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19th July 2017 @ 16:24

Scheme:

AcetylScheme.png

SM/P 

(Z)-2-(4-chlorophenyl)-3-methoxybut-2-enenitrile 

Guanidine chloride 

Sodium methoxide 

5-(4-chlorophenyl)-6-methylpyrimidine-2-diamine 

 

mw 

207.66 

95.53 

54.02 

234.69 
 

Equiv 

1.00 

3.00 

3.00 

1.00 

 

mmol 

2.72 

8.16 

8.16 

2.72 

 

g 

0.565 

0.78 

0.44 

0.6 
 

mL 

 
 

 

 
 

 

g/mL 

 

 

1.059 

 

Procedure:

7/17/17 Guanidine hydrochloride (0.78 g, 8.16 mmol, 3 eq) and sodium methoxide (0.44 g, 8.16 mmol, eq) were dissolved in ethanol (5.8 mL) and stirred for 5 min. A solution of (Z)-2-(4-chlorophenyl)-3-methoxy-3-phenylacrylonitrile (0.565 g, 2.72 mmol, 1 eq) in ethanol (1.9 mL) was added to the cloudy, white solution. The reaction mixture was heated under reflux for 6 hours. Some starting material was lost from syringe during transfer.  

AcetylTLC.jpg

30:70 EtOAc: Hexanes, silica gel plate. Starting material gone after 6 hours of reflux.

7/18/17 The reaction mixture was diluted with dichloromethane. Saturated NaCl was added and the mixture was extracted with dichloromethane and then washed with saturated NaCl. Organic extracts were dried over MgSO4. Solvent was removed under reduced pressure to yield a white/yellow solid (0.157 g,0.67 mmol26 % yield).  

Product was crystallized from methanol and placed in the freezer overnight to yield white crystals (0.0845 g, 0.36 mmol, 14% yield).  

JS_LTDaraprimAcetylS3Crystals_PROTON_cdcl3_01.jpeg

7/20/17 Product was derivitized in order to run GC-MS. A small sample of the crystals (1 mg) was placed in a microcentrifuge tube and a 40 mg/mL solution of methoxyamine hydrochloride in pyridine (10 μL) was added. The tube was heated to 30°C for 90 minutes in a standard heatblock. After 90 minutes, N-methyl-N-trimethylsilyltrifluoroacetamide (90 μL) was added and the samples were heated at 37°C for 30 minutes. (Fiehn, O.; Kind, T. Metabolomics 2009, 3–18.) This added TMS groups as shown:

Derivatization1.png

The GC-MS showed an mpeak at 378 and an [m-15]peak at 363. 

GCMS2.png
 
GCMS.png



Attached Files
17th July 2017 @ 20:12

Scheme:

Screen Shot 2017-07-17 at 2.58.36 PM.png

Compound

MW(m)

Mol(Mmol)

Mol(M)

Mass

Density

Volume(mL)

Equiv

LDA, 2.0 M in THF

 

20.78

2

 

 

10.4

1.05

4-chlorobenzeneacetonitrile

in 30mL dry THF

151.59

19.79

 

3000 mg

 

 

1

Acetyl chloride

in 5mL dry THF

77.99

19.79

 

 

 

 

1

2-(4-chlorophenyl)-3-oxopentanenitrile

 

193.63

19.79

 

3.83 g

1.104

1.41mL

1

5/26/17 – A dry 1-neck 250 mL 24/40 round bottom flask with septum was put under Argon while 10.4 mL 2.0 M LDA in 20 mL dry THF was added to give a light-med brown-orange solution, which was cooled to ~ -80 ˚C with dry ice/acetone.  A clear, colorless solution of 4-chlorobenzeneacetonitrile in 30 mL dry THF was then added to the solution at about 4 drops/sec down the cooled side of the 250 mL RB flask to give a clear light-med brown-orange solution, which was stirred for approximately 10 minutes at ~ -80 ˚C.  With dry ice/acetone bath still in place, a clear, colorless solution of propionyl chloride in 5 mL dry THF was added dropwise at about ~ 2 drops/second to give a cloudy light-medium tan suspension, which was allowed to stir overnight and warm to room temperature slowly as the cooling bath warmed.

