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US 6,551,591 B1Granted
Antibiotics from microbispora
AssigneeEssential Therapeutics, Inc.
InventorMay D. Lee
Life of this patent
Estimating the term…
Abstract
The present invention provides a novel strain of Microbispora corallina, means for identifying it, methods for culturing it at several different levels, two new antibiotics derived from the cultures and the physico-chemical characteristics of those compounds and their biological activity against a variety of microorganisms.
Claims10
Antibiotic MF-BA-1768α1, or a pharmaceutically acceptable salt thereof, having the following physicochemical characteristics in the non-salt form:
- a. molecular weight:
- 2214.880, calculated monoisotopic mass from [M+2H]2+ at m/z 1108.449, determined by high resolution ESI-FTMS using reference compound peptide mellitin (3+ charge);
- b. ultraviolet absorption spectrum shown in FIG. 1, comprising absorption maxima at λ1=223 nm (ε=73600, log ε=4.87) and λ2=270 nm (ε=17700, log ε=4.25);
- c. infrared absorption spectrum shown in FIG. 2, comprising absorption maxima at 3309 and 1669 (br) cm−1 (KBr disc);
- d. proton magnetic resonance spectrum shown in FIG. 3 (400 MHz, d6-DMSO, 55° C.;
- referenced to CD3SOCD2H quintet at 2.50 ppm);
- e. carbon-13 nuclear magnetic resonance signals comprising the following chemical shifts (in ppm, referenced to CD3SOCD2H heptet at 39.5 ppm, 55° C.), multiplicities (as the number of attached protons), chemical shifts of attached protons, and the COSY correlation of attached protons of select carbons:
- 13C (ppm, *Residue multiplicity) 1H (ppm) COSY (ppm) #Trp 120.8 (CH) 7.05 7.34 Trp 126.0 (CH) 7.19 10.89 #Trp 120.8 (CH) 7.05 7.54 Trp 117.2 (CH) 7.54 7.05 Trp 112.8 (CH) 7.34 7.05 Trp N—H 10.89 7.19 DHB 126.3 (CH) 6.21 1.72 DHB 13.0 (CH3) 1.72 6.21 DHA 103.7 (CH2) 5.98 5.40 AviCys 130.2 (CH) 6.83 8.38, 5.46 AviCys 102.5 (CH) 5.46 6.83 AviCys N—H 8.38 6.83 *Trp = tryptophan;
- DHB = dehydrobutyrine;
- DHA = dehydroalanine;
- AviCys = aminovinyl cysteine. #Signals overlapped
- f. retention time of 9.1 minutes when chromatographed under the following conditions:
- Column:
- 15.0×0.46 cm PLRPs (Polymer Labs, 5 um)
- Flow rate:
- 1 ml/min
- Detection:
- 254 nm
- Eluant:
- acetonitrile (Solvent A) 0.05% Formic Acid in Water (pH 2.83) (solvent B)
- Protocol:
- the following solvent ratios and gradient:
- Time Solvent mixture A/B, v/v 0 min 25/75 2 min 25/75 10 min 38/62 20 min 38/62.
20 more elements
Antibiotic MF-BA-1768β1, or a pharmaceutically acceptable salt thereof, having the following physicochemical characteristics in the non-salt form:
- a. molecular weight:
- 2180.805, calculated monoisotopic mass from [M+2H]2+ at m/z 1091.410, determined by high resolution ESI-FTMS using reference compound peptide mellitin (3+ charge);
- b. ultraviolet spectrum shown in FIG. 4, comprising absorption maxima at λ1 221 nm (ε=105300, log ε=5.02) and λ2 270 nm (ε=19300, log ε=4.28);
- c. infrared absorption spectrum shown in FIG. 5, comprising absorption maxima at 3309 and 1669 cm−1 (KBr disc);
- d. proton magnetic resonance spectrum shown in FIG. 6 (400 MHz, d6-dmso, 55° C.;
- referenced to CD3SOCD2H quintet at 2.50 ppm);
- e. carbon-13 nuclear magnetic resonance signals comprising the following chemical shifts (in ppm, referenced to CD3SOCD2H heptet at 39.5 ppm, 55° C.), multiplicities (as the number of attached protons), chemical shifts of attached protons, and COSY correlation of the attached protons of select carbons:
- 13C (ppm, *Residue multiplicity) 1H (ppm) COSY (ppm) Trp 120.9 (CH) 7.06 7.32 Trp 128.0 (CH) 7.12 10.71 Trp 118.2 (CH) 6.97 7.51 Trp 117.8 (CH) 7.51 6.97 Trp 111.2 (CH) 7.32 7.06 Trp N—H 10.71 7.12 DHB 125.7 (CH) 6.21 1.70 DHB 12.6 (CH3) 1.70 6.21 DHA 103.0 (CH2) 6.04 5.39 AviCys 130.1 (CH) 6.83 8.35, 5.45 AviCys 102.1 (CH) 5.45 6.83 AviCys N—H 8.35 6.83 *Trp = tryptophan;
- DHB = dehydrobutyrine;
- DHA = dehydroalanine;
- AviCys = aminovinyl cysteine.
