Patent Application
20210393520
The present disclosure relates to a method of treating a patient suffering from gram-negative folliculitis or an inflammation associated with said gram-negative folliculitis, by applying topical formulation of Besifloxacin hydrochloride to the affected skin area and its surrounding.
Gram negative folliculitis (GNF) is believed to be the consequence of prolong treatment with broad spectrum antibiotics. Longer duration of antibiotic regime suppresses the usual gram-positive bacterial flora of nasal and facial skin and latter facial skin is colonized by the gram-negative rod bacterial flora (Enterobacteriaceae family). A typical feature of gram-negative folliculitis includes presence of papules, pustules and sever seborrhea in perioral and perinasal area of face of the person who has undergone long time broad spectrum antibiotic treatment or of the patients who are not responding to the standard acne and rosacea treatment. Subsequent microbiological assay demonstrating predominance of gram-negative rod-shaped bacteria in facial skin and nose mucous membrane of the suspected patient confirms the diagonosis.1 Leyden et al2 observed that there are two types of gram-negative folliculitis based on their clinical symptoms and causative agent. Type I GNF is more predominant (80-90%), characterized by superficial papules and pustules present in and around the nose and mouth, caused by gram-negative, rod-shaped and lactose fermenting bacteria such as Escherichia coli, Klebsiella spp., Serratia spp. and Enterobacter spp. The Type II GNF is characterized by deep nodular and cystic infection and is caused by Proteus mirabilis.
The existing modes of treatment of GNF include antibiotics such as 3rd generation cephalosporins, trimethoprim-sulfamethoxazole and Ampicillin. However, these antibiotics have high MIC values and their overuse has led to evolution of bacteria resistant to antibiotics. Thus, there exists a need for formulations based on antibiotics, which can treat GNF at relatively lower MIC and can inhibit the causal organisms, regardless of their resistance to other conventionally employed antibiotics.
The present disclosure relates to a method for treating gram-negative folliculitis in a subject, said method comprising topically administering a therapeutically effective amount of a formulation of besifloxacin at a concentration ranging from about 0.5% to about 4%. The said formulation of besifloxacin is gel, cream, lotion, foam, emulgel, ointment or spray.
In embodiments herein, the formulation, in addition to besifloxacin, comprises excipients selected from a group comprising anti-acne agent, alkalizing agent, anti-oxidant, anti-microbial agent, chelating agent, conditioning agent, dispersing agent, emollient, emulsifier, humectant, moisturizer, isotonic agent, foam stabilizer, solubilizer, thickening agent, penetration enhancer, preservative, solvent, surfactant, stabilizer, lubricant, opacifier and viscosity modifier.
In some embodiments, the alkalizing agent is selected from a group comprising sodium hydroxide and triethanolamine or a combination thereof; the anti-oxidant is selected from a group comprising butylated hydroxytoluene (BHT) and D-α-tocopherol polyethylene glycol succinate (TPGS) or a combination thereof; the anti-microbial agent is phenonip; the chelating agent is selected from a group edetate disodium and edetate disodium dihydrate or a combination thereof; the conditioning agent is cyclopentasiloxane; the dispersing agent is selected from a group comprising poloxamer 407 and poloxamer 124 or a combination thereof; the emollient is selected from a group comprising behenyl alcohol, cyclomethicone, oleyl oleate, and light liquid paraffin or a combination thereof; the emulsifier is selected from a group comprising Brij 35, cetyl alcohol, glyceryl stearate, glyceryl monostearate, laureth 4, PEG-400 stearate, polysorbate 60, steareth 2, sodium palmitate and steareth 21 or any combination thereof; the humectant is selected from a group comprising glycerine, methyl gluceth-20, and propylene glycol or a combination thereof; the moisturizer is allantoin; the isotonic agent is sodium chloride; the foam stabilizer is cocamidopropylbetaine; the solubilizer is selected from a group comprising caproyl 90, diethylene glycol monoethyl ether, N-methyl 2-pyrrolidone and polyethylene glycol 400 or any combination thereof; the thickening agent is selected from a group comprising carbomer homopolymer type C, carbomer, carbopol 980, hydroxyethyl cellulose, pemulen, sepineo P600, sodium hyaluronate, stearyl alcohol, ultrez 21 and xanthan gum or any combination thereof; the preservative is selected from a group comprising phenoxyethanol and propyl paraben; the solvent is purified water; the lubricant is PEG-7 glycerylcocoate; the opacifier is titanium dioxide; the viscosity modifier is selected from a group comprising carbopolaqua SF-1 and petrolatum; and the surfactant is selected from a group comprising sodium lauryl sulphate, sodium C14-16 olefin sulfonate, sodium lauryl ether sulphate, polyquaternium-39, ammonium lauryl sulphate (30%), disodium laureth sulfosuccinate (39%), sorbitan stearate and tween 80 or a combination thereof.
In embodiments herein, the gram-negative folliculitis is caused by gram-negative bacteria selected from a group comprising Klebsiella pneumoniae, Pseudomonas aeruginosa, Acinetobacter baumannii, Enterobacter aerogenes and Proteus mirabilis.
In further embodiments, the said gram-negative bacteria is resistant to conventional antibiotics and fluoroquinolone other than besifloxacin. These include ampicillin, amoxicillin, cefotaxime, clindamycin, tetracycline and erythromycin.
In embodiments herein, the formulation is topically administered at least once a day to up to four times a day; and wherein each administration is in an amount ranging from about 2 finger-tip unit (FTU) to about 4.5 finger-tip unit (FTU), or from about 1 gram to about 2.5 grams.
In embodiments herein, the formulation, in addition to besifloxacin, comprises a second active agent selected from a group comprising retinoid derivative, sebum inhibitor, antibiotic and anti-inflammatory agent, or any combination thereof, at a concentration ranging from about 0.001% to about 10%.
The present disclosure further relates to the said formulation for treating gram-negative folliculitis in a subject, comprising besifloxacin at a concentration ranging from about 0.5% to about 4%.
In embodiments herein, the formulation is selected from a group comprising gel, cream, lotion, foam, emulgel, ointment and spray; and in addition to the besifloxacin comprises excipients selected from a group comprising anti-acne agent, alkalizing agent, anti-oxidant, anti-microbial agent, chelating agent, conditioning agent, dispersing agent, emollient, emulsifier, humectant, moisturizer, isotonic agent, foam stabilizer, solubilizer, thickening agent, penetration enhancer, preservative, solvent, surfactant, stabilizer, lubricant, opacifier and viscosity modifier
In further embodiments, the formulation, in addition to besifloxacin, comprises a second active agent selected from a group comprising retinoid derivative, sebum inhibitor, antibiotic and anti-inflammatory agent, or any combination thereof.
The present disclosure also relates to a method for treating inflammation associated with gram-negative folliculitis in a subject, said method comprising topically administering a therapeutically effective amount of a formulation of besifloxacin at a concentration ranging from about 0.5% to about 4%.
The present disclosure also relates to a corresponding formulation for treating inflammation associated with gram-negative folliculitis in a subject, comprising besifloxacin at a concentration ranging from about 0.5% to about 4%.
In order that the disclosure may be readily understood and put into practical effect, reference will now be made to exemplary embodiments as illustrated with reference to the accompanying figures. The figures together with a detailed description below, are incorporated in and form part of the specification, and serve to further illustrate the embodiments and explain various principles and advantages, in accordance with the present disclosure wherein:
In view of the drawbacks associated, and to remedy the need created by the art available in the field of medicine, it is an objective of the disclosure to provide a new topical composition and a method for treating gram negative folliculitis, hot tub folliculitis or any other inflammatory disease of pilo-sebaceous follicles of skin, which is caused as a result of a new infection or de novo proliferation of commensal gram-negative bacteria.
