2-(2-aminoethyldisulfanyl)ethanamine;dihydrochloride

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: 2.2 g of cystamine dihydrochloride were dissolved in 20 ml of water and adjusted to pH 10 with NaOH. 2.1 g of p-azidobenzenesulfonyl chloride were suspended in this solution and the suspension was stirred at room temperature for 5 hours. The precipitated N-(p-azidobenzenesulfonyl)cystamine was reacted with 2 g of succinic anhydride and stirred overnight. The resulting solution was acidified with HCl, subsequently filtered and washed with water. The residue was dried at room temperature in a high vacuum and gave 1.53 g of N-(p-azidobenzenesulfonyl) N'-(3-carboxypropionyl) cystamine. The IR showed bands at 3283 (amide NH) 2134 (azide), 1714 (acid carbonyl), 1650 (amide), 1589+1547 (aromatic), 1284 (COOH), 1328+1180 (arylsulfonyl ), 839 (p-disubst.benzene). TLC (silica gel--NH3 conc./EtOH=1%) Rf=0.7. m.p.: dec. at 163°. stirred overnight subsequently filtered washed with water The residue was dried at room temperature in a high vacuum

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: 2.2 g of cystamine dihydrochloride were dissolved in 20 ml of water and adjusted to pH 10 with NaOH. 2.1 g of p-azidobenzenesulfonyl chloride were suspended in this solution and the suspension was stirred at room temperature for 5 hours. The precipitated N-(p-azidobenzenesulfonyl)cystamine was reacted with 2 g of succinic anhydride and stirred overnight. The resulting solution was acidified with HCl, subsequently filtered and washed with water. The residue was dried at room temperature in a high vacuum and gave 1.53 g of N-(p-azidobenzenesulfonyl) N'-(3-carboxypropionyl) cystamine. The IR showed bands at 3283 (amide NH) 2134 (azide), 1714 (acid carbonyl), 1650 (amide), 1589+1547 (aromatic), 1284 (COOH), 1328+1180 (arylsulfonyl ), 839 (p-disubst.benzene). TLC (silica gel--NH3 conc./EtOH=1%) Rf=0.7. m.p.: dec. at 163°. stirred overnight subsequently filtered washed with water The residue was dried at room temperature in a high vacuum

Conditions: temp:75.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: A 25 mL round-bottom flask fitted with a magnetic stirrer was charged with 225 mg (1.0 mmol) of cystamine dihydrochloride, 409 mg (2.2 mmol) 2,4-dinitrofluorobenzene, 25 mg of K2CO3, and 10 mL of H2O. The mixture then stirred in a 75° C. oil bath for 18 h. After cooling, the solvent was decanted from a yellow insoluble sludge. The sludge crystallized on trituration with 10 mL of CH2Cl2. The yellow crystals were washed with additional CH2Cl2 and recrystallized from 0.5 mL of 15:85 H2O/DMF. The isolated product was washed with 1:1 H2O/DMF affording 77 mg of N,N'-bis-(2,4-dinitrophenyl) cystamine, mp 175°-176° C. Calcd for C16H16N6O8S2:C, 39.67; H, 3.31; N, 17.36; S, 13.22. Found: C, 39.74; H, 3.38; N, 17.26; S, 13.48. 200 MHz NMR (DMSO-d6): 3.08 (t, J =6.6, 4H), 3.80 (4 lines, 4H), 7.27 (d, J =9.6, 2H), 8.25 (dd, J =2.8, 9.6, 2H), 8.84 (dd, J =2.8, 0.8, 2H), 8.93 (t, J=5.6, 2H). A 25 mL round-bottom flask fitted with a magnetic stirrer After cooling the solvent was decanted from a yellow insoluble sludge The sludge crystallized on trituration with 10 mL of CH2Cl2 The yellow crystals were washed with additional CH2Cl2 recrystallized from 0.5 mL of 15:85 H2O/DMF The isolated product was washed with 1:1 H2O/DMF affording 77 mg of N,N'-bis-(2,4-dinitrophenyl) cystamine, mp 175°-176° C

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: m.p. 150°-151° C., 14.5 g., was prepared as in Example 1 using 33.6 g. of 4-trifluoromethoxybenzoyl chloride in 100 ml. of ethylene dichloride, 11.3 g. of cystamine dihydrochloride, 300 ml. of 10% sodium hydroxide solution, 400 ml. of ethylene dichloride and recrystallization from acetonitrile using decolorizing charcoal. , was prepared as in Example 1 using 33.6 g of ethylene dichloride and recrystallization from acetonitrile using