- f. retention time (Rt) of 10.6 minutes when chromatographed under the following conditions:
- Column:
- 15.0×0.46 cm PLRPs (Polymer Labs, 5 um)
- Flow rate:
- 1 ml/min
- Detection:
- 254 nm
- Eluant:
- acetonitrile (Solvent A) 0.05% Formic Acid in Water (pH 2.83) (solvent B)
- Protocol:
- the following solvent ratios and gradient:
- Time Solvent mixture A/B, v/v 0 min 25/75 2 min 25/75 10 min 38/62 20 min 38/62.
20 more elements
A process for producing antibiotic MF-BA-1768α1, comprising cultivating a strain of Microbispora corallina, NRRL 30420, or an antibiotic MF-BA-1768α1-producing mutant, variant or recombinant form thereof, in a culture medium containing assimilable sources of carbon, nitrogen and inorganic salts under aerobic fermentation conditions until antibiotic MF-BA-1768α1 is produced and then recovering the antibiotic therefrom.
A process for producing antibiotic MF-BA-1768β1, comprising cultivating a strain of Microbispora corallina, NRRL 30420, or an antibiotic MF-BA-1768β1-producing mutant, variant or recombinant form thereof, in a culture medium containing assimilable sources of carbon, nitrogen and inorganic salts under aerobic fermentation conditions until antibiotic MF-BA-1768β1 is produced and then recovering the antibiotic therefrom.
A composition comprising:
- a therapeutically effective amount of compound MF-BA-1768α1 or a pharmaceutically acceptable salt thereof;
- and,
- a pharmaceutically acceptable carrier,
- wherein compound MF-BA-1768α1 has the following physicochemical characteristics:
- a. molecular weight:
- 2214.880, calculated monoisotopic mass from [M+2H]2+ at m/z 1108.449, determined by high resolution ESI-FTMS using reference compound peptide mellitin (3+ charge);
- b. ultraviolet absorption spectrum shown in FIG. 1, comprising absorption maxima at λ1=223 nm (ε=73600, log ε=4.87) and λ2=270 nm (ε=17700, log ε=4.25);
- c. infrared absorption spectrum shown in FIG. 2, comprising absorption maxima at 3309 and 1669 (br) cm−1 (KBr disc);
- d. proton magnetic resonance spectrum shown in FIG. 3 (400 MHz, d6-DMSO, 55° C.;
- referenced to CD3SOCD2H quintet at 2.50 ppm);
- e. carbon-13 nuclear magnetic resonance signals comprising the following chemical shifts (in ppm, referenced to CD3SOCD2H heptet at 39.5 ppm, 55° C.), multiplicities (as the number of attached protons), chemical shifts of attached protons, and the COSY correlation of attached protons of select carbons:
- 13C (ppm, *Residue multiplicity) 1H (ppm) COSY (ppm) #Trp 120.8 (CH) 7.05 7.34 Trp 126.0 (CH) 7.19 10.89 #Trp 120.8 (CH) 7.05 7.54 Trp 117.2 (CH) 7.54 7.05 Trp 112.8 (CH) 7.34 7.05 Trp N—H 10.89 7.19 DHB 126.3 (CH) 6.21 1.72 DHB 13.0 (CH3) 1.72 6.21 DHA 103.7 (CH2) 5.98 5.40 AviCys 130.2 (CH) 6.83 8.38, 5.46 AviCys 102.5 (CH) 5.46 6.83 AviCys N—H 8.38 6.83 *Trp = tryptophan;
- DHB = dehydrobutyrine;
- DHA = dehydroalanine;
- AviCys = aminovinyl cysteine. #Signals overlapped
- f. retention time of 9.1 minutes when chromatographed under the following conditions:
- Column:
- 15.0×0.46 cm PLRPs (Polymer Labs, 5 um) Flow rate:
- 1 ml/min
- Detection:
- 254 nm
- Eluant:
- acetonitrile (Solvent A) 0.05% Formic Acid in Water (pH 2.83) (solvent B)
- Protocol:
- the following solvent ratios and gradient:
- Time Solvent mixture A/B, v/v 0 min 25/75 2 min 25/75 10 min 38/62 20 min 38/62.
