CN104490894A - Preparation method of spermacoce latifolia triterpenoids and application of spermacoce latifolia triterpenoid in preparation of glycosidase inhibitor medicine - Google Patents
Preparation method of spermacoce latifolia triterpenoids and application of spermacoce latifolia triterpenoid in preparation of glycosidase inhibitor medicine Download PDFInfo
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Abstract
The invention provides a preparation method of spermacoce latifolia triterpenoids and an application of the spermacoce latifolia triterpenoids in preparation of a glycosidase inhibitor medicine. Six compounds for intensively inhibiting the activity of alpha-glycosidase provided by the invention are natural compounds which are high in safety, and can be naturally degraded rapidly without a residue in the environment. The compounds can be separated from plant materials such as spermacoce latifolia; the plant materials are abundant in source; and the preparation process is easy to operate. Monomers of the six compounds are relatively stable and easy to store; the alpha-glycosidase inhibiting activity is obviously higher than that of clinical medicine acarbose; and the compounds are extremely likely to be further developed into the effective and safe alpha-glycosidase inhibitor medicines for preventing and treating type 2 diabetes mellitus, and have relatively good prospects.
Description
Technical field
The invention belongs to field of natural medicinal chemistry, be specifically related to six kinds of triterpenoid compound and mesembryanthemoidigenicacid (1), 29-hydroxyhederagenin (2), 3 β, 6 β-dihydroxy-olean-12-ene-28-oic acid (3), scutellaric acid (4), arjunic acid (5), 3 β, 6 β, the application in glycosidase inhibitor or medicine prepared by 23-trihydroxy-olean-12-en-28-oic acid (6) or its pharmaceutically useful salt, and their preparation method.
Background of invention
Diabetes are endocrine metabolism dysfunctional disease common clinically, and itself and cardiovascular disease and cancer etc. are occurred frequently year by year has important dependency, are the important killers of human health.At present, the sickness rate of diabetes improves in the world, has the prevalence more than 100,000,000 people especially in China, and presents the trend increased year by year.Diabetes cause more and more great loss just to our people's health and national economy.
Diabetes doctor trained in Western medicine be divided into type Ⅰ diabetes mellitus (or claim insulin-dependent, DM1) and type Ⅱdiabetes mellitus (or claim non-insulin-dependent, DM2), wherein type Ⅱdiabetes mellitus morbidity is with prevalence all far above type Ⅰ diabetes mellitus, thus endangers larger.Competitive alpha-glucosidase inhibitor has postpones glucide and digests and assimilates, controls blood glucose after meal and sharply raise and then make blood sugar concentration change the functions such as fluctuating margin reduction in one day, is to develop the potential drug being used for the treatment of type Ⅱdiabetes mellitus.The important alpha-glucosidase inhibitor having developed listing positive clinic trial do treatment type Ⅱdiabetes mellitus at present comprises a line medication acarbose (acarbose), voglibose, miglitol and emigliate etc.
Six kinds of triterpenoid mesembryanthemoidigenic acid (1), 29-hydroxyhederagenin (2), 3 β, 6 β-dihydroxy-olean-12-ene-28-oic acid (3), scutellaric acid (4), arjunic acid (5), 3 β, 6 β, 23-trihydroxy-olean-12-en-28-oic acid (6) was once separated to obtain respectively and (specifically saw document: Magina et al. in other plant, Quim Nova, 2012,35:1184-1188; Huu et al., J Chem, 2007,45:363-367; Kuo et al., Chem Pharm Bull, 1988,36:3619-3622; Ponouet al., Chem Biodivers, 2011,8:1301-1309; Lavaudet al., Phytochemistry, 2001,57:469-478; Khan et al., J Nat Prod, 1993,56:2163-2165), shown in their the following formula I of chemical structural formula, yet there are no the report that these compounds have glucosidase inhibitor activity up till now, and have no the report that it is separated acquisition in Spermacoce latifolia (Spermacoce latifolia) plant.
Summary of the invention
First object of the present invention is to provide compound mesembryanthemoidigenic acid (1), 29-hydroxyhederagenin (2), 3 β, 6 β-dihydroxy-olean-12-ene-28-oic acid (3), scutellaric acid (4), arjunic acid (5), 3 β, 6 β, 23-trihydroxy-olean-12-en-28-oic acid (6) or their officinal salt, preparing the application in alpha-glucosidase inhibitor medicine.
