetonitrile water solution, and the collection retention time is tRGetting the compound I R is R in 18.5min2The compound of (1).
5. Use of a diels-alder type compound of formula I according to claim 1 or a pharmaceutically acceptable salt thereof for the preparation of an α -glucosidase inhibitor.
6. Use of a diels-alder type compound of formula I according to claim 1 or a pharmaceutically acceptable salt thereof for the preparation of a medicament for lowering blood glucose.
7. A pharmaceutical composition comprising an effective amount of the diels-alder type compound of claim 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.
8. The pharmaceutical composition of claim 7, wherein said pharmaceutical composition further comprises an additional hypoglycemic active ingredient; the other active ingredients for reducing blood sugar are acarbose, voglibose or the combination thereof.
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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 |
|---|
| Diels–Alder adducts with PTP1B inhibition from Morus notabilis;Meng Wang,et al.;《Phytochemistry》;20141107;第109卷;第140-146页 * |
| 广东桑叶化学成分及其α-葡萄糖苷酶活性研究;唐本钦 等;《中草药》;20131130;第44卷(第22期);第3109-3113页 * |
| 桑属植物的化学成分及药理活性综述;何雪梅 等;《蚕业科学》;20141231;第40卷(第2期);第328-338页 * |
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| CN110218208A (en) | 2019-09-10 |
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