CN1535980A - New compound with cativity for inhibiting glycine transport factor - Google Patents

New compound with cativity for inhibiting glycine transport factor Download PDF

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Publication number
CN1535980A
CN1535980A CNA031176445A CN03117644A CN1535980A CN 1535980 A CN1535980 A CN 1535980A CN A031176445 A CNA031176445 A CN A031176445A CN 03117644 A CN03117644 A CN 03117644A CN 1535980 A CN1535980 A CN 1535980A
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China
Prior art keywords
glycine
new compound
methyl
chloroform
thiosulfuric acid
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褚以文
久
李俊英
一
户田喜久
哉
照井佑一
福永拓哉
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Sichuan Antobiotic Industry Research Inst State Medicine Supervison & Administ
Taisho Pharmaceutical Co Ltd
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Sichuan Antobiotic Industry Research Inst State Medicine Supervison & Administ
Taisho Pharmaceutical Co Ltd
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Abstract

The present invention discloses conjugated of glycine with glutamic acid receptor, and can activate the receptor, the number of glycines existed in the synaptic neural gap can be regulated by glycine transferase found in the neuroglial cell, so that the inhibitor of glycine transferase can increase the glycine in the synoptic neural gap, and can be used as medicine for activating glutamic acid receptor and for curing schizophrenia. Said invention also discloses a new compound with physiological activity, said new compound has inhibiting activity for glycine transferase of glycine cell and is produced by microbial fermentation.

Description

Glycine transporter had the active new compound of inhibition
Technical field
The present invention relates to that glycine transporter in the mammalian nervous spongiocyte is had the active novel cpd by microorganisms of inhibition.
Technical background
In recent years, studies have shown that the glutamate receptor quantity in the psychotic disorder patient brain obviously increases.The glycine that the activation of glutamate receptor need be present in the myelencephalon gap combines with glutamate receptor, and the quantity of glycine is regulated by the glycine transporter that is present in the neurogliocyte, therefore, the glycine transporter inhibitor can increase the quantity of glycine in the myelencephalon gap, thereby uses as the psychotic disorder medicine that can activate glutamate receptor.
Do not find and the The compounds of this invention similar as yet at present, and glycine transporter is had the active compound of inhibition.
Summary of the invention
The purpose of this invention is to provide a class neurogliocyte glycine transport enzyme is had the active novel physiologically active substance of inhibition.
The contriver carries out various researchs, found that some material has excellent inhibition activity to the glycine transporter of neurogliocyte, thereby has finished the present invention.
That is, the present invention is the compound with formula 1 expression
In the formula, R 1, R 2And R 3Be respectively hydrogen atom or methyl, R 4Be hydrogen atom, hydroxyl or methoxyl group, R 5Be hydrogen atom or methyl, the structure of concrete compound is as follows:
??R 1 ??R 2 ??R 3 ????R 4 ??R 5
?WSS2217 ??H ??H ??H ????OH ????H
?WSS2218 ??CH 3 ??CH 3 ??CH 3 ????H ????H
?WSS2219 ??CH 3 ??CH 3 ??H ????OH ????H
?WSS2220 ??H ??H ??SO 3H ????O＝ ????H
?WSS2221 ??H ??CH 3 ??H ????OH ????H
?WSS2222 ??CH 3 ??H ??H ????OH ????H
?WSS2227 ??CH 3 ??CH 3 ??H ????CH 3O ???CH 3
The contriver is in order to finish aforementioned purpose, from soil and plant, isolate multiple bacterial strain, and the meta-bolites of these bacterial strains various researchs have been carried out, the compound that the bacterial strain that found that produces has intensive inhibition activity to the glycine uptake of rat neurogliocyte, thereby has finished the present invention.
The bacteriology proterties of this bacterial strain outlines as follows.