Confirm through external pharmacological evaluation, compound mesembryanthemoidigenic acid (1) provided by the invention, 29-hydroxyhederagenin (2), 3 β, 6 β-dihydroxy-olean-12-ene-28-oic acid (3), scutellaric acid (4), arjunic acid (5) or 3 β, 6 β, 23-trihydroxy-olean-12-en-28-oic acid (6) all has the activity of potent Inhibiting α-glucosidase, the activity of their Inhibiting α-glucosidases is all significantly better than diabetes one line medication acarbose, therefore can develop and cause for the preparation of prevention and therapy alpha-glucosidase or the potential drug candidate molecules of relevant physiological change or disease.Wherein, to cause with alpha-glucosidase or relevant physiological change or disease include but not limited to type Ⅱdiabetes mellitus.
The pharmaceutically useful salt of six kinds of compounds of the present invention; it can be converted into the corresponding triterpenoid molecule shown in above formula I in people's digestive tract under the physiological conditions such as gastric acid; the active component of its substantive Inhibiting α-glucosidase is identical with six kinds of triterpenoid molecules shown in above, thus belongs to strict protection scope of the present invention.
Above-described six kinds of triterpenoid molecules of the present invention or its pharmaceutically useful salt can be combined with pharmaceutically conventional adjuvant or carrier, prepare the medicine that can be used for prevention and therapy type Ⅱdiabetes mellitus or the pharmaceutical composition of any one or the multiple suppression alpha-glucosidase activity with the above six kinds of compound molecule.This medicine or pharmaceutical composition can adopt the dosage forms such as wettable powder, tablet, granule, capsule, oral liquid, drop pill, also can adopt the known controlled release of pharmaceutical industry or slow release formulation or nanometer formulation.
The application in alpha-glucosidase inhibitor medicine is being prepared with one or more extracts comprising the plants such as Spermacoce latifolia being effective ingredient in above six kinds of triterpenoid molecules of the present invention; be be substantive effective ingredient with one or more in six kinds of compound molecules of the present invention because of it, thus belong to strict protection scope category of the present invention.
Second object of the present invention is to provide compound mesembryanthemoidigenic acid (1), 29-hydroxyhederagenin (2), 3 β, 6 β-dihydroxy-olean-12-ene-28-oic acid (3), scutellaric acid (4), arjunic acid (5), 3 β, 6 β, the preparation method of 23-trihydroxy-olean-12-en-28-oic acid (6), it is characterized in that, described compound mesembryanthemoidigenic acid (1), 29-hydroxyhederagenin (2), 3 β, 6 β-dihydroxy-olean-12-ene-28-oic acid (3), scutellaric acid (4), arjunic acid (5), 3 β, 6 β, 23-trihydroxy-olean-12-en-28-oic acid (6) is separated and obtains from Spermacoce latifolia (Spermacoce latifolia Aubl.K.Schum.) plant.
Preferably, concrete steps are:
Extract with ethanol or ethanol water after the Spermacoce latifolia Herb of drying is pulverized, extracting solution obtains extractum after concentrated, extractum adds water suspendible, uses petroleum ether and extraction into ethyl acetate successively, obtains petroleum ether portion and ethyl acetate extraction part respectively after concentrated; Petroleum ether portion is through purification on normal-phase silica gel column chromatography, with petroleum ether/acetone from volume ratio 100:0-0-100 gradient elution, collect the fraction E6 of petroleum ether-acetone volume ratio 10:1 eluting, fraction E6 is through MCI post removing pigment, methanol-eluted fractions E6-1 is obtained by methanol-eluted fractions, E6-1 is successively by anti-phase medium pressure column chromatography, methanol/water volume ratio is from 30:70-100:0 gradient elution, Sephadex LH-20 gel column, with methanol-eluted fractions, with purification on normal-phase silica gel chromatography, with chloroform/methanol volume ratio 100:1 eluting, obtain compound 1;
Ethyl acetate extraction part, through purification on normal-phase silica gel column chromatography, with chloroform/methanol from volume ratio 100:1-0:100 gradient elution, collects the fraction F of chloroform/methanol volume ratio 25:1 eluting
5, F
5through anti-phase medium pressure column chromatography, methanol/water is from volume ratio 30:70-100:0, and gradient elution, collects the fraction F of methanol/water volume ratio 80:20 eluting
5-4with the fraction F of methanol/water volume ratio 90:10 eluting
5-5; Fraction F
5-4through Sephadex LH-20 gel column with methanol-eluted fractions, then through the separation and purification of purification on normal-phase silica gel chromatographic column, with chloroform/methanol volume ratio 50:1 eluting, obtain compound 3,4,5 and 6; Fraction F
5-5through Sephadex LH-20 gel column with methanol-eluted fractions, obtain compound 2.
