Hypotonic hydrogel formulations for enhanced delivery of active drugs on mucosal surfaces

Abstract

To provide hypotonic hydrogel formulations for enhanced transport of active agents at mucosal surfaces.SOLUTION: Hypotonic gelling vehicles are used as solubilizing agents for drugs and as a means to provide sustained drug delivery to a mucosal tissue. Solubilizing drugs at higher concentrations enhances drug penetration into the tissues of the body, while the hypotonic gelling vehicle further improves distribution of the drug over a larger surface area for increased absorption and sustained release for reduced side effects and longer duration of action. The invention can provide improved compositions for delivery of active agents with greater efficacy and safety to mucosal surfaces that act as barriers to pathogen transport into the mucosa.SELECTED DRAWING: None

Description

Technical Field

[0001] Cross-reference to Related Applications This application claims the benefit of and priority to U.S. Provisional Application No. 62 / 596,578 filed on December 8, 2017 and U.S. Provisional Application No. 62 / 627,559 filed on February 7, 2018, which are incorporated herein by reference in their entireties.

[0002] Statement on Federally Sponsored Research or Development This invention was made with government support under Grant Number RO1DK107806 awarded by the National Institutes of Health of the United States. The government has certain rights in this invention.

[0003] Field of the Invention The present invention generally relates to the field of formulations for enhanced drug delivery, particularly drug delivery at mucosal surfaces.

Background Art

[0004] Background of the Invention Mucosa are membranes that line various cavities of the body, for example, the oral cavity, gastrointestinal tract, uterus, vagina, colon, anal canal, trachea, lungs, bladder and the like. As used herein, skin may be considered a mucosal surface. The mucosa consists of the epithelium itself and also a supporting loose connective tissue called the lamina propria immediately underlying the epithelium. The deeper connective tissue that supports the mucosa is called the submucosa. In the GI tract, there is a thin layer of smooth muscle, the muscularis mucosae, at the boundary between the mucosa and the submucosa.

[0005] The mucosal surfaces of the body are selectively permeable. Mucosal barrier injury (e.g., oral mucositis and gastrointestinal mucositis) is a common complication after cytoreductive cancer therapy and radiation therapy (Sonis et al., Cancer Supplement, 100(9):1995-2023, 2004). Some capsid viruses can spread through mucus as quickly as they would through water, even if they must spread "upstream" via continuously secreted mucus, thereby penetrating the epithelium. These viruses have a mucus mesh sense (mucus (Cone) It has a surface that is smaller than the mesh spacing and does not stick to mucus. RA, Adv. Drug Deliv Rev, 61(2):75-85, 2009). For example, women are disproportionately infected with HIV, partly due to the lack of preventative measures that women can control (Ndesendo et al., AAPS PharmSciTech, 9:505-520, 2008). Easily administered, thoughtful, and effective methods to protect women from vaginal HIV transmission could prevent millions of infections worldwide. However, vaginal folds, or "vaginal mucosal rugae," that adapt to dilation during intercourse and childbirth are typically folded by intraperitoneal pressure, and the surfaces of these folds make it difficult for drugs and drug carriers to access them (Alexander (et al., Sex Transm Dis, 29:655-664, 2004). Even after mimicking sexual intercourse, insufficient distribution to the vaginal folds has been cited as a significant factor in the inability to protect the susceptible vaginal surface from infection. Distribution across the entire susceptible target surface has been found to be crucial for preventing and treating infection. Furthermore, to enhance user accessibility, drugs delivered to the vagina should be retained in the vaginal area at effective concentrations over extended periods.

[0006] Achieving sustained local drug concentrations is difficult due to the high permeability of the vaginal epithelium to low molecules, as well as the fact that soluble drug dosage forms (gels, creams) can be expelled by intraperitoneal pressure and walking. Finally, the drug delivery method must be safe and non-toxic to the vaginal epithelium. Improvements in the distribution, retention, and safety profile of vaginal dosage forms can lead to a substantial increase in efficacy, as well as a reduction in side effects caused by largely ineffective systemic treatments for cervical vaginal infections and diseases (Thigpen T. Cancer J. 9:245-432, 2003; Robinson et al., Obstet Gynecol, 99:777-784, 2002).

[0007] Sustained drug delivery to the mucous membrane surface of the body has the potential to improve the treatment and prevention of many diseases (to name just a few, sexually transmitted infections, inflammatory bowel disease, pneumonia, and degenerative eye conditions). Achieving sustained concentrations of prophylactic or therapeutic drugs using conventional soluble dosage forms remains challenging due to drug degradation, rapid shedding, and rapid systemic absorption. There is still an unmet need for compositions for delivery to mucous membrane surfaces that provide a physical barrier against pathogen entry. There is also an unmet need for compositions for mucosal delivery that provide retention and sustained release of prophylactic, therapeutic, or diagnostic agents on the mucous membrane surface.

[0008] Therefore, an object of the present invention is to provide an improved composition for delivering an active drug with greater efficacy and safety to the mucosal surface, which acts as a barrier against pathogen transport to the mucous membrane.

[0009] A further object of the present invention is to provide an improved composition for delivery to a mucosal surface that enables retention and sustained release of prophylactic, therapeutic, or diagnostic agents on the mucosal surface.

[0010] A further object of the present invention is to provide a method for preparing an improved composition for delivery to the mucosal surface. [Prior art documents] [Non-patent literature]

[0011] [Non-Patent Document 1] Sonis et al., Cancer Supplement, 100(9):1995-2023, 2004 [Non-Patent Document 2] Cone RA, Adv. Drug Deliv Rev, 61(2):75-85, 2009 [Non-Patent Document 3] Ndesendo et al., AAPS PharmSciTech, 9:505-520, 2008 [Non-Patent Document 4] Alexander et al., Sex Transm Dis, 29:655-664, 2004 [Non-Patent Document 5] Thigpen T. Cancer J. 9:245-432, 2003; Robinson et al., Obstet Gynecol, 99:777-784, 2002 [Overview of the project] [Means for solving the problem]

[0012] Summary of the Invention Hypotonic gelling vehicles are used as solubilizers for drugs with low water solubility. This approach is in contrast to using nanoparticles or nanocrystals to improve the solubility / dissolution of hydrophobic drugs. While drug solubilization enhances mucosal permeability, hypotonic gelling vehicles further improve distribution and permeability. Many drugs have minimal water solubility, limiting their delivery to mucosal surfaces.

[0013] The solubilization of drugs in temperature-sensitive gelling vehicles improves the distribution, retention, and permeability of locally delivered drugs when administered in hypotonic solutions. Hypotonic formulations of hydrogel-forming polymers, preferably poloxamers (nonionic triblock copolymers consisting of a central hydrophobic chain of polyoxypropylene, such as poly(propylene oxide), with two hydrophilic chains of polyoxyethylene, such as poly(ethylene oxide), adjacent to each other) have been developed for the enhanced delivery of therapeutic, diagnostic, prophylactic, or other drugs to epithelial tissues, particularly tissues with mucosal coverings. These polymers are administered in hypotonic solutions at concentrations lower than their nominal critical gelling concentration (CGC). Typically, a poloxamer gel administered vaginally or colorectally at its CGC forms a gel "plug" in the lumen. In contrast, the fluid from the poloxamer solution administered at its hypotonic, sub-CGC level is absorbed by the epithelial surface, drawing the poloxamer into the mucus layer, upward against the epithelium, where the fluid is then concentrated sufficiently to gel, thereby enhancing and facilitating drug delivery to epithelial cells. As the poloxamer is concentrated at the tissue / mucosal interface, it mixes with the mucus and gels against the epithelial surface. The endogenous mucin glycopolymer influences the gelling properties of its hypotonic gelling agent, including the concentration of the gelling agent required for gelation and the concentration of pore structure in the resulting gel / mucin mixture. The hypotonic gelling vehicle coats the epithelium, including the folds, after vaginal and colorectal application.