The bacterial strain that produces WSS2217, WSS2218, WSS2219, WSS2220, WSS2221 and WSS2222 is that contriver etc. separates the actinomycetes that obtain from nature, below its bacteriology proterties of division.
1. proterties
By the aerial hyphae that forms on the living mycelia of branch-like base.Generate the short and small spore chain of forming by 4～15 spores of hook-shaped, curved shape or spirrillum (1～3 circle) on the aerial hyphae.The size of spore is 1.0～1.3 * 1.4～2.0 μ m, it is shaped as avette, surface wrinkle.Do not find sporangial formation of bacterioflora, sporocyst and class and the spore that moves about.
2. the character of substratum
In various substratum, 30 ℃ when cultivating for 3 weeks, the macroscopic table 1 that the results are shown in.
Table 1
Nutrient agar Growth conditions in the substratum Aerial hyphae The color of bacterium colony inner face The generation of soluble pigment
Form Color
Sucrose-nitrate Extremely - - - Do not have
Glucose-l-asparagine Extremely - - - Do not have
Glycerine-l-asparagine A little less than Seldom - Colourless Do not have
Starch-inorganic salt A little less than Do not have - Colourless Do not have
Tyrosine A little less than Seldom - Cream-colored Do not have
Common nutrition Moderate Do not have - Khaki color Do not have
Yeast-Fructus Hordei Germinatus Moderate protuberantia Well White Khaki color～filbert Do not have
Oat Moderate A little less than White Cream-colored Do not have
Peptone-yeast-molysite A little less than Do not have - Khaki color Do not have
3. physiological characteristics
1) growth temperature range
A. growth temperature: 16～39 ℃
B. optimum growth temp: 32～34 ℃
2) various character
The liquefaction of gelatin (30 ℃, 2 weeks): feminine gender
Solidifying (37 ℃, 2 weeks) of skimmed milk: feminine gender
The peptonizing of skimmed milk (30 ℃, 2 weeks): feminine gender
Produce melanochrome sample pigment (30 ℃, 2 weeks): feminine gender
Starch hydrolysis (30 ℃, 2 weeks): feminine gender
3) utilization of carbon source (30 ℃ of nutrient agars, 2 weeks)
L-arabinose: a little less than; The L-rhamnosyl: a little less than; The D-wood sugar :+; Honey mountain sugar :+; D-glucose :+; Inositol :+; D-fructose :+; The D-seminose :+; Sucrose :+
4. the character of chemotaxonomy
1) whether diaminopimelic acid exists and the optical siomerism type: the diaminopimelic acid that detects meso-form.
2) composition of methyl naphthoquinone: MK-9 (H 2), MK-9 (H 4) be detected as main component.
3) kind of thalline reducing sugar: detect ribose seminose and glucose.
4) phosphatide: detect the phosphatide that contains unknown glucosamine, do not detect phosphatidyl glycerol.
Above proterties is all retrieved according to actinomycetic separation and authentication method (Japanese actinomycetes learn to compile calendar year 2001), and the result shows that this bacterial strain can range Nonomuraea Pseudomonas actinomycetes, so be named as Nonomuraea sp.AT-0426.
This bacterial strain is preserved in the microorganism that application on August 28th, 2002 China Committee for Culture Collection of Microorganisms common micro-organisms center is used for patented procedure by Sichuan Inst of Antibiotic Industry National Medicine Supervision and Management, and deposit number is CGMCC No.0792.
According to the situation that produces general tunning, the generation of WS2217～WSS2222 should under good gas condition, be cultivated the TF-bacterial strain in containing the substratum of various nutritive substances.
The main liquid nutrient medium that uses, substratum is made of carbon source, nitrogenous source and inorganic salt, can add VITAMIN, precursor substance and foam killer as required, pH regulator to 7.0, carbon source such as glucose, glycerine and starch etc. can be used alone or as a mixture; Nitrogenous source such as meat soup, brose, yeast water, soyflour, peptone, urea and ammonium salt etc. can be used alone or as a mixture; Inorganic salt such as sodium phosphate, sal epsom, sodium chloride and salt of wormwood etc. can be used alone or as a mixture; Foam killer can use hydroxyl and silicon compound.
