CN101357988B - Polyethyleneglycol derivatives derivated by aminophenyl - Google Patents
Polyethyleneglycol derivatives derivated by aminophenyl Download PDFInfo
- Publication number
- CN101357988B CN101357988B CN2008101511282A CN200810151128A CN101357988B CN 101357988 B CN101357988 B CN 101357988B CN 2008101511282 A CN2008101511282 A CN 2008101511282A CN 200810151128 A CN200810151128 A CN 200810151128A CN 101357988 B CN101357988 B CN 101357988B
- Authority
- CN
- China
- Prior art keywords
- derivative
- water
- weight
- ether
- minutes
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Landscapes
- Indole Compounds (AREA)
Abstract
The invention provides a polyethylene glycol derivative A derived with phenalgin groups with the structure expressed by formula A, a preparation method thereof, and uses of the polyethylene glycol derivative A as a sub-bearer in liquid phase synthetic process of water-soluble unsymmetrical cyanine dyes.
Description
Technical field
The present invention relates to the polyethyleneglycol derivative of derivated by aminophenyl, and the preparation method of these derivatives and they purposes as Zhi Zaiti in the water-soluble unsymmetrical cyanine dye liquid phase building-up process.
Background technology
Polyoxyethylene glycol (PEG) is the macromolecular material of a class excellent performance, it is solvable in methylene dichloride, ethyl acetate equal solvent, and it is insoluble in ether, isopropyl ether equal solvent, this performance makes its Zhi Zaiti as reactant in many reactions, reaction can be carried out under homogeneous phase condition, and in reaction system, add after the organic solvent such as ether, polyglycol supported body is precipitated out, just can realize effective isolation and purification of product by simple filtering and washing.
Cyanine dyes (be called for short Cy) is the optical material of the important excellent performance of a class, and this compounds has that molar extinction coefficient is big, fluorescence property good, combine afterwards advantages such as fluorescence efficiency increase, absorbing wavelength wide ranges (about 540-1000nm), absorption and emmission spectrum scope be adjustable with matrix.These advantages make cyanine dyes be used as the recording medium of CD widely, solar cell photosensitizers, laserable material, the light collection part in the light compositing reaction; Particularly the asymmetric water-soluble indole cyanine dyes of structure has obtained important use as the fluorescent probe that fluorescent labeling reagent and target biological molecules (as antigen, antibody, protein, Nucleotide, oligonucleotide) are combined into biomolecules in fields such as biology, medical science.
The method of the asymmetric indole cyanine dyes of synthesizing water-solubility mainly is a liquid phase synthesizing method at present.Its preparation process at first is to obtain intermediate product with the 1-substituted indole reaction that has the nucleophilic active methyl on Electron Affinities condensing agent and the equimolar 2-position; And then the 1-substituted indole reaction that has active methyl on the 2-position that intermediate product is different with another structure obtains asymmetric indole cyanine dyes [Ratnakar B.Mujumdar, Lauren A.Ernst, Swati R.Mujumdar, Christopher J.Lewis, and Alan S.Waggoner.BioconjugateChemistry, 1993,4 (2), 105-111].This liquid-phase synthesis process products therefrom is the mixture of symmetrical configuration and asymmetric indole cyanine dyes, the separation and purification of product is very difficult, must utilize cumbersome chromatographic separation just can obtain the purity asymmetric product of structure preferably, and the productive rate of products obtained therefrom is lower.In order to solve the deficiency of water-soluble asymmetric indole cyanine dyes prior synthesizing method, it is synthetic that the inventor of this patent once with the polyoxyethylene glycol was that Zhi Zaiti carries out water-soluble asymmetric indole cyanine dyes, make the building-up reactions of polyglycol supported intermediate product and ideal compound all can in homogeneous system, carry out, and separating of intermediate and product only needs simple filtering and washing just can finish, successfully solved the deficiency [Li Baolin of prior synthesizing method, Dou Lifang, Jiang Linling. Chinese invention patent, application number: 200710017321.2, the applying date: 2007.1.30.].Used Zhi Zaiti is polyoxyethylene glycol connects an aminophenyl with each key of hydroxyl at molecule two ends a derivative in aforesaid method.Because this carrier has only two reaction site, its charge capacity is lower, causes the output of each unit operation lower.