[0014] The studies demonstrate the advantages of these hypotonic polymer gel formulations, particularly in relation to drug delivery via mucosal epithelium. The present invention provides, for example, the following items: (Item 1) A formulation for the delivery of therapeutic agents, prophylactic agents, diagnostic agents, or functional nutritional factors, wherein the formulation is Therapeutic agents, prophylactic agents, functional nutritional factors or diagnostic agents, A gel-forming polymer for application to mucosal tissue or skin, wherein the gel-forming polymer is formulated such that it is at a concentration below the critical gel concentration (CGC) of the polymer under isotonic conditions and at a temperature between room temperature and body temperature (about 25°C to about 37°C), and an excipient for forming a pharmaceutically acceptable hypotonic formulation of said polymer suitable for delivery to a subject in need thereof, A formulation comprising (Item 2) The formulation according to Item 1, which is in a dry form or a liquid form. (Item 3) The formulation according to any one of Items 1 to 2, wherein the gel-forming polymer is a temperature-sensitive gel-forming polymer. (Item 4) The formulation according to Item 3, wherein the temperature-sensitive gel-forming polymer has a lower critical solution temperature of less than 30°C, preferably less than 21°C. (Item 5) The formulation according to any one of Items 1 to 4, wherein the polymer is a poloxamer. (Item 6) The formulation according to any one of Items 1 to 5, wherein the polymer in combination with the excipient forms a gel on a mucosal surface or epithelial surface selected from the group consisting of the oral cavity, pharynx, esophagus, lung, eye, ear, nose, oral cavity, tongue, vagina, cervix, urogenital tract, gastrointestinal tract, anorectum, and the surface of the skin. (Item 7) The formulation according to Item 6, wherein the epithelial surface is on or in the eye. (Item 8) The formulation according to any one of Items 1 to 7, wherein the drug is water-soluble. (Item 9) The formulation according to any one of Items 1 to 7, wherein the drug has insufficient water solubility. (Item 10) The formulation according to any one of Items 1 to 9, wherein the formulation releases the therapeutic agent, prophylactic agent, or diagnostic agent at the epithelial surface over a period of at least 12 hours. (Item 11) The preparation according to item 10, wherein the preparation releases the therapeutic agent, prophylactic agent, or diagnostic agent on the epithelial surface over a period of at least 24 hours. (Item 12) The formulation according to any one of items 1 to 11, wherein the gel-forming polymer is F98 in an aqueous excipient at a concentration of more than 12% to less than 24%. (Item 13) The formulation according to any one of items 1 to 11, wherein the gel-forming polymer is F127 between 10 and 18%. (Item 14) The formulation according to any one of items 1 to 13, wherein the gel-forming polymer forms a uniformly thick layer when administered onto the epithelial surface. (Item 15) A formulation according to any one of items 1 to 14 for administration in the form of a dry powder, gel, or liquid. (Item 16) The formulation described in item 15 is provided in single-dose units or multiple-dose units with respect to administration. (Item 17) The preparation described in any one of items 1 to 16, wherein the drug is a protein or peptide, a small molecule, a sugar or polysaccharide, a lipid, a glycolipid, a glycoprotein, a nucleic acid, its oligomer or polymer, or a small molecule. (Item 18) The aforementioned drug is a preparation described in any one of items 1 to 15, selected from the group consisting of steroids, glaucoma drugs, tyrosine kinase inhibitors, immunosuppressants, antifibrotic drugs, anti-infective agents, hormones, and chemotherapeutic agents. (Item 19) A method for administering a drug to a mucosal surface or epithelial surface, the method comprising the step of administering a formulation described in any of items 1 to 18 to a site of need. (Item 20) The surface is located on or inside the eye, as described in item 19. [Brief explanation of the drawing]

[0015] [Figure 1] Figure 1 shows a scheme for the solubilization of hydrophobic drugs (e.g., cyclosporine A or budesonide) by solvent evaporation.

[0016] [Figure 2] Figure 2 shows a scheme for solubilizing hydrophobic drugs (e.g., moxifloxacin) by bead grinding.

[0017] [Figure 3] Figure 3 shows a scheme for solubilizing hydrophobic drugs (e.g., cyclosporine A or brimonidine) by dialysis.

[0018] [Figure 4] Figure 4 shows a scheme for the solubilization of hydrophobic drugs (e.g., brinzolamide) by solvent evaporation.

[0019] [Figure 5A] Figure 5A is a graph showing the effect of various concentrations of Pluronic F127 (10%, 12%, 15%, and 18%) on the cumulative increase in cyclosporine-induced tear production in healthy rabbit eyes over 12 hours after a single eye drop administration. Figure 5B is a graph showing tear production in healthy rabbit eyes at 10 hours after administration of a 5-day dose for each of the following: gelCsA, drug-free gel vehicle (gelCsA vehicle), Restasis, Restasis vehicle (Endura), or untreated. Figures 5C and 5D are line graphs showing the concentrations of cyclosporine in rat corneal (Figure 5C) and conjunctival (Figure 5D) tissue over an 8-hour period after administration of cyclosporine eye drops formulated as gelCsA (hypotonic), gelCsA (isotonic), gelCsA (conventional), or Restasis. [Figure 5B]Figure 5A is a graph showing the effect of various concentrations of Pluronic F127 (10%, 12%, 15%, and 18%) on the cumulative increase in cyclosporine-induced tear production in healthy rabbit eyes over 12 hours after a single eye drop administration. Figure 5B is a graph showing tear production in healthy rabbit eyes at 10 hours after administration of a 5-day dose for each of the following: gelCsA, drug-free gel vehicle (gelCsA vehicle), Restasis, Restasis vehicle (Endura), or untreated. Figures 5C and 5D are line graphs showing the concentrations of cyclosporine in rat corneal (Figure 5C) and conjunctival (Figure 5D) tissue over an 8-hour period after administration of cyclosporine eye drops formulated as gelCsA (hypotonic), gelCsA (isotonic), gelCsA (conventional), or Restasis. [Figure 5C] Figure 5A is a graph showing the effect of various concentrations of Pluronic F127 (10%, 12%, 15%, and 18%) on the cumulative increase in cyclosporine-induced tear production in healthy rabbit eyes over 12 hours after a single eye drop administration. Figure 5B is a graph showing tear production in healthy rabbit eyes at 10 hours after administration of a 5-day dose for each of the following: gelCsA, drug-free gel vehicle (gelCsA vehicle), Restasis, Restasis vehicle (Endura), or untreated. Figures 5C and 5D are line graphs showing the concentrations of cyclosporine in rat corneal (Figure 5C) and conjunctival (Figure 5D) tissue over an 8-hour period after administration of cyclosporine eye drops formulated as gelCsA (hypotonic), gelCsA (isotonic), gelCsA (conventional), or Restasis. [Figure 5D]Figure 5A is a graph showing the effect of various concentrations of Pluronic F127 (10%, 12%, 15%, and 18%) on the cumulative increase in cyclosporine-induced tear production in healthy rabbit eyes over 12 hours after a single eye drop administration. Figure 5B is a graph showing tear production in healthy rabbit eyes at 10 hours after administration of a 5-day dose for each of the following: gelCsA, drug-free gel vehicle (gelCsA vehicle), Restasis, Restasis vehicle (Endura), or untreated. Figures 5C and 5D are line graphs showing the concentrations of cyclosporine in rat corneal (Figure 5C) and conjunctival (Figure 5D) tissue over an 8-hour period after administration of cyclosporine eye drops formulated as gelCsA (hypotonic), gelCsA (isotonic), gelCsA (conventional), or Restasis.

[0020] [Figure 6A] Figures 6A and 6B are line graphs showing the change in intraocular pressure as a function of time (in units of time) under different treatments. Figure 6A is a graph showing the change in intraocular pressure over an 8-hour period when treated with a low concentration of (12% F127) gelBRZ, formulated as a conventional (18% F127) gelling substance, or as either hypotonic or isotonic. Figure 6B is a graph showing the change in intraocular pressure over an 8-hour period when treated with a hypotonic gelBRZ formulation, the commercially available eye drop AZOPT®, or a gelBRZ vehicle. [Figure 6B] Figures 6A and 6B are line graphs showing the change in intraocular pressure as a function of time (in units of time) under different treatments. Figure 6A is a graph showing the change in intraocular pressure over an 8-hour period when treated with a low concentration of (12% F127) gelBRZ, formulated as a conventional (18% F127) gelling substance, or as either hypotonic or isotonic. Figure 6B is a graph showing the change in intraocular pressure over an 8-hour period when treated with a hypotonic gelBRZ formulation, the commercially available eye drop AZOPT®, or a gelBRZ vehicle.