Cultural method uses good gas cultivations such as vibration cultivation, ventilation stir culture, pH=4～10, and 25～35 ℃ of temperature, 2～5 days time, pH=6～7 preferably, 25～28 ℃ of temperature were cultivated 4 days.
Compound of the present invention can be made with extra care from tunning with ordinary method and obtain.That is, after cultivation finishes, adopt centrifugation or filtration to obtain culturing filtrate, make it to be adsorbed in HP-20 polyvinyl resins such as (trade(brand)names, Mitsubishi Chemical Ind makes), with lower alcohol and acetone and other organic solvent wash-out.Thalline extracts with lower alcohol and acetone and other organic solvent.Subsequently, the extracting solution of thalline and the elutriant of polymeric adsorbent are merged, concentrating under reduced pressure is removed organic solvent, and residue with extracting in the water-insoluble organic solvents such as vinyl acetic monomer, chloroform and propyl carbinol, is condensed into pasty state with organic phase.This mashed prod is dissolved in once more in the organic solvents such as benzene, vinyl acetic monomer, acetone, methyl alcohol and chloroform.Carrying out column chromatography and high performance liquid chromatography by silica gel column chromatography, gel permeation chromatography post and anti-phase ODS packed column finishes the refining of The compounds of this invention and separates.
WSS8027 can be synthetic with the method for following record.That is, the Cha Naierao Buji, the method that JACS [J.Am.Chem.Soc. the 97th rolls up 3802 pages (1975)] is recorded is synthesized WSS2219 with the methyl iodide processing.
By to ultra-violet absorption spectrum through aforesaid method gained material WSS2217～WSS2222 and WSS8027 1H-NMR, 13C-NMR nuclear magnetic resonance spectrum etc. is resolved, and proves conclusively its chemical structure.
The WSS2217 physico-chemical property is as follows:
1, outward appearance: white powder;
2, molecular weight: 474;
3, molecular formula: C 25H 38N 4O 5
4, HR-TOF mass spectrograph;
Measured value: 497.2736; Theoretical value: 497.2740 (according to C 25H 38N 4O 5Na calculates);
5, specific optical rotation: [α] D25:-169.4 (c0.062 ， dioxs);
6, ultra-violet absorption spectrum: λ MAXNm (loge) diox: 206.5 (4.27), 223.5 (sh, 3.58);
7, 1The H-NMR nuclear magnetic resonance spectrum: measure with 500MHz in the deuterium thiosulfuric acid dimethyl ester, it the results are shown in Fig. 1.
8, 13The C-NMR nuclear magnetic resonance spectrum: measure with 125MHz in the deuterium thiosulfuric acid dimethyl ester, it the results are shown in Fig. 2.
9, solubleness: be dissolved in dioxane, methyl-sulfate, dimethyl formamide, be insoluble in chloroform, water insoluble, hexane, methyl alcohol, ether, acetone and vinyl acetic monomer.
10, potential of hydrogen: neutrality
The WSS2218 physico-chemical property is as follows:
1, outward appearance: white powder
2, molecular weight: 486
3, molecular formula: C 27H 42N 4O 4
4, HR-TOF mass spectrograph:
Measured value: 509.3099; Theoretical value: 509.3104 (according to C 27H 38N 4O 4Na calculates)
5, specific optical rotation: [α] D25:-46.2 (c0.081 ， dioxs)
6, ultra-violet absorption spectrum: λ MAXNm (loge) diox: 206.0 (4.26), 223.5 (sh, 3.58)
7, 1The H-NMR nuclear magnetic resonance spectrum: measure with 500MHz in the deuterated methanol, it the results are shown in Fig. 1.
