CN102050787B - 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 PDF

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CN102050787B
CN102050787B CN 200910201771 CN200910201771A CN102050787B CN 102050787 B CN102050787 B CN 102050787B CN 200910201771 CN200910201771 CN 200910201771 CN 200910201771 A CN200910201771 A CN 200910201771A CN 102050787 B CN102050787 B CN 102050787B
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guanidine derivatives
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王峥涛
耿放
孙虔
杨莉
祁萌
廖立平
季光
窦薇
郑秀棉
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Shanghai University of Traditional Chinese Medicine
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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

The purposes of guanidine derivatives and preparation, pharmaceutical composition and preparation treatment metabolic syndrome medicine
Technical field
The invention belongs to medical technical field, relate to a class and separate the guanidine derivatives that obtains from Semen Plantaginis and psyllium, 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, before Plantaginaceae plant car, the dry mature seed of Plantago asiatica L. and Plantago depressa Willd Plantago Depressa Willd. is Semen Plantaginis, dry herb is psyllium, be used for clearing away heat and promoting diuresis, excreting dampness is treating stranguria, the diseases such as oedema turgor.At present in bibliographical information and not mentioned its anti-diabetic and the treatment metabolic syndrome active.
Main component in Semen Plantaginis comprises polyose, phenylethyl alcohol glycoside, iridoids, flavonoid etc., and these compositions have the excretion that promotes water, sodium-chlor, urea and uric acid, the treatment oedema; Anti-ageing; Laxative; Relieving cough and asthma; Improve the effects such as cardiovascular status.The report that there is no at present Semen Plantaginis extract and treatment diabetes thereof, hypertension isoreactivity composition.
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 general formula I:
Figure G2009102017716D00021
(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 of replacing of 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; 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 pharmaceutically acceptable acid or the formed additive salt of alkali.
Preferred guanidine derivatives structural formula is as shown in structural formula I, II, III.
Figure G2009102017716D00031
Structural formula I
Figure G2009102017716D00032
Structural formula II
Figure G2009102017716D00033
Structural formula II I
The invention provides from Semen Plantaginis or psyllium the method for separating guanidine derivatives, as plant origin, obtain through separation and purification before the Plantago Plantaginaceae plant car.
Described plant origin comprises seed or the herb of (Plantago asiatica L.) before Plantaginaceae (plantaginaceae) Plantago (Plantago) 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; Solvent used 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, permeating extraction;
(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;
Again 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, acetic acid.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: take Plantaginaceae plant Semen Plantaginis or psyllium as raw material, after 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 water.
(2) enrichment: closely colourless to petroleum ether layer with sherwood oil (30～90 ℃) extraction, 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.Pass through 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, obtain medicinal extract, the water suspendible.By the ODS reversed phase column chromatography, use respectively 5%～25% methyl alcohol and 30%～90% methyl alcohol, 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 (Vanillin sulfuric acid test solution is aobvious blue), Fractional Collections; Again with the gel separated part, the reverse column chromatography of ODS, and wash-out, Fractional Collections, tlc is inspected, until obtain sterling.
Resulting guanidine derivatives can be used for the medicine of preparation treatment metabolic syndrome (such as diabetes, insulin resistance, impaired glucose tolerance, hypertension, high triglyceride, abnormalities of sugar/lipid metabolism, obesity, atherosclerosis etc.), and pharmaceutical dosage form can be 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 metabolic syndrome medicine such as treatment diabetes, insulin resistance, impaired glucose tolerance, hypertension, high triglyceride, dyslipidemias, obesity, atherosclerosis of this compounds.
The present invention for the drug provision of exploitation treatment diabetes and metabolic syndrome lead compound, for the new purposes of exploitation natural product and plant amedica significant.
Description of drawings
Fig. 1 is the main 2D NMR dependency structure schematic diagram of Plantagoamidinic acid A (Plantagoamidinic acid A), and wherein thick line is COSY; Solid line is relevant: HMBC; Dotted line is relevant: NOE
Fig. 2 is the chemical structural formula of element (Plantagoamidinic acid) A, B, C before car
Embodiment
Embodiment 1
Get Semen Plantaginis (dry mature seed) 1kg, add 4L 80% alcohol heating reflux to extract after pulverizing 4 hours, repeat 4 times, filter and united extraction liquid, after decompression recycling ethanol, medicinal extract is suspended in 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 successively methylene chloride-methanol-water 10: 1: 0.05,5: 1: 0.05 and 3: 1: 0.05 wash-outs, collect 3: 1: 0.05 elutriants, decompression and solvent recovery obtains medicinal extract, the water suspendible; By the ODS reversed phase column chromatography, use respectively 20% methyl alcohol, 40% methyl alcohol and 85% methanol-eluted fractions, 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 (Vanillin sulfuric acid test solution is aobvious blue), Fractional Collections;
With the gel separated part, with the reverse column chromatography 5%-25% of ODS methanol-eluted fractions, Fractional Collections, tlc is inspected (Vanillin sulfuric acid test solution is aobvious blue), until obtain Plantagoamidinic acid A, B, C sterling again.
Embodiment 2
Get psyllium (drying and ripening herb) 1kg, add 4L 80% ethanol heating and refluxing extraction 4 hours after pulverizing, repeat 4 times, filter and united extraction liquid, after decompression recycling ethanol, medicinal extract is suspended in water.
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 successively methylene chloride-methanol-water 10: 1: 0.05,5: 1: 0.05 and 3: 1: 0.05 wash-outs, collect 3: 1: 0.05 elutriants, decompression and solvent recovery obtains medicinal extract, the water suspendible; By the ODS reversed phase column chromatography, use respectively 20% methyl alcohol, 40% methyl alcohol and 85% methanol-eluted fractions, 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 (Vanillin sulfuric acid test solution is aobvious blue), Fractional Collections;
With the gel separated part, with the reverse column chromatography 5%-25% of ODS methanol-eluted fractions, Fractional Collections, tlc is inspected (Vanillin sulfuric acid test solution is aobvious blue), until obtain respectively Plantagoamidinic acid A, B, C sterling again.
By embodiment 1 and 2, separate from Semen Plantaginis (grass) and obtain before car element (Plantagoamidinic acid compounds) A, B and C-structure as shown in Figure 2, wherein 1 is Plantagoamidinic acid A (Plantagoamidinic acid A), 2 is Plantagoamidinic acid B (Plantagoamidinic acid B), 3 is Plantagoamidinic acid C (Plantagoamidinic acidC), and its spectroscopic data is:
Plantagoamidinic acid A (Plantagoamidinic acid A) is faint yellow oily thing, [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 schematic diagram of Plantagoamidinic acid A, and wherein thick line is COSY; Solid line is relevant: HMBC; Dotted line is relevant: NOE.
Plantagoamidinic acid B (Plantagoamidinic acid B) is faint yellow oily thing, [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.
Plantagoamidinic acid C (Plantagoamidinic acid C) is faint yellow oily thing, [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.
Above three compounds 1H NMR and 13C NMR data see Table 1.
Table 1. Plantagoamidinic acid A (1), B (2), C's (3) 1H NMR (500HZ) reaches 13C NMR (125MHZ) data
Figure G2009102017716D00091
Embodiment 3
Detect embodiment 1 and three guanidine constituents of 2 gained to the impact of 3T3-L1 cell and Hep G2 cell strain, result shows that Plantagoamidinic acid series derivates can be little on 3T3-L1 cell and Hep G2 cell proliferation impact, but can increase 3T3-L1 cell and Hep G2 grape cell sugar consumption amount, there is no cytotoxicity for 3T3-L1 and Hep G2 simultaneously.Illustrate that this compounds has effect and the better security that increases glucose utilization.
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, then with nutrient solution to the greatest extent, blot, 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.
Table 2 Plantagoamidinic acid A (1), B (2) and C (3) impact (x ± s, n=6) on the 3T3-L1 cell glucose metabolism
Figure G2009102017716D00101
Figure G2009102017716D00111
Table 3 Plantagoamidinic acid A (1), B (2) and C (3) impact (x ± s, n=6) on Hep G2 cell glucose metabolism
Figure G2009102017716D00112
Embodiment 3:
The present invention is take Plantagoamidinic acid A as representative, investigated the impact 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.
Table 4. Plantagoamidinic acid A causes the impact of blood glucose in diabetic mice on tetraoxypyrimidine
The present invention for the drug provision of exploitation treatment diabetes and metabolic syndrome lead compound, for the new purposes of exploitation natural product and plant amedica significant.