[0021] [Figure 7A] Figure 7A is a bar graph showing the cumulative change in IOP over 8 hours after administration of 0.15% brimonidine tartrate (gelBT) in 10%, 12%, 15%, and 18% F127. Figure 7B is a line graph showing the change in intraocular pressure over 10 hours in normotensive rabbits treated with hypotonic gelBT formulations, isotonic gelBT (12% F127), or conventional gelBT (18% F127). Figure 7C is a line graph showing the change in intraocular pressure over 10 hours in patients treated with hypotonic gelBT formulations, commercially available 0.15% brimonidine tartrate eye drops (ALPHAGAN®), or a gelBT vehicle. Figures 7D–7F show brimonidine levels in the cornea (Figure 7D), conjunctiva (Figure 7E), and aqueous humor (Figure 7F) of rats 1 hour, 4 hours, and 8 hours after the last dose (administered twice daily for 5 days). Figure 7G is a line graph showing the change in intraocular pressure when treated with mild hypotonic (gelBT 200 mOsm) or hypotonic gelBT. [Figure 7B] Figure 7A is a bar graph showing the cumulative change in IOP over 8 hours after administration of 0.15% brimonidine tartrate (gelBT) in 10%, 12%, 15%, and 18% F127. Figure 7B is a line graph showing the change in intraocular pressure over 10 hours in normotensive rabbits treated with hypotonic gelBT formulations, isotonic gelBT (12% F127), or conventional gelBT (18% F127). Figure 7C is a line graph showing the change in intraocular pressure over 10 hours in patients treated with hypotonic gelBT formulations, commercially available 0.15% brimonidine tartrate eye drops (ALPHAGAN®), or a gelBT vehicle. Figures 7D–7F show brimonidine levels in the cornea (Figure 7D), conjunctiva (Figure 7E), and aqueous humor (Figure 7F) of rats 1 hour, 4 hours, and 8 hours after the last dose (administered twice daily for 5 days). Figure 7G is a line graph showing the change in intraocular pressure when treated with mild hypotonic (gelBT 200 mOsm) or hypotonic gelBT. [Figure 7C]Figure 7A is a bar graph showing the cumulative change in IOP over 8 hours after administration of 0.15% brimonidine tartrate (gelBT) in 10%, 12%, 15%, and 18% F127. Figure 7B is a line graph showing the change in intraocular pressure over 10 hours in normotensive rabbits treated with hypotonic gelBT formulations, isotonic gelBT (12% F127), or conventional gelBT (18% F127). Figure 7C is a line graph showing the change in intraocular pressure over 10 hours in patients treated with hypotonic gelBT formulations, commercially available 0.15% brimonidine tartrate eye drops (ALPHAGAN®), or a gelBT vehicle. Figures 7D–7F show brimonidine levels in the cornea (Figure 7D), conjunctiva (Figure 7E), and aqueous humor (Figure 7F) of rats 1 hour, 4 hours, and 8 hours after the last dose (administered twice daily for 5 days). Figure 7G is a line graph showing the change in intraocular pressure when treated with mild hypotonic (gelBT 200 mOsm) or hypotonic gelBT. [Figure 7D] Figure 7A is a bar graph showing the cumulative change in IOP over 8 hours after administration of 0.15% brimonidine tartrate (gelBT) in 10%, 12%, 15%, and 18% F127. Figure 7B is a line graph showing the change in intraocular pressure over 10 hours in normotensive rabbits treated with hypotonic gelBT formulations, isotonic gelBT (12% F127), or conventional gelBT (18% F127). Figure 7C is a line graph showing the change in intraocular pressure over 10 hours in patients treated with hypotonic gelBT formulations, commercially available 0.15% brimonidine tartrate eye drops (ALPHAGAN®), or a gelBT vehicle. Figures 7D–7F show brimonidine levels in the cornea (Figure 7D), conjunctiva (Figure 7E), and aqueous humor (Figure 7F) of rats 1 hour, 4 hours, and 8 hours after the last dose (administered twice daily for 5 days). Figure 7G is a line graph showing the change in intraocular pressure when treated with mild hypotonic (gelBT 200 mOsm) or hypotonic gelBT. [Figure 7E]Figure 7A is a bar graph showing the cumulative change in IOP over 8 hours after administration of 0.15% brimonidine tartrate (gelBT) in 10%, 12%, 15%, and 18% F127. Figure 7B is a line graph showing the change in intraocular pressure over 10 hours in normotensive rabbits treated with hypotonic gelBT formulations, isotonic gelBT (12% F127), or conventional gelBT (18% F127). Figure 7C is a line graph showing the change in intraocular pressure over 10 hours in patients treated with hypotonic gelBT formulations, commercially available 0.15% brimonidine tartrate eye drops (ALPHAGAN®), or a gelBT vehicle. Figures 7D–7F show brimonidine levels in the cornea (Figure 7D), conjunctiva (Figure 7E), and aqueous humor (Figure 7F) of rats 1 hour, 4 hours, and 8 hours after the last dose (administered twice daily for 5 days). Figure 7G is a line graph showing the change in intraocular pressure when treated with mild hypotonic (gelBT 200 mOsm) or hypotonic gelBT. [Figure 7F] Figure 7A is a bar graph showing the cumulative change in IOP over 8 hours after administration of 0.15% brimonidine tartrate (gelBT) in 10%, 12%, 15%, and 18% F127. Figure 7B is a line graph showing the change in intraocular pressure over 10 hours in normotensive rabbits treated with hypotonic gelBT formulations, isotonic gelBT (12% F127), or conventional gelBT (18% F127). Figure 7C is a line graph showing the change in intraocular pressure over 10 hours in patients treated with hypotonic gelBT formulations, commercially available 0.15% brimonidine tartrate eye drops (ALPHAGAN®), or a gelBT vehicle. Figures 7D–7F show brimonidine levels in the cornea (Figure 7D), conjunctiva (Figure 7E), and aqueous humor (Figure 7F) of rats 1 hour, 4 hours, and 8 hours after the last dose (administered twice daily for 5 days). Figure 7G is a line graph showing the change in intraocular pressure when treated with mild hypotonic (gelBT 200 mOsm) or hypotonic gelBT. [Figure 7G]Figure 7A is a bar graph showing the cumulative change in IOP over 8 hours after administration of 0.15% brimonidine tartrate (gelBT) in 10%, 12%, 15%, and 18% F127. Figure 7B is a line graph showing the change in intraocular pressure over 10 hours in normotensive rabbits treated with hypotonic gelBT formulations, isotonic gelBT (12% F127), or conventional gelBT (18% F127). Figure 7C is a line graph showing the change in intraocular pressure over 10 hours in patients treated with hypotonic gelBT formulations, commercially available 0.15% brimonidine tartrate eye drops (ALPHAGAN®), or a gelBT vehicle. Figures 7D–7F show brimonidine levels in the cornea (Figure 7D), conjunctiva (Figure 7E), and aqueous humor (Figure 7F) of rats 1 hour, 4 hours, and 8 hours after the last dose (administered twice daily for 5 days). Figure 7G is a line graph showing the change in intraocular pressure when treated with mild hypotonic (gelBT 200 mOsm) or hypotonic gelBT.

[0022] [Figure 8A] Figure 8A is a bar graph showing the viscosity under low shear / no shear conditions for standard lubricating eye drops (GENTEAL®), 16% F127, and 16% F127 containing 0.5% HPMC. Figure 8B shows the shear viscosity reduction characteristics (decrease in viscosity when placed under shear) for GENTEAL®, 16% F127, and 16% F127 containing 0.5% HPMC. [Figure 8B] Figure 8A is a bar graph showing the viscosity under low shear / no shear conditions for standard lubricating eye drops (GENTEAL®), 16% F127, and 16% F127 containing 0.5% HPMC. Figure 8B shows the shear viscosity reduction characteristics (decrease in viscosity when placed under shear) for GENTEAL®, 16% F127, and 16% F127 containing 0.5% HPMC.

[0023] [Figure 9A]Figures 9A and 9B are histograms showing the concentrations of sunitinib or N-desethylsunitinib in various eye tissues and fluids of Dutch belted rabbits (Figure 13A) after daily topical administration of sunitinib malate (gelSUN) in a hypotonic gel vehicle for 14 days, and in domestic pigs after daily topical administration of sunitinib malate (gelSUN) in a hypotonic gel vehicle for 5 days. [Figure 9B] Figures 9A and 9B are histograms showing the concentrations of sunitinib or N-desethylsunitinib in various eye tissues and fluids of Dutch belted rabbits (Figure 13A) after daily topical administration of sunitinib malate (gelSUN) in a hypotonic gel vehicle for 14 days, and in domestic pigs after daily topical administration of sunitinib malate (gelSUN) in a hypotonic gel vehicle for 5 days. [Modes for carrying out the invention]

[0024] Detailed description of the invention I. Definition As used herein, “pharmaceutically acceptable” means those compounds, substances, compositions, and / or dosage forms suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, in proportion to a reasonable benefit / risk ratio, within the bounds of reasonable medical judgment.

[0025] As used herein, "biocompatible" and "biologically compatible" generally refer to a substance that, along with any metabolites or degradation products of the substance, is generally non-toxic to the recipient and does not cause any significant adverse effects to the recipient. Generally speaking, a biocompatible substance is one that, when administered to an individual, does not induce any significant inflammatory, immune, or toxic response.