23 more elements
A composition comprising:
- a therapeutically effective amount of compound MF-BA-1768β1 or a pharmaceutically acceptable salt thereof;
- and, a pharmaceutically acceptable carrier,
- wherein compound MF-BA-1768β1 has the following physicochemical characteristics:
- a. molecular weight:
- 2180.805, calculated monoisotopic mass from [M+2H]2+ at m/z 1091.410, determined by high resolution ESI-FTMS using reference compound peptide mellitin (3+ charge);
- b. ultraviolet spectrum shown in FIG. 4, comprising absorption maxima at λ1 221 nm (ε=105300, log ε=5.02) and λ2 270 nm (ε=19300, log ε=4.28);
- c. infrared absorption spectrum shown in FIG. 5, comprising absorption maxima at 3309 and 1669 cm−1 (KBr disc);
- d. proton magnetic resonance spectrum shown in FIG. 6 (400 MHz, d6-dmso, 55° C.;
- referenced to CD3SOCD2H quintet at 2.50 ppm);
- e. carbon-13 nuclear magnetic resonance signals comprising the following chemical shifts (in ppm, referenced to CD3SOCD2H heptet at 39.5 ppm, 55° C.), multiplicities (as the number of attached protons), chemical shifts of attached protons, and COSY correlation of the attached protons of select carbons:
- 13C (ppm, *Residue multiplicity) 1H (ppm) COSY (ppm) Trp 120.9 (CH) 7.06 7.32 Trp 128.0 (CH) 7.12 10.71 Trp 118.2 (CH) 6.97 7.51 Trp 117.8 (CH) 7.51 6.97 Trp 111.2 (CH) 7.32 7.06 Trp N—H 10.71 7.12 DHB 125.7 (CH) 6.21 1.70 DHB 12.6 (CH3) 1.70 6.21 DHA 103.0 (CH2) 6.04 5.39 AviCys 130.1 (CH) 6.83 8.35. 5.45 AviCys 102.1 (CH) 5.45 6.83 AviCys N—H 8.35 6.83 *Trp = tryptophan;
- DHB = dehydrobutyrine;
- DHA = dehydroalanine;
- AviCys = aminovinyl cysteine.
- f. retention time (Rt) of 10.6 minutes when chromatographed under the following conditions:
- Column:
- 15.0×0.46 cm PLRPs (Polymer Labs, 5 um)
- Flow rate:
- 1 ml/min
- Detection:
- 254 nm
- Eluant:
- acetonitrile (Solvent A) 0.05% Formic Acid in Water (pH 2.83) (solvent B)
- Protocol:
- the following solvent ratios and gradient:
- Time Solvent mixture A/B, v/v 0 min 25/75 2 min 25/75 10 min 38/62 20 min 38/62.
