WS7622A mono- or di- sulfate, process for preparation thereof and use thereof

WS7622A mono- or di- sulfate or their pharmaceutically acceptable salts, among which the di-sodium salt of WS7622A di-sulfate or the di-potassium salt of WS7622A di-sulfate have the following physico-chemical properties:

• ______________________________________ Di-sodium salt of WS7622A di-sulfate:
• Appearance:
• colorless crystals Solubility:
• soluble;
• water, methanol insoluble;
• chloroform, n-hexane Melting point:
• 257-263.degree. C. (dec.) Specific rotation:
• [.alpha]sub.D.sup.23 + 37.5.degree. (C = 1.0, methanol) Molecular formula:
• C.sub.47 H.sub.61 N.sub.9 O.sub.19 S.sub.2 Na.sub.2 Elemental analysis:
• calcd. (C.sub.47 H.sub.61 N.sub.9 O.sub.19 S.sub.2 Na.sub.2.6H.sub.2 O);
• C 44.30, H 5.77, N 9.89, S 5.03, Na 3.61% found:
• C 44.98, H 5.90, N 10.06, S 5.00, Na 3.98% Molecular weight:
• FAB-MS m/z 1188 (M + Na).sup.+ Thin layer chromatography:
• Stationary phase Developing solvent Rf value ______________________________________ silica gel CHCl.sub.3 --CH.sub.3 OH--H.sub.2 O 0.11 (Merck Art 5715) (65:25:4) n-butanol-acetic acid-water 0.29 (4:2:1) ______________________________________
• Infrared absorption spectrum:
• (attached FIG. 1):
• .nu..sub.max.sup.KBr 3360, 2960, 1735, 1660, 1640, 1530, 1500, 1380, 1250, 1200, 1060, 1030, 940, 890 cm.sup.-1
• ______________________________________ .sup.1 H Nuclear magnetic resonance spectrum (attached FIG. 2):
• (400 MHz, D.sub.2 O) .delta. 7.50 (1H, s) 7.27 (1H, s) 7.33-7.24 (3H, m) 6.94 (1H, q, J=7Hz) 6.85 (2H, br d, J=8Hz) 5.53 (1H, m) 5.37 (1H, m) 4.80 (1H, br s) 4.63-4.57 (2H, m) 4.53 (1H, m) 4.06 (1H, m) 3.99 (1H, d, J=10Hz) 3.56 (1H, br d, J=14Hz) 3.46 (1H, m) 2.97 (3H, s) 2.97-2.88 (2H, m) 2.72 (1H, m) 2.59 (1H, m) 2.51-2.38 (2H, m) 2.09-1.91 (4H, m) 1.82-1.60 (3H, m) 1.77 (3H, d, J=7Hz) 1.50 (3H, d, J=6.5Hz) 1.40 (1H, m) 1.11 (6H, d, J=7Hz) 0.99 (3H, d, J=6.5Hz) 0.97 (3H, d, J=6.5Hz) ______________________________________ .sup.13 C Nuclear magnetic resonance spectrum (attached FIG. 3):
• (100 MHz D.sub.2 O) .delta. 183.6 (s) 177.9 (s) 177.7 (s) 174.8 (s) 173.8 (s) 173.3 (s) 172.4 (s) 167.8 (s) 161.5 (s) 145.5 (s) 144.9 (s) 139.6 (d) 139.0 (s) 137.0 (s) 136.0 (s) 132.3 (d) .times. 2 131.0 (d) .times. 2 129.6 (d) 127.4 (d) 125.9 (d) 77.4 (d) 75.1 (d) 63.8 (d) 62.7 (d) 59.1 (d) 55.9 (d) 54.9 (d) 51.9 (d) 41.9 (t) 37.2 (d) 36.9 (t) 34.1 (q) 32.3 (d) 31.9 (t) 31.8 (t) 31.2 (t) 27.5 (t) 23.7 (t) 21.7 (q) 21.4 (q) .times. 2 21.3 (q) 21.1 (q) 15.5 (q) ______________________________________
• Amino acid analysis
• Di-sodium salt of WS7622A di-sulfate (1 mg) was hydrolyzed with 6N HCl (1 ml) at 110.degree. C. for 20 hours and the mixture was evaporated to dryness;
• the obtained mixture was analyzed on Hitachi 835 automatic amino acid analyzer;