[0026] As used herein, the terms "gel" and "hydrogel" refer to a swollen, water-containing network of finely dispersed, water-insoluble polymer chains, where the polymer molecules are external or in a dispersed phase, and water (or an aqueous solution) is internal or forms the dispersed phase. The chains may be chemically crosslinked (chemical gel) or physically crosslinked (physical gel). A chemical gel has polymer chains linked through covalent bonds, while a physical gel has polymer chains linked by non-covalent bonds or cohesive forces (e.g., van der Waals interactions, ionic interactions, hydrogen bonds, or hydrophobic interactions).

[0027] The polymer chain is typically hydrophilic or contains hydrophilic polymer blocks. The term "gel-forming polymer" is used to describe any biocompatible polymer (including homopolymers, copolymers, and combinations thereof) that can form a physical hydrogel in an aqueous medium when present at or above a critical gel concentration (CGC).

[0028] When used herein, “critical gel concentration” or “CGC” refers to the minimum concentration of a gel-forming polymer required for gel formation, such as the point at which a gel (sol-gel) transition occurs from solution. This critical gel concentration may depend on many factors, including the specific polymer composition, molecular weight, temperature, and / or the presence of other polymers or excipients.

[0029] The term "thermosensitive gel-forming polymer" refers to a gel-forming polymer that exhibits one or more changes in properties with respect to temperature. For example, some thermosensitive gel-forming polymers are water-soluble below a certain temperature but become water-insoluble as the temperature rises. The term "lower critical solution temperature (LCST)" refers to a temperature below which the gel-forming polymer and solvent are completely miscible and form a single phase. For example, "LCST of a polymer solution" means that the polymer is uniformly dispersed in the solution at that temperature (i.e., the LCST) or lower, but aggregates to form a second phase when the solution temperature rises above the LCST.

[0030] As used herein, "hydrophilic" refers to a molecule that has a greater affinity for and, therefore greater solubility in, water compared to an organic solvent. The hydrophilicity of a compound can be quantified by measuring its partition coefficient between water (or a buffered aqueous solution) and a water-immiscible organic solvent (e.g., octanol, ethyl acetate, methylene chloride, or methyl tert-butyl ether). After equilibration, if a greater concentration of the compound is present in water than in its organic solvent, the compound is considered hydrophilic.

[0031] As used herein, "hydrophobic" refers to a molecule that has a greater affinity for and, therefore greater solubility, to an organic solvent compared to water. The hydrophobicity of a compound can be quantified by measuring its partition coefficient between water (or a buffered aqueous solution) and a water-immiscible organic solvent (e.g., octanol, ethyl acetate, methylene chloride, or methyl tert-butyl ether). After equilibration, if a greater concentration of the compound is present in the organic solvent than in water, the compound is considered hydrophobic.

[0032] As used herein, the term “treating” includes inhibiting, alleviating, preventing or eliminating one or more symptoms or side effects associated with the disease, condition, or disorder being treated.

[0033] The terms “reduce,” “inhibit,” “alleviate,” or “decrease” may be used in relation to a control, no other treatment, or a treatment with a known degree of effectiveness. Those skilled in the art will readily identify appropriate controls for each experiment. For example, a reduced response in subjects or cells treated with a compound is compared to the response in subjects or cells not treated with that compound.

[0034] As used herein, the terms “effective amount” or “therapeutically effective amount” mean a dose sufficient to treat, inhibit, or alleviate one or more symptoms of the disease condition being treated, or to otherwise provide a desired pharmacological and / or physiological effect. The exact dose will vary depending on various factors such as subject-dependent variables (e.g., age, immune system health), the disease or disorder, and the treatment being administered. The effect of the effective amount may be relative to a control, which is known in the art and considered herein, and may be, for example, the subject’s condition before or in the absence of administration of a drug or drug combination, or, in the case of a drug combination, the effect of the combination may be compared to the effect of administration of one of the drugs alone.

[0035] The term "excipient" is used herein to include compounds that are not therapeutically or biologically active. Therefore, excipients should be pharmaceutically or biologically acceptable or relevant, and, for example, should generally be non-toxic to their subject. "Excipient" may include a single such compound, or it may include multiple such compounds.

[0036] The term "osmolarity," as used herein, generally refers to the total number of components dissolved per liter. Osmolarity is similar to molar concentration but includes the total number of moles of a species dissolved in a solution. An osmolarity of 1 Osm / L means that there are 1 mole of components dissolved per liter of solution. Some solutes (e.g., ionic solutes that dissolve in solution) give more than 1 mole of components dissolved per mole of solute in that solution. For example, NaCl in solution contains Na + and Cl - It dissociates into two components, thus providing 2 moles of dissolved components per mole of dissolved NaCl in solution. The physiological volumetric osmolality is typically in the range of approximately 280 to 310 mOsm / L.

[0037] The term "tonicity," as commonly used herein, refers to the osmotic gradient resulting from the separation of two solutions by a semipermeable membrane. In particular, tonicity is used to describe the osmotic pressure created across the cell membrane when a cell is exposed to an external solution. Solutes that can cross the cell membrane do not contribute to the final osmotic gradient. Only their dissolved species that do not cross the cell membrane contribute to the osmotic difference and, therefore, tonicity. The term "hypertonic," as commonly used herein, refers to a solution with a higher solute concentration present inside the cell. When a cell is immersed in a hypertonic solution, water tends to flow out of the cell to equilibrium with the concentration of its solute. The term "hypotonic," as commonly used herein, refers to a solution with a lower solute concentration present inside the cell. When a cell is immersed in a hypotonic solution, water flows into the cell to equilibrium with the concentration of its solute. The term "isotonic," as commonly used herein, refers to a solution in which the osmotic gradient across its cell membrane is essentially equilibrium. Isotonic preparations have essentially the same osmotic pressure as human blood. Isotonic preparations generally have an osmotic pressure of approximately 250–350 mOsm.

[0038] II. Hypotonic Gel-Forming Compositions Hypotonic formulations of hydrogel-forming polymers, preferably containing poloxamer, have been developed for enhanced mucosal delivery of therapeutic, diagnostic, prophylactic, or other drugs to epithelial tissue. These polymers are administered at concentrations lower than their normal critical gelling concentration (CGC). Poloxamer gels administered vaginally or colorectally at concentrations equal to or above their CGC form a gel "plug" in the lumen. In contrast, fluids from a hypotonic poloxamer solution, when the poloxamer is at a concentration below its CGC, are absorbed into the mucosal tissue, thereby pulling the poloxamer through the mucosal gel toward the epithelium, thereby enhancing and facilitating drug delivery to the body. As water is absorbed into the tissue, the poloxamer concentrates and gels near the surface of the epithelial tissue, thereby trapping the drug molecules in a sustained-release gel on the tissue surface (rather than a gel that primarily forms in the lumen, as occurs, for example, in conventional thermogelling methods, where the gelling polymer is administered at or above their CGC concentration). Endogenous mucin glycopolymers influence the gelling properties of the hypotonic gelling agent (including the concentration of the gelling agent required for gelation and the pore structure of the resulting gel / mucin mixture). After vaginal, colorectal, or ocular application, the hypotonic gelling vehicle coats the epithelium (including its folds or the inside of the eyelids).

[0039] This example demonstrates longer vaginal retention of model drugs administered with hypotonic gelling agents compared to a bolus of gel formed in the central part of the vaginal lumen, as observed with the gelling agent administered via CGC. In the case of ocular administration, the hypotonic gelling agent forms a uniform gel coating on the surface of the eye rather than the gelling agent in a bolus, which is rapidly removed by blinking.

[0040] A gel-forming composition is described herein that can form a uniform gel coating on an epithelial surface but does not gel under storage conditions. The gel-forming composition comprises one or more gel-forming polymers in a hypotonic carrier, optionally comprising one or more further excipients and / or one or more therapeutic, prophylactic, or diagnostic agents.

[0041] A. Hydrogel-forming polymers The hypotonic gel-forming composition comprises one or more gel-forming polymers. The gel-forming polymers are used at concentrations below their usual critical gel concentration (CGC), for example, at a concentration at which the polymer solution gels in a test tube when heated to 37°C.

[0042] A temperature-sensitive (also known as thermally responsive) hydrogel is a solution that undergoes a sol-gel transition when both of the following criteria are met: 1) At or above the critical gelling concentration (CGC), and 2) At or above the critical gelation temperature.

[0043] Temperature-sensitive gelling agents used in biomedical applications (at or above their CGC) are liquids at room temperature but form gels at body temperature. The increase in temperature induces rearrangement and alignment of their polymer chains, resulting in gelation into a three-dimensional structure. This phenomenon generally depends on the hydrophilic-to-hydrophobic ratio of their polymer chains. Common features include the presence of hydrophobic methyl, ethyl, or propyl groups. Temperature-sensitive polymers meeting these criteria can be topically administered to mucosal tissue in hypotonic solutions at concentrations below their CGC to form a uniform gel coating in vivo.

