IIIIIIIIIIIIIIIIIIIIIIIIIIIUS010071103B2 (12 ) United States Patent ( 10 ) Patent No. : US 10 ,071 , 103 B2 Sengupta et al. (45 ) Date of Patent: Sep . 11 , 2018 ( 54 ) TREATMENTS FOR RESISTANT ACNE ( 58 ) Field of Classification Search CPC .. A61K 31 /55 ; A61K 9 /0014 ; A61K 31/ 192 ; (71 ) Applicant: VYOME BIOSCIENCES PVT. LTD . , A61K 31 /4709 ; A61K 45 /06 ; A61K New Delhi ( IN ) 47 / 02 , A61K 47 / 08 ; A61K 47 / 10 ; A61K 47 / 183 ; A61K 47 /32 , A61K 47 / 36 ; A61K (72 ) Inventors : Shiladitya Sengupta , Delhi ( IN ) ; 47 /38 Suresh Rameshlal Chawrai, Pune USPC . .. .. .. 514 /210 . 16 ( IN ) ; Shamik Ghosh , Delhi ( IN ) ; See application file for complete search history . Sumana Ghosh , Delhi ( IN ) ; Nilu Jain , New Delhi ( IN ) ; Suresh Sadhasivam , ( 56 ) References Cited Salem ( IN ) ; Richard Buchta , Melbourne (AU ) ; Anamika U . S . PATENT DOCUMENTS Bhattacharyya , Delhi ( IN ) 4 , 668 ,664 A 5 / 1987 Rougier et al. 5 ,607 , 980 A 3 / 1997 McAtee et al . ( 73 ) Assignee: VYOME BIOSCIENNCES PVT. 6 , 255, 279 B1 7 / 2001 Christophers et al. LTD ., New Delhi ( IN ) 2005 / 0209157 Al 9 /2005 Owen 2005 /0245545 Al 11/ 2005 Kase et al . Subject to any disclaimer, the term of this 2005 /0282755 Al 12 /2005 Hart et al . ( * ) Notice : 2006 / 0008538 A1 1 / 2006 Wu et al . patent is extended or adjusted under 35 2007/ 0134729 AL 6 / 2007 Christensen et al. U . S . C . 154 ( b ) by 0 days. 2007 / 0197501 A1 8 / 2007 Schulz et al . 2011/ 0144329 A1 * 6 / 2011 Shawer . .. .. CO7D 223 / 12 ( 21 ) Appl. No .: 15 / 115 , 143 540 /605 2016 / 0058775 A1 * 3 / 2016 Prasad .. .. .. A61K 8 /342 ( 22 ) PCT Filed : Jan . 29 , 2015 424 / 401 ( 86 ) PCT No. : PCT/ IN2015 / 000057 FOREIGN PATENT DOCUMENTS $ 371 ( c ) ( 1 ), CN 103524492 A 1 / 2014 ( 2 ) Date : Jul. 28 , 2016 EP 0251330 A2 1 / 1998 JP 2009196934 A 9 / 2009 WO 2006 / 100495 A1 9 / 2006 (87 ) PCT Pub. No. : W02015 /114666 WO 2007 /070518 A2 6 / 2007 PCT Pub . Date : Aug. 6 , 2015 WO 2008 /035078 A1 3 / 2008 WO 2012 / 177770 A112 / 2012 wo 2012 / 177986 A2 12 / 2012 (65 ) Prior Publication Data WO 2014 / 167554 A2 10 / 2014 US 2016 /0346294 A1 Dec. 1 , 2016 WO 2014 / 195872 A112 / 2014 (30 ) Foreign Application Priority Data OTHER PUBLICATIONS Jan . 29 , 2014 ( IN ) . .. .. .. .. .. .. .. .. .. .. 269 /DEL /2014 Betriu et al . , “ Erythromycin and Clindamycin Resistance and Nov . 10 , 2014 ( IN ) .. .. .. .. 3247 /DEL / 2014 Telithromycin Susceptibility in Streptococcus agalactiae” , Antimi crobial Agents and Chemotherapy 47 ( 3 ) : 1112 - 1114 (2003 ) . (51 ) Int. CI. Bryskier et al. , “ Dual B - lactam - fluoroquinolone compounds: a novel A61K 31 /55 ( 2006 .01 ) approach to antibacterial treatment” , Expert Opinion on Investiga A61K 31/ 192 ( 2006 .01 ) tional Drugs 6 ( 10 ) : 1479 - 1499 ( 1997 ) . Cambau et al ., “ Target specificity of the new fluoroquinolone A61K 45 / 06 ( 2006 .01 ) besifloxacin in Streptococcus pneumoniae , Staphylococcus aureus A61K 47 / 02 ( 2006 . 01 ) and Escherichia coli” , Journal of Antimicrobial Chemotherapy A61K 47 / 08 ( 2006 .01 ) 63 : 443 -450 (2009 ) . A61K 47 / 10 ( 2017 .01 ) De Lucca et al. , “ Antifungal peptides : Origin , activity , and thera C07D 513 /04 ( 2006 .01 ) peutic potential" , Revista Iberoamericana de Micologia 17 : 116 - 120 A61K 31 / 4709 ( 2006 .01 ) ( 2000 ) . A61K 9 / 00 ( 2006 . 