Mucosal-adherent drug delivery system for intraocular administration of fluoroquinolone antibiotics

Abstract

The present disclosure relates to ocular mucoadhesive pharmaceutical compositions comprising a fluoroquinolone antibiotic, chitosan, polyvinyl alcohol, and polyvinylpyrrolidone, their uses, and methods of using such compositions.

Description

[Technical Field]

[0001] Cross-references to related applications This application claims priority to Indian Provisional Patent Application No. 201841012721, filed on 3 April 2018, which is incorporated herein by reference in its entirety.

[0002] Technical field The subject matter disclosed herein generally relates to compositions and methods for the delivery of therapeutic drugs to the eye. In particular, the subject matter relates to mucosal adhesive pharmaceutical compositions comprising fluoroquinolone antibiotics, chitosan, polyvinyl alcohol, and polyvinylpyrrolidone, their use, and methods for using such compositions. [Background technology]

[0003] Fluoroquinolone antibiotics are well-known for treating ocular infections such as keratitis, blepharitis, conjunctivitis, and endophthalmitis. However, despite the general efficacy of currently available ocular quinolone therapies, several limitations are associated with the formulation of the ocular drug delivery system. The main challenge is to provide an ocular delivery system with high therapeutic efficacy by achieving the optimal concentration of the drug at the active site for an appropriate duration. Due to physiological and anatomical constraints such as solution drainage, lacrimation, tear dynamics, tear dilution, tear turnover, conjunctival absorption, unproductive absorption, transient residence time in the blind duct, and the relative impermeability of the corneal epithelial membrane, only a small fraction of the drug (less than 5% of the injected dose) is absorbed into the eye.

[0004] Therefore, in this field, there is a continuous need to increase the bioavailability and duration of therapeutic effect of ophthalmic drugs. Furthermore, there is a need for improved compositions and therapeutic methods based on the use of antibiotics that are more effective than existing antibiotics against major ophthalmic pathogens and have a lower tendency to develop resistance in those pathogens.

[0005] Mucosal-adhering pharmaceutical compositions containing fluoroquinolone antimicrobial agents are described herein to address one or more problems in ocular drug delivery, including those disclosed above. [Overview of the Initiative]

[0006] This disclosure relates to an ocular mucosal adhesion pharmaceutical composition comprising a fluoroquinolone antibiotic, chitosan, polyvinyl alcohol, and polyvinylpyrrolidone. The composition is useful for the treatment and / or prevention of eye diseases, particularly eye infections and / or inflammatory conditions of the eye.

[0007] In another embodiment, the Disclosure also provides a kit comprising any of the pharmaceutical compositions disclosed herein. The kit may include instructions for use in the treatment and / or prevention of eye diseases.

[0008] In yet another aspect, the disclosure also provides a method for treating and / or preventing an eye disease in a subject, comprising administering a therapeutically effective amount of the pharmaceutical composition described herein to a subject in need thereof. [Brief explanation of the drawing]

[0009] [Figure 1] Figure 1 shows the in vitro release profiles of (a) besifloxacin and (b) levofloxacin from free drug solutions and fluoroquinolones incorporated into hydrogels. [Figure 2] Figure 2 shows the in vitro mucosal adhesion test. (a) Average fluorescence intensity of besifloxacin and (b) levofloxacin preparations. [Figure 3] Figure 3 shows the ex vivo corneal penetration profiles of (a) besifloxacin from Besix®, a hydrogel formulation and free drug solution, and (b) levofloxacin from Levotop®, a levofloxacin hydrogel and free drug solution, 24 hours after excision of human cornea. [Figure 4] Figure 4 shows the ex vivo corneal tissue concentrations of (a) besifloxacin and (b) levofloxacin preparations after 0.5 hours of incubation. [Figure 5] Figure 5 shows the antibacterial activity of besifloxacin and levofloxacin preparations against Staphylococcus aureus and ATCC 25923-infected corneas after 1.5 hours and 0.5 hours of incubation. [Modes for carrying out the invention]

[0010] Detailed explanation Before describing the method of this disclosure in more detail, it should be understood that the method is not limited to the specific embodiments described and is, of course, subject to change. It should also be understood that the terms used herein are intended solely to describe specific embodiments and are not intended to be limiting, as the scope of the method is limited only by the appended claims.

[0011] Where a range of values ​​is provided, unless the context explicitly indicates otherwise, it is understood that the method includes each intervening value between the upper and lower limits of that range, up to one-tenth of the lower limit unit, and any other listed or intervening values ​​within that listed range. The upper and lower limits of these smaller ranges may independently be included within their smaller ranges and, subject to any restrictions specifically excluded in the listed range, are also included in the method. If one or both of the restrictions are included in the listed range, the range excluding one or both of the included restrictions is also included in the method.

[0012] In this specification, a specific range is given numerically, preceded by the term “approximately.” The term “approximately” is used herein to provide literal support for the exact number it precedes, as well as for any number that is close to or nearly equal to the number it precedes. In determining whether a number is close to or nearly equal to a specifically listed number, a number that is close to or approximates an unlisted number may be a number that, in the context in which it is presented, is substantially equivalent to a specifically listed number.

[0013] Disclosed herein are ocular mucosal adhesion pharmaceutical compositions comprising a therapeutically effective amount of a fluoroquinolone antibiotic and a combination of polymers(s). The compositions disclosed herein exhibit enhanced corneal mucosal adhesion and permeability of besifloxacin and levofloxacin and are advantageous over currently available eye drops (Besix® and Levotop®) for the delivery of besifloxacin and levofloxacin to the eye.

[0014] The target fluoroquinolone antibiotics include valofloxacin, besifloxacin, cinoxacin, ciprofloxacin, clinafloxacin, danofloxacin, delafloxacin, difloxacin, enoxacin, enrofloxacin, fleroxacin, flumequin, gatifloxacin, gemifloxacin, grepafloxacin, ivafloxacin, JNJ-Q2, levofloxacin, lomefloxacin, marbofloxacin, moxifloxacin, and na This includes, but is not limited to, difloxacin, nalidixic acid, nemonoxacin, norfloxacin, ofloxacin, orbifloxacin, oxyphosphate, pazufloxacin, pefloxacin, pyromidic acid, pipemidic acid, prulifloxacin, losoxacin, rufloxacin, sarafloxacin, sparfloxacin, sitafloxacin, temafloxacin, tosufloxacin, and trovafloxacin, or pharmaceutically acceptable salts or stereoisomers thereof.