[0044] Examples of temperature-sensitive gel-forming materials that can be used include: polyoxyethylene-polyoxypropylene-polyoxyethylene triblock copolymer (e.g., designated poloxamer 407 by CTFA (CAS 9003-11-6)). Poloxamers include, but are not limited to, those with a molecular weight of 9,840–14,600 g / mol, representing approximately 70% polyoxyethylene by weight (commercially available from BASF as LUTROL® F127) and poloxamer 188 (CAS 9003-11-6, molecular weight 7680–9510 g / mol, representing approximately 80% polyoxyethylene by weight (commercially available from BASF as LUTROL® F68); poloxamers are also known by the trade name PLURONIC®, for example, PLURONIC® F98 (CAS 9003-11-6, molecular weight 13,000 g / mol, representing approximately 80% polyoxyethylene by weight (commercially available from BASF)). Tetronics tetrafunctional block copolymers based on ethylene oxide and propylene oxide, marketed by BASF as TETRONIC®; poly(N,N-diethylacrylamide); poly(N,N-dimethylacrylamide); poly(N-vinylcaprolactam); poly(N-alkylacrylamide); poly(N-vinylalkylamide); poly(N-isopropylacrylamide); polyethylene oxide methacrylate polymer; poly(lactic acid-co-glycolic acid) (PLGA)-polyethylene glycol triblock copolymer (PLGA-PEG-PLGA and PEG-PLGA-PEG); polycaprolactone (PCL)-polyethylene glycol triblock copolymer (PCL-PEG-PCL and PEG-PCL-PEG); chitosan; and combinations thereof.

[0045] The hydrogel can be formed from individual gel-forming materials or as a combination of gel-forming materials. For example, poloxamers and other gel-forming materials (e.g., tetronic polymers) can be used in combination to obtain desired properties. Furthermore, various forms of the same gel-forming material (e.g., poloxamer 188 and poloxamer 407) can be combined to obtain desired properties.

[0046] The polymer is supplied at a concentration lower than that of an aqueous solution that would form a gel in a test tube when heated to 37°C. The concentration must be sufficiently high, but below the CGC, so that the epithelium absorbs enough fluid to achieve its CGC in vivo, resulting in gelation preferentially occurring on / near its mucosal epithelial surface. The time range for gelation depends on the mucosal surface (water absorption capacity and rate), the tonicity of the administered solution (the more hypotonic the solution, the faster the fluid absorption), and the concentration of the administered polymer (if the polymer concentration is too low, there will be insufficient fluid absorption to concentrate the polymer into its CGC). However, gelation generally occurs within one hour in the vagina and colorectal.

[0047] The concentration of the polymer and the presence of further components (e.g., endogenous mucin) affect the scope, speed, and degree of gelation. An 18% F127 gel (1:1 ratio) mixed with purified porcine gastric mucin (1%) or human cervical-vaginal mucus did not capture virus-sized nanoparticles (polyethylene glycol-coated polystyrene nanoparticles, PSPEG) as effectively as the 18% F127 gel alone. In contrast, the 24% F98 gel, when mixed with mucin or human cervical-vaginal mucin, captured PSPEG particles more effectively. However, in vivo virus capture with hypotonic gelling agents was more effective in capturing viruses, including human immunodeficiency virus (HIV, approximately 120 nm) and herpes simplex virus (HSV, approximately 180 nm). Administration of a hypotonic solution containing 18% F98 and 24% CGC resulted in effective capture of subsequently administered HIV in the vagina. Similarly, hypotonic solutions containing 10% and 15% F127 and 18% CGC were effective in reducing HIV MSD, indicating capture. Furthermore, both 15% F127 and 18% F98 reduced the diffusion of HSV subsequently administered into mouse vaginal mucus. The distribution of individual viral MSDs on a 1-second timescale illustrated that the capture of the virus (shift to the left) was more uniform in the gel formed by the hypotonic 18% F98 vehicle compared to 15% F127. In further testing of viral capture by hypotonic gelling agents in the colorectal, 12% F98 (24% CGC) did not effectively capture PSPEG nanoparticles administered 30 minutes after its gelling vehicle, while 18% F98 was found to be effective in capturing PSPEG nanoparticles in the mouse colorectal. Importantly, these examples illustrate that there are differences in the gels formed when the hypotonic gelling agent is administered to different mucosal surfaces (in this case, when mixed with vaginal mucus compared to colorectal mucus before gelation).

[0048] B. Hypotonic carriers The gel-forming composition contains a hypotonic carrier. This hypotonic carrier is typically, preferably, a biocompatible carrier that causes little to no signs of irritation when administered to a human subject. The carrier may be naturally occurring or non-natural, and may include both synthetic and semi-synthetic carriers. A preferred carrier is water-based. Other solutions (including sugar-based solutions (e.g., glucose, mannitol)) and various buffers (phosphate buffers, Tris buffers, HEPES) may also be used.

[0049] When a hypotonic solution is applied to the epithelial surface, a fluid shift occurs, and water moves into the epithelial tissue. This can cause swelling of epithelial cells. In some cases, if the osmotic pressure difference is too large, epithelial cells may burst, causing tissue irritation or destruction of the epithelial membrane.

[0050] Hypotonic solutions refer to solutions that cause water absorption by the epithelial surface to which they are administered. Examples of hypotonic solutions include, but are not limited to, tris[hydroxymethyl]-aminomethane hydrochloride (Tris-HCl, 10-100 mM, pH 6-8), 4-(2-hydroxyethyl)-1-piperazine ethanesulfonic acid (HEPES, 10-100 mM, pH 6-8), and diluted PBS (e.g., a solution containing 0.2 g KCl, 0.2 g KH2PO4, 8 g NaCl, and 2.16 g Na2HPO4*7H2O in 1000 ml H2O).

[0051] A hypotonic carrier causes the dissolved gel-forming polymer to concentrate on the surface of the epithelium, resulting in uniform gel formation on that surface. This hypotonic carrier typically contains water as its primary component. While the hypotonic carrier can be water, a mixture of water and water-miscible organic solvents may also be used. Suitable water-miscible organic solvents include alcohols (e.g., ethanol, isopropanol); ketones (e.g., acetone); ethers (e.g., dioxane); and esters (e.g., ethyl acetate).

[0052] The hypotonic carrier may be distilled water containing one or more volumetric osmolality-modifying excipients. Sodium chloride is the most frequently used excipient to adjust the volumetric osmolality when the solution is hypotonic. Other excipients used to adjust hypotonic solutions include glucose, mannitol, glycerol, propylene glycol, and sodium sulfate. Possible volumetric osmolality-modifying excipients are pharmaceutically acceptable salts (e.g., sodium chloride, sodium sulfate, potassium chloride, and other salts that make up buffers, such as sodium phosphate dibasic, potassium phosphate monobasic, calcium chloride, and magnesium sulfate). Other excipients used to adjust tonicity include glucose, mannitol, glycerol, or propylene glycol.

[0053] The hypotonic carrier may have any volumetric osmolality lower than the effective isotonic point (the concentration at which the fluid is neither absorbed nor secreted by the underlying tissue) at its mucosal surface. This isotonic point varies for different mucosal surfaces and different buffers, depending on the active ion transport at its epithelial surface; for example, the inventors found that the isotonic point in the vagina for a sodium-based solution is approximately 300 mOsm / L, while in the colorectal it is approximately 450 mOsm / L. In some embodiments, the solution has a tonicity of 50 mOsm / L to 280 mOsm / L, 100 mOsm / L to 280 mOsm / L, 150 mOsm / L to 250 mOsm / L, 200 mOsm / L to 250 mOsm / L, 220 mOsm / L to 250 mOsm / L, 220 mOsm / L to 260 mOsm / L, 220 mOsm / L to 270 mOsm / L, or 220 mOsm / L to 280 mOsm / L.

[0054] The hypotonic carrier may include one or more pharmaceutically acceptable acids, one or more pharmaceutically acceptable bases, or salts thereof. Examples of pharmaceutically acceptable acids include hydrobromic acid, hydrochloric acid, and sulfuric acid, as well as organic acids (e.g., methanesulfonic acid, tartaric acid, and malic acid). Examples of pharmaceutically acceptable bases include hydroxides of alkali metals (e.g., sodium or potassium) and alkaline earth metals (e.g., calcium or magnesium), as well as organic bases (e.g., pharmaceutically acceptable amines). The hypotonic carrier may also include a pharmaceutically acceptable buffer (e.g., citrate buffer or phosphate buffer).

