CN102050787A - Guanidine derivatives and preparation, medicinal composition and application in preparation of medicaments for treating metabolic syndromes - Google Patents
Guanidine derivatives and preparation, medicinal composition and application in preparation of medicaments for treating metabolic syndromes Download PDFInfo
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- CN102050787A CN102050787A CN2009102017716A CN200910201771A CN102050787A CN 102050787 A CN102050787 A CN 102050787A CN 2009102017716 A CN2009102017716 A CN 2009102017716A CN 200910201771 A CN200910201771 A CN 200910201771A CN 102050787 A CN102050787 A CN 102050787A
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Abstract
The invention discloses guanidine derivatives with structure of a general formula I. R1 is hydrogen, hydroxy or C1-C5 alkoxy; R2 is C1-C20 saturated or unsaturated alkyl, hydroxy substituted C1-C20 saturated or unsaturated alkyl, or C1-C5 alkoxy substituted C1-C20 saturated or unsaturated alkyl groups; and the saturated or unsaturated alkyls are straight-chain or branch-chain. The compounds are prepared from plantago asiatica L. of plantago of plantaginaceae and genus plantago thereof serving as plant sources by separation and purification. The guanidine derivatives can be used for preparing medicaments for treating metabolic syndromes.
Description
Technical field
The invention belongs to medical technical field, relate to a class and from Semen Plantaginis and psyllium, separate the guanidine derivatives that obtains, and preparation method thereof and the application aspect the medicine of preparation treatment metabolic syndrome and diabetes.
Background technology
Semen Plantaginis and psyllium are conventional Chinese medicine, Chinese Pharmacopoeia (2005 editions) regulation, the dry mature seed of Plantago asiatica L. and Plantago depressa Willd Plantago Depressa Willd. is a Semen Plantaginis before the Plantaginaceae plant car, dry herb is a psyllium, be used for clearing away heat and promoting diuresis, excreting dampness is treating stranguria, diseases such as oedema turgor.At present in the bibliographical information and not mentioned its anti-diabetic and treatment metabolic syndrome activity.
Main component in the Semen Plantaginis comprises polyose, phenylethyl alcohol glycoside, iridoids, flavonoid etc., and these compositions have the drainage that promotes water, sodium-chlor, urea and uric acid, the treatment oedema; Anti-ageing; Laxative; Relieving cough and asthma; Improve effects such as cardiovascular status.The report that does not have Semen Plantaginis extract and treatment diabetes thereof, hypertension isoreactivity composition at present.
Summary of the invention
The present invention aims to provide a kind of new guanidine derivatives.
The present invention also provides the preparation method of above-mentioned guanidine derivatives.
Another object of the present invention is this guanidine derivatives is applied to prepare the medicine for the treatment of metabolic syndrome.
A kind of guanidine derivatives has the structure shown in the general formula I:
(general formula I)
Wherein, R1 is the alkoxyl group of hydrogen, hydroxyl or C1-C5;
R2 is the saturated or unsaturated alkyl group of C1～C20 that the alkoxyl group of the saturated or unsaturated alkyl of C1～C20 of replacing of the saturated or unsaturated alkyl of C1～C20, hydroxyl, C1-C5 replaces; Above hydrocarbyl group can be for straight or branched.
R1 is preferably hydroxyl.R2 is preferably 2-methyl-2-butene base, 2-methyl-2-hydroxyl-butyl or 2-methyl-2-methoxyl group-butyl.
Described guanidine derivatives also comprises above-mentioned all compounds and at pharmaceutically acceptable acid or the formed additive salt of alkali.
Preferred guanidine derivatives structural formula is shown in structural formula I, II, III.
Structural formula I
Structural formula II
Structural formula II I
The invention provides the method for from Semen Plantaginis or psyllium, separating guanidine derivatives,, obtain through separation and purification to be plant origin before the Plantago Plantaginaceae plant car.
Described plant origin comprises the seed or the herb of (Plantago asiatica L.) before Plantaginaceae (plantaginaceae) Plantago (Plantago) the plant car or congener (Genus Plantago).