The present disclosure thus provides a method of effectively treating gram-negative folliculitis or an inflammation associated with gram-negative folliculitis. The said treatment is carried out by topically administering a formulation that comprises besifloxacin at a specific concentration.
Accordingly, the present disclosure relates to a method for treating gram-negative folliculitis in a subject, said method comprising topically administering a therapeutically effective amount of a formulation of besifloxacin at a concentration ranging from about 0.5% to about 4%.
Similarly, the present disclosure relates to a method for treating inflammation associated with gram-negative folliculitis in a subject, said method comprising topically administering a therapeutically effective amount of a formulation of besifloxacin at a concentration ranging from about 0.5% to about 4%.
The besifloxacin employed in the present disclosure for treatment of gram-negative folliculitis or inflammation thereof is in different formulations including, but not limited to gel, cream, lotion, foam, emulgel, ointment and spray.
These formulations of besifloxacin are either aqueous or non-aqueous formulations. Accordingly, in embodiments of the present disclosure, when the formulation is non-aqueous, it is pH independent, whereas, when the formulation is an aqueous formulation, the pH of such formulation ranges from about 5 to about 8.
The said formulations of besifloxacin employed in the present disclosure for treatment of gram-negative folliculitis or an inflammation thereof comprises one or more components beyond besifloxacin itself. These components improve the activity and bioavailability of besifloxacin, and include anti-acne agent, alkalizing agent, anti-oxidant, anti-microbial agent, chelating agent, conditioning agent, dispersing agent, emollient, emulsifier, humectant, moisturizer, isotonic agent, foam stabilizer, solubilizer, thickening agent, penetration enhancer, preservative, solvent, surfactant, stabilizer, lubricant, opacifier and viscosity modifier.
In non-limiting embodiments of the present disclosure, the alkalizing agent is selected from a group comprising sodium hydroxide and triethanolamine or a combination thereof. When employed in a formulation of the present disclosure, the sodium hydroxide is at a concentration ranging from about 0.04% to about 1.2%, and the triethanolamine is at a concentration of about 1%.
In other non-limiting embodiments of the present disclosure, the anti-oxidant is selected from a group comprising butylated hydroxytoluene (BHT) and D-α-Tocopherol polyethylene glycol succinate (TPGS) or a combination thereof. When employed in a formulation of the present disclosure, the butylated hydroxytoluene (BHT) is at a concentration of about 0.1%, and the D-α-Tocopherol polyethylene glycol succinate (TPGS) is at a concentration ranging from about 3% to about 5%.
In further non-limiting embodiments of the present disclosure, the anti-microbial agent is phenonip. When employed in a formulation of the present disclosure, the phenonip is at a concentration ranging from about 0.3% to about 0.4%.
In further non-limiting embodiments of the present disclosure, the chelating agent is selected from a group edetate disodium and edetate disodium dihydrate or a combination thereof. When employed in a formulation of the present disclosure, the edetate disodium or the edetate disodium dihydrate is at a concentration of about 0.1%.
In further non-limiting embodiments of the present disclosure, the conditioning agent is cyclopentasiloxane. When employed in a formulation of the present disclosure, the cyclopentasiloxane is at a concentration of about 5%.
In further non-limiting embodiments of the present disclosure, the dispersing agent is selected from a group comprising poloxamer 407 and poloxamer 124 or a combination thereof. When employed in a formulation of the present disclosure, the poloxamer 407 or the poloxamer 124 is at a concentration ranging from about 0.5% to about 1%.
In further non-limiting embodiments of the present disclosure, the emollient is selected from a group comprising behenyl alcohol, cyclomethicone, oleyl oleate, and light liquid paraffin or a combination thereof. When employed in a formulation of the present disclosure, the behenyl alcohol is at a concentration ranging from about 1% to about 1.5%, the cyclomethicone is at a concentration ranging from about 1% to about 6%, the oleyl oleate is at a concentration of about 0.5%, and the light liquid paraffin is at a concentration ranging from about 2% to about 7%.
In further non-limiting embodiments of the present disclosure, the emulsifier is selected from a group comprising Brij 35, cetyl alcohol, glyceryl stearate, glyceryl monostearate, laureth 4, PEG-400 stearate, polysorbate 60, steareth 2, sodium palmitate and steareth 21 or any combination thereof. When employed in a formulation of the present disclosure, the Brij 35 is at a concentration of about 5.1%, the cetyl alcohol is at a concentration ranging from about 1% to about 2%, the glyceryl stearate or the glyceryl monostearate is at a concentration ranging from about 1.5% to about 3%, the laureth 4 is at a concentration of about 4%, the PEG-400 stearate is at a concentration ranging from about 2% to about 10%, the polysorbate 60 is at a concentration ranging from about 2% to about 4%, the sodium palmitate is at a concentration of about 94.2% and the steareth 2 or the steareth 21 is at a concentration ranging from about 2% to about 3%.
In further non-limiting embodiments of the present disclosure, the humectant is selected from a group comprising glycerin, methyl gluceth-20 and propylene glycol or a combination thereof. When employed in a formulation of the present disclosure, the glycerin is at a concentration ranging from about 1% to about 10%, methyl gluceth-20 is at a concentration ranging of about 0.3% to about 2.5% and the propylene glycol is at a concentration ranging from about 1% to about 22%.
In further non-limiting embodiments of the present disclosure, the moisturizer is allantoin. When employed in a formulation of the present disclosure, the allantoin is at a concentration of about 0.2%.
In further non-limiting embodiments of the present disclosure, the isotonic agent is sodium chloride. When employed in a formulation of the present disclosure, the sodium chloride is at a concentration of about 0.9%.
In further non-limiting embodiments of the present disclosure, the foam stabilizer is cocamidopropylbetaine. When employed in a formulation of the present disclosure, the cocamidopropylbetaine is at a concentration of about 0.5%.
In further non-limiting embodiments of the present disclosure, the solubilizer is selected from a group comprising caproyl 90, diethylene glycol monoethyl ether, N-methyl 2-pyrrolidone and polyethylene glycol 400 or any combination thereof. When employed in a formulation of the present disclosure, the caproyl 90 is at a concentration ranging from about 4% to about 5%, the diethylene glycol monoethyl ether is at a concentration ranging from about 1% to about 16%, the N-methyl 2-pyrrolidone is at a concentration of about 3%, and the polyethylene glycol 400 is at a concentration ranging from about 0.1% to about 8%.
In further non-limiting embodiments of the present disclosure, the thickening agent is selected from a group comprising carbomer homopolymer type C, carbomer, carbopol 980, hydroxyethyl cellulose, pemulen, sepineo P600, sodium hyaluronate, stearyl alcohol, ultrez 21 and xanthan gum or any combination thereof. When employed in a formulation of the present disclosure, the carbomer homopolymer type C is at a concentration ranging from about 0.3% to about 0.65%, the carbomer or the carbopol 980 is at a concentration ranging from about 0.1% to about 25%, the hydroxyethyl cellulose is at a concentration ranging from about 0.17% to about 1.75%, the pemulen is at a concentration ranging from about 1% to about 40%, the sepineo P600 is at a concentration ranging from about 4% to about 5%, the sodium hyaluronate is at a concentration ranging from about 0.1% to about 0.5%, the stearyl alcohol is at a concentration ranging from about 1% to about 2%, the ultrez 21 is at a concentration ranging from about 5% to about 20%, and the xanthan gum is at a concentration ranging from about 0.5% to about 0.6%.
