Cahuitamycins and methods of making and using the same

Patent metadata

Field	Value
Publication number	US-10239918-B2
Application number	US-201615576494-A
Country	US
Kind code	B2
Filing date	May 26, 2016
Priority date	May 27, 2015
Publication date	Mar 26, 2019
Grant date	Mar 26, 2019

Abstract

Provided herein are biofilm inhibitors obtained from marine microbial derived natural product extracts from Streptomyces gandocaensis , resulting in biofilm inhibitors, cahuitamycins. Also provided are mutant S. gandocaensis , methods of inhibiting biofilm formation, methods of producing, or increasing the production of, cahuitamycins, methods for synthesizing cahuitamycins, and methods of purifying cahuitamycins.

First claim

What is claimed is:

1. A compound having a structure:

or a pharmaceutically acceptable salt thereof.

2. The compound of claim 1 having a structure

3. The compound of claim 1 having a structure

4. A composition comprising (a) a cahuitamycin compound or pharmaceutically acceptable salt thereof and (b) a pharmaceutically acceptable carrier, wherein the cahuitamycin compound has a structure selected from:

5. A method of inhibiting biofilm formation comprising contacting a bacterium capable for forming a biofilm with the composition of claim 4 in an amount sufficient to inhibit the biofilm formation.

6. The method of claim 5 , wherein the biofilm is formed by a bacterium that is selected from the group consisting of: Acinetobacter baumannii, Aeromonas hydrophila, Aeromonas salmonicida, Agrobacterium tumefaciens, Brucella melitensis, Burkholderia cenocepacia, Burkholderia mallei, Burkholderia pseudomallei, Burkholderia vietnamiensis, Chromobacterium violaceum, Enterobacter agglomeran, Erwinia carotovora, Erwinia chrysanthemi, Escherichia coli, Nitrosomas europaea, Obesumbacterium proteus, Pantoea agglomerans, Pantoea stewartii, Pseudomonas aureofaciens, Pseudomonas aeruginosa, Pseudomonas fluorescens, Pseudomonas fuscovaginae, Pseudomonas syringae, Ralstonia solanacearum, Rhizobium etli, Rhizobium leguminosarum, Rhodobacter sphaeroides, Serratia liquefaciens, Serratia marcescens, Vibrio anguillarum, Vibrio fischeri, Vibrio parahaemolyticus, Vibrio salmonicida, Xanthomonas campestris, Xenorhabdus nematophilus, Yersinia enterocolitica, Yersinia pestis, Yersinia pseudotuberculosis, Yersinia medievalis, Yersinia ruckeri , and a combination thereof.

7. A method of treating a condition due to biofilm formation in a subject in need thereof comprising administering the composition of claim 4 to the subject.

8. The method of claim 7 , wherein the condition is selected from the group consisting of: cystic fibrosis, dental caries, periodonitis, otitis media, a muscular skeletal infection, pneumonia, necrotizing fasciitis, biliary tract infection, osteomyelitis, bacterial prostatitis, endocarditis, native valve endocarditis, cystic fibrosis pneumonia, meloidosis, a skin lesion associated with bullous impetigo, atopic dermatitis, pemphigus foliaceus, and an implanted device-related infection.

9. The method of claim 7 , further comprising administering a second therapeutic to the subject.

10. The method of claim 9 , wherein the second therapeutic comprises an antibiotic.

11. A method of producing a cahuitamycin compound comprising
(a) culturing Streptomyces gandocaensis
(i) in the presence of streptomycin to select for a mutated ribosomal protein, thereby generating a mutated S. gandocaensis,
(ii) under conditions wherein the activity of salicylate synthase is inhibited;
(iii) under conditions wherein the activity of 6-methylsalicylic acid synthase is inhibited;
(iv) in the presence of 2-hydroxy-5-methylbenzoic acid or 2-hydroxybenzoic acid and the mutant is Streptomyces gandocaensis lacking salicylate synthase activity; and

(b) obtaining the cahuitamycin compound from the culture.

12. The method of claim 11 , wherein the Streptomyces gandocaensis comprises a rpsL gene mutation, or a cahl mutation.

13. The method of claim 11 , wherein the S. gandocaensis is cultured in the present of streptomycin and the cahuitamycin compound is cahuitamycin D, cahuitamycin E or a mixture of cahuitamycins D and E.

14. The method of claim 11 , wherein the production of cahuitamycin A, cahuitamycin B, or a mixture of cahuitamycin A and B is increased due to culturing the Streptomyces gandocaensis under conditions wherein the activity of 6-methylsalicylic acid synthase is inhibited.

15. The method of claim 11 , comprising
culturing the Streptomyces gandocaensis in the presence of 2-hydroxy-5-methylbenzoic acid to form cahuitamycin D.

16. The method of claim 11 , comprising culturing the Streptomyces gandocaensis in the presence of 2-hydroxy-benzoic acid to form cahuitamycin E.

17. The method of claim 15 , further comprising isolating the cahuitamycin D.

18. The method of claim 17 , wherein the isolating comprises subjecting crude cahuitamycin D to extraction, chromatography, or both.

19. The method of claim 18 , wherein the extraction comprises contacting with an adsorbant resin, such as a styrene-divinylbenzene matrix.

20. The method of claim 18 , wherein the chromatography is fractionation, high performance liquid chromatography, reverse phase liquid chromatography or a combination thereof.

Assignees

• Univ Michigan Regents

Inventors

• Sherman David H
• Yu Fengan
• Xi Chuanwu
• Wu Jianfeng
• Schultz Pamela J
• Tripathi Ashootosh
• Park Sung Ryeol

Classifications

• C12Y402/99021

  Isochorismate lyase (4.2.99.21) · CPC title

• A01N37/46 Primary

  N-acyl derivatives · CPC title

• A01N43/76

  1,3-Oxazoles; Hydrogenated 1,3-oxazoles · CPC title

• C12N9/88

  Lyases (4.) · CPC title

• C07K5/081

  the side chain containing O or S as heteroatoms, e.g. Cys, Ser · CPC title