US2011286996A1PendingUtilityA1

Antibiotics ge 81112 factors a, b, b1, pharmaceutically acceptable salts and compositions, and use thereof

Assignee: SELVA ENRICOPriority: Nov 28, 2001Filed: Aug 5, 2011Published: Nov 24, 2011
Est. expiryNov 28, 2021(expired)· nominal 20-yr term from priority
A61P 31/04A61P 31/00C12P 1/04C12P 1/06C12P 1/00Y02A50/30
45
PatentIndex Score
0
Cited by
0
References
0
Claims

Abstract

The invention relates to an antibiotic substances of microbial origin, arbitrarily denominated antibiotic GE 81112 factor A, factor B1 and factor B, which is produced by fermentation of Streptomyces sp. DSMZ 14386, the pharmaceutically acceptable salts and compositions thereof, and their use as an antibacterial agent having inhibitory activity versus susceptible microbes.

Claims

exact text as granted — not AI-modified
1 . Antibiotic GE 81112 factor A having the following characteristics:
 A) Mass spectrum recorded from a 0.01 mg/ml solution in acetonitrile:water 50:50 giving a monoprotonated ion at 644 m/z on a Thermofinnigan LCQ deca instrument under the following electrospray conditions: Spray Voltage: 4.7 kV; Capillary temperature: 250° C.; Capillary Voltage: 9V; Infusion mode 41.1/min.   B) Infrared Spectrum ( FIG. 2 ) recorded as nujol mull with an IFS-48 Fourier Transform spectrophotometer showing the following main bands (cm −3 ):
 3356; 2956; 2855; 2256; 2128; 1663; 1596; 1455; 1378; 1345; 1123; 1050; 1026; 1002; 825; 764. 
   C)  1 H-NMR spectrum ( FIG. 3 ) recorded at 600 MHz at 25° C. in DMSO-d 6  and trifluoroacetic acid (drops) showing the following  1 H signals:   
       
         
           
                 
                 
               
                     
                 
                   Chemical 
                     
                 
                   shift 
                 
                   (ppm) 
                   Multiplicity 
                 
                     
                 
                   1.52 
                   br d 
                 
                   1.57-1.67 
                   m 
                 
                   1.81 
                   br d 
                 
                   1.88 
                   m 
                 
                   2.87 
                   dd 
                 
                   2.90 
                   m 
                 
                   3.12 
                   dd 
                 
                   3.15 
                   m 
                 
                   3.65 
                   m 
                 
                   3.83 
                   m 
                 
                   3.94 
                   br d 
                 
                   4.40 
                   br s 
                 
                   4.46 
                   m 
                 
                   4.50 
                   dd 
                 
                   4.70 
                   m 
                 
                   5.06 
                   d 
                 
                   6.46 
                   br 
                 
                   6.94 
                   s 
                 
                   7.29 
                   s 
                 
                   7.99 
                   d 
                 
                   8.17 
                   d 
                 
                   8.30 
                   m 
                 
                   8.57 
                   br d 
                 
                   8.95 
                   d 
                 
                   9.06 
                   br d 
                 
                   14.09  
                   br 
                 
                   14.27  
                   br 
                 
                   — 
                   — 
                 
                     
                 
             
                
                
                
                
                
               
               
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
         D)  13 C-NMR sprectrum ( FIG. 4 ) recorded at 150 MHz, at 5° C. in DMSO-d 6  and trifluoroacetic acids (drops) showing the following  13 C signals 
       
       
         
           
                 
                 
               
                     
                 
                   Chemical 
                     
                 
                   shift 
                 
                   (ppm) 
                   Multiplicity 
                 
                     
                 
                     
                 
                 
                 
               
                   15.9 
                   t 
                 
                   27.19 
                   t 
                 
                   28.41 
                   t 
                 
                   35.31 
                   t 
                 
                   42.86 
                   t 
                 
                   50.13 
                   d 
                 
                   51.30 
                   d 
                 
                   56.86 
                   d 
                 
                   59.71 
                   d 
                 
                   64.07 
                   d 
                 
                   65.11 
                   d 
                 
                   66.88 
                   d 
                 
                   67.76 
                   t 
                 
                   116.83 
                   d 
                 
                   118.49 
                   d 
                 
                   128.41 
                   s 
                 
                   129.03 
                   s 
                 
                   133.65 
                   d 
                 
                   139.66 
                   s 
                 
                   156.73 
                   s 
                 
                   167.39 
                   s 
                 
                   170.16 
                   s 
                 
                   171.29 
                   s 
                 
                   171.65 
                   s 
                 
                     
                 
             
                
                
                
                
                
               
               
                
               
            
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
         E) Retention time 14.1 min, determined by using an analytical HPLC Shimadzu LC10-AS, equipped with an “Altima” RP18, 5 μm, 250×4.6 mm i.d column; 
         isocratic elution perfomed at 1 ml/min. flow rate, with 40 mM ammonium formate buffer, brought to pH 4.5 by addition of formic acid; UV detection at 230 nm. 
       