8, 13The C-NMR nuclear magnetic resonance spectrum: measure with 125MHz in the deuterated methanol, it the results are shown in Fig. 2.
9, solubleness: be dissolved in dioxane, methyl-sulfate, dimethyl formamide, be insoluble in chloroform and methyl alcohol, water insoluble, hexane, ether, acetone and vinyl acetic monomer.
10, potential of hydrogen: neutrality
The WSS2219 physico-chemical property is as follows:
1, outward appearance: white powder
2, molecular weight: 502
3, molecular formula: C 27H 42N 4O 5
4, HR-TOF mass spectrograph:
Measured value: 525.3044; Theoretical value: 525.3053 (according to C 27H 42N 4O 5Na calculates)
5, specific optical rotation: [α] D25:-182.6 (c0.068 ， dioxs)
6, ultra-violet absorption spectrum: λ MAXNm (loge) diox: 206.5 (4.26), 223.0 (sh, 3.58)
7, 1The H-NMR nuclear magnetic resonance spectrum: measure with 500MHz in the deuterium thiosulfuric acid dimethyl ester, it the results are shown in Fig. 1.
8, 13The C-NMR nuclear magnetic resonance spectrum: measure with 125MHz in the deuterium thiosulfuric acid dimethyl ester, it the results are shown in Fig. 2.
9, solubleness: be dissolved in dioxane, methyl-sulfate, dimethyl formamide, be insoluble in chloroform, water insoluble, hexane, methyl alcohol, ether, acetone and vinyl acetic monomer.
10, potential of hydrogen: neutrality
The WSS2220 physico-chemical property is as follows:
1, outward appearance: white powder
2, molecular weight: 579
3, molecular formula: C 27H 39N 4O 8S
4, HR-TOF mass spectrograph:
Measured value: 625.2023; Theoretical value: 625.2284 (according to C 27H 39N 4O 8SNa 2Calculate)
5, specific optical rotation: [α] D25:-190.8 (c0.104 ， dioxs)
6, ultra-violet absorption spectrum: λ MAXNm (loge) diox: 207.5 (4.25), 223.5 (sh, 3.58)
7, 1The H-NMR nuclear magnetic resonance spectrum: measure with 500MHz in the deuterium thiosulfuric acid dimethyl ester, it the results are shown in Fig. 1.
8, 13The C-NMR nuclear magnetic resonance spectrum: measure with 125MHz in the deuterium thiosulfuric acid dimethyl ester, it the results are shown in Fig. 2.
9, solubleness:
Be dissolved in dioxane, methyl-sulfate, dimethyl formamide, be insoluble in chloroform, water insoluble, hexane, methyl alcohol, ether, acetone and vinyl acetic monomer.
10, potential of hydrogen: neutrality
The WSS2221 physico-chemical property is as follows:
1, outward appearance: white powder
2, molecular weight: 488
3, molecular formula: C 26H 40N 4O 5
4, HR-TOF mass spectrograph
Measured value: 511.6034; Theoretical value: 511.6155 (according to C 27H 38N 4O 5Na calculates)
5, specific optical rotation: [α] D25:-182.6 (c0.068 ， dioxs)
6, ultra-violet absorption spectrum: λ MAXNm (loge) diox: 206.5 (4.26), 223.0 (sh, 3.58)
7, 1The H-NMR nuclear magnetic resonance spectrum: measure with 500MHz in the deuterium thiosulfuric acid dimethyl ester, it the results are shown in Fig. 9.
8, 13The C-NMR nuclear magnetic resonance spectrum: measure with 125MHz in the deuterium thiosulfuric acid dimethyl ester, it the results are shown in Figure 10.
9, solubleness: be dissolved in dioxane, methyl-sulfate, dimethyl formamide, be insoluble in chloroform, water insoluble, hexane, methyl alcohol, ether, acetone and vinyl acetic monomer.