• Amino acid standard solution [Type H (Wako Chemicals 013-08391) and Type B (Wako Chemicals 016-08641) were used.] were used as reference;
• As the result, threonine, valine, phenylalanine, ornithine, NH.sub.3 and unknown ninhydrin-positive components were detected;
• or
• ______________________________________ Di-potassium salt of WS7622A di-sulfate:
• Appearance:
• colorless amorphous powder Solubility:
• soluble;
• water, methanol insoluble;
• chloroform, n-hexane Melting point:
• 230-237.degree. C. (dec.) Specific rotation:
• [.alpha]sub.D.sup.23 + 34.degree. (C = 1.0, methanol) Molecular formula:
• C.sub.47 H.sub.61 N.sub.9 O.sub.19 S.sub.2 K.sub.2 Elemental analysis:
• calcd. (C.sub.47 H.sub.61 N.sub.9 O.sub.19 S.sub.2 K.sub.2.6H.sub.2 O);
• C 43.21, H 5.63, N 9.65, S 4.91, K 5.99% found:
• C 43.96, H 5.44, N 9.97, S 5.09, K 4.49% Molecular weight:
• FAB-MS m/z 1236 (M + K).sup.+ Thin layer chromatography:
• Stationary phase Developing solvent Rf value ______________________________________ silica gel CHCl.sub.3 --CH.sub.3 OH--H.sub.2 O 0.13 (Merck Art 5715) (65:25:4) ______________________________________
• Melting point:
• 230.degree.-237.degree. C. (dec.)
• Specific rotation:
• [.alpha]sub.D.sup.23 +34.degree. (C=1.0, methanol)
• Molecular formula:
• C.sub.47 H.sub.61 N.sub.9 O.sub.19 S.sub.2 K.sub.2
• Elemental analysis:
• calcd. (C.sub.47 H.sub.61 N.sub.9 O.sub.19 S.sub.2 K.sub.2.6H.sub.2 O);
• C 43.21, H 5.63, N 9.65, S 4.91, K 5.99% found:
• C 43.96, H 5.44, N 9.97, S 5.09, K 4.49%
• Infrared absorption spectrum:
• (attached FIG. 4):
• .nu..sub.max.sup.KBr 3360, 2960, 1735, 1660, 1640, 1530, 1500, 1405, 1380, 1250, 1200, 1050, 1030, 940, 890 cm.sup.-1
• ______________________________________ .sup.1 H Nuclear magnetic resonance spectrum (attached FIG. ______________________________________ 5):
• (400 MHz, D.sub.2 O) .delta. 7.52 (1H, s) 7.28 (1H, s) 7.34-7.25 (3H, m) 6.96 (1H, q, J=7Hz) 6.87 (2H, br d, J=8Hz) 5.56 (1H, m) 5.40 (1H, m) 4.84 (1H, br s) 4.70-4.55 (3H, m) 4.10 (1H, m) 4.03 (1H, m) 3.60 (1H, br d, J=14Hz) 3.50 (1H, m) 3.00 (3H, s) 3.00-2.85 (2H, m) 2.76 (1H, m) 2.62 (1H, m) 2.55-2.40 (2H, m) 2.12-1.95 (4H, m) 1.90-1.65 (3H, m) 1.79 (3H, d, J=7Hz) 1.53 (3H, d, J=6.5Hz) 1.45 (1H, m) 1.14 (6H, d, J=7Hz) 1.02 (3H, d, J=6.5Hz) 1.00 (3H, d, J=6.5Hz) ______________________________________
• Amino acid analysis:
• Di-potassium salt of WS7622A di-sulfate (1 mg) was hydrolyzed with 6N HCl (1 ml) at 110.degree. C. for 20 hours and the mixture was evaporated to dryness;
• the obtained mixture was analyzed on Hitachi 835 automatic amino acid analyzer;
• Amino acid standard solution [Type H (Wako Chemicals 013-08391) and Type B (Wako Chemicals 016-08641) were used.] were used as reference;
• As the result, threonine, valine, phenylalanine, ornithine, NH.sub.3 and unknown ninhydrin-positive components were detected.