Claims (6)

1. a guanidine derivatives, is characterized in that, structural formula is as shown in general formula I:
Figure FDA00002847196200011
(general formula I)
R in formula 1Be hydroxyl; R 2Be 2-methyl-2-hydroxyl-amyl group or 2-methyl-2-methoxyl group-amyl group.
2. pharmaceutically acceptable acid or the formed additive salt of alkali of a guanidine derivatives formula of I compound claimed in claim 1.
3. the preparation method of the described guanidine derivatives of claim 1, is characterized in that, comprises the following 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; Solvent used is for arbitrarily than water-alcohol mixture; Alcohol is C 1～C 5Saturated or unsaturated monohydroxy-alcohol; Extracting method comprises that heating and refluxing extraction, supersound extraction, cold soaking extract, warm lixiviate is got or permeating extraction;
(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;
Again after the solvent extration extraction, with the organic phase concentrating under reduced pressure; Solvent for use is 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 C 1～C 10Halohydrocarbon, C 1～C 10Saturated or unsaturated alcohol, C 1～C 10Ketone, C 1～C 10Ester or its mixture, and the mixed solvent of the arbitrary proportion of above solvent and water, formic acid, acetic acid;
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.
4. the preparation method of guanidine derivatives claimed in claim 3, 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, and is aobvious blue with Vanillin sulfuric acid test solution, Fractional Collections; With the gel separated part, with the reverse column chromatography wash-out of ODS, tlc is inspected, until obtain sterling again.
5. pharmaceutical composition, comprising the described guanidine derivatives of at least a claim 1 is activeconstituents, separately or in conjunction with one or more pharmaceutically acceptable, inertia, nontoxic vehicle or carrier.
6. the described guanidine derivatives of claim 1 comprises purposes aspect diabetes, hypertension, abnormalities of sugar/lipid metabolism, obesity, atherosclerotic metabolic syndrome medicine in preparation treatment.
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CN102228517B (en) * 2011-06-23 2013-01-16 上海中医药大学 Plantain seed extract and application thereof
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Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
Yukihiro Goda et al..A guanidine derivative from seeds of Plantago asiatica.《Journal of Natural Medicines》.2008,(第63期),58-60. *
吴斐华,等.毛平车前活性部位调节血脂作用的研究.《中国药科大学学报》.2005,第36卷(第5期),448-452. *
吴斐华，等.毛平车前降血糖调血脂作用的研究.《中国中药杂志》.2005,第30卷(第15期),1179-1183. *

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