In further non-limiting embodiments of the present disclosure, the preservative is phenoxyethanol or propyl paraben. When employed in a formulation of the present disclosure, the phenoxyethanol is at a concentration ranging from about 0.3% to about 0.7%, and the propyl paraben is at a concentration of about 0.03%.
In further non-limiting embodiments of the present disclosure, the surfactant is selected from a group comprising sodium lauryl sulphate, sodium C14-16 olefin sulfonate, sodium lauryl ether sulphate, polyquaternium-39, ammonium lauryl sulphate (30%), disodium laureth sulfosuccinate (39%), sorbitan stearate and tween 80 or a combination thereof. When employed in a formulation of the present disclosure, the sodium lauryl sulphate is at a concentration of about 5%, the sodium C14-16 olefin sulfonate is at a concentration of about 35%, the sodium lauryl ether sulphate is at a concentration of about 2%, the polyquaternium-39 is at a concentration of about 1%, the ammonium lauryl sulphate (30%) is at a concentration of about 30%, the disodium laureth sulfosuccinate (39%) is at a concentration of about 2%, the sorbitan stearate is at a concentration of about 1.4% and the tween 80 is at a concentration of about 8%.
In further non-limiting embodiments of the present disclosure, the lubricant is PEG-7 glycerylcocoate. When employed in a formulation of the present disclosure, the PEG-7 glycerylcocoate is at a concentration of about 1%.
In further non-limiting embodiments of the present disclosure, the opacifier is titanium dioxide. When employed in a formulation of the present disclosure, the titanium dioxide is at a concentration of about 0.5%.
In further non-limiting embodiments of the present disclosure, the viscosity modifier is selected from a group comprising carbopolaqua SF-1 and petrolatum. When employed in a formulation of the present disclosure, the carbopolaqua SF-1 is at a concentration ranging from about 1% to about 6%, and the petrolatum is at a concentration of about 1%.
In further non-limiting embodiments of the present disclosure, the solvent employed for preparing the formulations is purified water.
While the formulations of the present disclosure primarily comprise besifloxacin, they may also comprise of a second active agent. These active agents in combination with besifloxacin help in enhancing the treatment of the gram-negative folliculitis or an inflammation thereof.
Accordingly, in embodiments of the present disclosure, the formulation in addition to besifloxacin, comprises a second active agent selected from a group comprising retinoid derivative, sebum inhibitor, antibiotic and anti-inflammatory agent, or any combination thereof. This second active agent is at a concentration ranging from about 0.001% to about 10%.
In non-limiting embodiments of the present disclosure, the retinoid derivative is selected from a group comprising isotretinoin, tretinoin, tazoretene and adapalene or any combination thereof. When employed in a formulation, the retinoid derivative is at a concentration ranging from about 0.025% to about 0.7%. In an exemplary embodiment, the isotretinoin is at a concentration ranging from about 0.025% to about 0.7%, the tretinoin is at a concentration ranging from about 0.025% to about 0.1%, and the adapalene is at a concentration of about 0.1%.
In non-limiting embodiments of the present disclosure, the sebum inhibitor is selected from a group comprising acetyle coenzyme A carboxylase inhibitor and olumacostat glasaretil or a combination thereof. When employed in a formulation, the sebum inhibitor is at a concentration ranging from about 0.1% to about 10%.
In non-limiting embodiments of the present disclosure, the antibiotic is selected from a group comprising sulphadiazine and metronidazole or a combination thereof. When employed in a formulation, the antibiotic is at a concentration ranging from about 0.1% to about 10%.
In non-limiting embodiments of the present disclosure, the anti-inflammatory agent is selected from a group comprising nonsteroidal anti-inflammatory drug, cox-2 inhibitor, JAK inhibitor, PDE4 inhibitor, anti-leukotriene, aspirin, ibuprofen, naproxen, diclofenac, and nimesulide, or any combination thereof. Further, the COX-2 inhibitor is selected from a group comprising Celecoxib, rofecoxib, and valdecoxib, or any combination thereof; the JAK inhibitor is selected from a group comprising tofacitinib, ruxolitinib, baricitinib, oclacitinib and baricitinib, or any combination thereof; the PDE4 inhibitor is selected from a group comprising apremilast, roflumilast, cilomilast, ibudilast, piclamilast and crisaborole, or any combination thereof, and the anti-leukotriene is a LTB4 pathway drug selected from a group comprising etalocib, amelubant, moxilubant, ubenimex, tosedostat and acebilustat.
When employed in a formulation, the anti-inflammatory agent is at a concentration ranging from about 0.1% to about 10%.
The besifloxacin based formulations of the present disclosure treat the gram-negative folliculitis or an inflammation thereof by inhibiting the causal organisms thereof. In non-limiting embodiments, the gram-negative folliculitis is caused by gram-negative bacteria selected from a group comprising Escherichia coli, Klebsiella spp., Pseudomonas spp., Serratia spp., Acinetobacter spp., Enterobacter spp. and Proteus spp.
In exemplary embodiments, the gram-negative folliculitis is caused by gram-negative bacteria selected from a group comprising Klebsiella pneumoniae, Pseudomonas aeruginosa, Acinetobacter baumannii, Enterobacter aerogenes and Proteus mirabilis.
Accordingly, the besifloxacin based formulations of the present disclosure inhibit the gram-negative bacteria causing the folliculitis, and the minimum inhibitory concentration (MIC) of the besifloxacin against said gram-negative bacteria is lower than the MIC of conventional antibiotics such as cefotaxime and ampicillin. Further, the besifloxacin based formulations of the present disclosure are also able to treat the gram-negative folliculitis or an inflammation, caused by a gram-negative bacteria which is resistant to conventional antibiotics and fluoroquinolone other than besifloxacin.
In embodiments of the present disclosure, the besifloxacin also inhibits gram-positive bacteria associated with acne vulgaris selected from a group comprising Propionibacterium acnes, Staphylococcus epidermidis, Staphylococcus haemolyticus, Bacillus megaterium, Dermabacter hominis, Kocuria spp., Microbacterium spp., and Blastococcus spp.
As mentioned previously, the besifloxacin based formulations of the present disclosure are administered or applied topically for curing the gram-negative folliculitis. In non-limiting embodiments of the present disclosure, the said administration or application is carried out at least once a day to up to four times a day, to a subject suffering from gram-negative folliculitis. in need thereof. In embodiments herein, the subject is a human being suffering from said gram-negative folliculitis.
The amount of the besifloxacin based formulations of the present disclosure that must be topically administered or applied each time, ranges from about 2 finger-tip unit (FTU) to about 4.5 finger-tip unit (FTU). In other words, and in terms of grams, the amount of the besifloxacin based formulations of the present disclosure that must be topically administered or applied each time, ranges from about 1 gram to about 2.5 grams.
Thus, the topical administration of the besifloxacin based formulation is carried out at least once a day to up to four times a day; and wherein each administration is in an amount ranging from about 2 finger-tip unit (FTU) to about 4.5 finger-tip unit (FTU), or from about 1 gram to about 2.5 grams.
Thus, in exemplary embodiments, the amount of the besifloxacin based formulations of the present disclosure that must be topically administered or applied, ranges from about 2 FTU per day to about 18 FTU per day. In other words, the total of amount of besifloxacin based formulations of the present disclosure that must be topically administered or applied, ranges from about 1 gram per day to about 10 grams per day.