3rd object of the present invention is to provide Spermacoce latifolia and is preparing compound mesembryanthemoidigenic acid (1), 29-hydroxyhederagenin (2), 3 β, 6 β-dihydroxy-olean-12-ene-28-oic acid (3), scutellaric acid (4), arjunic acid (5) or 3 β, application in 6 β, 23-trihydroxy-olean-12-en-28-oic acid (6).
The compound of six kinds of tool potent suppression alpha-glucosidase activity of the present invention is that safety is high and in the environment can the native compound of rapid and natural degraded noresidue, they can be separated acquisition in the vegetable materials such as Spermacoce latifolia, source of plant material is enriched, and preparation process is easy to operation.The monomer of six kinds of described compounds is all comparatively stable, easy to store, their alpha-glucoside inhibiting activity is significantly better than or is equivalent to clinical application acarbose, be further development of the alpha-glucosidase inhibitor class medicine of effective, safe prevention and therapy type Ⅱdiabetes mellitus most probably, there is better prospect.
Accompanying drawing illustrates:
Fig. 1 is compound mesembryanthemoidigenic acid (1)
13c NMR collection of illustrative plates.
Fig. 2 is compound 29-hydroxyhederagenin (2)
13c NMR collection of illustrative plates.
Fig. 3 is compound 3 β, 6 β-dihydroxy-olean-12-ene-28-oic acid's (3)
13c NMR collection of illustrative plates.
Fig. 4 is compound scutellaric acid (4)
13c NMR collection of illustrative plates.
Fig. 5 is compound a rjunic acid (5)
1h NMR collection of illustrative plates.
Fig. 6 is compound 3 β, 6 β, 23-trihydroxy-olean-12-en-28-oic acid's (6)
13c NMR collection of illustrative plates.
Detailed description of the invention:
Following examples further illustrate of the present invention, instead of limitation of the present invention, and essence according to the present invention all belongs to the scope of protection of present invention to the simple modifications that the present invention carries out.
Embodiment 1: the preparation of six kinds of triterpenoids in Spermacoce latifolia plant
1.1 plant origins and qualification
Spermacoce latifolia (Spermacoce latifolia Aubl.K.Schum.) plant Herb picked up from South China Botanical Garden, Guangzhou, Guangdong in 2012 09 month, identified through South China Botanical Garden Chinese Academy of Sciences Xing Fuwu researcher.
1.2 Extraction and isolation
Dry Spermacoce latifolia Herb (5.0Kg) is extracted in the ethanol water of room temperature by volume fraction 95% after pulverizing, and extracting solution merges afterwards that concentrating under reduced pressure must without alcohol extractum.Extractum adds water suspendible, uses petroleum ether and each 4 times of extraction into ethyl acetate successively; Petroleum ether portion (330g) and ethyl acetate extraction part (202g) is obtained after concentrating under reduced pressure.Institute's petroleum ether portion that obtains (330g) is through purification on normal-phase silica gel column chromatography (200-300 order, 1000 × 105mm i.d.), with petroleum ether-acetone (100:0-0:100, v/v, each 3.0L) gradient elution, through the flow point that TLC thin layer chromatography combining data detection principal point is identical, obtain 9 component (E altogether
1-E
9).E
6(5.0g) [fraction of petroleum ether-acetone (10:1, v/v) eluting] removes pigment through MCI post [200 × 40mm i.d.], obtains methanol-eluted fractions E by methanol-eluted fractions
6-1.E
6-1(3.0g) successively by anti-phase medium pressure column chromatography separation and purification, methanol/water (30:70-100:0, v/v, each 1.0L) for eluent gradient eluting, Sephadex LH-20 gel column (1550 × 13.4mm i.d.) are with methanol-eluted fractions and purification on normal-phase silica gel chromatographic column (200-300 order, 400 × 30mm i.d) separation and purification, with chloroform-methanol (100:1, v/v) constant gradient eluting, obtain compound 1 (3.4mg).Ethyl acetate extraction part (202g) is through purification on normal-phase silica gel column chromatography (200-300 order, 1000 × 105mm i.d.), with chloroform-methanol (100:1-0:100, v/v, each 3.0L) gradient elution, through the flow point that TLC thin layer chromatography combining data detection principal point is identical, obtain F
1-F
9totally 9 components.F
5(3.3g) [chloroform-methanol (25:1, v/v) fraction of eluting] continue through anti-phase medium pressure column chromatography separation and purification, methanol/water (30:70-100:0, v/v, each 1.0L) be eluent gradient eluting, the flow point that TLC thin layer combining data detection principal point is identical, obtains 6 subfraction (F
5-1-F
5-6).F
5-4(2.5g) [methanol/water (80:20, v/v) fraction of eluting] through Sephadex LH-20 gel column with methanol-eluted fractions, again through purification on normal-phase silica gel chromatographic column (200-300 order, 400 × 30mm i.d.) separation and purification, with chloroform-methanol (50:1, v/v) constant gradient eluting, obtain compound 3 (5.6mg), 4 (3.6mg), 5 (5.0mg) and 6 (4.0mg).F
5-5(1.0g) [fraction of methanol/water (90:10, v/v) eluting] is through Sephadex LH-20 gel column with methanol-eluted fractions, obtains compound 2 (4.2mg).