Its concrete preparation process is:
(1) with Chinese medicine Semen Plantaginis or psyllium, perhaps the Semen Plantaginis of fresh collection or psyllium are raw material, adopt solvent-extraction process, obtain extracting solution; Used solvent is preferably water, alcohol or arbitrarily than water-alcohol mixture; Alcohol is the saturated or unsaturated monohydroxy-alcohol of C1～C5; Extracting method is preferably that heating and refluxing extraction, supersound extraction, cold soaking are extracted, warm lixiviate is got, the diacolation extraction method;
(2) with after the solvent extration extraction, discard organic phase, obtain water; Solvent for use is preferably sherwood oil (30～90 ℃), hexanaphthene, normal hexane, chloroform, methylene dichloride, ethyl acetate;
Once more after the solvent extration extraction, with the organic phase concentrating under reduced pressure; Solvent for use is preferably propyl carbinol;
Products therefrom is also used solvent elution by column chromatography, and concentrating under reduced pressure reclaims solvent, obtains total guanidine derivatives; Solvent for use is selected from the haloalkane of C1～C10, saturated or unsaturated alcohol, C1～C10 ketone, C1～C10 ester or its mixture of C1～C10, and wherein haloalkane has following general formula: C
nH
2n+1R, C
nH
2nR
2, C
nH
2n-1R
3(n＜10; R=Cl, Br, I), alcoholic solvent has following general formula: C
nH
2n+1OH or C
nH
2n-1OH (n＜10), ketones solvent has following general formula: C
nH
2nO or C
nH
2n-2O (n＜10), esters solvent has following general formula: C
nH
2nO
2Or C
nH
2n-2O
2(n＜10), and the mixed solvent of the arbitrary proportion of above solvent and water, formic acid, acetate.Column chromatography solvent elution mode comprises the positive of single or mixed solvent, anti-phase or gradient elution; The column chromatography filler is one or more of following kind: silica gel, diatomite, aluminum oxide, polymeric adsorbent, dextrane gel or ODS.
Preferred preparation method is:
(1) extract: with Plantaginaceae plant Semen Plantaginis or psyllium is raw material, after the pulverizing, with the 50-95% ethanol (heating and refluxing extraction of 4-5 liter/kg) 4 hours, repeat to extract 2～6 times, filter and united extraction liquid, decompression recycling ethanol is suspended in the medicinal extract that obtains in the water.
(2) enrichment: closely colourless with sherwood oil (30～90 ℃) extraction to petroleum ether layer, discard petroleum ether layer; Water layer is used water saturated n-butanol extraction again, and is closely colourless to n-butanol layer; With the n-butanol layer reclaim under reduced pressure, get n-butanol portion.By silica gel column chromatography, be (7～12) with volume ratio successively: 1: (0～0.1) and (4～6): 1: (0～0.1) methylene chloride-methanol-water elution, collect (4～6): 1: (0～0.1) elutriant, decompression and solvent recovery obtains medicinal extract, the water suspendible.By the ODS reversed phase column chromatography, use 5%～25% methyl alcohol and 30%～90% methyl alcohol respectively, collect 5%～25% meoh eluate, decompression and solvent recovery obtains total guanidine derivatives.
(3) purifying: above-mentioned total guanidine derivatives, separate by gel filtration chromatography, use methanol-eluted fractions, tlc is inspected guanidine derivatives (it is blue that Vanillin sulfuric acid test solution shows), Fractional Collections; Again with the gel separation position, the reverse column chromatography of ODS, and wash-out, Fractional Collections, tlc is inspected, until obtaining pure product.
Resulting guanidine derivatives can be used for the medicine of preparation treatment metabolic syndrome (for example diabetes, insulin resistance, impaired glucose tolerance, hypertension, high triglyceride, abnormalities of sugar/lipid metabolism, obesity, atherosclerosis etc.), and pharmaceutical dosage form can be the interior injection and the exterior-applied formulation with formulation and non-oral administration of oral administration.
The present invention obtains in separation on the basis of guanidine derivatives, has confirmed the metabolism syndrome medicament such as treatment diabetes, insulin resistance, impaired glucose tolerance, hypertension, high triglyceride, dyslipidemias, obesity, atherosclerosis of this compounds.
