IL326320A - Improved formulation of polymyxin B/trimethoprim and rifampin and methods for ophthalmic use - Google Patents

Description

WO 2025/030169 PCT / US2024 / 0408 ENHANCED FORMULATION OF POLYMYXIN B / TRIMETHOPRIM AND RIFAMPIN AND METHODS FOR OPHTHALMIC USES [ 0001 ] Priority Claims and Related Patent Applications This application claims the benefit of priority to U.S. Provisional Application No. / 530,495 , filed August 3 , 2023 , the entire content of which is incorporated herein by reference . [ 0002 ] Technical Fields of the Invention The invention generally relates to pharmaceuticals and methods of preparation and use thereof . More particularly , the invention provides novel pharmaceutical compositions of polymyxin B / trimethoprim and rifampin and methods of their preparation and topical ophthalmic uses ( e.g. , treatment of ophthalmic infections ) .

Background of the Invention [ 0003 ] Bacterial keratitis is a vision - threatening disease caused by a bacterial infection of the cornea that is responsible for nearly 2 million cases of blindness annually . Many Gram - positive and Gram - negative bacteria can cause bacterial keratitis including but not limited to : Staphylococcus aureus , coagulase - negative staphylococci , Streptococcus pneumoniae , Pseudomonas aeruginosa , Moraxella marcesens , Corynebacterium spp . , Proprionibacterium spp . , Klebsiella , Proteus , and Serratia . The clinical presentation of bacterial keratitis frequently includes corneal ulceration , edema , immune infiltrates , and anterior chamber inflammation regardless of the causative agent . While clinicians may obtain a microbiology culture at initial diagnosis , it may take several days for the results to identify a specific causative agent and the tests only yield a positive result in 40-60 % of cases . Given the unknown causative agent at clinical presentation and the aggressive nature of bacterial keratitis , immediate and broad - spectrum ophthalmic antibiotic treatment that is effective toward both Gram - positive and Gram - negative organisms is required within days to prevent corneal perforation and / or loss of the eye . Prescribing an antibiotic which only targets either Gram - positive or Gram - negative bacteria for the treatment of bacterial keratitis is counter to established standard of care . 1 WO 2025/030169 PCT / US2024 / 0408 ( Whitcher et al . Corneal blindness : a global perspective . Bulletin of the World Health Organization 2001 ; 79 ( 3 ) : 214-21 ; Green et al . Cornea . 2008 ; 27 ( 1 ) : 22-7 . Epub 2008/02/05 ; Cabrera - Aguas et al . Clin Exp Ophthalmol . 2022 ; 50 ( 5 ) : 543-62 ; Ting et al . Eye ( Lond ) . 2021 ; 35 ( 4 ) : 1084-101 ; Al - Mujaini et al . Sultan Qaboos Univ Med J. 2009 ; 9 ( 2 ) : 184-95 ; Palioura et al . Clin Ophthalmol . 2016 ; 10 : 179-86 . McLeod et al . Ophthalmology . 1996 ; 103 ( 1 ) : 23-8 ; Levey et al . Cornea . 1997 ; 16 ( 4 ) : 383-6 . ) [ 0004 ] The fluoroquinolone class of antibiotics are the gold standard for bacterial keratitis treatment in the United States due to their broad - spectrum activity , rapid bacterial - killing kinetics , patient tolerability and low cost . In fact , there are only three FDA - approved ophthalmic solutions for managing bacterial keratitis and all are fluoroquinolones : ciprofloxacin 0.3 % ophthalmic solution or ointment , ofloxacin 0.4 % ophthalmic solution , and levofloxacin 1.5 % ophthalmic solution . [ 0005 ] While fluoroquinolones may present a number of side effects , more importantly , the efficacy of fluoroquinolones has declined with the rise of drug - resistant bacteria . This has resulted in increased disease severity and worse visual outcomes over the last decade . In fact , the most recently FDA approved antibiotic to treat bacterial keratitis , levofloxacin a third- generation fluoroquinolone , demonstrated resistance rates as high as 71 % among S. aureus ocular isolates . ( Lauren et al . US Pharmacist . 2018 ; ( 43 ) : 27-30 ; Lalitha et al . Clinical infectious diseases : an official publication of the Infectious Diseases Society of America . 2012 ; 54 ( 10 ) : 1381-7 ; Vazirani et al . Ophthalmology . 2015 ; 122 ( 10 ) ; Asbell et al . JAMA ophthalmology . 2015 ; 133 ( 12 ) . ) [ 0006 ] Due to the challenges with fluoroquinolones , clinicians often turn to off - label therapeutic agents or specialty compounded antibiotics such as fortified aminoglycosides , in combination with fortified vancomycin or a cephalosporin . While these drugs may be efficacious , variability can arise from the lack of standard formulation and may not be tolerated well by patients due to more severe side effects . Moreover , most patients and physicians lack access to a specialty compounding pharmacy , especially within the emergent timeframe of the disease , which limits these compounded antibiotics as a reliable therapy for bacterial keratitis . Furthermore , the duration of therapy for bacterial keratitis via specialty compounded antibiotics often requires frequent refills . 2 WO 2025/030169 PCT / US2024 / 0408 [ 0007 ] Additional to the availability and effectiveness of broad - spectrum antibiotic therapeutic options , any treatment of bacterial keratitis must be able to be efficiently delivered to the patient via a pharmacy and supply chain . Given the emergent nature of this disease , prescribed medications must be made available to the patient within hours of their initial diagnosis for effective and immediate treatment . [ 0008 ] There remains an ongoing need for improved pharmaceutical formulations and methods of use for treating ophthalmic infections . [ 0009 ] Summary of the Invention In one aspect , the invention generally relates to a kit or package comprising : an aqueous first composition comprising polymyxin B / trimethoprim ( PT ) , or pharmaceutically acceptable salts thereof , and a surfactant or polymer , wherein the first composition is characterized by a pH in the range from about 4.0 to about 6.5 ; and a second composition comprising Rif and a solid state polyol ( mannitol and / or sorbitol ) and optionally an antioxidant , wherein the first composition is a sterile aqueous solution , and the second composition is a sterile lyophilized solid or powdery composition . [ 0010 ] In another aspect , the invention generally relates to a method for treating a bacterial ocular infection , comprising administering to a subject in need thereof a therapeutically effective amount of a pharmaceutical composition disclosed herein .