Summary of the invention
The lower shortcoming of charge capacity when technical problem to be solved by this invention is to overcome above-mentioned polyethyleneglycol derivative as the Zhi Zaiti of water-soluble unsymmetrical cyanine dye building-up process, the polyethyleneglycol derivative A of the derivated by aminophenyl of the higher structure of a kind of charge capacity shown in figure A is provided
Wherein the molecular-weight average of peg moiety is 1000～8000.
Another object of the present invention is to provide the preparation method of derivative A.The preparation method of described derivative A carries out according to reaction equation as follows:
Its concrete preparation process is:
With 1,3-two (4-nitrophenoxy)-2-propyl alcohol (2) and the sodium hydride proportioning of 1:1.3～2.0 in molar ratio join in the dry tetrahydrofuran of 40～50 times of its gross weights, this solution are stirred at room temperature to add amount of substance after 30 minutes be the polyoxyethylene glycol (1) of the tosylation of 20.1～0.5 times of compounds; Then this reaction-ure mixture was reacted 24 hours under reflux state; Cooling obtains brown resistates with the Rotary Evaporators steaming except that behind the tetrahydrofuran (THF); The gained resistates is dissolved in the water of 5～10 times of its gross weights, with dichloromethane extraction three times, the consumption of each methylene dichloride is 5～6 times of used volume of water, the extraction back merges organic phase, and its volume is concentrated to is about 5% of original volume, add the ether of 5～10 times of reaction mixture gross weights again, separate out yellow mercury oxide after the vigorous stirring, filter solid derivative (3);
Derivative 3 is dissolved in the ethanol of its 9～15 times of weight, adds the activated carbon of derivative 3 weight 3～5%, 4～6% Iron(III) chloride hexahydrates are heated to backflow; Reflux and in this solution, slowly drip the hydrazine hydrate (concentration 80%) of 3～5 times of derivative 3 weight after 20 minutes; Dropwise the back and continue to reflux 24 hours, while hot suction filtration; Gained filtrate is concentrated the ether that the back adds 3～5 times of derivative 3 weight on Rotary Evaporators, stir, filter solid derivative A.
A further object of the present invention provides described derivative A as the purposes of structure suc as formula Zhi Zaiti in the water-soluble unsymmetrical cyanine dye building-up process shown in the I.
Among the formula I, R
1, R
2For-SO
3H ,-SO
- 3,-SO
3K ,-COOH ,-COO
-,-COOK ,-H ,-Ph a kind of, R
1, R
2Identical or inequality, R
1, R
2A place in the unitary 4-of indole structure, 5-, 6-, 7-position; R
3, R
4Be C
1～10Alkyl or m be 0～8-CH
2(CH
2)
mCOOH base or m be 0～8-CH
2(CH
2)
mSO
3The H base, R
3With R
4Be group inequality; N is 1 or 2.
Said herein Zhi Zaiti be meant structure suc as formula water-soluble unsymmetrical cyanine dye building-up process shown in the I in, generate polyglycol supported hemicyanine dye by the amino among the derivative A; When this quilt hemicyanine dye that carries and the reaction of another nucleophilicity heterogeneous ring compound, make hemicyanine dye when Zhi Zaiti dissociates, generate water-soluble unsymmetrical cyanine dye compound; Utilize this Zhi Zaiti of derivative A solvable in methylene dichloride, ethyl acetate, and in ether insoluble character, each step reaction in the water-soluble unsymmetrical cyanine dye building-up process can be carried out under homogeneous phase condition, by in reaction system, adding ether the intermediate product of reaction or derivative A are separated out, utilize to filter and realize separating of product with washing.
Embodiment
Further specify the present invention below in conjunction with embodiment, but be not any restriction of context of the present invention.
