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Chemical Structure| 79955-27-4 Chemical Structure| 79955-27-4
Chemical Structure| 79955-27-4

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5-CFDA is membrane-permeant and can be loaded into cells via incubation and hydrolyzed by intracellular esterases to 5-carboxyfluorescein and used for labeling human intervertebral disk cells in vitro for fluorescence microscopy.

Synonyms: 5-Carboxyfluorescein diacetate

4.5 *For Research Use Only! Not for Human Use. We Do Not Sell to Patients.

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Product Details of 5-CFDA

CAS No. :79955-27-4
Formula : C25H16O9
M.W : 460.39
SMILES Code : O=C(C1=CC2=C(C3(C4=C(OC5=C3C=CC(OC(C)=O)=C5)C=C(OC(C)=O)C=C4)OC2=O)C=C1)O
Synonyms :
5-Carboxyfluorescein diacetate
English Name :5-Carboxyfluorescein Diacetate
MDL No. :MFCD00036872
InChI Key :WPUZGNPQMIWOHE-UHFFFAOYSA-N
Pubchem ID :133314

Safety of 5-CFDA

Application In Synthesis of 5-CFDA

* All experimental methods are cited from the reference, please refer to the original source for details. We do not guarantee the accuracy of the content in the reference.

  • Downstream synthetic route of [ 79955-27-4 ]

[ 79955-27-4 ] Synthesis Path-Downstream   1~14

  • 1
  • [ 108-24-7 ]
  • [ 76823-03-5 ]
  • [ 79955-27-4 ]
YieldReaction ConditionsOperation in experiment
99% With sulfuric acid at 120℃;
5.17 g With pyridine at 85℃; for 0.25h;
With pyridine at 110℃; for 0.5h; 5(6)-Carboxyfluorescein diacetate General procedure: To a solution of 5(6)-carboxyfluorescein (15.0 g, 39.9 mmol) in Ac2O (180 mL) was added pyridine (18 mL, 22.3 mmol) and the reaction was stirred for 30 min at 110 °C. The clear solution was concentrated in vacuo. The crude mixture was dissolved in EtOAc (300 mL) and washed with aqueous KHSO4 (1 M, 2 x 300 mL) and brine (300 mL). The organic layer was dried over Na2SO4, filtered and concentrated under reduced pressure to give the desired compound as a light yellow solid (18.1 g, 98%). HPLC tR 7.5 and 7.6 min.
98 % With caesium carbonate In N,N-dimethyl-formamide at 0 - 20℃;