The Structural Identification of 1.3 compounds
Institute's compound that obtains 1 is white powder, ESI-MS m/z 495 [M+Na]
+, 471 [M-H]
–to should the molecular formula C of compound
30h
48o
4; Its
13c NMR collection of illustrative plates as shown in Figure 1,
1h-NMR (500MHz, CD
3oD): δ 5.25 (1H, t, J=3.5Hz, H-12), 3.19 (2H, s, H-29), 3.15 (1H, dd, J=11.4,4.7Hz, H-3), 2.88 (1H, dd, J=13.0,4.7Hz, H-18), 1.17 (3H, s), 0.97 (3H, s), 0.94 (3H, s), 0.93 (3H, s), 0.82 (3H, s), 0.78 (3H, s);
13c-NMR (125MHz, CD
3oD): δ 38.5 (CH
2, C-1), 27.6 (CH
2, C-2), 79.7 (CH, C-3), 38.9 (C, C-4), 56.7 (CH, C-5), 18.3 (CH
2, C-6), 32.9 (CH
2, C-7), 39.8 (C, C-8), 46.5 (CH, C-9), 36.8 (C, C-10), 23.2 (CH
2, C-11), 123.7 (CH, C-12), 145.2 (C, C-13), 42.7 (C, C-14), 28.7 (CH
2, C-15), 24.0 (CH
2, C-16), 47.6 (C, C-17), 40.6 (CH, C-18), 40.3 (CH
2, C-19), 35.5 (C, C-20), 29.6 (CH
2, C-21), 38.0 (CH
2, C-22), 28.3 (CH
3, C-23), 16.3 (CH
3, C-24), 15.9 (CH
3, C-25), 17.7 (CH
3, C-26), 25.4 (CH
3, C-27), 181.9 (C, C-28), 74.4 (CH
2, C-29), 19.5 (CH
3, C-30).Comprehensively analyze according to above spectral data, the chemical constitution parsing this compound 1 is mesembryanthemoidigenic acid, and its structural formula is as shown in 1 in formula I.
Institute's compound that obtains 2 is white powder, ESI-MS m/z 511 [M+Na]
+, 487 [M-H]
–, to should compound molecule formula C
30h
48o
5; Its
13c NMR collection of illustrative plates as shown in Figure 2,
1h-NMR (500MHz, CD
3oD): δ 5.26 (1H, t, J=3.5Hz, H-12), 3.60 (1H, dd, J=11.9,4.2Hz, H-3), 3.53,3.33, (each 1H, d, J=11.0Hz, H-23), 3.19 (2H, s, H-29), 2.88 (1H, dd, J=13.9,4.3Hz, H-18), 1.19 (3H, s), 0.98 (3H, s), 0.93 (3H, s), 0.82 (3H, s), 0.70 (3H, s);
13c-NMR (125MHz, CD
3oD): δ 38.2 (CH
2, C-1), 26.1 (CH
2, C-2), 75.3 (CH, C-3), 40.6 (C, C-4), 49.5 (CH, C-5), 18.7 (CH
2, C-6), 32.6 (CH
2, C-7), 39.7 (C, C-8), 47.4 (CH, C-9), 36.7 (C, C-10), 22.8 (CH
2, C-11), 122.8 (CH, C-12), 144.5 (C, C-13), 41.4 (C, C-14), 27.7 (CH
2, C-15), 23.2 (CH
2, C-16), 47.7 (C, C-17), 40.7 (CH, C-18), 40.7 (CH
2, C-19), 35.6 (C, C-20), 28.8 (CH
2, C-21), 32.1 (CH
2, C-22), 68.3 (CH
2, C-23), 11.6 (CH
3, C-24), 15.3 (CH
3, C-25), 16.8 (CH
3, C-26), 25.4 (CH
3, C-27), 182.7 (C, C-28), 73.4 (CH
2, C-29), 18.8 (CH
3, C-30).Comprehensively analyze according to above spectral data, the chemical constitution parsing this compound 2 is 29-hydroxyhederagenin, and its structural formula is as shown in 2 in formula I.