The present invention provides lead compound for the medicine of exploitation treatment diabetes and metabolic syndrome, for the new purposes of exploitation natural product and plant amedica significant.
Description of drawings
Fig. 1 is the main 2D NMR dependency structure synoptic diagram of plain A (Plantagoamidinic acid A) before the car, and wherein thick line is COSY; Solid line is relevant: HMBC; Dotted line is relevant: NOE
Fig. 2 is the chemical structural formula of plain (Plantagoamidinic acid) A, B, C before the car
Embodiment
Get Semen Plantaginis (dry mature seed) 1kg, pulverize the back and add 4L 80% alcohol heating reflux extraction 4 hours, repeat 4 times, filter and united extraction liquid, behind the decompression recycling ethanol medicinal extract is suspended in the water.
Repeatedly extract suspension with sherwood oil (60～90 ℃), closely colourless to petroleum ether layer, discard petroleum ether layer; Water layer is used water saturated n-butanol extraction again, and is closely colourless to n-butanol layer.N-butanol layer is merged reclaim under reduced pressure, get n-butanol portion.
By silica gel column chromatography, use methylene chloride-methanol-water 10: 1: 0.05,5: 1: 0.05 and 3: 1: 0.05 wash-outs successively, collect 3: 1: 0.05 elutriants, decompression and solvent recovery obtains medicinal extract, the water suspendible; By the ODS reversed phase column chromatography, use 20% methyl alcohol, 40% methyl alcohol and 85% methanol-eluted fractions respectively, collect 20% meoh eluate, decompression and solvent recovery obtains total guanidine derivatives;
Above-mentioned total guanidine derivatives separates by gel filtration chromatography, uses methanol-eluted fractions, and tlc is inspected guanidine derivatives (it is blue that Vanillin sulfuric acid test solution shows), Fractional Collections;
With the gel separation position, with the reverse column chromatography 5%-25% of ODS methanol-eluted fractions, Fractional Collections, tlc is inspected (it is blue that Vanillin sulfuric acid test solution shows), plain A, B, the pure product of C before obtaining car again.
Get psyllium (drying and ripening herb) 1kg, pulverize the back and add 4L 80% ethanol, repeats 4 times, filter also united extraction liquid, behind the decompression recycling ethanol medicinal extract is suspended in the water heating and refluxing extraction 4 hours.
Repeatedly extract suspension with ethyl acetate, closely colourless to ethyl acetate layer, discard ethyl acetate layer; Water layer is used water saturated n-butanol extraction again, and is closely colourless to n-butanol layer.N-butanol layer is merged reclaim under reduced pressure, get n-butanol portion.
By silica gel column chromatography, use methylene chloride-methanol-water 10: 1: 0.05,5: 1: 0.05 and 3: 1: 0.05 wash-outs successively, collect 3: 1: 0.05 elutriants, decompression and solvent recovery obtains medicinal extract, the water suspendible; By the ODS reversed phase column chromatography, use 20% methyl alcohol, 40% methyl alcohol and 85% methanol-eluted fractions respectively, collect 20% meoh eluate, decompression and solvent recovery obtains total guanidine derivatives;
Above-mentioned total guanidine derivatives separates by gel filtration chromatography, uses methanol-eluted fractions, and tlc is inspected guanidine derivatives (it is blue that Vanillin sulfuric acid test solution shows), Fractional Collections;
With the gel separation position, with the reverse column chromatography 5%-25% of ODS methanol-eluted fractions, Fractional Collections, tlc is inspected (it is blue that Vanillin sulfuric acid test solution shows), plain A, B, the pure product of C before obtaining car respectively again.
By embodiment 1 and 2, from Semen Plantaginis (grass), separate and obtain before the car plain (Plantagoamidinic acid compounds) A, B and C-structure as shown in Figure 2, wherein 1 is plain A (Plantagoamidinic acid A) before the car, 2 is plain B (Plantagoamidinic acid B) before the car, 3 is plain C (Plantagoamidinic acidC) before the car, and its spectroscopic data is:
Plain A (Plantagoamidinic acid A) is faint yellow oily thing before the car, [α]
D=+64.5 (MeOH), high resolution mass spectrum must this compound the accurate mass number be 226.1537, calculate that its molecular formula is C
11H
19N
3O
2Infrared spectra (KBr): 1776cm
-1, 1690cm
-1And1639cm
-1Fig. 1 is the 2D NMR dependency structure synoptic diagram of plain A before the car, and wherein thick line is COSY; Solid line is relevant: HMBC; Dotted line is relevant: NOE.