Brief Description of the Drawings [ 0011 ] FIG . 1 shows results indicating that Surfactants reduce PT's polymyxin B - associated rifampin destabilization . [ 0012 ] FIG . 2 shows results indicating that 1 % Polyvinyl alcohol ( PVA ) does not significantly impact Rifampin activity or PT + Rifampin synergy . [ 0013 ] FIG . 3 shows results indicating that 2 % PVP K30 does not significantly impact Rifampin activity or PT + Rifampin synergy . [ 0014 ] FIG . 4 shows results indicating that Poloxama 407 0.2 % does not significantly impact Rifampin activity or PT + Rifampin synergy . [ 0015 ] FIG . 5 shows results indicating that 1 % Tyloxapol does not significantly impact Rifampin activity or PT + Rifampin synergy . 3 WO 2025/030169 PCT / US2024 / 0408 [ 0016 ] FIG . 6 shows results indicating that greater than 0.033 % Polysorbate negatively impacts Rifampin activity .

Detailed Description of the Invention [ 0017 ] The present invention provides enhanced pharmaceutical formulations , efficient storage , and methods for ophthalmic uses ( e.g. , bacterial keratitis , surgical prophylaxis , bacterial conjunctivitis , keratoprosthesis ) . [ 0018 ] Pharmacies and the pharmacy supply chain must be able to ( 1 ) receive and transfer agents in a secure and durable method , ( 2 ) store component ingredients or the final products for a sufficient shelf - life , and ( 3 ) prepare any therapeutic agents quickly and repeatedly for effective and efficient delivery . [ 0019 ] A normal course of treatment for bacterial keratitis generally starts immediately upon diagnosis and continues for a number of weeks to months . Due to the potential rapid progression of the disease , treatment is dosed as frequently as once per hour for the first few days and then tapers to 8 or 12 times per day , then 4 times per day , and eventually to once or twice a day . Given the frequency of treatment , patients apply the solution , ointment , or other therapy to the effected eye wherever they are at that time : at their home , place of employment , etc. Patients must be able to easily and effectively prepare the treatment and dose themselves in a number of different locations and scenarios . Therefore , any steps taken by the patient must be extremely straightforward and easy to use . [ 0020 ] A significant number of peer - reviewed publications have documented and demonstrated the combination of polymyxin B / trimethoprim ( PT ) and rifampin ( Rif ) as a synergistic broad - spectrum antibiotic which rapidly kills bacteria , is effective towards established bacterial biofilms and can eradicate keratitis in animal models of Gram - positive and / or Gram - negative bacterial corneal infections . The use of polymyxin B / trimethoprim + rifampin ( PT + Rif ) as a synergistic antibiotic combination was disclosed in US 11,096,923 B2 . ( Laskey et al . Cornea . 2020 Oct ; 39 ( 10 ) : 1278-1284 ; Chojnacki et al . Antimicrob Agents Chemother . 2018 Dec 21 ; 63 ( 1 ) : e01929-18 ; Chojnacki et al . Antimicrob Agents Chemother . 20Sep 23 ; 63 ( 10 ) : e00777-19 ; Meijome et al . Transl Vis Sci Technol . 2022 Nov 1 ; 11 ( 11 ) : 12 ; Mei et al . Transl Vis Sci Technol . 2022 May 2 ; 11 ( 5 ) : 26 . ) 4 WO 2025/030169 PCT / US2024 / 0408 [ 0021 ] The present invention discusses details which are required for the preparation and formulation for the delivery and treatment of ophthalmic infections including but not limited to : bacterial keratitis , surgical prophylaxis , bacterial conjunctivitis , keratoprosthesis , and others . [ 0022 ] In one aspect , the invention generally relates to a kit or package comprising : an aqueous first composition comprising PT , or pharmaceutically acceptable salts thereof , and a surfactant or polymer , wherein the first composition is characterized by a pH in the range from about 4.0 to about 6.5 ; and a second composition comprising Rif and a solid state polyol ( mannitol and / or sorbitol ) and optionally an antioxidant , wherein the first composition is a sterile aqueous solution , the second composition is a sterile lyophilized solid or powdery composition . [ 0023 ] PT + Rif may be provided as a two - part formulation that is combined at point of care via a packaging system ( e.g. , a dual chamber bottle or eye dropper ) . Separating PT and Rif into two different chambers of a single dosing and packaging system enables a longer shelf - life of the non - mixed components as well as easier preparation and delivery at point of care ( patient use ) . Additional compounds are added to each part of the active ingredient components in order to further stabilize the solution when mixed together . PT is generally provided in a solution formulation while Rif can be added in either a powder / cake or solid - state formulation for reconstitution . Sterile powders can be produced by spray drying . [ 0024 ] In another aspect , the invention generally relates to a pharmaceutical composition comprising PT and a surfactant , wherein polymyxin B sulfate is present in the range of about 7,500 units / mL to about 12,500 units / mL ; trimethoprim sulfate is present in the range of about 0.75 mg / mL to about 1.25 mg / mL ; the surfactant is present in the range of about 0.5 mg / mL to about 5.0 mg / mL ; and the pharmaceutical composition is a sterile aqueous solution characterized by a pH in the range from about 4.0 to about 6.5 . [ 0025 ] In yet another aspect , the invention generally relates to a pharmaceutical composition comprising Rif , mannitol and an antioxidant , wherein Rif is present in the range of about 0.mg / mL to about 1.0 mg / mL ; mannitol is present in the range of about 10 mg / mL to about mg / mL ; the antioxidant is present in the range of about 0 mg / mL to about 1.0 mg / mL ; and the pharmaceutical composition is a sterile lyophilized solid or powdery mixture .

WO 2025/030169 PCT / US2024 / 0408 [ 0026 ] In yet another aspect , the invention generally relates to a reconstituted pharmaceutical composition which is prepared from combining a composition of PT and a surfactant composition and a composition of Rif , mannitol and an antioxidant . [ 0027 ] In yet another aspect , the invention generally relates to a method for treating a bacterial ocular infection , comprising administering to a subject in need thereof a therapeutically effective amount of a pharmaceutical composition disclosed herein . [ 0028 ] In yet another aspect , the invention generally relates to use of a composition disclosed herein to treat a disease or condition ( e.g. , bacterial ocular infection ) . [ 0029 ] The following examples are given for the purpose of illustrating the invention , but not for limiting the scope or spirit of the invention .