Institute's compound that obtains 3 is white powder, ESI-MS m/z 495 [M+Na]
+, 471 [M-H]
–, to should compound molecule formula C
30h
48o
4; Its
13c NMR collection of illustrative plates as shown in Figure 3,
1h-NMR (500MHz, CD
3oD): δ 5.30 (1H, t, J=3.6Hz, H-12), 4.51 (1H, br s, H-6), 3.10 (1H, dd, J=11.6,4.1Hz, H-3), 2.89 (1H, dd, J=13.7,4.2Hz, H-18), 1.32 (3H, s), 1.18 (3H, s), 1.15 (3H, s), 1.11 (3H, s), 1.06 (3H, s), 0.96 (3H, s), 0.93 (3H, s);
13c-NMR (125MHz, CD
3oD): δ 42.0 (CH
2, C-1), 28.1 (CH
2, C-2), 80.1 (CH, C-3), 40.1 (C, C-4), 57.1 (CH, C-5), 68.7 (CH, C-6), 41.6 (CH
2, C-7), 40.7 (C, C-8), 49.4 (CH, C-9), 37.7 (C, C-10), 24.5 (CH
2, C-11), 123.9 (CH, C-12), 144.5 (C, C-13), 43.4 (C, C-14), 28.7 (CH
2, C-15), 24.0 (CH
2, C-16), 47.7 (C, C-17), 40.6 (CH, C-18), 47.3 (CH
2, C-19), 30.5 (C, C-20), 33.6 (CH
2, C-21), 32.0 (CH
2, C-22), 28.3 (CH
3, C-23), 17.6 (CH
3, C-24), 17.3 (CH
3, C-25), 18.8 (CH
3, C-26), 26.4 (CH
3, C-27), 181.9 (C, C-28), 23.9 (CH
3, C-29), 31.8 (CH
3, C-30).Comprehensively analyze according to above spectral data, the chemical constitution parsing this compound 3 is 3 β, and 6 β-dihydroxy-olean-12-ene-28-oic acid, its structural formula is as shown in 3 in formula I.
Institute's compound that obtains 4 is white powder, ESI-MS m/z 495 [M+Na]
+, 471 [M-H]
–, to should compound molecule formula C
30h
48o
4; Its
13c NMR collection of illustrative plates as shown in Figure 4,
1h-NMR (500MHz, CD
3oD): δ 5.26 (1H, t, J=3.6Hz, H-12), 3.63 (1H, m, H-3), 3.70,3.44, (each 1H, d, J=11.0Hz, H-23), 2.87 (1H, dd, J=13.8,4.2Hz, H-18), 1.15 (3H, s), 1.10 (3H, s), 1.06 (3H, s), 0.93 (3H, s), 0.87 (3H, s), 0.72 (3H, s);
13c-NMR (125MHz, CD
3oD): δ 33.2 (CH
2, C-1), 26.1 (CH
2, C-2), 73.9 (CH, C-3), 40.6 (C, C-4), 43.5 (CH, C-5), 18.7 (CH
2, C-6), 33.6 (CH
2, C-7), 39.7 (C, C-8), 49.4 (CH, C-9), 37.7 (C, C-10), 23.5 (CH
2, C-11), 122.8 (CH, C-12), 144.5 (C, C-13), 42.4 (C, C-14), 28.7 (CH
2, C-15), 24.0 (CH
2, C-16), 47.7 (C, C-17), 42.1 (CH, C-18), 46.3 (CH
2, C-19), 31.5 (C, C-20), 34.2 (CH
2, C-21), 33.1 (CH
2, C-22), 68.3 (CH
2, C-23), 18.6 (CH
3, C-24), 15.3 (CH
3, C-25), 17.8 (CH
3, C-26), 26.4 (CH
3, C-27), 181.7 (C, C-28), 23.9 (CH
3, C-29), 33.8 (CH
3, C-30).Comprehensively analyze according to above spectral data, the chemical constitution parsing this compound 4 is scutellaric acid, and its structural formula is as shown in 4 in formula I.