10, potential of hydrogen: neutrality
The WSS2222 physico-chemical property is as follows:
1, outward appearance: white powder
2, molecular weight: 488
3, molecular formula: C 26H 40N 4O 5
4, HR-TOF mass spectrograph
Measured value: 511.6048; Theoretical value: 511.6155 (according to C 25H 38N 4O 5Na calculates)
5, specific optical rotation: [α] D25:-182.6 (c0.068 ， dioxs)
6, ultra-violet absorption spectrum: λ MAXNm (loge) diox: 206.5 (4.26), 223.0 (sh, 3.57)
7, 1The H-NMR nuclear magnetic resonance spectrum: measure with 500MHz in the deuterium thiosulfuric acid dimethyl ester, it the results are shown in Figure 11.
8, 13The C-NMR nuclear magnetic resonance spectrum: measure with 125MHz in the deuterium thiosulfuric acid dimethyl ester, it the results are shown in Figure 12.
9, solubleness:
Be dissolved in dioxane, methyl-sulfate, dimethyl formamide, be insoluble in chloroform, water insoluble, hexane, methyl alcohol, ether, acetone and vinyl acetic monomer.
10, potential of hydrogen: neutrality
The WSS8027 physico-chemical property is as follows:
1, outward appearance: white powder
2, molecular weight: 558
3, molecular formula: C 31H 50N 4O 5
4, HR-TOF mass spectrograph
Measured value: 581.7433; Theoretical value: 581.7496 (according to C 31H 50N 4O 5Na calculates)
5, specific optical rotation: [α] D25:-160.4 (c0.04 ， dioxs)
6, ultra-violet absorption spectrum: λ MAXNm (loge) diox: 206.5 (4.26), 223.0 (sh, 3.58)
7, 1The H-NMR nuclear magnetic resonance spectrum: measure with 500MHz in the deuterium thiosulfuric acid dimethyl ester, it the results are shown in Figure 13.
8, 13The C-NMR nuclear magnetic resonance spectrum: measure with 125MHz in the deuterium thiosulfuric acid dimethyl ester, it the results are shown in Figure 14.
9, solubleness: be dissolved in dioxane, methyl-sulfate, dimethyl formamide, be insoluble in chloroform, water insoluble, hexane, methyl alcohol, ether, acetone and vinyl acetic monomer.
10, potential of hydrogen: neutrality
The compounds of this invention is inhibited to the glycine transaminase of finding in neurogliocyte.So, useful to schizoid treatment.
Description of drawings
Fig. 1 is illustrated in deuterium thiosulfuric acid two formicesters WSS2217's that measures with 500MHz 1The H-NMR spectrum
Fig. 2 is illustrated in deuterium thiosulfuric acid two formicesters WSS2217's that measures with 125MHz 13The C-NMR spectrum
Fig. 3 is illustrated in deuterium thiosulfuric acid two formicesters WSS2218's that measures with 500MHz 1The H-NMR spectrum
Fig. 4 is illustrated in deuterium thiosulfuric acid two formicesters WSS2218's that measures with 125MHz 13The C-NMR spectrum
Fig. 5 is illustrated in deuterium thiosulfuric acid two formicesters WSS2219's that measures with 500MHz 1The H-NMR spectrum
Fig. 6 is illustrated in deuterium thiosulfuric acid two formicesters WSS2219's that measures with 125MHz 13The C-NMR spectrum
Fig. 7 is illustrated in deuterium thiosulfuric acid two formicesters WSS2220's that measures with 500MHz 1The H-NMR spectrum
Fig. 8 is illustrated in deuterium thiosulfuric acid two formicesters WSS2220's that measures with 125MHz 13The C-NMR spectrum
Fig. 9 is illustrated in deuterium thiosulfuric acid two formicesters WSS2221's that measures with 500MHz 1The H-NMR spectrum
Figure 10 is illustrated in deuterium thiosulfuric acid two formicesters WSS2221's that measures with 125MHz 13The C-NMR spectrum
Figure 11 is illustrated in deuterium thiosulfuric acid two formicesters WSS2222's that measures with 500MHz 1The H-NMR spectrum
Figure 12 is illustrated in deuterium thiosulfuric acid two formicesters WSS2222's that measures with 125MHz 13The C-NMR spectrum
Figure 13 is illustrated in the deuterated pyridine WSS8027's that measures with 500MHz 1The H-NMR spectrum
Figure 14 is illustrated in the deuterated pyridine WSS8027's that measures with 125MHz 13The C-NMR spectrum
Embodiment
Below enumerate embodiment and experimental example specifying to this patent.