     
     
         2 . Antibiotic GE 81112 Factor B having the following characteristics:
 A) Mass spectrum recorded from a 0.01 mg/ml solution in acetonitrile:water 50:50 giving a monoprotonated ion at 659 m/z on a Thermofinnigan LCQ deca instrument under the following electrospray conditions: Spray Voltage: 4.7 kV; Capillary temperature: 250° C.; Capillary Voltage: 9V; Infusion mode 5 μl/min.   B) Infrared spectrum ( FIG. 6 ) recorded in KBr with an IFS-48 Fourier Transform spectrophotometer showing the following main bands (cm −1 ):
 3359; 2958; 2852; 2258; 2129; 1681; 1591: 1450; 1385; 1347; 1122; 1072; 1025; 998; 765 
   C)  1 H-NMR spectrum ( FIG. 7 ) recorded at 600 MHz at 25° C. in DMSO-d 6  and trifluoroacetic acid (drops) showing the following  1 H signals:   
       
         
           
                 
                 
               
                     
                 
                   Chemical 
                     
                 
                   shift (ppm) 
                   Multiplicity 
                 
                     
                 
                   1.52 
                   br d 
                 
                   1.57-1.72 
                   m 
                 
                   1.83 
                   br d 
                 
                   1.90 
                   m 
                 
                   2.63 
                   dd 
                 
                   2.88 
                   dd 
                 
                   2.92 
                   m 
                 
                   3.13 
                   br d 
                 
                   3.64 
                   m 
                 
                   3.83 
                   m 
                 
                   3.95 
                   br d 
                 
                   4.35 
                   br s 
                 
                   4.46 
                   m 
                 
                   4.50 
                   dd 
                 
                   4.60 
                   m 
                 
                   5.05 
                   d 
                 
                   6.46 
                   br 
                 
                   6.48 
                   s 
                 
                   6.93 
                   s 
                 
                   7.40 
                   br s 
                 
                   7.84 
                   d 
                 
                   8.05 
                   d 
                 
                   8.33 
                   m 
                 
                   8.54 
                   d 
                 
                   9.00 
                   br d 
                 
                   11.72  
                   br s 
                 
                   11.75  
                   br s 
                 
                   — 
                   — 
                 
                     
                 
             
                
                
                
                
               
               
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
         D)  13 C-NMR sprectrum ( FIG. 8 ) recorded at 150 MHz, at 25° C. in DMSO-d 6  and trifluoroacetic acid (drops) showing the following  13 C signals 
       
       
         
           
                 
                 
               
                     
                 
                     13 C Chemical 
                     
                 
                   shift (ppm) 
                   Multiplicity 
                 
                     
                 
                     
                 
                 
                 
               
                   15.9 
                   t 
                 
                   27.36 
                   t 
                 
                   28.40 
                   t 
                 
                   35.47 
                   t 
                 
                   42.87 
                   t 
                 
                   50.1 
                   d 
                 
                   51.20 
                   d 
                 
                   56.78 
                   d 
                 
                   59.73 
                   d 
                 
                   64.09 
                   d 
                 
                   65.14 
                   d 
                 
                   66.90 
                   d 
                 
                   67.77 
                   t 
                 
                   110.3 
                   d 
                 
                   117.85 
                   d 
                 
                   122.5 
                   s 
                 
                   128.4 
                   s 
                 
                   139.66 
                   s 
                 
                   146.7 
                   s 
                 
                   156.7 
                   s 
                 
                   167.4 
                   s 
                 
                   170.5 
                   s 
                 
                   171.3 
                   s 
                 
                   171.38 
                   s 
                 
                     
                 
             
                
                
                
                
               
               
                
               
            
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
         E) Retention time 21.6 min, determined by using an analytical HPLC Shimadzu LC10-AS, equipped with an “Altima” RP18, 5 μm, 250×4.6 mm i.d column; isocratic elution perfomed at 1 ml/min. flow rate, with 40 mM ammonium formate buffer, brought to pH 4.5 by addition of formic acid; UV detection at 230 nm. 
       