01 ) A61K 47 / 18 ( 2017 .01 ) (Continued ) A61K 4732 ( 2006 . 01 ) Primary Examiner — Yevgeny Valenrod A61K 47 / 36 ( 2006 .01 ) ( 74 ) Attorney, Agent, or Firm — Nixon Peabody LLP ; A61K 47 /38 ( 2006 .01 ) David S . Resnick ; Ravinderjit Braich (52 ) U . S . CI. CPC . .. .. .. .. A61K 31/ 55 ( 2013 .01 ) ; A61K 9 / 0014 ( 2013 . 01 ) ; A61K 31 / 192 (2013 . 01 ) ; A61K (57 ) ABSTRACT 31 /4709 (2013 .01 ) ; A61K 45 /06 (2013 .01 ) ; The present disclosure relates generally to novel molecules , A61K 47 / 02 ( 2013 .01 ) ; A61K 47 /08 ( 2013 .01 ) ; compositions, and formulations for treatment of bacterial A61K 47 / 10 ( 2013. 01 ) ; A61K 47 / 183 infections in general and more specifically to bacterial ( 2013 .01 ) ; A61K 47 /32 ( 2013 .01 ) ; A61K 47 / 36 infections with antibiotic resistant pathogens. ( 2013 .01 ) ; A61K 47/ 38 ( 2013 .01 ) ; C07D 513 /04 ( 2013 .01 ) 16 Claims, 14 Drawing Sheets US 10 ,071 , 103 B2 Page 2 References Cited loproteinases in keratinocytes in response to Propionibacterium ( 56 ) acnes" , Archives of Dermatological Research 302 :745 - 756 ( 2010 ) . Liu et al. , “ Cutting Edge : All - trans Retinoic Acid Down -Regulates OTHER PUBLICATIONS TLR2 Expression and Function ” , The Journal of Immunology 174 :2467 - 2470 (2005 ) . Elitropi et al ., “ New Cephalosporins and 7a -Methoxy Cephalosporins Mouser et al ., “ Propionibacterium acnes- Reactive T Helper - 1 Cells in the Skin of Patients with Acne Vulgaris ” , The Journal of Chemistry and Biological Activities ” , The Journal of Antibiotics Investigative Dermatology 121 ( 5 ) : 1226 - 1228 (2003 ) . 32 ( 9 ) : 900 - 908 ( 1979 ) . Nagy et al ., “ Propionibacterium acnes and lipopolysaccharide induce Epand et al. , “ Diversity of antimicrobial peptides and their mecha the expression of antimicrobial peptides and proinflammatory cytokines/ nisms of action ” , Biochimica et Biophysica Acta 1462 : 11 -28 ( 1999 ) . chemokines in human sebocytes ” , Microbes and Infection 8 : 2195 Haas et al. , “ Besifloxacin , a Novel Fluoroquinolone, Has Broad 2205 ( 2006 ) . Spectrum In Vitro Activity against Aerobic and Anaerobic Bacte Regoes et al . , “ Pharmacodynamic Functions : a Multiparameter ria ” , Antimicrobial Agents and Chemotherapy 53 ( 8 ) :3552 - 3560 Approach to the Design of Antibiotic Treatment Regimens ” , Anti ( 2009 ) . microbial Agents and Chemotherapy 48 ( 10 ) : 3670 - 3676 ( 2004 ) . Jeremy et al. , “ Inflammatory Events Are Involved in Acne Lesion Schlunzen et al . , " Structural basis for the interaction of antibiotics Initiation ” , Journal of Investigative Dermatology 121 ( 1 ) :20 - 27 (2003 ) . with the peptidyl transferase centre in eubacteria ” , Nature 413 :814 Kabara et al. , “ Fatty Acids and Derivatives as Antimicrobial Agents ” , 821 (2001 ) . Antimicrobial Agents and Chemotherapy 2 ( 1 ) :23 - 28 ( 1972 ) . Taglietti et al . , " Novel Topical Drug Delivery Systems and Their Kato et al ., “ Comparison of In Vitro Activities of DU -6859a and Potential Use in Acne Vulgaris ” , Skin Therapy