[0015] In certain embodiments, the polymer is selected from the group comprising chitosan, polyvinyl alcohol (PVA), and polyvinylpyrrolidone (PVP). In further embodiments, the pharmaceutical composition comprises three polymers: chitosan, PVA, and PVP.

[0016] The composition may further comprise one or more permeation enhancers. In certain embodiments, the permeation enhancer is selected from the group consisting of lysophosphatidyl lipids (lysophosphatidylcholine, LPC), calcium chelating agents (EDTA), benzalkonium chloride, cetylpyridinium chloride, palmitoyl carnitine, nonionic surfactants (Brij 35, Brij 78, Brij 98, sodium deoxycholate, polyoxyethylene-9-lauryl ether), surfactant heteroglycosides, and bile salts (deoxycholate, taurodeoxycholate, and glycocholate), and glycosides (saponin, digitonin, caprylic acid, capric acid).

[0017] When a particular compound, e.g., levofloxacin or besifloxacin, is mentioned by name, the scope of the present disclosure is intended to encompass pharmaceutically acceptable salts, stereoisomers, esters, amides, or prodrugs of the designated compound. Further, when the designated compound contains a chiral center, the scope of the present disclosure includes compositions containing a racemic mixture of the two enantiomers, as well as compositions containing each enantiomer individually, but substantially free of the other enantiomer. In further embodiments, when the designated compound contains more than one chiral center, the scope of the present disclosure includes compositions containing a mixture of various diastereomers, as well as compositions containing each diastereomer, but substantially free of the other diastereomers. For example, when a commercially available fluoroquinolone antibiotic contains two stereocenters, the scope of the present disclosure includes pharmaceutical compositions containing all four diastereomers, pharmaceutical compositions containing a racemic mixture of the R,R and S,S isomers, pharmaceutical compositions containing a racemic mixture of the R,S and S,R isomers, pharmaceutical compositions containing the R,R enantiomer but substantially free of the other diastereomers, pharmaceutical compositions containing the S,S enantiomer but substantially free of the other diastereomers, pharmaceutical compositions containing the R,S enantiomer but substantially free of the other diastereomers, and pharmaceutical compositions containing the S,R enantiomer but substantially free of the other diastereomers.

[0018] In certain embodiments, a composition containing the R enantiomer substantially does not contain the S enantiomer, or a composition containing the S enantiomer substantially does not contain the R enantiomer. In this context, "substantially does not contain" means that the composition contains less than about 20%, or less than about 15%, or less than about 10%, or less than about 5%, or less than about 3%, or less than about 3%, or less than about 1% of the minor enantiomer.

[0019] In certain embodiments, the present disclosure discloses a mucoadhesive pharmaceutical composition comprising a therapeutically effective amount of a fluoroquinolone antibiotic, chitosan, PVA, PVP, and LPC.

[0020] In certain embodiments, the fluoroquinolone antibiotic is present in an amount of about 0.01 to about 1% by weight. In further embodiments, the fluoroquinolone antibiotic is present in an amount of about 0.3 to about 0.6% by weight.

[0021] In certain embodiments, chitosan is present in an amount of about 0.2 to about 3% by weight. In further embodiments, chitosan is present in an amount of about 0.5 to about 2% by weight.

[0022] In certain embodiments, PVA is present in an amount of about 0.2 to about 8% by weight. In further embodiments, PVA is present in an amount of about 0.5 to about 5% by weight.

[0023] In certain embodiments, PVP is present in an amount of about 0.5 to about 8% by weight. In further embodiments, PVP is present in an amount of about 0.5 to about 5% by weight.

[0024] In certain embodiments, LPC is present in an amount of about 0.01 to about 2% by weight. In further embodiments, LPC is present in an amount of about 0.05 to about 1% by weight.

[0025] In certain embodiments, the pharmaceutical composition comprises: · a fluoroquinolone antibiotic in an amount of about 0.3 to about 0.6% w / v; • Chitosan in an amount of approximately 0.5 to 2% w / v; • Approximately 0.5 to 5% w / v of PVA; • PVP is approximately 0.5 to 5% w / v; and • LPC in an amount of approximately 0.05 to 0.5% w / v.

[0026] The compositions can be formulated as topical solutions, suspensions, gels, emulsions, ointments, inserts, or films. In certain embodiments, the compositions disclosed herein may be administered in the form of eye drops, eye washes, eye ointments, eye creams, ophthalmic gels, and the like. The compositions may be in a single-dose dosage form or in a dosage form comprising multiple doses.

[0027] In certain embodiments, disclosed herein are hydrogel compositions comprising a therapeutically effective amount of a fluoroquinolone antibiotic, chitosan, PVA, and PVP. The hydrogel composition may further comprise one or more permeabilizing agents.

[0028] Fluoroquinolone antibiotics and permeabilis enhancers are as defined above. In certain embodiments, the fluoroquinolone antibiotic is selected from the group including becifloxacin, ciprofloxacin, ofloxacin, levofloxacin, gatifloxacin, and moxifloxacin. In further embodiments, the fluoroquinolone antibiotic is becifloxacin or levofloxacin, or a combination thereof.

[0029] In certain embodiments, the hydrogel composition comprises a therapeutically effective amount of besifloxacin, chitosan, PVA, PVP, and optionally LPC.

[0030] In certain embodiments, the hydrogel composition comprises a therapeutically effective amount of levofloxacin, chitosan, PVA, PVP, and optionally LPC.

[0031] In certain embodiments, the hydrogel composition contains a fluoroquinolone antibiotic in an amount of about 0.01 to about 1% w / v. In further embodiments, the fluoroquinolone antibiotic is present in an amount of about 0.1 to about 0.8% w / v, for example, about 0.3 to about 0.6% w / v, for example, about 0.35% w / v, or about 0.4% w / v, or about 0.45% w / v, or about 0.5% w / v, or about 0.55% w / v. In some cases, the fluoroquinolone antibiotic is becifloxacin or levofloxacin, or a combination thereof.