The polyoxyethylene glycol of used tosylation is pressed document [Alexander Fishman among the embodiment, Mohammad Elmi Farrah, Jian-Hua Zhong, Subakar Paramanantham, ClaudiaCarrera, and Edward Lee-Ruff.J.Org.Chem., 2003,68 (25), 9843-9846] the method preparation; 1,3-two (4-nitrophenoxy)-2-propyl alcohol press document [lucky antidimmer, Chen Huating. chemical research and application, 1999,11 (6), 673-675] method preparation; 1,2,3,3-tetramethyl-indoles-5-sulfonate and 1-(ε-carboxylic amyl group)-2,3,3-tri-methyl indole-5-sulfonate is pressed document [Ratnakar B.Mujumdar, Lauren A.Ernst, Swati R.Mujumdar, Christopher J.Lewis, and Alan S.Waggoner.Bioconjugate Chemistry, 1993,4 (2), 105-111] the method preparation; Other chemical reagent is analytical pure.Compound structure identifies that used nuclear magnetic resonance analyser is a Bruker AVANCE300 NMR spectrometer with superconducting magnet, and TMS is as interior mark; Infrared spectra adopts Nicolet170SX FT-IR determination of infrared spectroscopy, and mass spectrum adopts Bruker Esquire3000plu mass spectrograph to measure.
Embodiment 1
The preparation of the polyethyleneglycol derivative A of derivated by aminophenyl
Get 1; 3-two (4-nitrophenoxy)-2-propyl alcohol 0.651g (1.9mmol); sodium hydride 0.062g (2.6mmol) is dissolved in the 30mL exsiccant tetrahydrofuran (THF), this mixing solutions is at room temperature stirred the polyoxyethylene glycol-40001.39g (0.3mmol) that adds tosylation after 30 minutes.Then this reaction-ure mixture was reacted 24 hours under reflux state; Cooling obtains brown resistates with the Rotary Evaporators steaming except that behind the tetrahydrofuran (THF).The gained resistates is dissolved in the 10mL water, and (3 * 50mL) extractions, extraction back merges organic phase, and its volume is concentrated to is about 5% of original volume, adds the 10mL ether again, has yellow mercury oxide to separate out after the vigorous stirring with methylene dichloride.Filtration obtains the derivative 3 of 1.45g polyoxyethylene glycol, productive rate 90.6%.
1H-NMR(300MHz，CDCl
3)：δ8.22(d，8H，J＝8.8Hz)，7.01(d，8H，J＝8.8Hz)，4.2-3.4(m，PEG)，3.89(m，8H)。IR(KBr)：2884，1607，1513，1467，1358，1094cm
-1。Spectral data shows that the structure of derivative 3 is:
The derivative 3 of 3.15g (0.68mmol) is dissolved in the 30mL ethanol, adds the 0.122g activated carbon again, the 0.143g Iron(III) chloride hexahydrate, heating makes its backflow.Reflux and in this solution, slowly drip 10mL hydrazine hydrate (concentration 80%) after 20 minutes.Dropwise the back and continue to reflux 24 hours, while hot suction filtration; After gained filtrate concentrated with Rotary Evaporators, add the 10mL ether, stir, filter the polyethyleneglycol derivative A2.80g of solid ammonia phenyl derivative, productive rate 96.5%.
1H-NMR(300MHz，CDCl
3)：δ6.78(d，8H，J＝8.2Hz)，6.58(d，8H，J＝8.2Hz)，4.2-3.4(m，PEG)，3.89(m，8H)。IR(KBr)：3430，3386，3318，1511，1463，1237，1032cm
-1。Spectral data shows that the structure of derivative A is:
Embodiment 2
The application of derivative A in synthesizing water-solubility unsymmetrical cyanine dye Cy3 '
Step 1 derivative A props up the preparation of the two methyne hemicyanine dyes that carry
Get derivative A1.05g (0.23mmol), triethyl orthoformate 3.25mL (20mmol) is dissolved in the 9mL Glacial acetic acid, induction stirring following 55 ℃ the reaction 5.5 hours.Be chilled to room temperature, add the 50mL ether in reaction solution, induction stirring left standstill after 5 minutes 10～20 minutes, filter faint yellow solid 0.951g, yield 95.4%.