Plain B (Plantagoamidinic acid B) is faint yellow oily thing before the car, [α]
D=+75.1 (MeOH), high resolution mass spectrum must this compound the accurate mass number be 224.1643, calculate that its molecular formula is C
11H
22N
3O
3Infrared spectra (KBr) has shown 1690cm
-1And 3300cm
-1Characteristic signal.
Plain C (Plantagoamidinic acid C) is faint yellow oily thing before the car, [α]
D=+78.2 (MeOH), high resolution mass spectrum must this compound the accurate mass number be 258.1801, calculate that its molecular formula is C
12H
24N
3O
3Infrared spectra (KBr) has shown 1690cm
-1Characteristic signal.
More than three compounds
1H NMR and
13C NMR data see Table 1.
Table 1. car preceding plain A (1), B (2), C's (3)
1H NMR (500HZ) reaches
13C NMR (125MHZ) data
Detect the influence of embodiment 1 and three guanidine constituents of 2 gained to 3T3-L1 cell and Hep G2 cell strain, the result shows that Plantagoamidinic acid series derivates can be little to 3T3-L1 cell and Hep G2 cell proliferation influence, but can increase 3T3-L1 cell and Hep G2 grape cell sugar consumption amount, not have cytotoxicity for 3T3-L1 and Hep G2 simultaneously.Illustrate that this compounds has effect and the better security that increases the glucose consumption amount.
What experiment was adopted is mtt assay.Consuming experiment when closing to an end, nutrient solution is shifted out, change the nutrient solution that contains 0.5g/L MTT and continue to hatch 4h, with nutrient solution to the greatest extent, blot then, every hole adds 200 μ l methyl-sulphoxides again, on the rearmounted microplate reader of mixing in 570nm place survey absorbancy.The results are shown in Table 2 and table 3.
Plain A (1) before table 2 car, B (2) and C (3) to the influence of 3T3-L1 cell glucose metabolism (x ± s, n=6)
Plain A (1) before table 3 car, B (2) and C (3) to the influence of Hep G2 cell glucose metabolism (x ± s, n=6)
Embodiment 3:
The present invention is representative with Plantagoamidinic acid A, investigated the influence of the diabetic mice fasting plasma glucose that guanidine compound causes tetraoxypyrimidine.After the tetraoxypyrimidine modeling, the mouse blood sugar value obviously increases.After 2 weeks of administration, the administration group has shown the remarkable blood sugar reducing function identical with positive control drug.Proved the effect aspect the treatment diabetes of Semen Plantaginis and guanidine compound.The results are shown in Table 4.
Plain A causes the influence of blood glucose in diabetic mice before table 4. car to tetraoxypyrimidine
The present invention provides lead compound for the medicine of exploitation treatment diabetes and metabolic syndrome, for the new purposes of exploitation natural product and plant amedica significant.
Claims (9)
1. guanidine derivatives is characterized in that structural formula is shown in general formula I:
(general formula I)
R1 is the alkoxyl group of hydrogen, hydroxyl or C1～C5 in the formula; R2 is the saturated or unsaturated alkyl group of C1～C20 that the alkoxyl group of the saturated or unsaturated alkyl of C1～C20 of replacing of the saturated or unsaturated alkyl of C1～C20, hydroxyl, C1～C5 replaces;
Above-mentioned saturated or unsaturated alkyl is a straight or branched.
2. the described guanidine derivatives of claim 1 is characterized in that, R1 is a hydroxyl; R2 is 2-methyl-2-butene base, 2-methyl-2-hydroxyl-butyl or 2-methyl-2-methoxyl group-butyl.
3. the described guanidine derivatives of claim 1 is characterized in that, comprises that compound of Formula I is at pharmaceutically acceptable acid or the formed additive salt of alkali.
4. the preparation method of the described guanidine derivatives of claim 1 is characterized in that, to be plant origin before the Plantago Plantaginaceae plant car, obtains through separation and purification.