[0032] In certain embodiments, the hydrogel composition contains chitosan in an amount of about 0.2 to about 3% w / v. In further embodiments, the chitosan is present in an amount of about 0.5 to about 2% w / v, for example, about 0.5 to about 1.5% w / v, for example, about 0.6% w / v, or about 0.7% w / v, or 0.8% w / v, or about 0.9% w / v, or about 1.0% w / v, or about 1.1% w / v, or about 1.2% w / v, or about 1.3% w / v, or about 1.4% w / v. In some cases, chitosan can be used with any suitable molecular weight, e.g., about 1 to 1000 kDa, e.g., 1 to 500 kDa, e.g., about 2 kDa, or about 4 kDa, or about 5 kDa, or about 8 kDa, or about 9 kDa, or about 10 kDa, or about 12 kDa, or about 15 kDa, or about 20 kDa, or about 25 kDa, or about 30 kDa, or about 40 kDa, or about 45 kDa, or about 50 kDa, or about 55 kDa, or about 60 kDa, or about 65 kDa, or about 70 kDa, or about 75 kDa, or about 80 kDa, or about 85 kDa, or about 90 kDa. a, or about 95 kDa, or about 100 kDa, or about 110 kDa, or about 120 kDa, or about 130 kDa, or about 140 kDa, or about 150 kDa, or about 160 kDa, or about 170 kDa, or about 180 kDa, or about 190 kDa, or about 200 kDa, or about 220 kDa, or about 240 kDa, or about 260 kDa, or about 280 kDa, or about 300 kDa, or about 320 kDa, or about 340 kDa, or about 360 kDa, or about 380 kDa, or about 400 kDa, or about 420 kDa, or about 480 kDa. In certain embodiments, the hydrogel composition may contain chitosan polymers of different molecular weights. The chitosan used may be deacetylated chitosan. In certain embodiments, the degree of deacetylation may be, but is not limited to, about 50% or more deacetylation, for example, 50-99% deacetylation, for example, about 50-90% deacetylation, or about 55-85% deacetylation, or about 60-85% deacetylation, or about 70-85% deacetylation, or about 80-85% deacetylation.

[0033] In certain embodiments, the hydrogel composition contains polyvinyl alcohol (PVA) in an amount of about 0.2 to about 8% w / v. In further embodiments, the PVA is about 0.2 to about 7% w / v, for example, about 0.5 to about 5% w / v, for example, about 0.6% w / v, or about 0.7% w / v, or about 0.8% w / v, or about 0.9% w / v, or about 1.0% w / v, or about 1.2% w / v, or about 1.4% w / v, or about 1.6% w / v, or about 1.8% w / v, or about 2.0% It is present in amounts of w / v, or approximately 2.2% w / v, or approximately 2.4% w / v, or approximately 2.6% w / v, or approximately 2.8% w / v, or approximately 3.0% w / v, or approximately 3.2% w / v, or approximately 3.6% w / v, or approximately 3.8% w / v, or approximately 4.0% w / v, or approximately 4.2% w / v, or approximately 4.4% w / v, or approximately 4.6% w / v, or approximately 4.8% w / v. In some cases, PVA may have any suitable molecular weight, for example, about 1 to 1000 kDa, for example, 1 to 500 kDa, for example, about 2 kDa, or about 3 kDa, or about 4 kDa, or about 5 kDa, or about 8 kDa, or about 9 kDa, or about 10 kDa, or about 12 kDa, or about 15 kDa, or about 20 kDa, or about 25 kDa, or about 30 kDa, or about 40 kDa, or about 45 kDa, or about 50 kDa, or about 55 kDa, or about 60 kDa, or about 65 kDa, or about 70 kDa, or It may have approximately 75kDa, or approximately 80kDa, or approximately 85kDa, or approximately 90kDa, or approximately 95kDa, or approximately 100kDa, or approximately 110kDa, or approximately 120kDa, or approximately 130kDa, or approximately 140kDa, or approximately 150kDa, or approximately 160kDa, or approximately 170kDa, or approximately 180kDa, or approximately 190kDa, or approximately 200kDa, or approximately 210kDa, or approximately 250kDa, or approximately 300kDa, or approximately 350kDa, or approximately 400kDa, or approximately 450kDa.

[0034] In certain embodiments, the hydrogel composition contains polyvinylpyrrolidone (PVP) in an amount of about 0.5 to about 8% w / v. In further embodiments, PVP is present in an amount of about 0.5 to about 5% w / v. In yet another embodiment, PVP is present in an amount of about 1% w / v, or about 1.5% w / v, or about 2.0% w / v, or about 2.5% w / v, or about 3.0% w / v, or about 3.5% w / v, or about 4.0% w / v, or about 4.5% w / v. In some cases, PVP can have any suitable molecular weight, e.g., about 1 to 1000 kDa, e.g., 1 to 500 kDa and 1 to 250 kDa, e.g., about 2 kDa, or about 4 kDa, or about 5 kDa, or about 8 kDa, or about 9 kDa, or about 10 kDa, or about 12 kDa, or about 15 kDa, or about 20 kDa, or about 25 kDa, or about 30 kDa, or about 40 kDa, or about 45 kDa, or about 50 kDa, or about 55 kDa, or about 60 kDa, or about 65 kDa. , or may have about 70 kDa, or about 75 kDa, or about 80 kDa, or about 85 kDa, or about 90 kDa, or about 95 kDa, or about 100 kDa, or about 110 kDa, or about 120 kDa, or about 130 kDa, or about 140 kDa, or about 150 kDa, or about 160 kDa, or about 170 kDa, or about 180 kDa, or about 190 kDa, or about 200 kDa, or about 210 kDa, or about 220 kDa, or about 230 kDa, or about 240 kDa. In certain embodiments, the PVP polymer may be homopolymer PVP and copolymer vinyl acetate vinylpyrrolidone. Homopolymer PVP is known in the pharmaceutical industry by various names including povidone, polyvidone, polyvidonum, polyvidonum soluble, and poly(1-vinyl-2-pyrrolidone). The copolymer vinyl acetate vinylpyrrolidone is known in the pharmaceutical industry as copolyvidon, copolyvidone, and copolyvidonum.