  • 2
  • [ 754172-97-9 ]
  • [ 79955-27-4 ]
  • [ 690958-19-1 ]
YieldReaction ConditionsOperation in experiment
56% Stage #1: 5-Carboxyfluorescein diacetate With benzotriazol-1-ol; N-(3-dimethylaminopropyl)-N-ethylcarbodiimide In tetrahydrofuran for 0.5h; cooling; Stage #2: aminothioacetic acid S-benzyl ester In tetrahydrofuran; N,N-dimethyl-formamide at 20℃; for 6h; Further stages.;
  • 3
  • [ 1004524-64-4 ]
  • [ 3348-03-6 ]
  • [ 79955-27-4 ]
  • [ CAS Unavailable ]
  • [ CAS Unavailable ]
YieldReaction ConditionsOperation in experiment
With triethanolamine In N,N-dimethyl-formamide at 31℃; 5 Example 5:; N-[4-(4-Aminopyrimidin-2-yloxymethyl)-benzyl]-diacetylfluorescein-6-carboxamide (6) and N-[4-(4-Aminopyrimidin-2-yloxymethyl)-benzyl]-diacetylfluorescein-5-carboxamide (7); [Show Image] 2-(4-(Aminomethyl)benzyloxy)pyrimidin-4-amine (2) (4.1 mg, 0.018 mmol) and 5(6)-carboxyfluorescein diacetate succinimidyl ester (10 mg, 0.018 mmol) are dissolved in 800 µL DMF with 2.7 µL TEA and heated overnight at 31 °C. The solvent is evaporated under vacuum and the compounds isolated by reversed phase MPLC on a C18 column using a linear gradient of water:acetonitrile (from 95:5 to 20:80 in 20 min, 0.08% TFA). MS (ESI) m/z 673 [M+H]+.
  • 4
  • [ CAS Unavailable ]
  • [ 79955-27-4 ]
  • [ CAS Unavailable ]
  • [ CAS Unavailable ]
  • [ CAS Unavailable ]
YieldReaction ConditionsOperation in experiment
With benzotriazol-1-yloxyl-tris-(pyrrolidino)-phosphonium hexafluorophosphate; N-ethyl-N,N-diisopropylamine In N,N-dimethyl-formamide at 20℃; for 2h;
  • 5
  • [ CAS Unavailable ]
  • [ 79955-27-4 ]
  • [ CAS Unavailable ]
YieldReaction ConditionsOperation in experiment
86% Stage #1: C37H51N3O6S; 5-Carboxyfluorescein diacetate With benzotriazol-1-yloxyl-tris-(pyrrolidino)-phosphonium hexafluorophosphate; N-ethyl-N,N-diisopropylamine In N,N-dimethyl-formamide at 20℃; for 2h; Stage #2: With sodium methylate In methanol; dichloromethane at 20℃; for 8h; Further stages.;
  • 6
  • [ 108-46-3 ]
  • [ 79955-27-4 ]
YieldReaction ConditionsOperation in experiment
Multi-step reaction with 2 steps 1: methanesulfonic acid / 12 h / 80 - 85 °C 2: 5.17 g / pyridine / 0.25 h / 85 °C
  • 7
  • [ 552-30-7 ]
  • [ 79955-27-4 ]
YieldReaction ConditionsOperation in experiment
Multi-step reaction with 2 steps 1: methanesulfonic acid / 12 h / 80 - 85 °C 2: 5.17 g / pyridine / 0.25 h / 85 °C
Multi-step reaction with 3 steps 1.1: methanesulfonic acid / 80 °C 1.2: 80 °C 2.1: methanesulfonic acid / 20 °C 3.1: caesium carbonate / N,N-dimethyl-formamide / 2 h / 0 - 20 °C
  • 8
  • [ 79955-27-4 ]
  • [ 500533-95-9 ]
YieldReaction ConditionsOperation in experiment
With N-(2,6-diisopropylphenyl)-N'-(2-(N-succinimidyl)carboxyethyl)-1,6,7,12-tetra(4-sulfophenoxy)perylene-3,4:9,10-tetracarboxydiimide labelled phospholipase A1; water; Triton X-100 Enzymatic reaction;
YieldReaction ConditionsOperation in experiment