The preparation of embodiment 1 WSS2217, WSS2218, WSS2219 and WS2220.
1, the liquid nutrient medium that will contain 2.5% Zulkovsky starch, 1% glucose, 0.5% fish meal, 0.3% cottonseed meal, 0.3%NZ CASE, 0.2% yeast water, 0.2% lime carbonate placed triangular flask, in 121 ℃, sterilization 20 minutes.Then, inoculation Nonomuraea sp.TA-0426 bacterial strain on this aseptic culture medium was cultivated three days in 28 ℃, 200rpm jolting, as seed culture fluid.
Then, will pack in the triangular flask of 500ml by the 100ml aseptic liquid nutrient medium that the lime carbonate of 2% Semen Maydis powder, 1.0% glycerine, 0.5% glucose, 0.5% cottonseed meal, 0.5% wort, peptone more than 0.3%, 0.05% sal epsom and 0.3% is formed.Aforementioned seed culture fluid 4ml is added wherein, prepare 100 bottles, 28 ℃, the cultivation of 180rpm shake 14 days.
After cultivating end, gained nutrient solution 10L is added the 5L propyl carbinol, stirring, centrifugation, concentrating under reduced pressure propyl carbinol component obtains brown oil matter 6.03 grams.
2, aforementioned brown oil matter 6.03 grams are dissolved in the 10ml chloroform, last silicagel column [SilicaGel60 (manufacturing of Merck company) 200ml: chromatography column f=40mm] absorption.With 400ml chloroform wash-out, carry out gradient elution with the mixed solvent of chloroform-methyl alcohol (98: 2～50: 50) after, wherein the component of chloroform-methyl alcohol (95: 5) merges, and is evaporated to driedly, obtains the cut 1 of 272.8mg.The component of chloroform-methyl alcohol (90: 10) is merged, be evaporated to driedly, obtain the cut 2 of 172.5mg.Again the component of chloroform-methyl alcohol (80: 20) is merged, be evaporated to driedly, obtain the cut 3 of 398.0mg.
3, preceding paragraph cut 1 is dissolved in a spot of methyl alcohol, trifluoroacetic acid aqueous solution (65: 35) with acetonitrile-0.02% carries out high performance liquid chromatography separation [instrument] as moving phase: the watt corporate system, chromatography column: Wa Aimu corporate system ODS-AM (Φ 10 * 250mm)], detect in 210nm, flow velocity 2.5ml/min, the collection retention time is 13～16 minutes a cut.This cut of concentrating under reduced pressure, remove acetonitrile after, divide 2 extractings with isopyknic ethyl acetate.Merge organic phase, after the anhydrous sodium sulphate dehydration, be evaporated to driedly, obtain the WSS2218 of 15.2mg.
4, preceding paragraph cut 2 is dissolved in a spot of methyl alcohol, trifluoroacetic acid aqueous solution (45: 65) with acetonitrile-0.01% carries out high performance liquid chromatography separation [instrument: watt corporate system as moving phase, chromatography column: Wa Aimu corporate system ODS-AM (Φ 10 * 250mm)], detect in 210nm, flow velocity 2.5ml/min, the cut of collect retention time 9.5～10 minutes and 15～16 minutes.Merge isolating two components respectively, concentrating under reduced pressure, remove acetonitrile after, divide 2 extractings with isopyknic ethyl acetate.Merge organic phase, after the anhydrous sodium sulphate dehydration, be evaporated to driedly, obtain the WSS2217 of 3.5mg and the WSS2219 of 13.4mg.