While the present disclosure provides formulations such as gel, cream, lotion, foam, emulgel, ointment and spray, to treat the gram-negative folliculitis or an inflammation thereof, it also provides corresponding processes to obtain the formulations. Since the formulations comprise of various components other than besifloxacin in different concentrations, different formulations may require slightly different processes to prepare them. However, the processes provided by this disclosure are not absolute, as a person skilled in the art will understand that once the components of a formulation are listed, how to best combine them for an effective formulation. Thus, in that regard, the importance of the present disclosure lies on providing the formulations and the components that make them.
Accordingly, the present disclosure provides a gel formulation of besifloxacin for treatment of gram-negative folliculitis or an inflammation thereof. Such a formulation at least comprises components selected from a group comprising besifloxacin, humectant, chelating agent, thickening agent, preservative, solubilizer and alkalizing agent.
In an exemplary embodiment, such a formulation comprises glycerin, besifloxacin HCl, purified water, edetate disodium, hydroxyethyl cellulose, sodium hyaluronate, carbomer 980, phenoxyethanol, polyethylene glycol 400, diethylene glycol monoethyl ether, and sod. hydroxide (18% w/w).
In another exemplary embodiment, such a formulation comprises besifloxacin.HCl, edetate disodium dihydrate, carbomer homopolymer type c, diethylene glycol monoethyl ether, polyethylene glycol 400, hydroxyethyl cellulose, sepineo p600, sodium hyaluronate, phenoxyethanol, glycerin, sodium hydroxide solution and purified water.
The gel formulations herein, in addition to above components, may further comprise moisturizer, dispersing agent, surfactant, retinoid derivative and isotonic agent.
In an exemplary embodiment, such a formulation comprises purified water, edetate disodium, allantoin, hydroxyethyl cellulose, carbomer 980, purified water, sodium hyaluronate, poloxamer 407, phenoxyethanol, sodium hydroxide (18% w/v), glycerin, besifloxacin HCl equivalent to besifloxacin, purified water, sodium hydroxide solution, polyethylene glycol 400, diethylene glycol monoethyl ether and sodium hydroxide (18% w/v).
In another exemplary embodiment, such a formulation comprises purified water, allantoin, carbomer 980, sodium hyaluronate, poloxamer 407, besifloxacin HCl, glycerin, polyethylene glycol 400, edetate disodium, phenoxyethanol, triethanolamine and diethylene glycol monoethyl ether.
In yet another exemplary embodiment, such a formulation comprises besifloxacin.HCl equivalent to besifloxacin, sodium lauryl sulphate (30%), tween 80, diethylene glycol monoethyl ether, propylene glycol, disodium edetate, sodium hydroxide (10%), carbomer 980 (3%), water, phenoxyethanol and triethanolamine.
In yet another exemplary embodiment, such a formulation comprises glycerin, besifloxacin HCl, purified water, edetate disodium, hydroxyethyl cellulose, sodium hyaluronate, carbomer 980, phenoxyethanol, isotretinoin, tretinoin, poloxamer 407, polyethylene glycol 400, diethylene glycol monoethyl ether and sod. hydroxide (18% w/w).
In yet another exemplary embodiment, such a formulation comprises glycerin, besifloxacin HCl, purified water, edetate disodium, hydroxyethyl cellulose, sodium hyaluronate, carbomer 980, phenoxyethanol, polyethylene glycol 400, diethylene glycol monoethyl ether, sodium chloride and sod. hydroxide solution.
Similarly, the present disclosure provides a cream formulation of besifloxacin for treatment of gram-negative folliculitis or an inflammation thereof. Such a formulation at least comprises components selected from a group comprising besifloxacin, thickening agent, emulsifier, humectant, alkalizing agent and anti-oxidants. In addition, such formulations can also comprise further components such as conditioning agent, anti-microbial agent, emollient, retinoid derivative, dispersing agent, solubilizer and preservative.
In an exemplary embodiment, such a formulation comprises pemulen TR1, cyclopentasiloxane, cetyl alcohol, stearyl alcohol, steareth 2, besifloxacin.HCl equivalent to besifloxacin, propylene glycol, steareth 21, water, sodium hydroxide (10%), BHT and phenonip.
In another exemplary embodiment, such a formulation comprises cyclomethicone, cetyl alcohol, stearyl alcohol, steareth 2, steareth 21, light liquid paraffin, besifloxacin HCl equivalent to besifloxacin, sodium hydroxide solution (10%), water, glycerol, carbopol 980 (2%), sodium hydroxide solution (10%), water, isotretinoin, tretinoin, poloxamer 407, water, PEG 400, butylated hydroxy toluene and phenoxyethanol.
The present disclosure also provides a lotion formulation of besifloxacin for treatment of gram-negative folliculitis or an inflammation thereof. Such a formulation at least comprises components selected from a group comprising besifloxacin, thickening agent, humectant, alkalizing agent and preservative. In addition, such formulations can also comprise further components such as anti-oxidant, emollient, emulsifier, chelating agent, solubilizer and isotonic agent.
In an exemplary embodiment, such a formulation comprises PEG-400 stearate, glyceryl stearate, light liquid paraffin, behenyl alcohol, glycerol, propylene glycol, besifloxacin HCl equivalent to besifloxacin, water, sodium hydroxide, carbopol 980, xanthan gum, sepineo P600, water, BHT and phenoxyethanol.
In another exemplary embodiment, such a formulation comprises glycerin, besifloxacin HCl, purified water, edetate disodium, hydroxyethyl cellulose, sodium hyaluronate, carbomer 980, phenoxyethanol, polyethylene glycol 400, diethylene glycol monoethyl ether, sodium chloride and sod. hydroxide solution.
In yet another exemplary embodiment, such a formulation comprises glycerin, besifloxacin HCl, purified water, edetate disodium, hydroxyethyl cellulose, sodium hyaluronate, carbomer 980, phenoxyethanol, polyethylene glycol 400, diethylene glycol monoethyl ether, sodium chloride and sod. hydroxide solution.
The present disclosure further provides a form formulation of besifloxacin for treatment of gram-negative folliculitis or an inflammation thereof. Such a formulation comprises components selected from a group comprising besifloxacin, thickening agent, emulsifier, isotonic agent, humectant, alkalizing agent, anti-oxidant, retinoid derivative and preservative.
In an exemplary embodiment, such a formulation comprises cetyl alcohol, glyceryl monostearate, cocamidopropylbetaine, steareth 2, steareth 21, polysorbate 60, glycerol, propylene glycol, besifloxacin.HCl equivalent to besifloxacin, water, sodium hydroxide, carbomer 980, sodium hydroxide, water, isotretinoin, tretinoin, BHT and phenoxyethanol.
The present disclosure further provides a spray formulation of besifloxacin for treatment of gram-negative folliculitis or an inflammation thereof. Such a formulation comprises components selected from a group comprising besifloxacin, humectant, thickening agent, preservative and alkalizing agent.
In an exemplary embodiment, such a formulation comprises glycerin, besifloxacin HCl, sodium carboxy methyl cellulose, phenoxyethanol, sodium hydroxide and purified water.
Similarly, the present disclosure also provides an emulgel formulation of besifloxacin for treatment of gram-negative folliculitis or an inflammation thereof. Such a formulation at least comprises components selected from a group comprising besifloxacin, emulsifier, anti-oxidant, humectant, thickening agent, solubilizer, alkalizing agent and dispersing agent.
In an exemplary embodiment, such a formulation comprises caproyl 90, cetyl alcohol, steareth 2, laureth 4, BRIJ 35, besifloxacin.HCl equivalent to besifloxacin, steareth 21, D-α-tocopherol polyethylene glycol 1000 succinate (tpgs), poloxamer 407, poloxamer 124, propylene glycol, purified water, propylene glycol, purified water, ultrez 21 (0.5% w/w) and triethanolamine.