[0055] C. Further medications The hypotonic gel-forming composition may contain one or more agents to be delivered. Examples include therapeutic agents, prophylactic agents, diagnostic agents, and / or functional nutritional factors. Biologically active agents are substances used for treatment (e.g., therapeutic agents), prevention (e.g., prophylactic agents), diagnosis (e.g., diagnostic agents), or to bring about the cure or alleviation of a disease or illness, or to qualitatively alter the structure or function of the body, or prodrugs that are biologically active or become more active after being placed in a given physiological environment. These may be small molecule drugs (e.g., molecular weights of less than 2000, less than 1500, less than 1000, less than 750, or less than 500 atomic mass units (amu)), peptides or proteins, sugars or polysaccharides, nucleotides or oligonucleotides (e.g., aptamers, siRNA, and miRNA), lipids, glycoproteins, lipoproteins, or combinations thereof.

[0056] The drug may contain one or more of those listed in Martindale: The Complete Drug Reference, 37th edition (Pharmaceutical Press, London, 2011).

[0057] In one embodiment, the drug to be delivered is poorly soluble in water but soluble in a carrier containing its gelling polymer. In another embodiment, the drug is water-soluble. The data also show that benefits can be obtained with water-soluble drugs (e.g., brimonidine tartrate) that are soluble down to approximately 1 mg / mL.

[0058] The hypotonic gel-forming formulation may contain a therapeutically effective amount of a therapeutic agent to treat, inhibit, or alleviate one or more symptoms of the disease condition being treated. The hypotonic gel-forming composition may contain a therapeutically effective amount of a prophylactic agent to prevent one or more symptoms of a disease or disorder.

[0059] The drugs may be anti-infective agents (antibiotics, antivirals, antifungals) for the treatment of eye disorders (glaucoma, dry eye), anti-inflammatory agents (inhibiting angiogenesis and fibrosis), contraceptives, agents for the treatment of metabolic disorders, agents for the treatment of heartburn or ulcers, agents for the treatment of cardiovascular disorders (e.g., hypertension and atherosclerosis), neuroactive agents, or chemotherapeutic agents for the treatment of diseases such as cancer.

[0060] Examples of drugs include brinzolamide, cyclosporine A, brimonidine tartrate, moxifloxacin, budesonide, sunitinib, and acriflavin.

[0061] Examples of useful proteins include hormones (e.g., insulin, growth hormone including somatomedin, and reproductive hormones). Examples of useful drugs include neurotransmitters (e.g., L-DOPA), antihypertensives or saluretics (e.g., Metolazone from Searle Pharmaceuticals), carbonic anhydrase inhibitors (e.g., Acetazolamide from Lederle Pharmaceuticals), insulin-like drugs (e.g., glybride), sulfonylurea class hypoglycemic agents, and synthetic hormones (e.g., Android F from Brown Pharmaceuticals and TESTRED® (methyltestosterone) from ICN Pharmaceuticals). Representative antiproliferative (anti-cancer or endometriotic) drugs include, but are not limited to, the following: alkylating agents (e.g., cisplatin, carboplatin, oxaliplatin, mechloretamine, cyclophosphamide, chlorambucil, dacarbazine, lomustine, carmustine, procarbazine, chlorambucil, and ifosfamide), antimetabolites (e.g., fluorouracil (5-FU), gemcitabine, methotrexate, cytosine arabinoside, fludarabine, and phloxuridine), antimitotic agents (taxanes (e.g., paclitaxel and docetaxel) and vinca alkaloids (e.g., vincristine) Examples include vinblastine, vinorelbine, and vindesine), anthracyclines (examples include doxorubicin, daunorubicin, barurubicin, idarubicin, and epirubicin), as well as actinomycin (e.g., actinomycin D), cytotoxic antibiotics (examples include mitomycin, plicamycin, and bleomycin), topoisomerase inhibitors (examples include camptothecin (e.g., camptothecin, irinotecan, and topotecan)), as well as derivatives of epipodophyllotoxin (e.g., amsacrin, etoposide, etoposide phosphate, and teniposide), and combinations thereof.Other suitable anticancer agents include: angiogenic inhibitors (antibodies against vascular endothelial growth factor (VEGF) (e.g., bevacizumab (AVASTIN®), other anti-VEGF compounds); thalidomide (THALOMID®) and its derivatives (e.g., lenalidomide (REVLIMID®)); endostatins; angiostatins; receptor tyrosine kinase (RTK) inhibitors (e.g., sunitinib (SUTEN)); T(registered trademark)); tyrosine kinase inhibitors (e.g., sorafenib (NEXAVAR(registered trademark)), erlotinib (TARCEVA(registered trademark)), pazopanib, axitinib, and lapatinib); inhibitors of transforming growth factor-α or transforming growth factor-β, and antibodies against epidermal growth factor receptors (e.g., panituzumab (VECTIBIX(registered trademark)) and cetuximab (ERBITUX(registered trademark))).

[0062] Regarding imaging, radioactive materials (for example, technetium-99) 99m Tc)) or magnetic materials (e.g., labeled Fe2O3) may be used. Examples of other materials include radiopaque compounds.

[0063] III. Methods and Preparations of Hypotonic Gel-Forming Compositions Drug solubilization offers potential benefits including enhanced physical stability during storage, increased drug penetration into the body, and more reproducible drug doses when administered to patients.

[0064] Drugs insoluble in water are particularly difficult to deliver to mucosal surfaces (e.g., mucosal surfaces of the eyes, gastrointestinal tract, female reproductive organs, and respiratory tract) due to their lack of absorption. Their formulations are particularly suitable for delivering therapeutic agents that are poorly water-soluble. Water-soluble drugs are also difficult to deliver to mucosal surfaces in a sustained manner. Therefore, their gelling agents can also be used as vehicles to improve the mucosal absorption of water-soluble drugs, for example, by providing more sustained drug absorption that can reduce side effects and offer longer-lasting efficacy.

[0065] Furthermore, the mucosal surface is rapidly cleansed and regenerated as a normal defense mechanism against infection and foreign particles. Improved drug solubilization can improve mucosal permeability, and improved distribution, permeability, and retention of hydrophobic drugs are promising strategies for improving therapeutic efficacy. The following examples demonstrate the solubilization of various drugs and drug conjugates with low water solubility into substances that also possess temperature-sensitive gelling properties.

[0066] Figures 1-4 show schemes for achieving direct drug solubilization of hydrophobic drugs and drug complexes into gelling materials.

[0067] Figure 1 shows a scheme for the solubilization of hydrophobic drugs (e.g., cyclosporine A or budesonide) by solvent evaporation.

[0068] Figure 2 shows a scheme for solubilizing hydrophobic drugs (e.g., moxifloxacin) by bead grinding.

[0069] Figure 3 shows a scheme for solubilizing hydrophobic drugs (e.g., cyclosporine A or brimonidine) by dialysis.

[0070] Figure 4 shows a scheme for the solubilization of hydrophobic drugs (e.g., brinzolamide) by solvent evaporation.

[0071] The formulation may be prepared as described in the following example.

[0072] The formulation may be prepared as a liquid for administration. The gel-forming liquid or polymer solubilizes insoluble drugs by forming micelles. The powder may be produced by lyophilization and reconstituted at the time of use.

[0073] The formulation may also contain pharmaceutically acceptable diluents, preservatives, solubilizers, stabilizers, emulsifiers, auxiliary substances, and / or carriers. Stabilizers (e.g., SPAN® 20 (sorbitan laurate, CAS number 1338-39-2)) may promote dissolution and prevent re-aggregation. Other exemplary stabilizers include polysorbates or TWEENS®, e.g., polysorbate 20, polysorbate 60, polysorbate 65, and polysorbate 80, as well as polyglycerol esters (PGE), polyoxyethylene alkyl ethers, polyoxyl stearate, fatty acids (e.g., oleic acid), and propylene glycol monostearate (PGMS). In some cases, the composition may contain one or more stabilizers.

[0074] Increased viscosity without shear can result in increased residence time on the mucous membrane surface of the eye, while shear reduction during blinking maximizes comfort and lubrication. Therefore, gelling materials having these properties can have favorable lubrication properties on the surface of the eye while resisting removal during blinking. In some examples, one or more polymeric materials are incorporated into the gelling vehicle to increase viscosity without shear and / or shear reduction during blinking. In one example, hydroxypropyl methylcellulose (HPMC) is incorporated into the F127 solution to achieve these desirable properties. In some examples, shear-reducing polymers are also included in the composition in a concentration range in which they exhibit conventional shear-reducing properties. Exemplary shear-reducing polymers include cellulose derivatives (e.g., methylcellulose, carboxymethylcellulose, hydroxypropylcellulose, methylcellulose, etc.).