Get the faint yellow solid 1.10g (0.23mmol) that makes, 1,2,3,3-tetramethyl-indoles-5-sulfonate 0.132g (0.52mmol) and triethyl orthoformate 0.5mL (3.05mmol) are dissolved in the 5mL dehydrated alcohol, feed nitrogen, and the flow velocity of nitrogen is 0.2～0.3L/min.Back flow reaction is 2.5 hours under the induction stirring.Be chilled to room temperature and add the 50mL ether, induction stirring 5 minutes left standstill 10～20 minutes, filtered, and got orange solids material-derivative A and propped up the two methyne hemicyanine dye 1.21g that carry, yield 93.1%.IR(KBr)：3433，2884，1671，1639，1516，1463，1183，1110cm
-1。Reaction formula is as follows:
The preparation of the water-soluble unsymmetrical cyanine dye Cy3 ' of step 2
Get derivative A and prop up two methyne hemicyanine dye 0.833g (0.15mmol), the 1-(ε-carboxylic amyl group)-2 that carries, 3,3-tri-methyl indole-5-sulfonate 0.058g (0.16mmol) is dissolved in the mixed solvent that is formed by 2mL acetic anhydride and 1mL pyridine, feed nitrogen, the flow velocity of nitrogen is 0.2～0.3L/min, the following 110 ℃ of reactions of induction stirring 15 minutes.Adding 40mL ethyl acetate gets the scarlet precipitation after being chilled to room temperature.This precipitation with washed with dichloromethane to methylene dichloride washing lotion be after colourless the water-soluble unsymmetrical cyanine dye Cy3 ' of 0.021g red solid, yield 20.8%.
1H?NMR(300MHz，D
2O)：δ8.38(t，1H，βproton?of?thebridge，J＝11.5Hz)，7.78-7.70(m，4H，4-H，4’-H，6-H，6’-H)，7.23-7.71(m，2H，7-H，7’-H)，6.27-6.19(m，2H，α，α’proton?of?the?bridge)，3.95(t，2H，α-CH
2)，3.46(s，3H，CH
3)，2.12(t，2H，-CH
2COOH，J＝6.9Hz)，1.97-1.18(m，18H，3CH
2groups?and?2(CH
3)
2groups).
13C?NMR(300MHz，D
2O)：δ182.2，176.4，176.3，151.9，144.7，144.6，144.0，141.5，141.3，139.7，126.8，119.8，111.5，111.3，103.6，49.3，44.2，36.5，31.3，27.0，26.6，25.9，25.2.MS：m/z?for?C
30H
36S
2O
8N
2[M-1]
-calcd615.2，found615.1。Reaction formula is as follows:
Embodiment 3
The application of derivative A in synthesizing water-solubility unsymmetrical cyanine dye Cy5 '
Step 1 derivative A props up the preparation of the four methyne hemicyanine dyes that carry
Get derivative A1.02g (0.22mmol), 1,1,3,3-tetramethoxy propane 3.7mL (22.2mmol) is dissolved in the 9ml Glacial acetic acid, the following 55 ℃ of reactions of induction stirring 5.5 hours.Be chilled to room temperature, add the 50mL ether in reaction solution, induction stirring left standstill after 5 minutes 10～20 minutes, filter yellow solid matter 0.996g, yield 91.7%.
Get the yellow solid 1.01g (0.21mmol) that makes, 1,2,3,3-tetramethyl-indoles-5-sulfonate 0.111g (0.42mmol) joins in the 9mL Glacial acetic acid, feeds nitrogen, and the flow velocity of nitrogen is 0.2～0.3L/min.80 ℃ were reacted 1 hour under induction stirring.Add the 50mL ether after being chilled to room temperature, induction stirring 5 minutes left standstill 10～20 minutes, filtered to such an extent that brown solid material-derivative A props up the four methyne hemicyanine dye 1.15g that carry, yield 95.8%.IR(KBr):3429，2917，1679，1645，1600，1446，1180，1108cm
-1。Reaction formula is as follows:
The preparation of the water-soluble unsymmetrical cyanine dye Cy5 ' of step 2
Get derivative A and prop up four methyne hemicyanine dye 0.601g (0.10mmol), the 1-(ε-carboxylic amyl group)-2 that carries, 3,3-tri-methyl indole-5-sulfonate 0.038g (0.11mmol) is dissolved in the mixed solvent that is formed by 2mL acetic anhydride and 1mL pyridine, feed nitrogen, the flow velocity of nitrogen is 0.2～0.3L/min, the following 110 ℃ of reactions of induction stirring 15 minutes.Adding 40mL ethyl acetate gets blue precipitation after being chilled to room temperature.This precipitation with washed with dichloromethane to methylene dichloride washing lotion be after colourless the water-soluble unsymmetrical cyanine dye Cy5 ' of 0.015g blue solid, yield 21.3%.