Institute's compound that obtains 5 is white powder, ESI-MS m/z 511 [M+Na]
+, 487 [M-H]
–, to should compound molecule formula C
30h
48o
5; Its
1h NMR collection of illustrative plates as shown in Figure 5,
1h-NMR (500MHz, CD
3oD): δ 5.31 (1H, t, J=3.6Hz, H-12), 3.62 (1H, m, H-2), 3.24 (1H, d, J=3.7Hz, H-19), 3.05 (1H, d, J=3.7Hz, H-18), 2.90 (1H, d, J=9.6Hz, H-3), 1.31 (3H, s), 1.03 (3H, s), 1.00 (3H, s), 0.96 (3H, s), 0.93 (3H, s), 0.84 (3H, s), 0.76 (3H, s);
13c-NMR (125MHz, CD
3oD): δ 48.2 (CH
2, C-1), 69.6 (CH, C-2), 84.9 (CH, C-3), 40.6 (C, C-4), 57.0 (CH, C-5), 19.7 (CH
2, C-6), 34.0 (CH
2, C-7), 40.9 (C, C-8), 49.4 (CH, C-9), 39.7 (C, C-10), 25.1 (CH
2, C-11), 124.8 (CH, C-12), 144.9 (C, C-13), 42.7 (C, C-14), 29.7 (CH
2, C-15), 28.6 (CH
2, C-16), 46.7 (C, C-17), 45.1 (CH, C-18), 82.3 (CH, C-19), 36.2 (C, C-20), 29.5 (CH
2, C-21), 34.1 (CH
2, C-22), 29.3 (CH
3, C-23), 17.5 (CH
3, C-24), 17.1 (CH
3, C-25), 17.8 (CH
3, C-26), 25.4 (CH
3, C-27), 181.7 (C, C-28), 25.2 (CH
3, C-29), 28.8 (CH
3, C-30).Comprehensively analyze according to above spectral data, the chemical constitution parsing this compound 5 is arjunic acid, and its structural formula is as shown in 5 in formula I.
Institute's compound that obtains 6 is white powder, ESI-MS m/z 511 [M+Na]
+, 487 [M-H]
–, to should compound molecule formula C
30h
48o
5; Its
13c NMR collection of illustrative plates as shown in Figure 6,
1h-NMR (500MHz, CD
3oD): δ 5.29 (1H, t, J=3.6Hz, H-12), 4.39 (1H, br s, H-6), 3.56 (1H, m, H-3), 3.60,3.47, (each 1H, d, J=11.0Hz, H-23), 2.87 (1H, dd, J=13.7,3.9Hz, H-18), 1.33 (3H, s), 1.14 (3H, s), 1.10 (3H, s), 1.06 (3H, s), 0.96 (3H, s), 0.91 (3H, s);
13c-NMR (125MHz, CD
3oD): δ 41.2 (CH
2, C-1), 28.1 (CH
2, C-2), 73.5 (CH, C-3), 44.0 (C, C-4), 49.5 (CH, C-5), 68.6 (CH, C-6), 41.0 (CH
2, C-7), 39.3 (C, C-8), 48.7 (CH, C-9), 36.9 (C, C-10), 23.7 (CH
2, C-11), 122.8 (CH, C-12), 144.5 (C, C-13), 42.4 (C, C-14), 28.7 (CH
2, C-15), 24.0 (CH
2, C-16), 46.7 (C, C-17), 42.1 (CH, C-18), 46.3 (CH
2, C-19), 31.0 (C, C-20), 34.2 (CH
2, C-21), 33.1 (CH
2, C-22), 66.7 (CH
2, C-23), 14.6 (CH
3, C-24), 17.5 (CH
3, C-25), 18.8 (CH
3, C-26), 26.4 (CH
3, C-27), 181.9 (C, C-28), 23.7 (CH
3, C-29), 33.2 (CH
3, C-30).Comprehensively analyze according to above spectral data, the chemical constitution parsing this compound 6 is 3 β, and 6 β, 23-trihydroxy-olean-12-en-28-oic acid, its structural formula is as shown in 6 in formula I.
Embodiment 2: the alpha-glucoside inhibiting activity of described six kinds of triterpenoids detects
2.1 instruments and reagent
Experimental apparatus: microplate reader Genois microplate reader (Tecan GENios, Swizerland).
Reagent sample: alpha-glucosidase is purchased from Sigma Chemical Co. (Sigma-Aldrich, St.Louis, USA); Acarbose (Acarbose) is purchased from Tokyo Chemical Industry Co., Ltd. (Japan); 4-nitrophenols-alpha-D-glucose pyrans glycosides (PNPG) is purchased from Tokyo Chemical Industry Co., Ltd. (Japan); Six kinds of triterpenoid mesembryanthemoidigenic acid (compound 1), 29-hydroxyhederagenin (compound 2), 3 β, 6 β-dihydroxy-olean-12-ene-28-oic acid (compound 3), scutellaric acid (compound 4), arjunic acid (compound 5), 3 β, 6 β, 23-trihydroxy-olean-12-en-28-oic acid (compound 6) is prepared by the method for above experimental example 1, also can according to document Magina et al., Quim Nova, 2012,35:1184-1188; Huu et al., J Chem, 2007,45:363-367; Kuo et al., Chem Pharm Bull, 1988,36:3619-3622; Ponouet al., Chem Biodivers, 2011,8:1301-1309; Lavaud et al., Phytochemistry, 2001,57:469-478; Khan et al., J Nat Prod, the method in 1993,56:2163-2165 prepares.