5, preceding paragraph cut 3 is dissolved in a spot of methyl alcohol, acetonitrile with 30% carries out high performance liquid chromatography as moving phase and separates [instrument: watt corporate system, chromatography column: Wa Aimu corporate system ODS-AM (Φ 10 * 250mm)], detect in 210nm, flow velocity 10ml/min collects 8.0～9.0 minutes cut of retention time.Separate fraction is evaporated to dried, obtains the WSS2220 of 8.8mg.
The preparation of embodiment 2 WSS2221 and WSS2222
1, the liquid nutrient medium 60ml that will contain 2.5% Zulkovsky starch, 1% glucose, 0.5% fish meal, 0.3% cottonseed meal, 0.3%NZ CASE, 0.2% yeast water and 0.2% lime carbonate adds in the triangular flask, 121 ℃, sterilization 20 minutes.Then, inoculation Nonomuraea sp.TA-0426 bacterial strain on this aseptic culture medium was cultivated three days in 28 ℃, 200rpm jolting, as seed culture fluid.
Then, will pack into by the aseptic liquid nutrient medium 100ml that the lime carbonate of 2% Semen Maydis powder, 1.0% glycerine, 0.5% glucose, 0.5% cottonseed meal, 0.5% wort, peptone more than 0.3%, 0.05% sal epsom and 0.3% is formed in the triangular flask of 500ml.Aforementioned seed culture fluid 4ml is added wherein, prepare 150 bottles, 28 ℃, 180rpm jolting cultivation 14 days.
After cultivating end, with gained nutrient solution 15L adding 5L propyl carbinol, stirring, centrifugation with propyl carbinol component concentrating under reduced pressure, add the water 500ml that transfers pH3.0 with acetic acid, and twice of 500ml ethyl acetate extraction used in stirring.This ethyl acetate extraction component is merged, and concentrating under reduced pressure gets brown oil matter 2.83g.
2, preceding paragraph oily mater 2.83 grams are dissolved in the 10ml chloroform, last silicagel column [SilicaGel60 (manufacturing of Merck company) 200ml: chromatography column f=40mm] absorption.With 400ml chloroform wash-out, carry out gradient elution with chloroform-methyl alcohol (99: 1～50: 50).Will be wherein merge with the component of chloroform-methanol (90: 10) component with (80: 20), be evaporated to dried, brown oil matter 864mg.
3, preceding paragraph brown material 864mg is dissolved in a spot of methyl alcohol, trifluoroacetic acid aqueous solution (45: 55) with acetonitrile-0.01% carries out high performance liquid chromatography separation [instrument: watt corporate system as moving phase, chromatography column: Wa Aimu corporate system ODS-AM (Φ 20 * 250mm)], detect in 210nm, flow velocity 10ml/min collect to merge the cut of retention time 12～12.5 minutes and 13～14 minutes respectively.Be evaporated to driedly respectively, promptly get the WSS2221 of 5mg and the WSS2222 of 3.7mg.
The preparation of embodiment 3 WSS8027
Under nitrogen gas stream, in sodium hydroxid (100.6mg), add the 1ml dimethyl formamide, cooling, stirring.To wherein adding the dimethyl formamide 1ml that is dissolved with WSS2219 (12.9mg), add methyl iodide (300ml) again, stirred 1 hour under the room temperature.After reaction finishes, add entry and vinyl acetic monomer, concentrating under reduced pressure vinyl acetic monomer layer under the ice bath cooling.Enriched material high performance liquid chromatography [moving phase: acetonitrile-0.02% trifluoroacetic acid aqueous solution (80: 20), device: watt company, chromatography column: Wa Aimu corporate system ODS-AM (Φ 10 * 150mm)] system's separation, detect at the 210nm place, flow velocity is 2.5ml/min.Isolating component is evaporated to dried, promptly gets the WSS8027 of 4.4mg.