Letter 1 - 5 (2008 ) . Other Fluoroquinolones Against Japanese Isolates of Anaerobic Thiboutot et al. , “ IL - 17 : A Key Player in the P . acnes Inflammatory Bacteria ” , Clinical Infectious Diseases 23 ( Suppl 1 ) : S31 - S35 ( 1996 ) . Cascade ? " , Journal of Investigative Dermatology 134 : 307 -310 ( 2014 ) . Keren et al ., " Specialized Persister Cells and the Mechanism of Toyoda et al ., “ An Overview of Topical Antibiotics for Acne Multidrug Tolerance in Escherichia coli” , Journal of Bacteriology Treatment " , Dermatology 196 ( 1 ) : 130 -134 ( 1998 ) . 186 ( 24 ) :8172 - 8180 ( 2004 ) . Zasloff M . , “ Antimicrobial peptides of multicellular organisms” , Kim et al. , " Activation of Toll - Like Receptor 2 in Acne Triggers Nature 415 : 389 -395 ( 2002 ) . Inflammatory Cytokine Responses ” , The Journal of Immunology Miller et al. , “ SOS Response Induction by B - Lactams and Bacterial 169 : 1535 - 1541 ( 2002 ) . Defense Against Antibiotic Lethality ” , Science 305 : 1629 - 1631 ( 2004 ) . Lau et al. , “ Scope and Limitations of The Co - Drug Approach to Ye et al . , " Synthesis and antibacterial activity of pyridonecarboxylic Topical Drug Delivery ” , Current Pharmaceutical Design 14 ( 8 ) :794 acid derivatives containing 2 -methyl - 5 - nitroimidazol ” , Acta 802 ( 2008 ) . Pharmaceutica Sinica 38 ( 4 ) : 260 - 263 (2003 ) . Lee et al. , “ Protease - activated receptor - 2 mediates the expression of inflammatory cytokines , antimicrobial peptides , and matrix metal * cited by examiner U . S . Patent Sep . 11, 2018 Sheet 1 of 14 US 10 ,071 , 103 B2 + Cephalothin + Nadifloxacin * Roxythromycin %viablecells - Besifloxacin * Prulifloxacin + Clindamycin + Cefoxitin # Ulifloxacin FIG . IA 150 + Cephalothin * Nadifloxacin 100k * Roxithromycin - Besifloxacin %viablecells + Ulifloxacin Prulifloxacin + Clindamycin Leo + Cefoxitin FIG . IB U . S . Patent Sep . 11, 2018 Sheet 2 of 14 US 10, 071 , 103 B2 DARTmolecules Dose response Curve Against P . acnes MTCC 1951 Ourwww sabansa WMSpH %cellsviability I 1116 115 öppna 0. 00 0. 05 0 .100 .150 .200 .250 . 30 MA Antibiotic Concentration (ugimi ) . FIG . IC DARTmolecules Dose Response Curve against P . acnes CCARM 9010 113 wwwwwwwwwwwwwwwwwwwwwwwwwwwwww trement alertow %Cellsviability 116 } 0. 00 0. 05 0. 10 0 .16 0. 20 0. 25 0 .30 . Antibiotic Concentration (ug / ml ) FIG . ID U . S . Patent Sep . 11, 2018 Sheet 3 of 14 US 10 ,071 , 103 B2 Lanes 3 - 7 Lane1 Comp. 91 conc. (um ) Lane8 IA 0 . 25come 0 .50 1one .00 come 2 . 50 come5 .00 .. , Controls Lane 1 - Relaxed DNA (Negative Control) Lane 2 - DNA + Gyrase (Positive Control) Lane 3 - 7 Comp. 91 with increasing conc . Lane 8 - Supercoiled DNA marker D ALS. TO P WUNSTAR - DL + Ito + CompoundGyrase 91 0 .25 0 . 50 1 .00 2 . 50 5 .00 Compound Conc . (um ) FIGS. 2A and 2B U . S . Patent Sep . 11, 2018 Sheet 4 of 14 US 10 ,071 , 103 B2 Vun Compound 113 AWW Compound 94 3 Compound 115 og B Compound 90 werth Um Compound 116 w HP Compound 91 Percentageofsupercoiling % Ô ? HUMIHITTITUTITI 0.211111111111 III. o ?????????????? 1uM Compound 2 . 5 uM Compound FIG . 3A AW Nadifloxacin E Compound 91 Percentageofsupercoiling Estadoe . o 1?M 2 . 5 PM FIG . 3B U . S