5/30/17 -  In the morning a light-medium tan suspension was present.  The reaction was quenched at RT with saturated aqueous ammonium chloride which afforded a clear, light-medium red-orange solution with a white/colorless solid in the bottom.

5/30/17.  The reaction mixture was transferred to a separatory funnel, and the organic layer was washed with H2O (the NH4Cl solid dissolved with addition of H2O), brine. The aqueous layer was extracted 2 x EtOAc, and the combined organic layers were dried over MgSO4 and concentrated to a dark red-brown liquid. TLC taken.

AcetylS1.png

5/31/17 -   To the resulting crude product was added a minimal amount of CH2Cl2, followed by a minimal amount of dry silica.  The volatiles were removed. The resulting silica was dry transferred onto a silica column. The product was purified with a column with the starting eluent 50:50 EtOAc/Hexanes until the last product began to come off column, followed by EtOAc to completely remove it.

6/1/17 - Fractions 45-60 and Erlenmeyer’s 1-4 were collected and concentrated on high vacuum. NMR was obtained. 

AcetylNMRCDCl3.png
Attached Files
17th July 2017 @ 19:03

Scheme: 

DaraprimS3Scheme.png

SM/P

3-(4-chlorophenyl)-4-methoxyhexanenitrile

Guanidine chloride

Sodium methoxide

product

mw

221.68

95.53

54.02

248.71

Equiv

1.00

3.00

3.00

1.00

mmol

0.50

1.45

1.49

0.50

g

0.11

0.143

0.080

0.124

mL

 

 

 

 

g/mL

 

 

1.059

 

Procedure: 

7/7/17 Guanidine hydrochloride (0.143 g, 1.45 mmol, 3 eq) and sodium methoxide (0.080 g, 1.49 mmol, 3 eq) were dissolved in ethanol (1.13 mL) and stirred for 5 minutes. A solution of 3-(4-chlorophenyl)-4-methoxyhexanenitrile (o.11 g, 0.50 mmol, 1 eq) in ethanol (0.37 mL) was added. The reaction mixture was heated under reflux for 18 h.

7/8/17 Reaction removed from reflux at 2:30 pm.

TLC in 30:70 EtOAc in Hexanes. Spots are UV active.

SM (left)= 3-(4-chlorophenyl)-4-methoxyhexanenitrile

Cospot (Middle)

Product (Right) 

DaraprimS3_TLC.jpg

7/10/17 The reaction mixture was diluted with dichloromethane. Saturated NaCl was added and the mixture was extracted three times with dichloromethane. Organic extracts were combined and dried over MgSO4. Solvent was removed under reduced pressure. NMR obtained. 

JSLT_Daraprim_NMRcrude_CDCl3.png

7/11/17 Product was crystallized from methanol. Too much methanol was added the first time and no crystals formed.

7/12/17 Solvent was removed under reduced pressure. Product was crystallized from methanol and placed in the freezer overnight.

7/13/17 Solvent was removed carefully with a pipette, and crystals were washed with cold methanol. NMR obtained. 

JSLT_DaraprimS3_NMR_CDCl3.jpg

7/20/17 Crystals were derivitized in order to run GC-MS. A small sample of the crystals (1 mg) was placed in a microcentrifuge tube and a 40 mg/mL solution of methoxyamine hydrochloride in pyridine (10 μL) was added. The tube was heated to 30°C for 90 minutes in a standard heatblock. After 90 minutes, N-methyl-N-trimethylsilyltrifluoroacetamide (90 μL) was added and the samples were heated at 37°C for 30 minutes. (Fiehn, O.; Kind, T. Metabolomics 2009, 3–18.

This added TMS groups as shown:

 

Derivatization.png

 The GC-MS showed an mpeak at 392 and an [m-15]peak at 377


pyrimethamineGCMS2.png
pyrimethamineGCMS.png
Attached Files