Experimental example 1
Inhibition experiment to rat neurogliocyte glycine transport enzyme
A corpse or other object for laboratory examination and chemical testing
WSS2217, WSS2218, WSS2219, WSS2220, WSS2221, WSS2222 and WSS8027 be dissolved in dimethyl sulfoxide (DMSO) respectively and be mixed with the concentration of 10mg/ml, it is stand-by to be diluted to desired concn with aqua sterilisa.
The test cell
Rat neurogliocyte C6.
Test method
Containing the flat 96 hole flat boards of liquid scintillation solution (Cytostar-T: Armagh west corporate system), with 5 * 10 4Cell/ware inoculation C6 cell places 5%CO in the substratum that contains 10% calf serum (Yi Buke corporate system) 2Cultivated 24 hours in 37 ℃ in the incubator.Then, will use by the 10mM phosphoric acid buffer and compare the buffer reagent replacement medium that this (pH7.4), 150mM sodium chloride, 1mM calcium chloride, 1mM sodium chloride, 1mM magnesium chloride and 10mM glucose are formed.Adding concentration drug solution 0.01ml and 375K Bq/ml [ 14C] glycine 0.01ml, cultivated 50 minutes in 30 ℃.After the cultivation, discard damping fluid, drying is 5 hours under the room temperature.Use (TopCount then; Crust card corporate system) measures its radioactivity.As 0%, the test group of adding excessive glycine (10mM) is calculated inhibiting rate of each group as 100% with following formula with the radioactivity of control group.
Inhibiting rate (%)=[(data of the data of drug test group-excessive glycine test group)/(not adding the data of the data-excessive glycine test group of drug test group)] * 100
To cause that the compound concentration that shows 50% inhibiting rate is as IC 50(μ M), the IC of each compound and glycine 50List in table 2.Obtain the inhibiting rate of Glycine simultaneously.
Table 2
Compound name ????IC 50
????WSS2217 ????0.019
????WSS2218 ????0.452
????WSS2219 ????0.007
????WSS2220 ????0.025
????WSS2221 ????0.013
????WSS2222 ????0.028
????WSS8027 ????20
????Glycine ????123

Claims (4)

1, a kind ofly suppress active new compound suc as formula glycine transporter is had shown in the I,
Figure A031176440002C1
In the formula, R 1, R 2, and R 3Be respectively hydrogen atom or methyl, R 4Be hydrogen atom, hydroxyl or methoxyl group, R 5Be hydrogen atom or methyl.
2, a kind ofly suppress active new compound suc as formula glycine transporter is had shown in the II
3, according to claim 1 and 2 described new compounds; The pharmaceuticals of selecting.
4, according to claim 1 and 2 described new compounds; Inhibitor as the glycine transporter of effective constituent.
CNA031176445A 2003-04-08 2003-04-08 New compound with cativity for inhibiting glycine transport factor Pending CN1535980A (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2013075624A1 (en) 2011-11-22 2013-05-30 北京哈三联科技股份有限公司 Glycine reuptake inhibitor and use thereof
CN103374057A (en) * 2012-04-16 2013-10-30 中国医药集团总公司四川抗菌素工业研究所 New compound with inhibiting activity to glycine transporters

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2013075624A1 (en) 2011-11-22 2013-05-30 北京哈三联科技股份有限公司 Glycine reuptake inhibitor and use thereof
CN103374057A (en) * 2012-04-16 2013-10-30 中国医药集团总公司四川抗菌素工业研究所 New compound with inhibiting activity to glycine transporters
CN103374057B (en) * 2012-04-16 2015-08-19 中国医药集团总公司四川抗菌素工业研究所 A kind of compound glycine transporter to inhibit activities

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