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: 2.2 g of cystamine dihydrochloride were dissolved in 20 ml of water and adjusted to pH 10 with NaOH. 2.1 g of p-azidobenzenesulfonyl chloride were suspended in this solution and the suspension was stirred at room temperature for 5 hours. The precipitated N-(p-azidobenzenesulfonyl)cystamine was reacted with 2 g of succinic anhydride and stirred overnight. The resulting solution was acidified with HCl, subsequently filtered and washed with water. The residue was dried at room temperature in a high vacuum and gave 1.53 g of N-(p-azidobenzenesulfonyl) N'-(3-carboxypropionyl) cystamine. The IR showed bands at 3283 (amide NH) 2134 (azide), 1714 (acid carbonyl), 1650 (amide), 1589+1547 (aromatic), 1284 (COOH), 1328+1180 (arylsulfonyl ), 839 (p-disubst.benzene). TLC (silica gel--NH3 conc./EtOH=1%) Rf=0.7. m.p.: dec. at 163°. stirred overnight subsequently filtered washed with water The residue was dried at room temperature in a high vacuum

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: Cystamine dihydrochloride (68 mg, 0.301 mmol) and DIEA (110 μL, 2.1 eq.) were dissolved in DMF (10 mL), followed by addition of NHS-fluorescein, a mixture of 5- and 6-carboxyfluorescein (300 mg, 0.634 mmol) and the resulting reaction mixture was stirred overnight at room temperature. Then it was diluted with ethyl acetate and washed three times with water and one time with brine. The extract was dried over anhydrous sodium sulfate, concentrated under reduced pressure, redissolved in small amount of methanol and ethyl acetate, and then triturated with diethyl ether to obtain 140 mg of fluorescein-cystamine adduct as a bright orange solid. the resulting reaction mixture washed three times with water and one time with brine The extract was dried over anhydrous sodium sulfate concentrated under reduced pressure redissolved in small amount of methanol and ethyl acetate triturated with diethyl ether

Conditions: temp:-15.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: Arachidonic acid (152 mg, 0.5 mmol) and triethylamine (52 mg, 0.51 mmol) were dissolved in 3 ml of dry acetonitrile and chilled to −15° C., and 70 mg (0.51 mmol) of butyl chloroformate was added. After 30 min, the mixture free of the precipitated triethylamine hydrochloride was pipetted in a solution of cystamine dihydrochloride (56 mg, 0.25 mmol) and triethylamine (52 mg, 0.51 mmol) in 1 ml of methanol, stirring was continued for 15 min at −15° C., then the mixture obtained was allowed to warm to room temperature. After 2 h, 0.5 M HCl was added, and the mixture was extracted with ether (20 ml). The extract was washed with water, then dried with Na2SO4, and evaporated under reduced pressure. The residue was dissolved in 2 ml of chloroform and purified by column (2×2 cm) chromatography on aluminium oxide (basic, Brockmann II). Elution of the column with chloroform-methanol (9:1 v/v) and evaporation of the appropriate fractions gave 181 mg (75%) of the desired N,N′-diarachidonoylcystamine as a waxy solid: TLC, Merck silica gel 60 pre-coated plates [system A: benzen-dioxan-acetic acid (25:5:1 v/v/v)] Rf0.5. was continued for 15 min at −15° C. the mixture obtained to warm to room temperature After 2 h the mixture was extracted with ether (20 ml) The extract was washed with water dried with Na2SO4 evaporated under reduced pressure The residue was dissolved in 2 ml of chloroform purified by column (2×2 cm) chromatography on aluminium oxide (basic, Brockmann II) Elution of the column with chloroform-methanol (9:1 v/v) and evaporation of the appropriate fractions

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: Cystamine dihydrochloride (68 mg, 0.301 mmol) and DIEA (110 μL, 2.1 eq.) were dissolved in DMF (10 mL), followed by addition of NHS-fluorescein, a mixture of 5- and 6-carboxyfluorescein (300 mg, 0.634 mmol) and the resulting reaction mixture was stirred overnight at room temperature. Then it was diluted with ethyl acetate and washed three times with water and one time with brine. The extract was dried over anhydrous sodium sulfate, concentrated under reduced pressure, redissolved in small amount of methanol and ethyl acetate, and then triturated with diethyl ether to obtain 140 mg of fluorescein-cystamine adduct as a bright orange solid. the resulting reaction mixture washed three times with water and one time with brine The extract was dried over anhydrous sodium sulfate concentrated under reduced pressure redissolved in small amount of methanol and ethyl acetate triturated with diethyl ether