[0035] In certain embodiments, the hydrogel composition contains LPC in an amount of about 0.01 to about 2% w / v. In further embodiments, the LPC is present in an amount of about 0.01 to about 1.5% w / v, for example, about 0.01 to about 1.0% w / v, for example, about 0.05 to about 0.5% w / v, for example, 0.1% w / v, or about 0.2% w / v, or about 0.3% w / v, or about 0.4% w / v, or about 0.5% w / v, or about 0.6% w / v, or about 0.7% w / v, or about 0.8% w / v, or about 0.9% w / v.

[0036] In a particular embodiment, the hydrogel composition includes: • Approximately 0.3-0.6% w / v of fluoroquinolone antibiotics; • Chitosan in an amount of approximately 0.5 to 2% w / v; • Approximately 0.5 to 5% w / v of PVA; • PVP is approximately 0.5 to 5% w / v; and • LPC in an amount of approximately 0.05 to 0.5% w / v.

[0037] The compositions disclosed herein may further comprise pharmaceutically acceptable carriers or excipients. Examples of pharmaceutically acceptable excipients include, but are not limited to, solvents such as water, pH adjusters such as sodium hydroxide and hydrochloric acid, isotonic adjusters such as glycerol, mannitol, sorbitol and xylitol, gelling agents such as cellulose derivatives (e.g., hydroxyethylcellulose, hydroxypropylmethylcellulose, etc.) and polymers from the Pluronic® series, viscosity adjusters such as hydroxyethylcellulose and hydroxypropylmethylcellulose, preservatives such as benzalkonium chloride, parabens, chlorohexidine, chlorohexidine gluconate and chlorohexidine acetate, chelating agents such as EDTA, and stabilizers / antioxidants such as tocopherol, tocopherol acetate, butylhydroxytoluene and butylhydroxyanisole.

[0038] The compositions disclosed herein can be used as pharmaceuticals (pharmaceuticals). In certain embodiments, the compositions are useful for the treatment and / or prevention of eye diseases, particularly eye infections and / or inflammatory conditions of the eye. In certain embodiments, the compositions disclosed herein are useful for treating ophthalmic infections such as keratitis, blepharitis, conjunctivitis and endophthalmitis.

[0039] In certain embodiments, the Disclosure also provides a kit comprising any of the pharmaceutical compositions disclosed herein. The kit may include instructions for use in the treatment and / or prevention of eye diseases.

[0040] This disclosure also provides a mucosal-adhesion-based ophthalmic drug delivery system comprising a hydrogel composition as disclosed herein. A mucosal-adhesion drug delivery system is useful in a method for delivering one or more pharmacologically active drugs via target ocular tissue. In certain embodiments, the ocular tissue is corneal tissue. In certain embodiments, the method involves adhering the mucosal-adhesion drug delivery system to the target corneal tissue.

[0041] This disclosure also describes methods for treating and / or preventing ocular diseases or conditions, diseases, or conditions in a subject, comprising administering a therapeutically effective amount of a pharmaceutical composition as described herein to a subject in need thereof. In certain embodiments, this disclosure describes methods for treating ocular conditions, diseases, or conditions in a subject, comprising administering a composition as described herein to a target site of the eye in the subject. Ocular conditions are selected from the group including blepharitis, conjunctivitis, keratitis, endophthalmitis, and conditions caused by Gram-positive and Gram-negative bacteria such as Staphylococcus aureus, Staphylococcus epidermidis, and Pseudomonas aeruginosa.

[0042] The composition can be administered to various different types of subjects, for example, as described herein. Subjects include, but are not limited to, both human and non-human mammals, including Carnivora (e.g., cats and dogs), Rodentia (e.g., mice, guinea pigs, and rats), Lagomorpha (e.g., rabbits), and Primates (e.g., humans, chimpanzees, and monkeys). In certain embodiments, the subject (e.g., patient) is human.

[0043] In certain embodiments, the compositions disclosed herein may inhibit the growth of at least one pathogen selected from the group including Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus pneumoniae, Pseudomonas aeruginosa, and Haemophilus influenzae.

[0044] In certain embodiments, the mucosal adhesion of besifloxacin and levofloxacin hydrogel was 3.5 times and 8 times higher, respectively, compared to commercially available eye drops and free drug solutions (p<0.001).

[0045] In certain embodiments, the cumulative corneal permeability of the hydrogel formulation is higher compared to that of commercially available besifloxacin and levofloxacin formulations.

[0046] In certain embodiments, the hydrogel compositions / formulations disclosed herein showed enhanced mucosal adhesion and superior permeability of fluoroquinolone antibiotics to the human cornea compared to commercially available formulations. In further embodiments, hydrogel compositions such as those disclosed herein showed excellent antimicrobial activity.

[0047] In certain embodiments, the hydrogel composition of a fluoroquinolone antibiotic is formulated such that the hydrogel composition exhibits up to 100 times higher mucosal adhesion and superior cumulative corneal permeability compared to commercially available ophthalmic formulations and free drug solutions. In further embodiments, the hydrogel compositions / formulations of fluoroquinolone antibiotics of the present disclosure may exhibit mucosal adhesion and superior cumulative corneal permeability about 1 to 50 times, for example, about 1 time, or about 1.5 times, or about 2 times, or about 2.5 times, or about 3 times, or about 3.5 times, or about 4 times, or about 4.5 times, or about 5 times, or about 5.5 times, or about 6 times, or about 6.5 times, or about 7.5 times, or about 8 times, or about 8.5 times, or about 9 times, or about 9.5 times, or about 10 times, or about 10.5 times, or about 11 times, or about 12 times, or about 12.5 times, or about 13 times, or about 13.5 times, or about 14 times, or about 14.5 times, or about 15 times higher than commercially available ophthalmic formulations and free drug solutions.

[0048] In certain embodiments, hydrogel compositions / formulations of besifloxacin and levofloxacin were found to have 3.5- and 8-fold higher (p<0.001) mucosal adhesion and superior cumulative corneal permeability compared to commercially available ophthalmic formulations and free drug solutions. The formulations exhibited excellent in vitro anti-infective properties. When besifloxacin and levofloxacin formulations were incubated with a corneal model infected with Staphylococcus aureus for 0.5 hours, the hydrogel formulations showed higher efficacy compared to commercially available eye drops and standard solutions.