1H?NMR(300MHz，D
2O)：δ7.88-7.66(m，6H，4-H，4’-H，6-H，6’-H，β，β’protons?of?the?bridge)，7.21-7.18(m，2H，7-H，7’-H)，6.38(dd，1H，γproton?of?the?bridge，J＝12.2，12.5Hz)，6.11-5.98(m，2H，α，α’proton?of?thebridge)，3.93(t，2H，α-CH
2)，3.49(s，3H，CH3group)，2.07(t，2H，-CH
2COOH)，1.79-1.31(m，18H，3CH
2groups，and2(CH
3)
2groups).
13C?NMR(300MHz，D
2O)：δ180.1，173.8，173.7，154.2，144.1，143.7，141.8，141.7，139.3，139.2，126.6，119.8，111.1，111.0，104.1，103.9，49.3，49.0，44.1，37.0，26.3，25.7，25.2，22.9，18.9.MS：m/z?for?C
32H
38S
2O
8N
2[M-1]
-calcd641.2，found641.1。Reaction formula is as follows:
Claims (3)
1. the aminophenyl derivative A of a polyoxyethylene glycol (PEG), its structure is suc as formula shown in the A:
Wherein the molecular-weight average of peg moiety is 1000～8000.
2. the preparation method of the described derivative A of claim 1; Carry out according to reaction equation as follows;
Concrete preparation process is:
With 1, the proportioning of 3-two (4-nitrophenoxy)-2-propyl alcohol 2 and sodium hydride 1: 1.3 in molar ratio～2.0 joins in the dry tetrahydrofuran of 40～50 times of its gross weights, this solution is stirred at room temperature to add amount of substance after 30 minutes be the polyoxyethylene glycol 1 of the tosylation of 20.1～0.5 times of compounds; Then this reaction mixture was reacted 24 hours under reflux state; Cooling obtains brown resistates with the Rotary Evaporators steaming except that behind the tetrahydrofuran (THF); The gained resistates is dissolved in the water of 5～10 times of its gross weights, with dichloromethane extraction three times, the consumption of each methylene dichloride is 5～6 times of used volume of water, the extraction back merges organic phase, and its volume is concentrated to 5% of original volume, add the ether of 5～10 times of reaction mixture gross weights again, separate out yellow mercury oxide after the vigorous stirring, filter solid derivative 3;
Derivative 3 is dissolved in the ethanol of its 9～15 times of weight, adds the activated carbon of derivative 3 weight 3～5%, 4～6% Iron(III) chloride hexahydrates are heated to backflow; The concentration that slowly drips 3～5 times of derivative 3 weight in this solution of refluxing after 20 minutes is 80% hydrazine hydrate; Dropwise the back and continue to reflux 24 hours, while hot suction filtration; Gained filtrate is concentrated the ether that the back adds 3～5 times of derivative 3 weight on Rotary Evaporators, stir, filter solid derivative A.