The emulgel formulations herein, in addition to above components, may further comprise preservative, retinoid derivative and emollient.
In an exemplary embodiment, such a formulation comprises besifloxacin.HCl equivalent to besifloxacin, caproyl 90, cetyl alcohol, steareth 2, steareth 21, D-α-tocopherol polyethylene glycol 1000 succinate (tpgs), poloxamer 407, poloxamer 124, propylene glycol, purified water, ultrez 21 (2% w/w), isotretinoin, tretinoin, PEG 500, water, triethanolamine and phenoxyethanol.
In another exemplary embodiment, such a formulation comprises besifloxacin.HCl equivalent to besifloxacin, steareth 21, D-α-tocopherol polyethylene glycol 1000 succinate (tpgs), poloxamer 407, poloxamer 124, propylene glycol, purified water, caproyl 90, cetyl alcohol, steareth 2, cyclomethicone, ultrez 21 (1% w/w) and triethanolamine.
In another embodiment herein, a formulation of the present disclosure comprises Water, Carbopol 940, Allantoin, Besifloxacin HCl, (equivalent to besifloxacin), Adapalene, Triethanolamine, Glycerol, Propylene Glycol, PEG 400, Poloxamer 407, Disodium EDTA and Phenoxyethanol.
In yet another embodiment herein, a formulation of the present disclosure comprises Water, Sodium hydroxide (18% aq.), PEG 1450, Methyl Gluceth-20, Glycerin, Besifloxacin, Adapalene, Isopropyl alcohol, diethylene glycol monethyl ether, Propylene glycol, N-methyl 2-pyrrolidone, Sodium hydroxide, Phenoxyethanol and Fragrance
In still another embodiment herein, a formulation of the present disclosure comprises Water, Carbopol aqua SF-1, Sodium C14-16 Olefin Sulfonate, Sodium lauryl ether sulphate, Sodium hydroxide (18% aq.), Cocamidopropylbetaine (30%), Disodium EDTA, Glycerin, Besifloxacin, Adapalene, N-methyl 2-pyrrolidone, PEG-7 glycerylcocoate and Citric Acid (50%)
In still another embodiment herein, a formulation of the present disclosure comprises Sodium palmitate, Sodium lauryl ether sulphate, Polyquaternium-39, Methyl Gluceth-20, Titanium dioxide, Besifloxacin, Adapalene, Oleyl oleate and BHT (Butylated Hydorxy Toluene)
In still another embodiment herein, a formulation of the present disclosure comprises Water, Disodium EDTA, Carbopolaqua SF-1, Ammonium lauryl sulphate (30%), Propylene glycol, Besifloxacin, Adapalene, N-methyl 2-pyrrolidone, Ethanol, Propyl paraben, Methyl gluceth-10, Disodium laureth sulfosuccinate (39%), Fragrance and Triethanolamine
In still another embodiment herein, a formulation of the present disclosure comprises Water, Disodium EDTA, Carbopol aqua SF-1, Petrolatum, Cyclomethicone, Sorbitan stearate, Polysorbate 60, Methyl gluceth-20, Cetyl alcohol, Tocopheryl acetate, Besifloxacin, Adapalene, N-methyl 2-pyrrolidone, Propylene glycol, Glycerin, Ethanol, Phenoxyethanol and Sodium hydroxide
In still another embodiment herein, a formulation of the present disclosure comprises Besifloxacin.HCl Equivalent to Besifloxacin, Adapalene, Allantoin, Diethylene glycol monoethyl ether, Edetate disodium dihydrate, Glycerin, Hyaluronate Sodium, Hydroxy ethylcellulose, Phenoxyethanol, Poloxamer, Polyethylene glycol 400, and Purified water.
In still another embodiment herein, a formulation of the present disclosure comprises Adapalene, Besifloxacin.HCl equivalent to besifloxacin, Allantoin, Citric acid solution, Diethylene glycol monoethyl ether, Edetate disodium dehydrate, Glycerin, Hyaluronate Sodium, Hydroxy ethyl cellulose, Phenoxyethanol, Poloxamer 407, Polyethylene glycol 400, Sodium Hydroxide Solution and Purified Water.
In still another embodiment herein, a formulation of the present disclosure comprises Besifloxacin.HCl equivalent to besifloxacin, Adapalene, Allantoin, Carbomer homopolymer type C, Diethylene glycol monoethyl ether, Edetate disodium dehydrate, Glycerin, Hyaluronate sodium, Phenoxyethanol, Poloxamer 407, Polyethylene glycol 400, Sodium hydroxide solution and Purified water.
As a person skilled in the art understands, while the concentrations of the components employed in different formulations differ slightly, the overall concentration of every component employed in formulations of the present disclosure is governed by the ranges provided herein. Accordingly, the sodium hydroxide is at a concentration ranging from about 0.04% to about 1.2%, the triethanolamine is at a concentration of about 1%, the butylated hydroxytoluene (BHT) is at a concentration of about 0.1%, the D-α-Tocopherol polyethylene glycol succinate (TPGS) is at a concentration ranging from about 3% to about 5%, the phenonip is at a concentration ranging from about 0.3% to about 0.4%, the edetate disodium or the edetate disodium dihydrate is at a concentration of about 0.1%, the cyclopentasiloxane is at a concentration of about 5%, the poloxamer 407 or the poloxamer 124 is at a concentration ranging from about 0.5% to about 1%, the behenyl alcohol is at a concentration ranging from about 1% to about 1.5%, the cyclomethicone is at a concentration ranging from about 1% to about 6%, the oleyl oleate is at a concentration of about 0.5%, the light liquid paraffin is at a concentration ranging from about 2% to about 7%, the Brij 35 is at a concentration of about 5.1%, the cetyl alcohol is at a concentration ranging from about 1% to about 2%, the glyceryl stearate or the glyceryl monostearate is at a concentration ranging from about 1.5% to about 3%, the laureth 4 is at a concentration of about 4%, the PEG-400 stearate is at a concentration ranging from about 2% to about 10%, the polysorbate 60 is at a concentration ranging from about 2% to about 4%, the sodium palmitate is at a concentration of about 94.2%, the steareth 2 or the steareth 21 is at a concentration ranging from about 2% to about 3%, the glycerin is at a concentration ranging from about 1% to about 10%, Methyl Gluceth-20 is at a concentration ranging of about 0.3% to about 2.5%, the propylene glycol is at a concentration ranging from about 1% to about 22%, the allantoin is at a concentration of about 0.2%, the sodium chloride is at a concentration of about 0.9%, the cocamidopropylbetaine is at a concentration of about 0.5%, the caproyl 90 is at a concentration ranging from about 4% to about 5%, the diethylene glycol monoethyl ether is at a concentration ranging from about 1% to about 16%, the N-methyl 2-pyrrolidone is at a concentration of about 3%, the polyethylene glycol 400 is at a concentration ranging from about 0.1% to about 8%, the carbomer homopolymer type C is at a concentration ranging from about 0.3% to about 0.65%, the carbomer or the carbopol 980 is at a concentration ranging from about 0.1% to about 25%, the hydroxyethyl cellulose is at a concentration ranging from about 0.17% to about 1.75%, the pemulen is at a concentration ranging from about 1% to about 40%, the sepineo P600 is at a concentration ranging from about 4% to about 5%, the sodium hyaluronate is at a concentration ranging from about 0.1% to about 0.5%, the stearyl alcohol is at a concentration ranging from about 1% to about 2%, the ultrez 21 is at a concentration ranging from about 5% to about 20%, the xanthan gum is at a concentration ranging from about 0.5% to about 0.6%, the phenoxyethanol is at a concentration of about 0.5%, the propyl paraben is at a concentration of about 0.03%, glycerylcocoate is at a concentration of about 1%, the titanium dioxide is at a concentration of about 0.5%, the carbopolaqua SF-1 is at a concentration ranging from about 1% to about 6%, the petrolatum is at a concentration of about 1%, the sodium C14-16 olefin sulfonate is at a concentration of about 35%, the sodium lauryl ether sulphate is at a concentration of about 2%, the polyquaternium-39 is at a concentration of about 1%, the ammonium lauryl sulphate (30%) is at a concentration of about 30%, the disodium laureth sulfosuccinate (39%) is at a concentration of about 2%, the sorbitan stearate is at a concentration of about 1.4%, the isotretinoin is at a concentration ranging from about 0.025% to about 0.7%, the tretinoin is at a concentration ranging from about 0.025% to about 0.1%, sodium lauryl sulphate is at a concentration of about 5%, and the tween 80 is at a concentration of about 8%.