[0049] In certain embodiments, 0.5-hour incubation of the besifloxacin and levofloxacin hydrogel formulations / compositions of the present disclosure with Staphylococcus aureus-infected corneas showed higher efficacy compared to commercially available eye drops and standard solutions.

[0050] The compositions disclosed herein can be prepared by any method known in the art. In certain embodiments, a process is disclosed for preparing a hydrogel composition comprising a fluoroquinolone antibiotic, chitosan, PVA, PVP, and optionally a permeation enhancer. This process includes preparing a multicomponent hydrogel comprising chitosan, polyvinyl alcohol, and polyvinylpyrrolidone; and adding a fluoroquinolone antibiotic to the multicomponent hydrogel. The process for preparing the multicomponent hydrogel includes treating chitosan with polyvinyl alcohol in an aqueous solution containing an acid; and adding polyvinylpyrrolidone followed by lysophosphatidylcholine. Any acid (organic or inorganic) suitable for the process can be used in the process. In certain embodiments, the chitosan is treated with polyvinyl alcohol in an acetic acid solution at a temperature of about 30–80°C.

[0051] Unless otherwise defined, all technical and scientific terms used herein have the same meaning, and the meaning of such terms is independent of each instance and is generally understood by one of the persons skilled in the art to which the subject matter herein belongs. Nevertheless, unless otherwise specified, the following definitions apply throughout the specification and claims.

[0052] As used herein, the terms “comprise”, “comprises”, and “comprising” are generally used to mean “to include,” that is, to allow the presence of one or more additional (unspecified) features or components.

[0053] As used herein, the term "including," as well as other forms such as "include," "includes," and "included," are not limited to this definition.

[0054] The phrase "pharmaceutically acceptable" is used herein to mean a compound, material, composition, and / or dosage form that is suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, provided that the benefits / risk ratio is reasonable, within the bounds of sound medical judgment.

[0055] The term "pharmaceutically acceptable salt" refers to a product obtained by the reaction of the fluoroquinolone antibiotics of this disclosure with a suitable acid or base. pharmaceutically acceptable salts of the compounds of the present invention include those derived from suitable inorganic bases such as Li, Na, K, Ca, Mg, Fe, Cu, Al, Zn, and Mn salts. Examples of pharmaceutically acceptable non-toxic acid addition salts are salts of amino groups formed by inorganic acids such as hydrochlorides, hydrobroms, hydroiodides, nitrates, sulfates, bisulfates, and phosphates, as well as isonicotinates, acetates, lactates, salicylates, citrates, tartrates, pantothenates, tartrates, ascorbic acidates, succinates, maleates, gentisinates, fumarates, glucons, glucaronates, sugarates, formates, benzoates, glutamates, methanesulfons, ethanesulfons, benzenesulfons, 4-methylbenzenesulfons, or p-toluenesulfons. Fluoroquinolone antibiotics can also form pharmaceutically acceptable salts with various organic bases such as lysine, arginine, guanidine, diethanolamine, or metformin. Suitable base salts include, but are not limited to, salts of aluminum, calcium, lithium, magnesium, potassium, sodium, and zinc.

[0056] The phrase “pharmaceutically acceptable carrier” is recognized in the art and includes, for example, pharmaceutically acceptable materials, compositions, or vehicles such as liquid or solid fillers, diluents, solvents, or encapsulating materials involved in the transport or delivery of any subject composition from one organ or part of the body to another organ or part of the body. Each carrier must be “acceptable” in the sense that it is compatible with the other components of the subject composition and is not harmful to the patient. In certain embodiments, a pharmaceutically acceptable carrier is non-pyrogenic. Some examples of materials that may serve as pharmaceutically acceptable carriers include: (1) Sugars such as lactose, glucose, and sucrose; (2) Starches such as corn starch and potato starch; (3) Cellulose, and its derivatives such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; (4) Powdered tragacanth; (5) Malt; (6) Gelatin; (7) Talc; (8) Cocoa butter and suppository waxes; (9) Oils such as peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, and soybean oil; (10) Propylene glycol, etc. (11) Glycols; polyols such as glycerin, sorbitol, mannitol, and polyethylene glycol; (12) esters such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) phosphate buffer; and (21) other non-toxic, suitable substances used in pharmaceutical formulations.

[0057] The term "prodrug" is intended to encompass compounds that, under physiological conditions, are converted into therapeutically active agents of the present invention. A common method for constructing a prodrug is to include a selected portion that, under physiological conditions, is hydrolyzed to reveal the desired molecule. In other embodiments, the prodrug is converted by the enzymatic activity of a host animal.

[0058] The terms “preventive” or “curative” or “therapeutic” treatments are recognized in the art and involve the administration of one or more subject compositions to a host. A treatment is preventive if it is administered before clinical signs of an undesirable condition (e.g., a disease or other undesirable condition in the host animal), i.e., it protects the host from the onset of the undesirable condition, but a treatment is therapeutic if it is administered after signs of an undesirable condition (i.e., it aims to reduce, improve or stabilize an existing undesirable condition or its side effects).

[0059] As used herein, the term “stereoisomer” is used for all isomers of the individual compounds of this disclosure that differ only in the orientation of their atoms in space. The term stereoisomer includes enantiomers of the compounds of this disclosure, mixtures of enantiomers of the compounds of this disclosure (racemates, racemic mixtures), geometric isomers of the compounds of the present invention (cis / trans or E / Z, R / S), and isomers of the compounds of this disclosure having two or more chiral centers that are not mirror images of each other (diastereoisomers).

[0060] The phrase “therapeutic effective dose” is a term recognized in the art. In certain embodiments, this term refers to the amount of a solvate or hydrate or composition disclosed herein that produces some desired effect with a reasonable benefit / risk ratio applicable to any medical treatment. In certain embodiments, this term refers to the amount necessary or sufficient to eliminate or alleviate a medical symptom for a given period of time. The effective dose may vary depending on factors such as the disease or condition being treated, the specific target construct administered, the size of the subject, or the severity of the disease or condition. Those skilled in the art can empirically determine the effective dose of a particular composition without requiring excessive experimentation.