Examples Solution stability [ 0030 ] The ophthalmic formulation includes trimethoprim sulfate ( 1 mg / mL ) , polymyxin B sulfate ( 10,000 units / mL ) , and rifampin ( 0.5 mg / mL ) . A combination product of trimethoprim sulfate and polymyxin B sulfate is currently marketed ( Polytrim ) and is stable at room temperature . Rif is marketed as an oral capsule and as a lyophilized powder for reconstitution and IV administration . The IV solution of Rif must be used shortly after reconstitution due to the poor solution stability of the compound . [ 0031 ] Addition of a surfactant resulted in improved solution stability . Surfactants approved for ophthalmics include : octoxynol - 40 , nonoxyl - 9 , poloxamer 188 , poloxamer 407 , PEG - castor oil , PEG - 40 hydrogenated castor oil , PEG - 40 stearate , polysorbate 80 , and tyloxapol . The surfactant levels are in the 0.05-0.3 % w / w range . Approved polymers include polyvinylprrolidone ( povidone , or PVP ) , polyvinyl alcohol ( PVA ) , and modified celluloses . The polymer levels can be up to 2 % w / w . PVA and PVP did not inhibit rifampin activity .

Lyophilization [ 0032 ] The marketed lyophilized formulation of Rif includes sodium formaldehyde sulfoxylate and NaOH to adjust pH . Sodium formaldehyde sulfoxylate may improve dissolution of the lyophilized cake by forming a salt with Rif , but it is not approved for ophthalmics . 6 WO 2025/030169 PCT / US2024 / 0408 [ 0033 ] During lyophilization experiments , it was found that adding mannitol to the lyophilized formulation ( 1 : 1 w / w to 10 : 1 w / w mannitol rifampin ) improved the reconstitution of the lyophilized cake significantly compared to a neat lyophilized rifampin cake . [ 0034 ] An antioxidant , such as ascorbic acid ( or its salts and esters ) can be included in the lyophilized formulation . This may improve the solid - state stability of rifampin in the lyophilized cake or other solid - state formulation and its solution stability after reconstitution . [ 0035 ] During excipient screening studies , the surfactant polysorbate 80 was found to inhibit the antimicrobial activity of rifampin . Surprisingly , it was found that the surfactants tyloxapol and poloxamer 407 did not inhibit rifampin activity . [ 0036 ] Exemplary reconstituted formulation compositions : [ 0037 ] Ingredient Polymyxin B Sulfate Active Trimethoprim Sulfate Active Rifampin Active Benzalkonium Chloride Mannitol Function Concentration 10,000 units / mL +/- 1,0units / mL Antimicrobial preservative Lyophilization aid / solid state bulking agent ( solid ) / Osmolyte 1 mg / mL +/- .1 mg / mL 0.5 mg / mL +/- .2 mg / mL 0.02-0.04 mg / mL +/- .0mg / mL 10-60 mg / mL Glycerin Tyloxapol Osmolyte 0-20 mg / mL Surfactant / polymer 1-3 mg / mL Sodium ascorbate Antioxidant 0-1 mg / mL Sodium hydroxide or pH adjustment q.s. to pH 4.0-6.sulfuric acid Sterile Water Diluent to 1 mL +/- .5 mL Additional potential excipients : Poloxamer ( surfactant ) , 0-5 mg / mL Povidone ( viscosity modifier ) , 0-60 mg / mL 7 WO 2025/030169 PCT / US2024 / 0408 [ 0038 ] Polyvinyl alcohol ( viscosity modifier ) , 0-60 mg / mL Disodium edetate ( chelating agent ) , 0-1 mg / mL Likely Solid Formulation ( amount per vial for 1 mL when reconstituted ) : 0.5 mg rifampin 10-50 mg mannitol 0-1 mg ascorbic acid 0-30 mg of povidone - .5 mg of poloxamer 407 or 10-60 mg of polyvinyl alcohol [ 0039 ] FIG . 