With caesium carbonate In N,N-dimethyl-formamide at 20℃; for 1h; 1.2 General procedure: First, compound (f) to be used as a starting material was obtained by being purchased from SIGMA. Compound (f) was then reacted in DMF with Ac2O and Cs2CO3 at room temperature for 1 hour to give compound (g) (yield: 96%). Compound (g) was then reacted in TsCl and triethylamine with NH4Cl supported on silica gel at room temperature for 10 minutes to give compound (h). Compound (h) was further reacted in MeOH with MeONa at room temperature for 10 minutes to give compound (i). Finally, compound (i) was reacted in CsF and DMF with 1 equivalent of CH3I at room temperature for 1 hour to give compounds 3 and 4 (yield: 2.3%).
  • 10
  • [ 79955-27-4 ]
  • [ 76823-03-5 ]
YieldReaction ConditionsOperation in experiment
With 3-(4-acetoxyphenyl)acrylic acid; water In dimethyl sulfoxide at 20℃; Irradiation; Enzymatic reaction;
  • 11
  • [ 2462011-32-9 ]
  • [ 79955-27-4 ]
  • [ 76-05-1 ]
  • [ 1610377-63-3 ]
YieldReaction ConditionsOperation in experiment
6.5 mg Stage #1: N-(2-(2-(2-aminoethoxy)ethoxy)ethyl)-4-(4-(6-(4-methylpiperazin-1-yl)-1H,3'H-[2,5'-bibenzo[d]imidazol]-2'-yl)phenoxy)butanamide; 5-Carboxyfluorescein diacetate With 1-ethyl-(3-(3-dimethylamino)propyl)-carbodiimide hydrochloride; N-ethyl-N,N-diisopropylamine In dimethyl sulfoxide at 20℃; for 2h; Stage #2: trifluoroacetic acid In water; acetonitrile
  • 12
  • [ 6066-82-6 ]
  • [ 79955-27-4 ]
  • [ 150206-05-6 ]
YieldReaction ConditionsOperation in experiment
25% With dicyclohexyl-carbodiimide In dichloromethane for 0.5h; 5 and 6-Carboxyfluorescein diacetate N-hydroxysuccinimide ester General procedure: 5(6)-Carboxyfluorescein diacetate (15.0 g, 32.6 mmol) was dissolved in DCM (225 mL) and N-hydroxysuccinimide (4.5 g, 39.1 mmol) was added and stirred until dissolved. DIC (6.06 mL, 39.1 mmol) was added and the reaction was stirred for 30 min. The organic layer was washed with water (2 50 mL), brine (50 mL), and dried over Na2SO4. The crude mixture was dissolved in toluene (50 mL), loaded onto an equilibrated column (7 15 cm bed of silica) and eluted with 20% EtOAc in toluene to afford the pure individual isomers (5-isomer 4.5 g, 25%; 6-isomer 6.4 g, 35%). 5-Carboxyfluorescein diacetate N-hydroxysuccinimide ester: Rf 0.37 (50:50 EtOAc/Toluene), HPLC tR 8.6 min (<1%of 6-isomer). 6-Carboxyfluorescein diacetate N-hydroxysuccinimide ester: Rf 0.53 (50:50 EtOAc/Toluene), HPLC tR 7.8 min (<1% of 5-isomer).
With 1-ethyl-(3-(3-dimethylamino)propyl)-carbodiimide hydrochloride In dichloromethane at 20℃; Inert atmosphere;
With 1-ethyl-(3-(3-dimethylamino)propyl)-carbodiimide hydrochloride In dichloromethane at 20℃; for 3h; 9 diacetyl-5-(2-carboxyethylaminocarbonyl) fluorescein 22 5-Carboxyfluorescein diacetate (17) (28 mg) was stirred with NHS (8.3 mg, 1.2 equiv) and EDAC (14 mg, 1.2 equiv) in DCM (2 mL) at room temperature for 3 h and then beta-alanine (54 mg, 10 equiv) and pyridine (2 mL) were added into the mixture which was further stirred at 40 °C overnight. The solvents were removed in vacuo and the residue was purified by preparative HPLC using a Luna 5 mih C18 column (isocratic, 50% MeCN in 0.1% formic acid) to yield diacetyl-5-(2-carboxyethylaminocarbonyl) fluorescein (22) (20 mg).
  • 13
  • [ 108-24-7 ]
  • [ 108-46-3 ]
  • [ 552-30-7 ]
  • [ 3348-03-6 ]
  • [ 79955-27-4 ]
YieldReaction ConditionsOperation in experiment