[0061] The term “to treat” (or “to treat”) is recognized in the art and includes preventing the development of a disease, disorder or condition in a subject who is susceptible to a disease, disorder or condition but has not yet been diagnosed with it; inhibiting a disease, disorder or condition, e.g., preventing its progression; and mitigating a disease, disorder or condition, e.g., causing regression of the disease, disorder or condition. Treatment of a disease or condition includes improving at least one symptom of a particular disease or condition, even if the underlying pathophysiology is not affected, such as treating keratitis, blepharitis, conjunctivitis and endophthalmitis, and other related diseases or other medical conditions, and includes administering a composition that reduces the frequency of symptoms of a medical condition or delays its onset in a subject compared to a subject not administered the composition, as is well understood in the art.

[0062] The present invention is described in the following embodiments, but this should not be construed as limiting. Other objects, features, advantages and aspects of the present invention will be apparent to those skilled in the art from this description and the embodiments. However, it should be understood that the description and the specific embodiments below, while illustrating preferred embodiments of the present invention, are given only as examples. Various changes and modifications within the spirit and scope of the disclosed invention will be readily apparent to those skilled in the art from reading this disclosure and other parts thereof.

[0063] The sources of all materials, reagents, and compounds used in this disclosure are as follows: Besifloxacin HCl was provided by the RV Northland Institute in India as unconditional support for the study. Levofloxacin, poly(N-vinylpyrrolidone) (PVP K-30, MW: 40 kDa), chitosan (80-85% deacetylated, MW: 100-300 kDa), polyvinyl alcohol (PVA, MW: 140 kDa), potassium dihydrogen phosphate, dipotassium hydrogen phosphate, lysophosphatidylcholine, sodium chloride, and potassium chloride were obtained from Sigma Aldrich in India. Milli-Q (Millipore, USA) water was used in all experiments. Commercial formulations of besifloxacin (Besix®) and levofloxacin (Levotop®) were used as control formulations. Excised human corneas were obtained from the Ramayamma International Eye Bank, LV Prasad Eye Institute in Hyderabad, India. The corneas were transported in Dulbecco's Medium Eagle medium and stored at -20°C. [Examples]

[0064] example Preparation of hydrogel formulations: Step A: Preparation of a multi-component hydrogel formulation containing chitosan, polyvinyl alcohol, and polyvinylpyrrolidone polymer. In the first step, chitosan (0.5-1% w / w) was added to deionized water containing glacial acetic acid (0.5% v / v) with continuous stirring. Polyvinyl alcohol (1-3% w / w) was slowly added to the above solution. The temperature of the preparation was maintained at 55°C to dissolve the PVA. Polyvinylpyrrolidone (0.5-1% w / w) was added to this solution. In the final step, lysophosphatidylcholine (0.05% w / w) was added. Step B: To prepare the fluoroquinolone-containing hydrogel, 0.4% w / v besifloxacin or levofloxacin was added to the blank hydrogel while continuously stirring. Table 1 shows the final composition of the formulation.

[0065] Hydrogels were prepared using the components listed in Table 1 by a conventional stirring method. The polymer concentrations were optimized to 5% (3% PVA:1% PVP:1% chitosan) for besifloxacin and 2% (1% PVA:0.5% PVP:0.5% chitosan) for levofloxacin. The pH values ​​of the besifloxacin and levofloxacin hydrogels were 6.29±0.001, 6.14±0.002, 5.78±0.005, and 5.75±0.002 with and without LPC, respectively. The viscosities of the hydrogels were 139.93±1.70, 142.80±4.25, 21.41±0.52, and 22.09±1.79 for the besifloxacin and levofloxacin formulations, respectively. The transparency of the hydrogel was >95% with besifloxacin and >98% with levofloxacin preparations.

[0066] Characterization of hydrogel formulations: The hydrogels were characterized for their physicochemical properties. pH was measured using a digital pH meter (Elico Ltd., India). The viscosity of the sample preparations was measured using a cup-and-bob rheometer (Brookfield Inc., USA) after applying various shear rates at 25±1°C. The transmittance of the hydrogels was recorded at wavelengths of 400–700 nm using a UV-Vis spectrophotometer (Jasco Instruments, USA). Drug content was evaluated by dissolving 20 μl of drug-loaded hydrogel in 980 μl of distilled water, and the concentration was determined using high-performance liquid chromatography (HPLC).

[0067] HPLC method HPLC analysis was performed using a Shimadzu Prominence UFLC, LC-20 system (Shimadzu, Japan) equipped with a C18 column (Shimpack GIST column, 5 μm, 4.6 × 250 mm). The mobile phase used for besifloxacin analysis consisted of a mixture of 0.5% triethylamine solution (pH adjusted to 3.0 with 10% orthophosphate) and acetonitrile (ACN) (74:26, v / v) (Costa et al, 2014). For levofloxacin analysis, 20 mM potassium dihydrogen phosphate (KH2PO4) containing 1.0 ml of triethylamine solution (pH adjusted to 2.5 with 10% orthophosphate) and acetonitrile (77:23, v / v) was used (Dabhi et al, 2013). The injection volume was 50 μl for all standard and test solutions. The flow rate and column temperature were maintained at 1.0 ml / min and 25°C, respectively. Besifloxacin and levofloxacin are, respectively, λ max(ベシフロキサシン) =295 and λ max(レボフロキサシン) It was detected at =294. The amount of drug release was determined using the respective standard calibration curves. The composition and physicochemical properties of the hydrogel formulation are shown in Table 1 below.

[0068] [Table 1]

[0069] In vitro drug release studies In vitro drug release was performed using a Franz diffusion cell apparatus (Permegear Inc., USA). A dialysis membrane (molecular weight cutoff 2.5–3.0 kDa, Himedia, India) was placed between the donor and receptor compartments of the diffusion cell. Phosphate-buffered saline (100 mM, pH 7.4) was used as the receptor medium, and the temperature was maintained at 33 ± 1°C using a heated water circulator. The donor compartment was loaded with 2 mg of drug, including a polymer hydrogel, a standard drug solution, or a commercially available formulation. Samples (300 μl) were withdrawn from the sampling port at predetermined time intervals of 0.5, 1, 2, 4, 6, 12, and 24 hours and replaced with fresh medium. The samples were analyzed using the HPLC method described above.