1 shows results indicating that Surfactants reduce PT's polymyxin B - associated rifampin destabilization . Shown in panel A are the rifampin degradation profiles of rifampin , rifampin in the presence of trimethoprim , and rifampin in the presence of polymyxin B at 25 ° C , establishing that rifampin is destabilized in the presence of polymyxin B. Shown in panel B are the rifampin degradation profiles of rifampin , rifampin in the presence of polymyxin B , rifampin in the presence of PT ( polymyxin B + trimethoprim ) supplemented with polysorbate 80 , and rifampin in the presence of PT ( polymyxin B + trimethoprim ) supplemented with tyloxapol , establishing that surfactants such as tyloxapol and polysorbate stabilize rifampin at 5 ° C . As shown in Figures 5 and 6 , rifampin remains active in tyloxapol but loses antimicrobial activity in the presence of polysorbate . [ 0040 ] FIG . 2 shows results indicating that 1 % Polyvinyl alcohol ( PVA ) does not significantly impact Rifampin activity or PT + Rifampin synergy . Shown are the antimicrobial properties of increasing concentrations ( 0 , 0.0006 , 0.0012 , 0.0025 , 0.005 , 0.01 , 0.02 , 0.4 , 0.08 , 0.16 , 0.32 and 64 gµ / ml ) of Rifampin loaded into columns 1-12 of a microtiter plate containing media supplemented with 1 % PVA and inoculated with 10,000 viable cells of the bacterial pathogen , Staphylococcus aureus . Also shown are the antimicrobial properties of increasing concentrations of PT ( 0 , 0.26 , 0.53 , 1.10 , 2.10 , 4.20 , and 8.40 gµ / ml ) applied to each row of the plate . In the absence of PT ( row A ) as little as 0.0025 gµ / ml rifampin inhibits S. aureus growth ( well is highlighted by green in row A ) . In the absence of Rifampin ( column 1 ) as little as 0.gµ / ml PT inhibits S. aureus growth ( highlighted by green in column 1 ) . In combination 0.00gµ / ml rifampin and 0.26 gµ / ml PT inhibits S. aureus growth ( highlighted in red ) , establishing 8 WO 2025/030169 PCT / US2024 / 0408 that the combination of PT + rifampin displays synergistic antimicrobial activity in the presence of % PVA . [ 0041 ] FIG . 3 shows results indicating that 2 % PVP K30 does not significantly impact Rifampin activity or PT + Rifampin synergy . Shown are the antimicrobial properties of increasing concentrations ( 0 , 0.0006 , 0.0012 , 0.0025 , 0.005 , 0.01 , 0.02 , 0.4 , 0.08 , 0.16 , 0.32 and gµ / ml ) of Rifampin loaded in columns 1-12 of a microtiter plate containing media supplemented with 2 % PVP K30 and inoculated with 10,000 viable cells of the bacterial pathogen , Staphylococcus aureus . Also shown are the antimicrobial properties of increasing concentrations of PT ( 0 , 0.26 , 0.53 , 1.10 , 2.10 , 4.20 , and 8.40 gµ / ml ) applied to each row of the plate . In the absence of PT ( row A ) as little as 0.0025 gµ / ml rifampin inhibits S. aureus growth ( well is highlighted by green in row A ) . In the absence of Rifampin ( column 1 ) as little as 0.gμ / ml PT inhibits S. aureus growth ( highlighted by green in column 1 ) . In combination 0.00gµ / ml rifampin and 0.53 gµ / ml PT inhibits S. aureus growth ( highlighted in red ) , establishing that the combination of PT + rifampin displays synergistic antimicrobial activity in the presence of % PVP K30 . [ 0042 ] FIG . 4 shows results indicating that Poloxama 407 0.2 % does not significantly impact Rifampin activity or PT + Rifampin synergy . Shown are the antimicrobial properties of increasing concentrations ( 0 , 0.0006 , 0.0012 , 0.0025 , 0.005 , 0.01 , 0.02 , 0.4 , 0.08 , 0.16 , 0.32 and gμ / ml ) of Rifampin loaded in columns 1-12 of a microtiter plate containing media supplemented with 0.2 % Poloxama 407 and inoculated with 10,000 viable cells of the bacterial pathogen , Staphylococcus aureus . Also shown are the antimicrobial properties of increasing concentrations of PT ( 0 , 0.26 , 0.53 , 1.10 , 2.10 , 4.20 , and 8.40 gµ / ml ) applied to each row of the plate . In the absence of PT ( row A ) as little as 0.005 gµ / ml rifampin inhibits S. aureus growth ( well is highlighted by green in row A ) . In the absence of Rifampin ( column 1 ) as little as 1.gμ / ml PT inhibits S. aureus growth ( highlighted by green in column 1 ) . In combination 0.00gμ / ml rifampin and 0.53 gµ / ml PT inhibits S. aureus growth ( highlighted in red ) , establishing that the combination of PT + rifampin displays synergistic antimicrobial activity in the presence of 0.2 % Poloxama 407 . [ 0043 ] FIG . 