In a preferred embodiment, the present disclosure provides a besifloxacin formulation for treatment of gram-negative folliculitis or inflammation thereof, comprising: about 0.5 to about 4 (% w/w) besifloxacin.HCl (Equivalent to besifloxacin); about 2 to about 7 (% w/w) diethylene glycol monoethyl ether; about 0.1 (% w/w) Edetate disodium dihydrate (EDTA); about 2 to about 10 (% w/w) glycerin; about 0.9 to about 1.75 (% w/w) hydroxyethyl cellulose; 0 to about 0.8 (% w/w) carbomer; about 0.3 to about 0.7 (% w/w) phenoxyethanol; about 2 to about 7 (% w/w) polyethylene glycol 400; 0 to about 0.5 (% w/w) sodium hyaluronate; sodium hydroxide; and purified water.
In another preferred embodiment, the present disclosure provides a besifloxacin formulation for treatment of gram-negative folliculitis or inflammation thereof, comprising: about 1 to about 4 (% w/w) besifloxacin.HCl (Equivalent to Besifloxacin); about 5 (% w/w) diethylene glycol mono ethyl ether; about 0.1 (% w/w) Edetate disodium dihydrate (EDTA); about 5 (% w/w) glycerin; about 0.5 to about 1.5 (% w/w) hydroxyethyl cellulose; about 0.3 to about 1.2 (% w/w) carbomer; about 0.7 (% w/w) phenoxyethanol; about 5 (% w/w) polyethylene glycol 400; 0 to about 1 (% w/w) sodium hyaluronate; sodium hydroxide; and purified water.
As mentioned previously, each of these formulations comprise besifloxacin at a concentration ranging from about 0.5% to about 4%, thereby making the formulation effective against microorganisms that cause gram-negative folliculitis. The MIC of besifloxacin against such microorganism ranges from about 0.125 μg/ml to about 16 μg/ml. This MIC is better by at least 50% when compared with other conventional antibiotic such as cefotaxime, ampicillin, clindamycin, tetracycline and erythromycin.
Further, the formulations of the present disclosure also showcase enhanced drug release properties. Drug release is an important property of a therapeutic agent and is a prerequisite for its absorption and penetration of drug to the site of action on the skin layer. Demonstration of drug release from a formulation attests to its availability on the hair follicles and upper layer of skin where pathogen responsible for disease resides, thus resulting in its activity against the target disease/condition. The data from the present disclosure suggests that amount of drug released and retained onto the skin from the besifloxacin based formulations of the present disclosure, is sufficient to exert its therapeutic activity.
Due to long time antibiotic treatment, acne patients often show gram-negative bacterial colonization on anterior nares. Usually the gram-negative bacterial population in normal subjects is below 1% of nasal bacterial population but, in the case of gram-negative folliculitis, it increases to more than 3-4%. While ampicillin is choice of antibiotic conventionally used against gram-negative bacteria, due to increasing antibiotic resistance in gram negative bacterial strains towards multiple drugs, cure of disease caused by such resistant organism is difficult. Accordingly, when tested, besifloxacin shows promising results.
Thus, further, and equally importantly, the MIC of besifloxacin is also considerably low against such gram-negative bacteria which are resistant or susceptible to other conventional antibiotic such as cefotaxime, ampicillin, amoxicillin, cephalosporin, clindamycin, tetracycline and erythromycin.
Therefore, all the bacterial species which are known to be involved in gram-negative folliculitis, are susceptible to besifloxacin, irrespective of their response against other conventional antibiotics or quinolones.
The MIC of besifloxacin against gram-negative bacteria selected from Escherichia coli, Klebsiella spp., Pseudomonas spp. Serratia spp., Acinetobacter spp., Enterobacter spp. and Proteus spp., is lower by at least 50% when compared with
MIC of at least one antibiotic selected from cefotaxime, ampicillin, clindamycin, tetracycline and erythromycin.
Further, none of the isolates of skin resident bacteria in acne patients or healthy individuals demonstrate resistance to besifloxacin in micro broth dilution assay. Moreover, as the MIC values are low and within a narrow range, the chance of future resistance generation due to sub optimal drug delivery in skin is also substantially lower.
Additional embodiments and features of the present disclosure will be apparent to one of ordinary skill in art based upon description provided herein. The embodiments herein and the various features and advantageous details thereof are explained with reference to the nonlimiting embodiments in the description. Descriptions of well-known/conventional methods and techniques are omitted so as to not unnecessarily obscure the embodiments herein. The examples used herein are intended merely to facilitate an understanding of ways in which the embodiments herein may be practiced and to further enable those of skill in the art to practice the embodiments herein. Accordingly, the following examples should not be construed as limiting. The examples are illustrative only, and are not intended to limit, in any manner, any of the aspects described herein. The following examples do not in any way limit the invention.
MIC of besifloxacin and other antibiotics molecules were determined by micro broth dilution method as per the Clinical and Laboratory Standards Institute (CLSI) guideline. MIC assay is the determinant of the inherent anti-microbial potency of a particular molecule. For preparing inoculum, bacterial strains were cultured in Brain Heart Infusion Agar (BHIA) at 37° C. for 24 hours. For MIC test, sterile BHI broth (100 μl) was added into all 96 wells and 100 μl of broth containing drug was added to first well (1A to 1H) and serial (double) dilution was carried out for up to 10 wells (column 1 to column 10 of 96 well plate). For bacterial inoculum, bacterial culture turbidity was adjusted to 0.1 OD at 600 nm in UV-Visual spectrophotometer (approximately 1.5×108 cells/ml) and further diluted (100 times with sterile BHI broth). Diluted bacterial suspension (100 μl) was added to each well except sterility control wells (column 12 of 96 well plate). Column 11 of 96 well plate was used as growth control and vehicle control. The plates were incubated at 37° C. for 24 h. The MIC of the test compounds were determined by observing the lowest concentration of test compound that prevented the visual bacterial growth. Interestingly all the bacterial species which are known to be involved in gram negative folliculitis, are susceptible to besifloxacin irrespective of their response against other conventional antibiotics or Quinolones (Table 1).