     
     
         3 . Antibiotic GE 81112 factor B1 having the following characteristics:
 A) Mass sprectrum recorded from a 0.01 mg/ml solution in acetonitrile:water 50:50 giving a monoprotonated ion at 658 m/z ib a Termofinnigan LCQ deca instrument under the following electrospray conditions; Spray Voltage: 47 kV; Capillary Temperature: 250° C.; Capillary Voltage: 9V; Infusion Mode: δ 1/min.   B) H-NMR spectrum ( FIG. 10 ) recorded at 600 MH at 25° C. in DMSO-d 6  showing the following  1 H signals:   
       
         
           
                 
                 
               
                     
                 
                   Chemical 
                     
                 
                   shift (ppm) 
                   Multiplicity 
                 
                     
                 
                   1.38 
                   br d 
                 
                   1.56-1.64 
                   m 
                 
                   1.76 
                   m 
                 
                   2.70 
                   m 
                 
                   2.82 
                   br s 
                 
                   2.92 
                   m 
                 
                   3.01-3.06 
                   m 
                 
                   3.49 
                   br m 
                 
                   3.57 
                   br s 
                 
                   4.20 
                   br dd 
                 
                   4.26 
                   br 
                 
                   4.40 
                   br m 
                 
                   4.46 
                   m 
                 
                   5.05 
                   d 
                 
                   5.60 
                   br s 
                 
                   6.21 
                   br 
                 
                   6.52 
                   br 
                 
                   6.78 
                   br s 
                 
                   7.13 
                   br 
                 
                   7.80 
                   br d 
                 
                   7.90 
                   br d 
                 
                   8.44 
                   br 
                 
                     
                 
             
                
                
                
                
               
               
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
         C)  13 C-NMR spectrum ( FIG. 11 ) recorded at 150 MHz, at 25° in DMSO-d 6  showing the following  13 C signals: 
       
       
         
           
                 
                 
               
                     
                 
                   Chemical 
                     
                 
                   shift (ppm) 
                   Multiplicity 
                 
                     
                 
                     
                 
                 
                 
               
                   17.95 
                   t 
                 
                   27.97 
                   t 
                 
                   29.8 
                   t 
                 
                   36.54 
                   t 
                 
                   43.75 
                   t 
                 
                   45.65 
                   d 
                 
                   50.52 
                   d 
                 
                   51.9 
                   d 
                 
                   57.75 
                   d 
                 
                   61.0 
                   d 
                 
                   64.03 
                   d 
                 
                   66.7 
                   d 
                 
                   66.88 
                   d 
                 
                   111.07 
                   d 
                 
                   118.97 
                   d 
                 
                   122.47 
                   s 
                 
                   127.88 
                   s 
                 
                   148.08 
                   s 
                 
                   159.18 
                   s 
                 
                   169.44 
                   s 
                 
                   171.53 
                   s 
                 
                   171.81 
                   s 
                 
                   173.08 
                   s 
                 
                   — 
                   — 
                 
                     
                 
             
                
                
                
                
               
               
                
               
            
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
         D) Retention time 18.4 min, determined by using an analytical HPLC Shimadzu LC10-AS, equipped with an “Altima” RP18, 5 μm, 250×4.6 mm i.d column; isocratic elution perfomed at 1 ml/min. flow rate, with 40 mM ammonium formate buffer brought to pH 4.5 by addition of formic acid; UV detection at 230 nm. 
       
     
     
         4 . A process for producing a GE 81112 antibiotic according to any of  claims 1 - 3 , which comprises:
 cultivating  Streptomyces  sp. DSMZ 14386 or a variant or mutant thereof maintaining the genetic ability to produce the GE 81112 antibiotics under aerobic conditions in an aqueous nutrient medium comprising assimilable sources of carbon, nitrogen and inorganic salts;   recovering the resulting antibiotic mixture from the fermentation broth;   purifying the recovered antibiotic mixture and separating the three antibiotic GE 81112 factors A, B, and B1.   
     
     
         5 . A mixture of two or more antibiotic GE 81112 factors according to  claim 1  in any proportion and the pharmaceutically acceptable salts thereof. 
     
     
         6 . A mixture of two or more antibiotic GE 81112 factors according to  claim 2  in any proportion and the pharmaceutically acceptable salts thereof. 
     
     
         7 . A mixture of two or more antibiotic GE 81112 factors according to  claim 3  in any proportion and the pharmaceutically acceptable salts thereof.

Join the waitlist — get patent alerts

Track US2011286996A1 — get alerts on status changes and closely related new filings.

We store only your email — no account needed. See our privacy policy.