5 shows results indicating that 1 % Tyloxapol does not significantly impact Rifampin activity or PT + Rifampin synergy . Shown are the antimicrobial properties of increasing concentrations ( 0 , 0.0006 , 0.0012 , 0.0025 , 0.005 , 0.01 , 0.02 , 0.4 , 0.08 , 0.16 , 0.32 and 9 WO 2025/030169 PCT / US2024 / 0408 64 gμ / ml ) of Rifampin loaded in columns 1-12 of a microtiter plate containing media supplemented with 1 % Tyloxapol and inoculated with 10,000 viable cells of the bacterial pathogen , Staphylococcus aureus . Also shown are the antimicrobial properties of increasing concentrations of PT ( 0 , 0.26 , 0.53 , 1.10 , 2.10 , 4.20 , and 8.40 gµ / ml ) applied to each row of the plate . In the absence of PT ( row A ) as little as 0.005 gµ / ml rifampin inhibits S. aureus growth ( well is highlighted by green in row A ) . In the absence of Rifampin ( column 1 ) as little as 1.gμ / ml PT inhibits S. aureus growth ( highlighted by green in column 1 ) . In combination 0.00gμ / ml rifampin and 0.53 gµ / ml PT inhibits S. aureus growth ( highlighted in red ) , establishing that the combination of PT + rifampin displays synergistic antimicrobial activity in the presence of 1 % Tyloxapol . [ 0044 ] FIG . 6 shows results indicating that greater than 0.033 % Polysorbate negatively impacts Rifampin activity : Less than 22.5 ug / ml BAC has no impact on Rifampin activity , whereas greater than 45 ug / ml exhibits antibacterial activity . Shown are the antimicrobial properties of 0.005 gµ / ml rifampin ( loaded in columns 1-12 of a microtiter plate containing media and inoculated with 10,000 viable cells of the bacterial pathogen , Staphylococcus aureus ) in the presence of 0 , 0.0001 , 0.0003 , 0.0006 , 0.0017 , 0.0033 , 0.0075 , 0.015 , 0.033 , 0.075 , 0.15 , and 0.3 % polysorbate . Also shown are the antimicrobial properties of 0.005 gµ / ml rifampin in the presence of 0 , 0.703 , 1.406 , 2.813 , 5.625 , 11.25 , 22.5 and 45 gµ / ml Benzalkonium Chloride ( rows ) . Results show that 0.005 gµ / ml rifampin displays antimicrobial effects and inhibits the growth of S. aureus in the presence of 330.0≤ % polysorbate ( highlighted in green ) , whereas higher concentrations of polysorbate interfere with the antimicrobial effects of rifampin . Further , polysorbate - associated inactivation of rifampicin occurs in the presence of 5.22≤ gµ / ml of the common ophthalmic preservative benzalkonium chloride . Finally , 5.22≤ % benzalkonium chloride has no effect on the effect of rifampin . [ 0045 ] The described features , structures , or characteristics of Applicant's disclosure may be combined in any suitable manner in one or more embodiments . In the description , herein , numerous specific details are recited to provide a thorough understanding of embodiments of the invention . One skilled in the relevant art will recognize , however , that Applicant's composition and / or method may be practiced without one or more of the specific details , or with other methods , components , materials , and so forth . In other instances , well - known structures , WO 2025/030169 PCT / US2024 / 0408 materials , or operations are not shown or described in detail to avoid obscuring aspects of the disclosure . [ 0046 ] In this specification and the appended claims , the singular forms " a , " " an , " and " the " include plural reference , unless the context clearly dictates otherwise . [ 0047 ] The term " comprising " , when used to define compositions and methods , is intended to mean that the compositions and methods include the recited elements , but do not exclude other elements . The term " consisting essentially of " , when used to define compositions and methods , shall mean that the compositions and methods include the recited elements and exclude other elements of any essential significance to the compositions and methods . For example , " consisting essentially of " refers to administration of the pharmacologically active agents expressly recited and excludes pharmacologically active agents not expressly recited . The term consisting essentially of does not exclude pharmacologically inactive or inert agents , e.g. , pharmaceutically acceptable excipients , carriers or diluents . The term " consisting of " , when used to define compositions and methods , shall mean excluding trace elements of other ingredients and substantial method steps . Embodiments defined by each of these transition terms are within the scope of this invention . [ 0048 ] Unless specifically stated or obvious from context , as used herein , the term " about " is understood as within a range of normal tolerance in the art , for example within 2 standard deviations of the mean . About can be understood as within 10 % , 9 % , 8 % , 7 % , 6 % , 5 % , 4 % , 3 % , % , 1 % , 0.5 % , 0.1 % , 0.05 % , or 0.01 % of the stated value . Unless otherwise clear from context , all numerical values provided herein can be modified by the term about . [ 0049 ] The terms " disease " , " disorder " and " condition " are used interchangeably unless indicated otherwise . [ 0050 ] As used herein , the terms " effective amount " or " therapeutically effective amount " refer to that amount of compound ( s ) or pharmaceutical composition described herein that is sufficient to effect the intended application including , but not limited to , disease treatment , as illustrated below . [ 0051 ] The therapeutically effective amount can vary depending upon the intended application , or the subject and disease condition being treated , e.g. , the desired biological endpoint , the pharmacokinetics of the compound , the disease being treated , the mode of administration , and the weight and age of the patient , which can readily be determined by one of 11 WO 2025/030169 PCT / US2024 / 0408 ordinary skill in the art . The term also applies to a dose that will induce a particular response . The specific dose will vary depending on , for example , the particular compounds chosen , the species of subject and their age / existing health conditions or risk for health conditions , the dosing regimen to be followed , the severity of the disease , whether it is administered in combination with other agents , timing of administration , the tissue to which it is administered , and the physical delivery system in which it is carried . [ 0052 ] Unless defined otherwise , all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art . Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present disclosure , the preferred methods and materials are now described . Methods recited herein may be carried out in any order that is logically possible , in addition to a particular order disclosed . [ 0053 ] Incorporation by Reference References and citations to other documents , such as patents , patent applications , patent publications , journals , books , papers , web contents , have been made in this disclosure . All such documents are hereby incorporated herein by reference in their entirety for all purposes . Any material , or portion thereof , that is said to be incorporated by reference herein , but which conflicts with existing definitions , statements , or other disclosure material explicitly set forth herein is only incorporated to the extent that no conflict arises between that incorporated material and the present disclosure material . In the event of a conflict , the conflict is to be resolved in favor of the present disclosure as the preferred disclosure .