[0075] Drug formulations are typically prepared as single or multiple liquid or dry dose units in a suitable applicator. Those skilled in the art are aware of the many options for drug storage and application (e.g., dual-chamber devices that can be used to keep various components separated during storage). Multiple dosage forms typically involve a cylinder loaded with powder and a plunger therein having a dose increment. These are typically sterilized and packaged in sealed, sterile packages for storage and distribution. See also Remington: The Science and Practice of Pharmacy, 22nd edition.

[0076] The dose-unit dispenser (administrator) may be designed to fit the anatomical location where the drug is to be delivered (e.g., rectally, vaginally, nasally, or orally).

[0077] IV. Method for administering a hypotonic gel-forming composition The hypotonic gel-forming composition can, in principle, be applied to any water-absorbing surface (including skin and mucous membrane tissue) to form a gel. Preferably, the formulation can be applied as a liquid to mucosal coverings on the epithelial surface of a subject requiring therapeutic, preventive, diagnostic, or nutritional effects. The gel-forming composition can be applied in any number of ways known to those skilled in the art, insofar as the hypotonic solution, or the reagent that forms the hypotonic solution, comes into contact with the surface.

[0078] By applying the gel-forming composition as a hypotonic formulation, water is absorbed into the epithelial tissue. This water absorption provides a concentration of the gel-forming polymer on its surface, resulting in uniform gel formation. The gel can act as a barrier, reservoir, or a combination thereof. The agents or excipients in the gel-forming composition may be trapped within the gel and released on or to its surface.

[0079] Epithelial surfaces include the oral cavity, pharynx, esophagus, lungs, eyes, ears, nose, inside of the mouth, tongue, vagina, cervix, genitourinary tract, digestive tract, anorectal surface, and / or skin surface.

[0080] In some cases, the hypotonic gel-forming composition maintains an effective concentration of one or more active agents on one or more mucosal surfaces for extended periods, such as over 6 hours, over 12 hours, over 1 day, over 2 days, over 3 days, over 4 days, over 5 days, over 6 days, or over 1 week.

[0081] In some cases, the hypotonic gel-forming composition increases the concentration of one or more active agents at or near its application site by a factor of 2, 3, 4, 5, 6, 7, 8, 9, 10, or more than 10 compared to the active agent delivered in, for example, physiological saline solution, without a gel-forming vehicle. When the hypotonic gel-forming composition is applied to the eye, the mucous membrane sites with increased concentrations of the active agent include one or more of the cornea, aqueous humor, sclera, conjunctiva, iris, lens, retina, and retinal pigment epithelium.

[0082] In some cases, the hypotonic gel-forming composition delivers active agents (e.g., acriflavin and sunitinib malate) to the retina and / or choroid at 10%, 20%, 30%, 40%, 50%, or more than 50% of an amount effective to reduce neovascularization of the retina and / or choroid, compared to the active agent delivered in, for example, physiological saline solution, without a gel-forming vehicle.

[0083] In other examples, the hypotonic gel-forming composition increases the survival of retinal ganglion cells after optic nerve injury and / or delivers an active neuroprotective agent (e.g., sunitinib malate) to the retina in an amount effective to double, triple, quadruple, quintuple, or more than quintuple the expression of γ-synuclein and / or βIII tubulin in retinal ganglion cells after optic nerve injury compared to the active agent delivered in, for example, physiological saline solution, without a gel-forming vehicle.

[0084] In a further example, the hypotonic gel-forming composition delivers an active drug (e.g., brinzolamide) to the eye in an amount effective to lower intraocular pressure (IOP) by 10%, 20%, 30%, 40%, 50%, or more than 50% in a range of less than 2 hours, less than 4 hours, less than 6 hours, less than 8 hours, less than 10 hours, less than 12 hours, or less than 24 hours, compared to the active drug delivered in, for example, physiological saline solution, without a gel-forming vehicle.

[0085] In other examples, the hypotonic gel-forming composition delivers an active agent (e.g., cyclosporine A) to the eye in an amount effective to increase tear production by 10%, 20%, 30%, 40%, 50%, or more than 50% in the range of less than 2 hours, less than 4 hours, less than 6 hours, less than 8 hours, less than 10 hours, less than 12 hours, or less than 24 hours, compared to the active agent delivered in, for example, physiological saline solution, without a gel-forming vehicle.

[0086] The present invention can be further understood by referring to the following non-limiting embodiments. [Examples]

[0087] Example 1: Cyclosporine A eye drops Rapid removal of drugs from the ocular surface is a major obstacle to local drug delivery, and as a result, many eye drops are prescribed for application several times a day.

[0088] material and method The first example embodies the solubilization of cyclosporine A (used for dry eye) (gelCsA) in a temperature-sensitive gel vehicle prepared using the method schematically shown in Figure 3. Cyclosporine A and Pluronic F127 (10-18% w / w) were dissolved together in DMSO and dialyzed for 2 days against water equilibrated with salt (sodium chloride, final osmolality 200 mOsm) to remove the DMSO. The CsA concentration in the total preparation was 0.05%. Tear production was measured using a Schirmer test piece held in contact with the ocular surface. Drug concentration was measured using LC-MS.

[0089] result The literature has reported that cyclosporine increases tear production in the eyes of healthy animals. Figure 5A shows the cumulative increase in cyclosporine-induced tear production over 12 hours in the eyes of healthy rabbits after a single instillation of cyclosporine A formulated in various concentrations of Pluronic F127 (including 10%, 12%, 15%, and 18% w / w). The hypotonic 12% F127 vehicle of cyclosporine provided the greatest increase in tear production.

[0090] Figure 5B is a graph of tear production in healthy rabbit eyes 10 hours after each of five daily doses: gelCsA, gel vehicle (no drug), RESTASIS®, RESTASIS® vehicle (Refresh Endura), or no treatment. Only with gelCsA was tear production attributable to cyclosporine delivery observed 10 hours after administration of the eye drops. This further suggests the possibility of administering gelCsA once daily compared to RESTASIS®, which is prescribed twice daily.

[0091] Figures 5C and 5D show cyclosporine levels in corneal and conjunctival tissue of rats after administration of various cyclosporine eye drops. This demonstrates that its hypotonic gelCsA provides superior cyclosporine delivery compared to isotonic and conventional gel vehicles, as well as RESTASIS®.

[0092] Example 2: Brinzolamide eye drops for the treatment of glaucoma material and method Brinzolamide is used to lower intraocular pressure (IOP) as a treatment for glaucoma. Brinzolamide was formulated in its gel vehicle (gelBRZ) using the method shown in Figure 4. Pluronic F127 was present in concentrations ranging from 10 to 18%, and salt was added to adjust the osmolality by weight until isotonic (300 mOsm / kg) was achieved, if necessary. The effects of the vehicle itself (gelBRZ vehicle) and brinzolamide eye drops on IOP in normotensive rabbits were tested after administering a single 50 μL dose of eye drops at 10 mg / mL.

[0093] result Figure 6A is a graph showing gelBRZ formulated as either a conventional (18% F127) gelling agent or a low-concentration (12% F127) formulation that is either hypotonic or isotonic. Figure 6B shows that the gelBRZ vehicle was ineffective against IOP in the treated eye or the contralateral eye in rabbits with normal blood pressure. This further compares the hypotonic gelBRZ formulation with the commercially available eye drop AZOPT® (1% brinzolamide clinical ophthalmic suspension). One dose of 1% brinzolamide is compared to gelBRZ (12%, When delivered as hypotonic gel BRZ (35 mOsm), it had a more pronounced effect on IOP reduction up to 8 hours after treatment. Furthermore, hypotonic gel BRZ (35 mOsm) reduced IOP more significantly than isotonic gel BRZ (300 mOsm) at an early stage of 4 hours.

[0094] Example 3: Brimonidine tartrate ophthalmic preparation material and method Brimonidine tartrate is a water-soluble drug that can be directly dissolved in a gel vehicle (gelBT). We first tested whether there was an optimal F127 concentration for ocular administration of brimonidine tartrate in a hypotonic gel vehicle, as measured by reduction of IOP in normotensive rabbits. Next, we compared IOP reduction for F127 concentrations ranging from 12–18% in both hypotonic (without salt) and isotonic (adjusted to 300 mOsm / kg with salt) solutions in normotensive rabbits. Drug concentrations were measured by LC-MS. result

[0095] Figure 7A shows the cumulative change in IOP over 8 hours after administration of 0.15% brimonidine tartrate (gelBT) in F127 (10-18%). In normotensive rabbits, the cumulative reduction in IOP over 8 hours after a single instillation of hypotonic gelBT was highest with respect to gelBT containing 12% F127. Figure 7B shows that hypotonic gelBT was more effective in reducing IOP over an 8-hour period in normotensive rabbits compared to isotonic (12% F127) and conventional (18% F127) eye drops. Figure 7C shows that the gelBT vehicle had no effect on IOP, and that hypotonic gelBT was more effective than commercially available Alphagan eye drops in reducing IOP over 8 hours. Figures D-F show BT levels in the cornea, conjunctiva, and aqueous humor of rats 1, 4, and 8 hours after the final dose (2x administration per day over 5 days). Drug levels were higher with hypotonic gelBT than with all other formulations. Figure 7G shows that even mild hypotonic gelBT (200 mOsm) was effective in lowering IOP.