2.2 method of testings:
A) compounding pharmaceutical solution: solution testing compound and acarbose being prepared respectively 10mg/ml by dimethyl sulfoxide (DMSO), and prepare the phosphate buffer (ultra-pure water preparation) of 67mmol/L, the NaCO of PNPG substrate solution (5mM, phosphate buffer is prepared) and 0.2M
3solution (phosphate buffer preparation).
B) adopt colorimetry, measured with regard to the half-inhibition concentration of testing compound to alpha-glucosidase by 96 orifice plates.First the alpha-glucosidase (0.8U) of 20 μ l is added sample well, then sample solution phosphate buffer is diluted in proportion, every hole adds sample solution 120 μ l, the ultimate density of test sample (comprising testing compound and acarbose) is made to be: 500 μ g/mL, 250 μ g/mL, 125 μ g/mL, 62.5 μ g/mL, 31.25 μ g/mL, 15.625 μ g/mL, finally add reaction substrate 4-nitrophenols-α-D-pyranglucoside 20 μ l (5mM) again.After 37 DEG C of water-bath 15min, in each sample well, add the Na of 80 μ l
2cO
3(0.2M) cessation reaction, in the place's colorimetric determination of 405nm wavelength.The phosphate buffer of same volume replaces enzymatic solution.Compound suppression ratio is calculated for blank and contrast OD value by sample OD value, and computing formula is as follows: suppression ratio (%)=(OD
control– OD
neg)-(OD
test– OD
test control)/(OD
control– OD
neg) × 100%.Wherein test six kinds of compounds respectively to the half-inhibition concentration (IC of alpha-glucosidase
50) obtained by dose effect curve.
2.3 experimental datas are see table 1:
The a-glucosidase inhibitory active of table 1. six kinds of triterpenoids
2.4 experiment conclusion:
A-glucosidase is the index tested enzyme of a-glycosidase inhibitor class type Ⅱdiabetes mellitus medicine screening.This experimental result shows, compound mesembryanthemoidigenic acid (compound 1) provided by the invention, 29-hydroxyhederagenin (compound 2), 3 β, 6 β-dihydroxy-olean-12-ene-28-oic acid (compound 3), scutellaric acid (compound 4), arjunic acid (compound 5) and 3 β, 6 β, 23-trihydroxy-olean-12-en-28-oic acid (compound 6) all has the activity being significantly better than diabetes one line medication acarbose Inhibiting α-glucosidase, therefore can develop and cause for the preparation of prevention and therapy alpha-glucosidase or the potential drug candidate molecules of relevant physiological change or disease, there is stronger application and development potential quality, be expected the medication that can further develop the prevention and therapy type Ⅱdiabetes mellitus becoming new, application potential quality is extensive.
Claims (6)
1. six kinds of triterpenoid mesembryanthemoidigenic acid, 29-hydroxyhederagenin, 3 β, 6 β-dihydroxy-olean-12-ene-28-oic acid, scutellaric acid, arjunic acid or 3 β, 6 β, 23-trihydroxy-olean-12-en-28-oic acid, or their officinal salt, preparing the application in alpha-glucosidase inhibitor or medicine, described six kinds of triterpenoid structures are as shown in formula I:
Wherein compound 1 is mesembryanthemoidigenic acid; Compound 2 is 29-hydroxyhederagenin; Compound 3 is 3 β, 6 β-dihydroxy-olean-12-ene-28-oic acid; Compound 4 is scutellaric acid; Compound 5 is arjunic acid; Compound 6 is 3 β, 6 β, 23-trihydroxy-olean-12-en-28-oic acid.
2. application according to claim 1, is characterized in that: described alpha-glucosidase inhibitor or medicine, and its dosage form is wettable powder, tablet, granule, capsule, oral liquid, drop pill, controlled release or slow release formulation or nanometer formulation.
3. an alpha-glucosidase inhibitor or medicine, it is characterized in that, include triterpenoid mesembryanthemoidigenic acid, 29-hydroxyhederagenin, 3 β of effective amount, 6 β-dihydroxy-olean-12-ene-28-oic acid, scutellaric acid, arjunic acid or 3 β, 6 β, one or more in 23-trihydroxy-olean-12-en-28-oic acid, or their officinal salt, and pharmaceutically acceptable carrier or adjuvant.