[0070] In vitro release of besifloxacin and levofloxacin Figure 1 shows the diffusion of free drugs and drug release from the hydrogel. Free besifloxacin completely diffused across the membrane within 10 hours. In contrast, 100% release of besifloxacin from the hydrogel formulation was achieved in 24 hours (Figure 1a). The release of besifloxacin from the hydrogel formulation was initially higher than that of the free drug (up to 2 hours). In the case of levofloxacin, 100% of the drug concentration diffused within 2 hours and 24 hours in the free drug and the drug-loaded hydrogel, respectively (Figure 1b).

[0071] Mucosal adhesion test The adhesion strength of the hydrogel to the mucosa was qualitatively investigated using methods known in the art (Choy et al., 2008). A dialysis membrane (PVDF membrane, pore size 0.45 μm) was immersed in an aqueous mucin solution (0.1% mucin from pig stomach, type II conjugated with sialic acid) for 2 hours. A hydrogel (20 μl) containing rhodamine B (50 μg / ml) was applied drop by drop to the center of each membrane. The membrane was then immediately washed with PBS for 5 minutes. The number of remaining particles was measured using a fluorescence microscope and a spectrofluorometer.

[0072] In vitro mucosal adhesion test Mucosal adhesion of besifloxacin and levofloxacin formulations was investigated in vitro using a dialysis membrane immersed in salicylic acid-bound mucin to simulate the surface of the eye. The mucin surface was washed with PBS to simulate tear secretion. As shown in Figure 2, mucosal adhesion of besifloxacin hydrogel was 3.5 times higher than that of commercially available eye drops, and levofloxacin hydrogel was 8 times higher (p<0.001). Fluorescence microscopy images show greater adhesion of the hydrogel formulations to the cornea compared to free dyes.

[0073] Antibiotic ex vivo corneal permeability Corneal thickness was measured using a digital micrometer (Baker Gauges India Pvt. Ltd., India). Corneal resistance (R) was measured by applying direct current (I, 1 mA) to the cornea using a DC power supply unit, and voltage drop (V) was measured using a digital multimeter (Fluke Corporation, USA). Resistance (R) was calculated using Ohm's law (V=IR).

[0074] Ex vivo corneal permeabilization of besifloxacin and levofloxacin was performed using a Franz diffusion cell apparatus (PearmGear Corp., USA). Excised human corneas were washed with PBS and placed between the donor and receptor compartments with the epithelial side facing the donor chamber. The receptor medium contained PBS maintained at a temperature of 37±1°C using a warm water circulator unit. The donor compartment was filled with 0.4% of the drug, including polymer hydrogel, standard drug solution, or commercially available formulations. Samples (300 μl) were withdrawn from the receptor chamber at predetermined time intervals, including 0.5, 1, 2, 4, 6, 12, and 24 hours. Samples were analyzed by the HPLC method described above. The amount of permeated drug was determined using the respective standards (0.1, 1, 5, 10, 50, and 100 μg / ml) and the R of besifloxacin was determined. 2The R2 value for levofloxacin was 0.999, and the R2 value for levofloxacin was 0.997. Corneal permeability parameters were calculated from the cumulative dose-time profile of the permeated drug. Flux (J) was calculated from the slope of the linear portion of the cumulative dose-time profile of the permeated drug. Delay time (t lag The transmission coefficient (Kp) was calculated by extrapolating the linear portion of the curve onto the time axis. The transmission coefficient (Kp) was calculated using Equation 1.

number

[0075] In vitro corneal permeability of besifloxacin and levofloxacin preparations The average corneal thickness of human corneas used in in vitro studies was found to be 597.3 ± 40.8 μm. The average transepithelial electrical resistance (TEER) of intact corneas was 2.6 ± 0.4 kΩ. Table 2 shows the corneal TEER values ​​after application of various formulations. Table 3 shows the permeability parameters of various formulations of besifloxacin and levofloxacin. In general, application of hydrogels significantly reduced the delay time compared to commercially available products. The delay times for Besix® and Levotop® were 1.25 ± 0.09 hours and 1.42 ± 0.04 hours, respectively. The hydrogel formulations of besifloxacin and levofloxacin reduced the delay time to 0.92 ± 0.16 hours and 0.56 ± 0.01 hours, respectively. The hydrogel formulations of besifloxacin and levofloxacin were compared with Besix® (1.86 ± 0.74 μg / cm³). 2 / h) and Levotop(registered trademark) (7.85±1.14μg / cm³) 2 Compared to ( / h), the values ​​were 84.08±13.50 and 77.02±3.71 μg / cm³, respectively. 2The flux was significantly (p<0.05) greater at / h. Addition of lysophosphotidylcholine improved the flux by up to 10% with besifloxacin and up to 20% with levofloxacin hydrogel. Figure 3 shows the cumulative corneal permeation of besifloxacin and levofloxacin from the hydrogel formulations. The cumulative amount of besifloxacin permeating the cornea after application using the hydrogel formulation, Besix®, and free drug solution was found to be 1747±95, 55±12, and 1267±38 μg, respectively, after 24 hours. Similarly, the corneal tissue concentrations were found to be 258±37, 16±1, and 398±50 μg per gram of tissue for the hydrogel formulation, Besix®, and free drug solution.

[0076] In the case of levofloxacin, the cumulative corneal permeation from the hydrogel formulation, Levotop®, and free drug solution was found to be 1369±98, 259±13, and 1019±144 μg, respectively. The corneal tissue concentration was found to be 305±38, 130±17, and 251±13 μg per gram of tissue, respectively, for the hydrogel formulation, Levotop®, and free drug solution. Furthermore, the hydrogel formulation was found to have significantly higher diffusion and permeation coefficients (p<0.05) compared to commercially available products.

[0077] [Table 2]

[0078] [Table 3]

[0079] Evaluation of antibacterial activity in an ex vivo corneal infection model Human corneas were incubated overnight at 37°C in antibiotic-free DMEM medium. The following day, the corneas were washed three times with sterile PBS and tested for Staphylococcus aureus. 8Individual cells were injected into the stroma (using a 26-gauge needle). After incubation at 37°C for 24 hours, the infected corneas were washed again with PBS and used for further studies. Non-infected corneas were used as controls (Pinnock, et.al, 2017).