Stage #1: recorcinol; trimellitic Anhydride With zinc dibromide at 180℃; for 1.33333h; Stage #2: acetic anhydride for 2h; Reflux; Overall yield = 4.3 g;
1: 161 mg 2: 152 mg Stage #1: recorcinol; trimellitic Anhydride With methanesulfonic acid at 80℃; for 24h; Stage #2: acetic anhydride With pyridine at 80℃; for 0.0833333h; 5 4. SYNTHESIS OF 5-CARBOXYFLUORESCEIN DIPIVALATE (16) Trimellitic anhydride (436 mg, 1 equiv) and resorcinol (500 mg, 2 equiv) were stirred in methanes ulfonic acid (12 mL) at 80 °C for 24 h. The resulted mixture was poured into ice water (thO). The precipitate was collected and washed with water to yield the crude 5 (6) -carboxy fluorescein (15) (820 mg).5(6)-carboxyfluorescein (15): yellow powder; 'H NMR (500 MHz, methanol -d 8.59 (d, J = 1.5 Hz, 1H), 8.37 (dd, J = 8.0, 1.5 Hz, 1H), 8.31 (dd, J = 8.0, 1.5 Hz, 1H), 8.09 (d, J = 8.0 Hz, lH), 7.74 (s, 1H), 7.30 (d, J = 8.0 Hz, 1H), 6.70-6.72 (d, J = 9.7 Hz, 4H), 6.60- 6.42 (br s, 4H), 6.53-6.57 (m, 4H). 5. SYNTHESIS OF 5-CARBOXYFLUORESCEIN DIACETATE (17)5(6)-carboxyfluorescein (15) (320 mg) and pyridine (260 pL) were stirred in acetic anhydride (5 mL) at 80 °C for 5 min. The solvents were removed in vacuo and the residue was purified by preparative HPLC using a Luna 5 pm C18 column (isocratic, 50% MeCN in 0.1% formic acid) to yield 5-carboxyfluorescein diacetate (17) (161 mg) and 6- carboxy fluorescein diacetate (152 mg).[0487] 5-carboxyfluorescein diacetate (17): yellow powder; 'H NMR (400 MHz, methanol-iL): d 8.61 (s, lH), 8.37 (d, J = 8.0 Hz, lH), 7.32 (d, J = 8.0 Hz, lH), 7.18 (br s, 2H), 6.88 (br s, 4H), 2.28 (s, 6H).
  • 14
  • [ 1802991-75-8 ]
  • [ 79955-27-4 ]
  • [ 1802991-71-4 ]
YieldReaction ConditionsOperation in experiment
110 mg Stage #1: 5-Carboxyfluorescein diacetate With N-ethyl-N,N-diisopropylamine; HATU In N,N-dimethyl-formamide at 0℃; for 0.5h; Stage #2: bis(acetoxymethyl) 2,2'-((4-amino-2-methoxyphenyl)azanediyl)diacetate In N,N-dimethyl-formamide at 20℃; for 2h; 1F Example 1F Example 1F To a solution of commercially available 5-carboxyfluoroscein diacetate (174 mg, 0.43 mmol) in DMF (4 mL) at 0° C. was added DIPEA (0.26 mL, 1.52 mmol) followed by HATU (198 mg, 0.52 mmol). After stirring 30 min at 0° C., the aminobenzene product of Example 1E (200 mg, 0.43 mmol) was added to the reaction mixture and stirred for 2 h at room temperature. The mixture was then poured into water and resulting orange solid was filtered off. The residue was purified by column chromatography using Hex/EtOAc (gradient: 0 to 100% EtOAc with 1% of triethylamine) to afford the desired amide product (110 mg, 30%) as a white solid: 1H NMR (CDCl3, 400 MHz) δ (ppm) 8.19-8.06 (m, 3H), 7.57 (s, 1H), 7.34 (s, 1H), 7.04 (s, 2H), 6.86 (dd, J=8.8, 1.6 Hz, 1H), 6.81-6.73 (m, 5H), 5.75 (s, 4H), 4.13 (s, 4H), 3.72 (s, 3H), 2.32 (s, 6H), 2.08 (s, 6H); 13C NMR (CDCl3, 100 MHz) δ 170.0, 169.4, 169.1, 167.9, 163.2, 153.0, 152.1, 151.5, 151.2, 141.3, 135.0, 132.9, 129.8, 128.8, 128.2, 125.6, 122.1, 119.4, 117.9, 115.6, 112.5, 110.5, 105.1, 81.6, 79.2, 55.4, 53.5, 21.0, 20.6; LCMS (ESI) tR: 0.856 min (>99%, ELSD), m/z: 841.2 [M+1]+
 

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