4. a compound mesembryanthemoidigenic acid, 29-hydroxyhederagenin, 3 β, 6 β-dihydroxy-olean-12-ene-28-oic acid, scutellaric acid, arjunic acid, 3 β, 6 β, the preparation method of 23-trihydroxy-olean-12-en-28-oic acid, it is characterized in that, described compound mesembryanthemoidigenic acid, 29-hydroxyhederagenin, 3 β, 6 β-dihydroxy-olean-12-ene-28-oic acid, scutellaric acid, arjunic acid, 3 β, 6 β, 23-trihydroxy-olean-12-en-28-oic acid is separated and obtains from Spermacoce latifolia (Spermacoce latifolia Aubl.K.Schum.) plant.
5. preparation method according to claim 4, is characterized in that, concrete steps are:
Extract with ethanol or ethanol water after the Spermacoce latifolia Herb of drying is pulverized, extracting solution obtains extractum after concentrated, extractum adds water suspendible, uses petroleum ether and extraction into ethyl acetate successively, obtains petroleum ether portion and ethyl acetate extraction part respectively after concentrated; Petroleum ether portion is through purification on normal-phase silica gel column chromatography, with petroleum ether/acetone from volume ratio 100:0-0-100 gradient elution, collect the fraction E6 of petroleum ether-acetone volume ratio 10:1 eluting, fraction E6 is through MCI post removing pigment, methanol-eluted fractions E6-1 is obtained by methanol-eluted fractions, E6-1 is successively by anti-phase medium pressure column chromatography, methanol/water volume ratio is from 30:70-100:0 gradient elution, Sephadex LH-20 gel column, with methanol-eluted fractions, with purification on normal-phase silica gel chromatography, with chloroform/methanol volume ratio 100:1 eluting, obtain compound 1;
Ethyl acetate extraction part, through purification on normal-phase silica gel column chromatography, with chloroform/methanol from volume ratio 100:1-0:100 gradient elution, collects the fraction F of chloroform/methanol volume ratio 25:1 eluting
5, F
5through anti-phase medium pressure column chromatography, methanol/water is from volume ratio 30:70-100:0, and gradient elution, collects the fraction F of methanol/water volume ratio 80:20 eluting
5-4with the fraction F of methanol/water volume ratio 90:10 eluting
5-5; Fraction F
5-4through Sephadex LH-20 gel column with methanol-eluted fractions, then through the separation and purification of purification on normal-phase silica gel chromatographic column, with chloroform/methanol volume ratio 50:1 eluting, obtain compound 3,4,5 and 6; Fraction F
5-5through Sephadex LH-20 gel column with methanol-eluted fractions, obtain compound 2.
6. Spermacoce latifolia is preparing compound mesembryanthemoidigenic acid, 29-hydroxyhederagenin, 3 β, 6 β-dihydroxy-olean-12-ene-28-oic acid, scutellaric acid, arjunic acid or 3 β, application in 6 β, 23-trihydroxy-olean-12-en-28-oic acid.
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| CN105949191A (en) * | 2016-07-18 | 2016-09-21 | 唐翔 | Technology for extracting borreria stricta alkali |
| CN106265681A (en) * | 2016-07-20 | 2017-01-04 | 中国科学院华南植物园 | Compound 2 α, 3 β dihydroxy 23 aldehyde radical olive 12 alkene 28 acid application in preparing glycosidase inhibitor |
| CN108314616A (en) * | 2017-01-17 | 2018-07-24 | 浙江工业大学 | Triterpene compound and its preparation and application |
| CN110563794A (en) * | 2019-09-27 | 2019-12-13 | 桂林电子科技大学 | Myrtle triterpene lactone A as well as extraction method and application thereof |
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| CN103622972A (en) * | 2013-12-04 | 2014-03-12 | 中国科学院华南植物园 | Preparation method for compound 3 alpha-Akebonolic acid and application of compound to preparation of glycosidase inhibitor drug |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105949191A (en) * | 2016-07-18 | 2016-09-21 | 唐翔 | Technology for extracting borreria stricta alkali |
| CN106265681A (en) * | 2016-07-20 | 2017-01-04 | 中国科学院华南植物园 | Compound 2 α, 3 β dihydroxy 23 aldehyde radical olive 12 alkene 28 acid application in preparing glycosidase inhibitor |
| CN108314616A (en) * | 2017-01-17 | 2018-07-24 | 浙江工业大学 | Triterpene compound and its preparation and application |
| CN108314616B (en) * | 2017-01-17 | 2020-10-27 | 浙江工业大学 | Triterpenoid and preparation and application thereof |
| CN110563794A (en) * | 2019-09-27 | 2019-12-13 | 桂林电子科技大学 | Myrtle triterpene lactone A as well as extraction method and application thereof |
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| CN104490894B (en) | 2019-06-28 |
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