[0080] The infected corneas were washed and a metal ring was placed on the corneal-scleral button to create a watertight seal. The drug formulation (0.4%) was added to the center of the ring and incubated at 37°C for 0.5 hour and 1.5 hours. Then, the corneas were washed thoroughly with PBS to remove the excess formulation and incubated at 37°C for 24 hours. Next, the corneas were homogenized and the resulting suspension was serially diluted. To count the number of colonies, 10 μl of the sample was spotted onto agar plates at dilutions of 10 -5 、10 -6 、10 -7 、10 -8 The colonies were visually counted and expressed as colony-forming units (CFU) using Equation 2.

Number

[0081] Antibacterial Activity of Besifloxacin and Levofloxacin Hydrogel Formulations The antibacterial activity of the drugs loaded in the hydrogel formulation was evaluated against human corneas (n = 3) infected with Staphylococcus aureus (ATCC 25923). Figures 5 and Table 4 show the concentration-dependent inhibition of bacterial growth after incubating the infected corneas with the drug formulation for 30 minutes. Both hydrogel formulations of besifloxacin and levofloxacin were effective in antibacterial activity, with 0.1×10 9 and 2.8×10 9 CFU / ml compared to 8.0×10 9Only CFU / ml was recorded.

[0082] [Table 4]

[0083] Each embodiment is provided as a description of the invention, not as a limitation of the invention. Indeed, it will be apparent to those skilled in the art that various modifications and variations can be made to the compounds, compositions, and methods described herein without departing from the scope or spirit of the invention. For example, features described or mentioned as part of one embodiment can be applied to another embodiment to produce yet another embodiment. Thus, this disclosure is intended to include such modifications and variations, as well as their equivalents. Other objects, features, and aspects of the invention are disclosed or apparent therefrom in the following detailed description. It will be understood by those skilled in the art that this discussion is merely a description of exemplary embodiments and should not be construed as limiting broader aspects of this disclosure.

Claims

1. A pharmaceutical composition for treating a target eye condition, comprising a therapeutically effective amount of a fluoroquinolone antibiotic, chitosan with a molecular weight of 100 to 300 kDa, polyvinyl alcohol, and polyvinylpyrrolidone.

2. The pharmaceutical composition according to claim 1, wherein a fluoroquinolone antibiotic is present in an amount of about 0.01 to about 1% w / v.

3. The pharmaceutical composition according to claim 1, wherein chitosan is present in an amount of approximately 0.2 to approximately 3% w / v.

4. The pharmaceutical composition according to claim 1, wherein the polyvinyl alcohol is present in an amount of about 0.2 to about 8% w / v.

5. The pharmaceutical composition according to claim 1, wherein polyvinylpyrrolidone is present in an amount of about 0.5 to about 8% w / v.

6. A pharmaceutical composition for treating a target eye condition, comprising a therapeutically effective amount of a fluoroquinolone antibiotic, chitosan having a molecular weight of 100 to 300 kDa, polyvinyl alcohol and polyvinylpyrrolidone, and one or more permeabilizing agents.

7. The pharmaceutical composition according to claim 6, wherein the permeation enhancer is a lysophosphatidyl lipid.

8. The pharmaceutical composition according to claim 6, wherein the permeation enhancer is lysophosphatidylcholine.

9. The pharmaceutical composition according to claim 8, wherein lysophosphatidylcholine is present in an amount of about 0.01 to about 2% w / v.

10. A pharmaceutical composition for treating a target eye condition, comprising the following: Approximately 0.3 to 0.6% w / v of fluoroquinolone antibiotics; Chitosan in an amount of approximately 0.5 to 2% w / v; Polyvinyl alcohol in an amount of approximately 0.5 to 5% w / v; Polyvinylpyrrolidone in an amount of approximately 0.5 to approximately 5% w / v; and Lysophosphatidylcholine in an amount of approximately 0.05 to 0.5% w / v.

11. Fluoroquinolone antibiotics include balofloxacin, besifloxacin, cinoxacin, ciprofloxacin, clinafloxacin, danofloxacin, delafloxacin, difloxacin, enoxacin, enrofloxacin, fleroxacin, flumequin, gatifloxacin, gemifloxacin, grepafloxacin, ivafloxacin, JNJ-Q2, levofloxacin, lomefloxacin, marbofloxacin, moxifloxacin, nadifloxacin, and nalidic. A pharmaceutical composition according to any one of claims 1 to 10, selected from the group comprising sic acid, nemonoxacin, norfloxacin, ofloxacin, orbifloxacin, oxyphosphate, pazufloxacin, pefloxacin, pyromidic acid, pipemidic acid, prulifloxacin, losoxacin, rufloxacin, sarafloxacin, sparfloxacin, sitafloxacin, temafloxacin, tosufloxacin, trovafloxacin, or pharmaceutically acceptable salts or stereoisomers thereof.

12. The pharmaceutical composition according to any one of claims 1 to 10, wherein the fluoroquinolone antibiotic is becifloxacin or levofloxacin.

13. A pharmaceutical composition for treating a target eye condition, comprising a therapeutically effective amount of a fluoroquinolone antibiotic, chitosan having a molecular weight of 100 to 300 kDa, polyvinyl alcohol and polyvinylpyrrolidone, one or more permeabilisers, and a pharmaceutically acceptable carrier or excipient.

14. A pharmaceutical composition according to any one of claims 1 to 13, wherein the composition is a hydrogel.

15. The pharmaceutical composition according to claim 1, wherein the eye condition is selected from the group including blepharitis, conjunctivitis, keratitis, endophthalmitis, and conditions caused by Gram-positive and Gram-negative bacteria such as Staphylococcus aureus, Staphylococcus epidermidis, and Pseudomonas aeruginosa.

16. A mucosal-adhering ophthalmic drug delivery system comprising the pharmaceutical composition according to any one of claims 1 to 14.

17. A mucosal-adherent ophthalmic drug delivery system according to claim 16, for use in a method of delivering a therapeutically effective amount of fluoroquinolone antibiotic through target ocular tissue.

18. The mucosal-adhering ophthalmic drug delivery system according to claim 17, wherein the ocular tissue is corneal tissue.

19. A kit comprising the pharmaceutical composition according to any one of claims 1 to 14.

Images