5. the preparation method of the described guanidine derivatives of claim 4 is characterized in that, described plant origin comprises before the Plantaginaceae Plantago plant car or the seed or the herb of congener.
6. the preparation method of the described guanidine derivatives of claim 4 is characterized in that, may further comprise the steps:
(1) with Chinese medicine Semen Plantaginis or psyllium, perhaps the Semen Plantaginis of fresh collection or psyllium are raw material, adopt solvent-extraction process, obtain extracting solution; Used solvent is water, alcohol or arbitrarily than water-alcohol mixture; Alcohol is the saturated or unsaturated monohydroxy-alcohol of C1～C5; Extracting method comprises that heating and refluxing extraction, supersound extraction, cold soaking extract, warm lixiviate is got or the diacolation extraction method;
(2) with after the solvent extration extraction, discard organic phase, obtain water; Solvent for use is sherwood oil, hexanaphthene, normal hexane, chloroform, methylene dichloride or ethyl acetate;
Once more after the solvent extration extraction, with the organic phase concentrating under reduced pressure; Solvent for use is a propyl carbinol;
(3) products therefrom is also used solvent elution by column chromatography, concentrating under reduced pressure reclaims solvent, obtains total guanidine derivatives;
Solvent for use is selected from the halohydrocarbon of C1～C10, saturated or unsaturated alcohol, C1～C10 ketone, C1～C10 ester or its mixture of C1～C10, reaches the mixed solvent of the arbitrary proportion of above solvent and water, formic acid, acetate;
Wherein the halohydrocarbon general formula is C
nH
2n+1R, C
nH
2nR
2Or C
nH
2n-1R
3, n＜10, R=Cl, Br or I; The alcoholic solvent general formula is C
nH
2n+1OH or C
nH
2n-1OH, n＜10; The ketones solvent general formula is C
nH
2nO or C
nH
2n-2O, n＜10; The esters solvent general formula is C
nH
2nO
2Or C
nH
2n-2O
2
Column chromatography solvent elution mode comprises the positive of single or mixed solvent, anti-phase or gradient elution; The column chromatography filler is one or more of following kind: silica gel, diatomite, aluminum oxide, polymeric adsorbent, dextrane gel or ODS reverse phase filler.
7. the preparation method of the described guanidine derivatives of claim 6 is characterized in that, the resulting total guanidine derivatives of step (3) is separated by gel filtration chromatography, use methanol-eluted fractions, tlc is inspected guanidine derivatives, shows blue with Vanillin sulfuric acid test solution, Fractional Collections; With the gel separation position, with the reverse column chromatography wash-out of ODS, tlc is inspected, until obtaining pure product again.
8. pharmaceutical composition comprises at least a claim 1 or 2 described guanidine derivatives are activeconstituents, independent or pharmaceutically acceptable in conjunction with one or more, inert, nontoxic vehicle or carrier.
9. claim 1 or 2 described guanidine derivatives comprise purposes aspect diabetes, hypertension, high triglyceride, abnormalities of sugar/lipid metabolism, obesity, the atherosclerotic metabolic syndrome medicine in preparation treatment.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102228517A (en) * | 2011-06-23 | 2011-11-02 | 上海中医药大学 | Plantain seed extract and application thereof |
| CN102258581A (en) * | 2011-08-23 | 2011-11-30 | 北京科莱博医药开发有限责任公司 | Plantain seed extract, preparation method and application thereof |
-
2009
- 2009-11-09 CN CN 200910201771 patent/CN102050787B/en active Active
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102228517A (en) * | 2011-06-23 | 2011-11-02 | 上海中医药大学 | Plantain seed extract and application thereof |
| CN102228517B (en) * | 2011-06-23 | 2013-01-16 | 上海中医药大学 | Plantain seed extract and application thereof |
| CN102258581A (en) * | 2011-08-23 | 2011-11-30 | 北京科莱博医药开发有限责任公司 | Plantain seed extract, preparation method and application thereof |
| CN102258581B (en) * | 2011-08-23 | 2013-04-24 | 北京科莱博医药开发有限责任公司 | Plantain seed extract, preparation method and application thereof |
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