E. coli MTCC 1687 (wild type)
E. coli ATCC BAA 196 (multi-drug resistant strain)
K. pneumoniae ATCC 13883 (wild type)
P. aeruginosa ATCC 27853 (wild type)
P. aeruginosa CCARM 2204 (multi-drug resistant,
A. haumannii ATCC 19606 (wild type)
A. baumannii CCARM 12020 (multi-drug resistant,
A. baumannii CCARM 12001 (Quinolone
E. aerogenes NCIM 5139 (wild type)
E. aerogenes M2-1 (clinical isolate)
Besifloxacin HCL was tested against ampicillin and cefotaxime (3rd generation cephalosporin) susceptible and resistant gram-negative bacterial strains involved in GNF (gram negative folliculitis) by broth dilution method as per the Clinical & Laboratory Standards Institutes (CLSI) guidelines. MIC assay was performed in 96 well plates by micro broth dilution method, in 96 well plates, all the wells were added with 100 μl of sterilized Muller Hinton broth No: 2 cation broth (HiMedia, India). The initial stock of test compounds (1 mg/ml) were prepared in dimethyl sulfoxide (DMSO) and further dilutions were made in culture broth to get the desirable concentration. Then 100 μl of broth containing test compounds were initially added to the first column's wells and then serial dilution was performed up to well of 10th column. Wells of 11th and 12th column served as a positive and negative control respectively. Finally, 100 μl of inoculum (0.5 McFarland equal bacterial suspension was 100 times diluted in culture broth) was added to all the wells. Plates were incubated at 37° C. for 18-24 hrs. After incubation, MIC of each test compound was determined by observing the lowest concentration of test compound that has no visual growth compared to growth control. Minimum inhibitory concentration of besifloxacin, ampicillin and cefotaxime against gram negative bacterial strains were shown in Table 1. Besifloxacin was very effective against both susceptible (S) and resistant (R) (ampicillin and cefotaxime) gram negative bacterial strains (Table 1B).
Escherichia coli 591
Escherichia coli 592
Klebsiella sp. 512
Klebsiella sp. 498
Pseudomonas sp. 556
Pseudomonas sp. 537
Enterobacter sp. 523
Enterobacter sp. 532
Enterobacter sp. 1
Enterobacter sp. 2
Acinetobacter sp. 488
Escherichia coli
Klebsiella sp.
Pseudomonas sp.
Enterobacter sp.
Acinetobacter sp.
Due to long time antibiotic treatment, acne patients often show gram negative bacterial colonization on anterior nares. Usually the gram-negative bacterial population in normal people is below 1% of nasal bacterial population but, in the case of gram-negative folliculitis, it increases to more than 3-4%. Gram-negative bacteria include Escherichia coli, Klebsiella, Enterobacter, Proteus and Pseudomonas, which were isolated from infective persons. Ampicillin is the antibiotic of choice used for GNF treatment. However, due to increasing antibiotic resistance in gram negative bacterial strains towards multiple drugs, GNF is difficult to cure. Hence besifloxacin has been tested against various gram-negative bacterial strains susceptible and resistant to ampicillin and cefotaxime. As per the break points provided by EUCAST, some of these isolates showed higher MICs against ampicillin and cefotaxime. Besifloxacin was found to be efficacious when tested against these isolates.
Different skin resident bacteria were isolated from acne lesions of moderate to severe acne patients. As expected, P. acnes and S. epidermidis were two most prevalent species. Members of the skin microbiomes were of diverse nature. Most of the genus were gram positive, but few of them, like Klebsiella, were gram negative. Most of them were aerobic, but few, like P. acnes, were anaerobic in nature. Irrespective of these differences and their prevalence, it was important to evaluate the MIC of besifloxacin vis a vis other conventionally used antibiotics in acne (clindamycin, tetracycline, erythromycin) and fluoroquinolones against these species. Even if they are present in low numbers in microbiomes, they can act as reservoirs of resistance against antibiotic of choice which may in the log run affect the prognosis of GNF. Besifloxacin has shown potent antibacterial activity against all of them (Table 2).
P. acnes V3-1
S. epidermidis S1-1
P. avidum S3-1
K. pneumoniae S6-1
Dermatobacter hominis
Kocuria sp. S18-1
Microbacterium sp. S20-2
Blastococcus sp. S21A-2
Staphylococcus
haemolyticus M1-1
Bacillus megaterium M1-2.
Enterobacter aerogenes
Gel formulations containing suspended besifloxacin is prepared using different concentrations of thickening agents as per the compositions shown in Table 3. The formulations had pH of 5.0-6.5.
Gel formulations containing suspended besifloxacin using various thickening agents are prepared as per the compositions shown in Table 4. The formulations had pH of 5.0-6.5.
Gel formulations containing suspended besifloxacin are prepared as per the compositions shown in Table 5. The formulations prepared is off-white to pale yellow in appearance with a pH of 5.0-6.5.
Gel formulations containing suspended besifloxacin are prepared using carbomer 980 as per the compositions shown in Table 6. The formulations prepared is off-white to pale yellow in appearance with a pH of 5.0-6.5. Method of preparation is same as given in example 3.
Cream formulations containing suspended besifloxacin are prepared as per the compositions shown in Table 7. The formulations had pH of 5.0-6.5.
Cream formulations containing suspended actives such as besifloxacin alone and/or combination of besifloxacin and isotretinoin and/or besifloxacin and tretinoin are prepared as per the compositions shown in Table 8. The formulations had pH of 5.0-6.5.
Gel formulations containing soluble besifloxacin is prepared as per compositions shown in Table 9.
Lotion formulations of besifloxacin HCl are prepared as per compositions shown in Table 10.
Foam formulations of besifloxacin HCl are prepared as per compositions shown in Table 11.
Emulgel formulations of besifloxacin are prepared as per compositions shown in Table 12.
Emulgel formulations are prepared by addition of besifloxacin in oil phase as per compositions shown in Table 13.
Emulgel formulations of besifloxacin HCl are prepared by addition of besifloxacin in water phase as per compositions shown in Table 14.
Gel formulations containing besifloxacin and isotretinoin or tretinoin are prepared using different concentrations of thickening agents as per the compositions shown in Table 15. The formulations had pH of 5.0-6.5.
Gel formulations containing suspended besifloxacin was prepared using isotonic agent such as sodium chloride as per the compositions shown in Table 1. The formulations had pH of 5.5-6.5 and other characterization details such as viscosity, assay of besifloxacin and phenoxyethanol are given below (Table 16).
Lotion containing suspended besifloxacin was prepared using isotonic agent such as sodium chloride, as per the compositions shown in Table 2. The formulations had a pH of 5.0-6.5 and other characterization details such as viscosity, assay of besifloxacin and phenoxyethanol are given below (Table 17).
Lotion containing suspended besifloxacin was prepared using homogenization of drug phase, as per the compositions shown in Table 3. The particle size of active phase was found to be 2.3 μm with PDI:0.326. The final formulations had a pH of 5.1 and other characterization details such as viscosity, assay of besifloxacin and phenoxyethanol are given below (Table 18).
Besifloxacin ointment containing suspended besifloxacin was prepared as per the compositions shown in Table 4. The formulations had a pH of 5.5-6.5 and other characterization details such as viscosity, assay of besifloxacin are given below (Table 19).
Besifloxacin spray containing suspended besifloxacin was prepared as per the composition shown in Table 5. The formulations had pH of 5.0-6.5 and other characterization details such as viscosity, is given below (Table 20).