Conditions: temp:-15.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: Arachidonic acid (152 mg, 0.5 mmol) and triethylamine (52 mg, 0.51 mmol) were dissolved in 3 ml of dry acetonitrile and chilled to −15° C., and 70 mg (0.51 mmol) of butyl chloroformate was added. After 30 min, the mixture free of the precipitated triethylamine hydrochloride was pipetted in a solution of cystamine dihydrochloride (56 mg, 0.25 mmol) and triethylamine (52 mg, 0.51 mmol) in 1 ml of methanol, stirring was continued for 15 min at −15° C., then the mixture obtained was allowed to warm to room temperature. After 2 h, 0.5 M HCl was added, and the mixture was extracted with ether (20 ml). The extract was washed with water, then dried with Na2SO4, and evaporated under reduced pressure. The residue was dissolved in 2 ml of chloroform and purified by column (2×2 cm) chromatography on aluminium oxide (basic, Brockmann II). Elution of the column with chloroform-methanol (9:1 v/v) and evaporation of the appropriate fractions gave 181 mg (75%) of the desired N,N′-diarachidonoylcystamine as a waxy solid: TLC, Merck silica gel 60 pre-coated plates [system A: benzen-dioxan-acetic acid (25:5:1 v/v/v)] Rf0.5. was continued for 15 min at −15° C. the mixture obtained to warm to room temperature After 2 h the mixture was extracted with ether (20 ml) The extract was washed with water dried with Na2SO4 evaporated under reduced pressure The residue was dissolved in 2 ml of chloroform purified by column (2×2 cm) chromatography on aluminium oxide (basic, Brockmann II) Elution of the column with chloroform-methanol (9:1 v/v) and evaporation of the appropriate fractions

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: 10,10-dimethyl-8-(1-methyl-1H-indol-3-yl)-10H-pyrido-[1,2-a]indolium-2-carboxylate 0.37 g was solubilized in 5 mL N-methylpyrrolidinone (NMP), a solution of HCl in dry dioxane and diethyl ether (1 mL; 1N; 1:3) was added and 1,1′-carbonyldiimidazole (320 mg) was progressively added. After 3 h, 200 mg of cystamine dihyrochloride in mL diisopropylethylamine (DIPEA) were added to the mixture. After 24 h, the reaction mixture was poured in 100 mL acetone. The resulting solid was filtered, washed with acetone and dried under vacuum (340 mg, yellow powder). Analyses were in accordance with the expected structure. After 24 h The resulting solid was filtered washed with acetone dried under vacuum (340 mg, yellow powder)

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: 2.2 g of cystamine dihydrochloride were dissolved in 20 ml of water and adjusted to pH 10 with NaOH. 2.1 g of p-azidobenzenesulfochloride were suspended in the solution and the mixture was stirred at room temperature for 5 hours. The precipitated N-(p-azidobenzenesulfonyl)cystamine was reacted with 2 g of succinic anhydride and stirred overnight. The resulting solution was acidified with HCl and the product was subsequently filtered off and washed with water. The residue was dried at room temperature in a high vacuum and gave 1.53 g of N-(p-azidobenzenesulfonyl)-N'-(3-carboxypropionyl)cystamine. The IR showed bands at 3283 (amide NH), 2134 (azide), 1714 (acid carbonyl), 1650 (amide), 1589+1547 (aromatic), 1284 (COOH), 1328+1180 (arylsulfonyl), 839 (p- disubst. benzene); TLC (silica gel: conc. NH3 (EtOH=1%). Rf=0.7; m.p. 163° (dec.). stirred overnight the product was subsequently filtered off washed with water The residue was dried at room temperature in a high vacuum

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: 11.9g. of calcium pantothenate, 6.8g. of cystamine dihydrochloride and 7.7g. of 1-hydroxylbenzotrizole were dissolved in 50ml. of pyridine and 10ml. of water, and thereafter pyridine (25ml.) solution of 10.5g. of dicyclohexylcarbodiimide was added thereto at a time, and the reaction was effected at a temperature below 10° C for 1 hour and then for 5 hours at a room temperature. Next the solvent was removed under a reduced pressure, the residue thereof was added with 50ml. of water, and the deposited crystals were removed by filtration. Pyridine left in the filtered mother liquor was completely removed azetropically with water. The residue thereof was added with 50ml. of water and after cooling, the deposited crystals were filtration removed and extracted with 20ml. of chloroform two times. The water layer was treated under a reduced pressure to remove the remaining chloroform, and thereafter like Example 1, passed through the ion exchange resin, and the flowed out liquid was concentrated under a reduced pressure and thereby colorless transparent viscous liquid of pantethine which was the objective compound was obtained. Drying was carried out by means of a vacuum dryer for one night and as a result 9.6g. of pantethine (yield rate 69.2%) was obtained. of 1-hydroxylbenzotrizole were dissolved in 50ml for 5 hours at a room temperature Next the solvent was removed under a reduced pressure the residue thereof was added with 50ml of water, and the deposited crystals were removed by filtration Pyridine left in the filtered mother liquor was completely removed azetropically with water The residue thereof was added with 50ml of water and after cooling filtration removed extracted with 20ml The water layer was treated under a reduced pressure to remove the remaining chloroform was concentrated under a reduced pressure was obtained Drying was carried out by means of a vacuum dryer for one night and as a result 9.6g