23 more elements
A method for treating a bacterial infection comprising administering to a patient in need thereof a therapeutically effective amount of compound MF-BA-1768α1 which has the following physicochemical characteristics:
- a. molecular weight:
- 2214.880, calculated monoisotopic mass from [M+2H]2+ at m/z 1108.449, determined by high resolution ESI-FTMS using reference compound peptide mellitin (3+ charge);
- b. ultraviolet absorption spectrum shown in FIG. 1, comprising absorption maxima at λ1=223 nm (ε=73600, log ε=4.87) and λ2=270 nm (ε=17700, log ε=4.25);
- c. infrared absorption spectrum shown in FIG. 2, comprising absorption maxima at 3309 and 1669 (br) cm−1, (KBr disc);
- d. proton magnetic resonance spectrum shown in FIG. 3 (400 MHz, d6-DMSO, 55° C.;
- referenced to CD3SOCD2H quintet at 2.50 ppm);
- e. carbon-13 nuclear magnetic resonance signals comprising the following chemical shifts (in ppm, referenced to CD3SOCD2H heptet at 39.5 ppm, 55° C.), multiplicities (as the number of attached protons), chemical shifts of attached protons, and the COSY correlation of attached protons of select carbons:
- 13C (ppm, *Residue multiplicity) 1H (ppm) COSY (ppm) #Trp 120.8 (CH) 7.05 7.34 Trp 126.0 (CH) 7.19 10.89 #Trp 120.8 (CH) 7.05 7.54 Trp 117.2 (CH) 7.54 7.05 Trp 112.8 (CH) 7.34 7.05 Trp N—H 10.89 7.19 DHB 126.3 (CH) 6.21 1.72 DHB 13.0 (CH3) 1.72 6.21 DHA 103.7 (CH2) 5.98 5.40 AviCys 130.2 (CH) 6.83 8.38, 5.46 AviCys 102.5 (CH) 5.46 6.83 AviCys N—H 8.38 6.83 *Trp = tryptophan;
- DHB = dehydrobutyrine;
- DHA = dehydroalanine;
- AviCys = aminovinyl cysteine. #Signals overlapped
- f. retention time of 9.1 minutes when chromatographed under the following conditions:
- Column:
- 15.0×0.46 cm PLRPs (Polymer Labs, 5 um)
- Flow rate:
- 1 ml/min
- Detection:
- 254 nm
- Eluant:
- acetonitrile (Solvent A) 0.05% Formic Acid in Water (pH 2.83) (solvent B)
- Protocol:
- the following solvent ratios and gradient:
- Time Solvent mixture A/B, v/v 0 min 25/75 2 min 25/75 10 min 38/62 20 min 38/62.
20 more elements
A method for treating a bacterial infection comprising administering to a patient in need thereof a therapeutically effective amount of compound MF-BA-1768β1 which has the following physicochemical characteristics:
- a. molecular weight:
- 2180.805, calculated monoisotopic mass from [M+2H]2+ at m/z 1091.410, determined by high resolution ESI-FTMS using reference compound peptide mellitin (3+ charge);
- b. ultraviolet spectrum shown in FIG. 4, comprising absorption maxima at λ1 221 nm (ε=105300, log ε=5.02) and λ2 270 nm (ε=19300, log ε=4.28);
- c. infrared absorption spectrum shown in FIG. 5, comprising absorption maxima at 3309 and 1669 cm−1 (KBr disc);
- d. proton magnetic resonance spectrum shown in FIG. 6 (400 MHz, d6-dmso, 55° C.;
- referenced to CD3SOCD2H quintet at 2.50 ppm);
- e. carbon-13 nuclear magnetic resonance signals comprising the following chemical shifts (in ppm, referenced to CD3SOCD2H heptet at 39.5 ppm, 55° C.), multiplicities (as the number of attached protons), chemical shifts of attached protons, and COSY correlation of the attached protons of select carbons:
- 13C (ppm, *Residue multiplicity) 1H (ppm) COSY (ppm) Trp 120.9 (CH) 7.06 7.32 Trp 128.0 (CH) 7.12 10.71 Trp 118.2 (CH) 6.97 7.51 Trp 117.8 (CH) 7.51 6.97 Trp 111.2 (CH) 7.32 7.06 Trp N—H 10.71 7.12 DHB 125.7 (CH) 6.21 1.70 DHB 12.6 (CH3) 1.70 6.21 DHA 103.0 (CH2) 6.04 5.39 AviCys 130.1 (CH) 6.83 8.35, 5.45 AviCys 102.1 (CH) 5.45 6.83 AviCys N—H 8.35 6.83 *Trp = tryptophan;
- DHB = dehydrobutyrine;
- DHA = dehydroalanine;
- AviCys = aminovinyl cysteine.
- f. retention time (R1) of 10.6 minutes when chromatographed under the following conditions:
- Column:
- 15.0×0.46 cm PLRPs (Polymer Labs, 5 um)
- Flow rate:
- 1 ml/min
- Detection:
- 254 nm
- Eluant:
- acetonitrile (Solvent A) 0.05% Formic Acid in Water (pH 2.83) (solvent B)
- Protocol:
- the following solvent ratios and gradient:
- Time Solvent mixture A/B, v/v 0 min 25/75 2 min 25/75 10 min 38/62 20 min 38/62.
20 more elements
Dependencies are inferred from the claim text.