Equivalents [ 0054 ] The representative examples are intended to help illustrate the invention , and are not intended to , nor should they be construed to , limit the scope of the invention . Indeed , various modifications of the invention and many further embodiments thereof , in addition to those shown and described herein , will become apparent to those skilled in the art from the full contents of this document , including the examples and the references to the scientific and patent literature included herein . The examples contain important additional information , exemplification and 12 WO 2025/030169 PCT / US2024 / 0408 guidance that can be adapted to the practice of this invention in its various embodiments and equivalents thereof . 13

Claims (31)

WO 2025/0301 What is Claimed is :

1. A kit or package comprising : CLAIMS PCT / US2024 / 0408 an aqueous first composition comprising polymyxin B / trimethoprim , or pharmaceutically acceptable salts thereof , and a surfactant , wherein the first composition is characterized by a pH in the range from about 4.0 to about 6.5 ; and a second composition comprising rifampin and mannitol and optionally an antioxidant , wherein the first composition is a sterile aqueous solution , and the second composition is a sterile lyophilized solid or powdery composition .

2. The kit or package of claim 1 , wherein polymyxin B is polymyxin B sulfate .

3. The kit or package of claim 1 or 2 , wherein trimethoprim is trimethoprim sulfate .

4. The kit or package of any one of claims 1-3 , wherein the surfactant is selected from tyloxapol and poloxamer 407 .

5. The kit or package of any one of claims 1-3 , wherein the surfactant is tyloxapol .

6. The kit or package of any one of claims 1-5 , wherein the second composition comprises ascorbic acid , or a salt or ester thereof , as an antioxidant .