3. the described derivative A of claim 1 is as the purposes of structure suc as formula the Zhi Zaiti in the water-soluble unsymmetrical cyanine dye building-up process shown in the I;
Among the formula I, R
1, R
2For-SO
3H ,-SO
3 -,-SO
3K ,-COOH ,-COO
-,-COOK ,-H ,-Ph a kind of, R
1, R
2Inequality, R
1, R
2A place in the unitary 4-of indole structure, 5-, 6-, 7-position; R
3, R
4Be C
1～10Alkyl or m be 0～8-CH
2(CH
2)
mCOOH base or m be 0～8-CH
2(CH
2)
mSO
3The H base, R
3With R
4Be group inequality; N is 1 or 2.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2008101511282A CN101357988B (en) | 2008-09-26 | 2008-09-26 | Polyethyleneglycol derivatives derivated by aminophenyl |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2008101511282A CN101357988B (en) | 2008-09-26 | 2008-09-26 | Polyethyleneglycol derivatives derivated by aminophenyl |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101357988A CN101357988A (en) | 2009-02-04 |
| CN101357988B true CN101357988B (en) | 2010-09-15 |
Family
ID=40330596
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2008101511282A Expired - Fee Related CN101357988B (en) | 2008-09-26 | 2008-09-26 | Polyethyleneglycol derivatives derivated by aminophenyl |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101357988B (en) |
-
2008
- 2008-09-26 CN CN2008101511282A patent/CN101357988B/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CN101357988A (en) | 2009-02-04 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108033907A (en) | A kind of Heptamethine cyanines active fluoro probe and preparation method and application | |
| CN103073909B (en) | Water-soluble asymmetric indocyanine fluorescent dye and preparation method thereof | |
| CN104449669A (en) | Multi-aryl substituted imidazole fluorescent probe, as well as preparation method and application thereof in detecting G-quadruplex structure | |
| CN111718365B (en) | Trimeric indenyl conjugated tri-BODIPY near-infrared fluorescent dye and preparation method thereof | |
| CN100569861C (en) | The solid phase synthesis process of asymmetric indole cyanine dyes | |
| CN104910124B (en) | The preparation method and application of 9,9 ' (4,4 ' xenyl) double fluorone brominated reagents | |
| CN104710977B (en) | A kind of double function probe and preparation method thereof and the application in detecting positive parallel conformation G-tetra-serobila | |
| CN106831719B (en) | A kind of more pyridine naphthalimide fluorescence dendrimers and its preparation method and application | |
| CN111793371B (en) | 3, 5-asymmetrically modified BODIPY near-infrared fluorescent dye and preparation method thereof | |
| CN106433186A (en) | Fluorescent dichroic dye containing 4-biphenyl acetylene acenaphtho[1,2-b]pyrazine-8,9-dinitrile, preparation method of fluorescent dichroic dye and application of fluorescent dichroic dye | |
| CN101357988B (en) | Polyethyleneglycol derivatives derivated by aminophenyl | |
| CN104478984B (en) | The preparation method and application of amphipathic Tb (III) complex and preparation method thereof and helical form fluorescent nano-fiber | |
| CN101357987B (en) | Hexamine type polyethyleneglycol derivatives | |
| EP2157088A1 (en) | Indicyanine dyes and the derivatives thereof for analysing biological micromolecules | |
| CN107090190A (en) | One class acridine salt dyestuff and its preparation method and application | |
| CN114773875B (en) | Azaindole-squaraine dye, and synthetic method and application thereof | |
| CN103183696A (en) | Nitrogen heterocyclic BODIPY compound and synthesis method thereof | |
| CN114133387B (en) | Fluorescent probe with viscosity sensing property and capable of targeting multiple organelles | |
| CN114106024B (en) | Fluorescent probe and preparation method and application thereof | |
| CN112480025B (en) | Compound with aggregation-induced emission function and preparation method and application thereof | |
| CN110669350B (en) | Piperidyl BODIPY red-light fluorescent dye and preparation method and application thereof | |
| CN111607248B (en) | Quinoline type lyotropic color-changing fluorescent dye, preparation method thereof and application thereof in organic solvent water content measurement | |
| CN109206405B (en) | Triazolyl quinoline copper complex with AIE property and preparation method thereof | |
| CN101768112B (en) | Two-photon induced luminous compound with symmetrical both ends and synthesis method and purposes thereof | |
| WO2019227524A1 (en) | Fluorescent dye, preparation method therefor, and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20100915 Termination date: 20130926 |