Formulations comprising besifloxacin in combination with adapalene were prepared. Tables 21 to 26 below provide the said formulations:
Gel formulations containing suspended besifloxacin were tested for in-vitro release testing. Briefly, franz diffusion cell assembly was used for in-vitro release testing of besifloxacin formulations. Receptor compartment was filled with citrate buffer pH 4.0 and maintain temperature at 32° C.±1° C. while stirring at 700 rpm. The Polysulfone membrane were fixed between donor and receptor compartment of Franz diffusion cell. The set assembly was left completely undisturbed for 30 min to equilibrate membrane. Then the stirring of the instrument was turned off just before gel application to the membrane. The besifloxacin gel was evenly applied over membrane with spatula to the exposed area fixed on Franz diffusion cell. Then stirring of the Franz diffusion cell was turned on and recorded initial time point. Release of besifloxacin from donor compartment to receptor compartment was monitored at predetermined time points of 0.5, 1, 2, 4 and 6 h. Turned off stirring before sampling. At defined time points, release medium aliquot (1 ml) of the receptor compartment was collected and replaced with fresh release medium, so that the lower surface of the membrane remains in contact with the receptor compartments over the experimental time period. At the end, release profile of besifloxacin from formulation was determined, and observed as in
Besifloxacin topical gel (2%) containing suspended besifloxacin were tested for in-vitro release testing. Release testing procedure was similar to example 21. Release profile is given in
Besifloxacin ointment formulations containing suspended besifloxacin were tested for in-vitro release testing. Release testing procedure was similar to example 21, where release medium, membrane used for study were ethanolic water (20.0%) and Start-M®, respectively. Release profile is given in
Besifloxacin emulgel formulations containing suspended besifloxacin were tested for in-vitro release testing. Release testing procedure was similar to example 21, where release medium, dose of besifloxacin formulation applied and membrane used for study were ethanolic water (20.0%), 25 mg/cm2 and Start-M®, respectively. Release profile is given below in
Besifloxacin cream containing suspended besifloxacin was tested for skin retention study on pig ear skin. Franz diffusion cell assembly was used for ex-vivo skin retention study. The receptor compartment was filled with Phosphate buffer saline with 0.005% albumin, pH 7.4 and maintain temperature at 32° C.±1° C. while stirring at 700 rpm. The processed pig ear skin was equilibrated with receptor medium for 30 minutes and then fixed between donor and receptor compartment of Franz diffusion cell. Then the stirring of the instrument was turned off just before gel application to the membrane. The besifloxacin gel was evenly applied over skin with spatula to the exposed area fixed on Franz diffusion cell. Then stirring of the Franz diffusion cell was turned on and recorded initial time point. At the end of study (5 hrs to 6 hrs), the excess formulation remaining on the surface of pig ear skin was removed with the help of 3 cotton balls (weight of each cotton ball=0.5 g). Drug was extracted from pig ear skin and the extract samples were submitted for HPLC analysis. HPLC results of release samples, skin extracts and cotton ball extract were used for calculation of release profile, skin retention and recovery of drug from cotton ball, respectively. Skin retention results are shown in
Besifloxacin gel containing suspended besifloxacin was tested for skin retention study on pig ear skin. Testing procedure was similar to example 26. Skin retention results are shown in
Besifloxacin gel and cream containing suspended besifloxacin were tested for skin retention study on human abdominal skin. Besifloxacin gel and cream were applied over a 12 hr application period. Heat-separated epidermal membranes from a single female donor were mounted in horizontal Franz-type diffusion cells (exposed surface area 1.33 cm2) over a receptor solution of phosphate buffered saline, pH 7.4 containing 5% ethanol to facilitate solubilisation of any material penetrating the skin. Cells were maintained at 35° C. and receptor solution was stirred throughout. Formulations containing approximately 1.0% besifloxacin w/w and applied at a dose of approximately 10 mg/cm2. The samples of the receptor solution were taken at times up to 12 hrs and extracts from surface swabs, stratum corneum tape strips and residual epidermis were obtained at the end of the experiment at 12 hrs. All samples were analysed by validated LCMS.
The results suggest that negligible amounts of besifloxacin were recovered from receptor solution with formulation. However, after application of cream, significantly greater amounts of besifloxacin were recovered from the viable epidermis and the entire epidermal membrane (Subcutaneous+epidermis) (
Besifloxacin gel containing suspended besifloxacin was tested for skin retention study on human abdominal skin. Testing procedure was similar to example 27. Skin retention results are shown in
Drug release is an important property of a therapeutic agent and is a prerequisite for its absorption and penetration of drug to the site of action on the skin layer. Demonstration of drug release from a formulation attests to its availability on the hair follicles and upper layer of skin where pathogen responsible for disease resides, thus resulting in its activity against the target disease/condition. The above generated data suggests that amount of drug released and retained onto the skin should be sufficient to exert its therapeutic activity.
Besifloxacin ointment (2%) and gel (2%) formulations were tested on E. coli MTCC 1687 (wild type strain) and E. coli ATCC BAA196 (multi-drug resistant strain), and the results are as provided in table 30 below:
E. coli MTCC1687
E. coli BAA196
As seen, both the formulations showed similar ZOI value against susceptible and multi drug resistant E. coli strains, confirming that the formulations of the present disclosure are effective against gram negative organisms.
All patents and other publications identified in the specification and examples are expressly incorporated herein by reference for all purposes. These publications are provided solely for their disclosure prior to the filing date of the present application. Nothing in this regard should be construed as an admission that the inventors are not entitled to antedate such disclosure by virtue of prior invention or for any other reason. All statements as to the date or representation as to the contents of these documents is based on the information available to the applicants and does not constitute any admission as to the correctness of the dates or contents of these documents.
Although preferred embodiments have been depicted and described in detail herein, it will be apparent to those skilled in the relevant art that various modifications, additions, substitutions, and the like can be made without departing from the spirit of the invention and these are therefore considered to be within the scope of the invention as defined in the claims which follow. Further, to the extent not already indicated, it will be understood by those of ordinary skill in the art that any one of the various embodiments herein described and illustrated can be further modified to incorporate features shown in any of the other embodiments disclosed herein.
Additional embodiments and features of the present disclosure will be apparent to one of ordinary skill in art based on the description provided herein. The embodiments herein provide various features and advantageous details thereof in the description. Descriptions of well-known/conventional methods and techniques are omitted so as to not unnecessarily obscure the embodiments herein.
The foregoing description of the specific embodiments will so fully reveal the general nature of the embodiments herein that others can, by applying current knowledge, readily modify and/or adapt for various applications such specific embodiments without departing from the generic concept, and, therefore, such adaptations and modifications should and are intended to be comprehended within the meaning and range of equivalents of the disclosed embodiments. It is to be understood that the phraseology or terminology employed herein is for the purpose of description and not of limitation. Therefore, while the embodiments in this disclosure have been described in terms of preferred embodiments, those skilled in the art will recognize that the embodiments herein can be practiced with modification within the spirit and scope of the embodiments as described herein.
Any discussion of documents, acts, materials, devices, articles and the like that has been included in this specification is solely for the purpose of providing a context for the disclosure. It is not to be taken as an admission that any or all of these matters form a part of the prior art base or were common general knowledge in the field relevant to the disclosure as it existed anywhere before the priority date of this application.
While considerable emphasis has been placed herein on the particular features of this disclosure, it will be appreciated that various modifications can be made, and that many changes can be made in the preferred embodiments without departing from the principles of the disclosure. These and other modifications in the nature of the disclosure or the preferred embodiments will be apparent to those skilled in the art from the disclosure herein, whereby it is to be distinctly understood that the foregoing descriptive matter is to be interpreted merely as illustrative of the disclosure and not as a limitation.
| Number | Date | Country | Kind |
|---|---|---|---|
| 201711033747 | Feb 2018 | IN | national |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/IB2019/051119 | 2/12/2019 | WO | 00 |