7. The kit or package of claim 6 , wherein the antioxidant is sodium ascorbate .

8. The kit or package of any one of claims 1-7 , one or both of the first and second compositions further comprises an antimicrobial agent .

9. The kit or package of claim 8 , wherein the antimicrobial agent is benzalkonium chloride . 14

10.

11.

12. WO 2025/030169 PCT / US2024 / 0408 The kit or package of any one of claims 1-9 , one or both of the first and second compositions further comprises an osmolyte . The kit or package of claim 10 , wherein the osmolyte is glycerin . The kit or package of any one of claims 1-11 , wherein the first and / or the second composition further comprises one or more pharmaceutically acceptable excipient , carrier , or diluent .

13.

14.

15. A pharmaceutical composition comprising polymyxin B / trimethoprim and a surfactant , wherein polymyxin B is present in the range of about 7,500 units / mL to about 12,5units / mL ; trimethoprim is present in the range of about 0.75 mg / mL to about 1.25 mg / mL ; the surfactant is present in the range of about 0.5 mg / mL to about 5.0 mg / mL ; and the pharmaceutical composition is a sterile aqueous solution characterized by a pH in the range from about 4.0 to about 6.5 . The pharmaceutical composition of claim 13 , wherein polymyxin B is present in the range of about 9,000 units / mL to about 11,0units / mL ; trimethoprim is present in the range of about 0.9 mg / mL to about 1.1 mg / mL ; and the surfactant is present in the range of about 1.0 mg / mL to about 3.0 mg / mL . The pharmaceutical composition of claim 13 or 14 , wherein polymyxin B is polymyxin B sulfate .

16. The pharmaceutical composition of any one of claims 13-15 , wherein trimethoprim is trimethoprim sulfate . 15 WO 2025/030169 PCT / US2024 / 0408

17.

18.

19. The pharmaceutical composition of any one of claims 13-16 , wherein the surfactant is selected from tyloxapol and poloxamer 407 . The pharmaceutical composition of any one of claims 13-17 , wherein the surfactant is tyloxapol . The pharmaceutical composition of any one of claims 13-18 , wherein the pharmaceutical composition further comprises one or more pharmaceutically acceptable excipient , carrier , or diluent .

20.

21.

22. A pharmaceutical composition comprising rifampin , mannitol and an antioxidant , wherein rifampin is present in the range of about 0.2 mg / mL to about 1.0 mg / mL ; mannitol is present in the range of about 10 mg / mL to about 60 mg / mL ; the antioxidant is present in the range of about 0 mg / mL to about 1.0 mg / mL ; and the pharmaceutical composition is a sterile lyophilized solid or powdery mixture . The pharmaceutical composition of claim 20 , wherein rifampin is present in the range of about 0.4 mg / mL to about 0.6 mg / mL ; mannitol is present in the range of about 15 mg / mL to about 50 mg / mL ; and the antioxidant is present in the range of about 0.1 mg / mL to about 1.0 mg / mL . The pharmaceutical composition of claim 20 or 21 , wherein the antioxidant is ascorbic acid , or a salt or ester thereof .

23. The pharmaceutical composition of claim 22 , wherein the antioxidant is sodium ascorbate .

24. The pharmaceutical composition of any one of claims 13-23 , further comprising an antimicrobial agent . 16

25. WO 2025/030169 PCT / US2024 / 0408 The pharmaceutical composition of claim 24 , wherein the antimicrobial agent is benzalkonium chloride .

26.

27.

28. The pharmaceutical composition of any one of claims 13-25 , further comprising an osmolyte . The pharmaceutical composition of claim 26 , wherein the osmolyte is glycerin . The pharmaceutical composition of any one of claims 13-27 , wherein the pharmaceutical composition further comprises one or more pharmaceutically acceptable excipient , carrier , or diluent .

29.

30.

31. A pharmaceutical composition comprising the composition of any one of claims 13-and the composition of any one of claims 20-28 . A method for treating a bacterial ocular infection , comprising administering to a subject in need thereof a therapeutically effective amount of the composition of any one of claims 13-19 and / or the composition of any one of claims 20-28 . Use of the composition of any one of claims 13-19 and / or the composition of any one of claims 20-28 for treating a disease or condition . 17