[0096] Example 4: Incorporation of polymer material into a gelling vehicle material and method The polymer substance was incorporated into its gelling vehicle, hydroxypropyl methylcellulose (HPMC), into an F127 solution (due to its shear-thinning properties), which may be advantageous for in-vitro (ocular administration) or lubrication (rectal administration). The polymer substance may also increase the viscosity of the gel at rest (under no shear) and improve its retention time on the mucosal surface.

[0097] result As shown in Figure 8A, the addition of 0.5% HPMC to F127 increases the viscosity under low shear / no shear conditions, which is already several thousand times higher than that of a standard lubricating eye drop (GENTEAL®). Figure 8B shows that the shear-thinning properties (decrease in viscosity when placed under shear) are similar for the F127 solution containing HPMC compared to a standard lubricating eye drop (GENTEAL®). Thus, the addition of hydroxypropyl methylcellulose (HPMC) to the F127 solution increases the viscosity under low shear / no shear conditions while maintaining the low viscosity observed in a standard lubricating eye drop (e.g., GENTEAL®) under high shear conditions (similar to the shear rate during blinking).

[0098] The increased viscosity without shearing can result in a longer residence time on the mucous membrane surface of the eye, while shearing viscosity reduction during blinking maximizes comfort and lubrication. Therefore, such gelling materials can have desirable lubrication properties on the surface of the eye while resisting removal during blinking. Examples of such materials include shearing viscosity-reducing polymers in the conventional concentration range that exhibit shearing viscosity-reducing properties, such as cellulose derivatives (e.g., methylcellulose, carboxymethylcellulose, hydroxypropylcellulose, methylcellulose, etc.).

[0099] Example 5: No eye irritation in the presence of hypotonic gelling vehicles with various gravimetric osmolal concentrations. New Zealand white rabbits were given 50 μL of the test drug topically twice daily for 5 weeks to evaluate potential eye irritation. The test drug contained a 12% F127 solution with sodium chloride to produce final osmolality by weight ranging from no added salt to 300 mOsm / kg. The negative control was a group of animals that received equilibrated salt solution (BSS) and no treatment. There were no differences in corneal staining (lisamine green 1%) or blink rate (number of blinks in 3 minutes) for any formulation compared to untreated animals.

[0100] Example 6: Drug delivery to the retina / choroid via local administration C57 / B6 mice were used to test the local delivery of sunitinib malate (5 μL at 4 mg / mL) and acriflavin (5 μL at 5 mg / mL) to prevent choroidal angiogenesis (CNV) in mice after laser-induced rupture of Bruch's membrane at three sites. Both drugs, dissolved in either 12% F127 or an aqueous vehicle control (saline or water), were administered daily for 7 days after laser treatment. Only formulations containing F127 were effective in suppressing angiogenesis in the choroid.

[0101] Example 7: Long-term melanin-binding therapeutic effect of sunitinib administered locally in a hypotonic gel vehicle after optic nerve injury. Rats were topically administered sunitinib malate (5 μL at 4 mg / mL) in 12% F127 daily to one eye for 5 days. The optic nerve heads were then exposed, one eye was crushed, and the contralateral eye was used as a control for comparison. Fourteen days after nerve crushing (without further topical administration), the tissue was isolated for qPCR for genes expressed by retinal ganglion cells (RGCs). This administration scheme resulted in protection of RGCs in pigmented rats (Brown Norway) and non-pigmented rats (Wistar). This suggests that binding to melanin in the eye contributed to the long-term therapeutic effect.

[0102] Example 8: Treatment of optic nerve injury using a hypotonic gel vehicle Sunitinib malate in 12% F127 or saline was administered topically to one eye once a week in pigmented rats (Brown Norway) (5 μL at 4 mg / mL). The optic nerve head was exposed, and one eye was crushed on day 8, with the contralateral eye used as a control for comparison. Fourteen days after nerve crushing (day 22), the tissue was isolated for qPCR for genes expressed by retinal ganglion cells (RGCs). RGC protection was observed only when sunitinib malate was administered in 12% F127, and not when sunitinib malate was administered in saline.

[0103] Example 9: Distribution of therapeutic agent to ocular tissue delivered in a hypotonic gel vehicle material and method Sunitinib malate was dissolved in 12% F127 at a concentration of 4 mg / ml and administered topically once daily to pigmented rabbits (Dutch breed) and pigmented pigs (young domestic pigs) for a total of 14 days or 5 days, respectively. Administration to both rabbits and pigs was performed without anesthesia, with 50 μL or 50–100 μL of eye drops administered to each. However, administration to pigs was performed while they were distracted with food. Tissue was collected 6 hours after the last dose for rabbits and 1 hour after the last dose for pigs. The drug concentration was measured by LC-MS.

[0104] result Sunitinib malate was administered topically to Dutch rabbits (Figure 9A) daily for 14 days and to domestic pigs (Figure 9B) daily for 5 days in a hypotonic gel vehicle. Tissues were isolated, and sunitinib and its main metabolite (N-desethylsunitinib) levels were quantified in various ocular tissues and fluids. Therapeutically relevant levels of the drug were found in the anterior and posterior segments of the eye.

[0105] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art to which the disclosed invention pertains. Publications cited herein and materials from which they are cited are specifically incorporated herein by reference.

[0106] Those skilled in the art can recognize or confirm many equivalents to specific embodiments of the inventions described herein simply by using conventional experimental methods. Such equivalents are intended to be encompassed by the following claims.

Claims

1. A formulation for the delivery of a therapeutic agent, prophylactic agent, diagnostic agent, or functional nutritional factor, wherein the formulation maintains an effective concentration of the therapeutic agent, prophylactic agent, diagnostic agent, or functional nutritional factor on the epithelial surface or mucous membrane surface inside or over the eye for treating, preventing, or diagnosing an eye disease, and the formulation is Therapeutic agents, prophylactic agents, functional nutritional factors or diagnostic agents, A gel-forming polymer for application to the eye, wherein the gel-forming polymer is formulated such that it is at a concentration below the critical gel concentration (CGC) of the polymer under isotonic conditions and at a temperature between 25°C and 37°C, and Excipients for forming a pharmaceutically acceptable hypotonic formulation of the polymer suitable for delivery to the eyes of individuals in need, Includes, Here, the polymer is poloxamer, the poloxamer is in a concentration of more than 10% to 15%, and the formulation is hypotonic. The formulation is characterized by being administered to the epithelial surface or mucous membrane surface inside or over the eye, the therapeutic agent, prophylactic agent, diagnostic agent or functional nutritional factor being retained inside or over the eye for a period longer than one week, and the formulation is effective in delivering the therapeutic agent, prophylactic agent, diagnostic agent or functional nutritional factor to the posterior segment of the eye. formulation.

2. The formulation according to claim 1, wherein the gel-forming polymer is a temperature-sensitive gel-forming polymer.

3. The formulation according to claim 2, wherein the temperature-sensitive gel-forming polymer has a lower critical solution temperature of less than 30°C.

4. The formulation according to claim 3, wherein the temperature-sensitive gel-forming polymer has a lower critical solution temperature of less than 21°C.

5. The formulation according to claim 1, wherein the polymer, in combination with an excipient, forms a gel on the mucous membrane surface or epithelial surface in or over the eye.

6. The formulation according to claim 1, wherein the therapeutic agent, preventive agent, diagnostic agent, or functional nutritional factor is water-soluble.

7. The formulation according to claim 1, wherein the therapeutic agent, prophylactic agent, diagnostic agent, or functional nutritional factor is insufficiently water-soluble.

8. The formulation according to claim 1, wherein the gel-forming polymer is PLURONIC® F98 in an aqueous excipient in an amount between 12% and 15%.

9. The formulation according to claim 1, wherein the gel-forming polymer is approximately 12% PLURONIC® F127.

10. The formulation according to claim 1, wherein the gel-forming polymer forms a uniformly thick layer when administered onto the epithelial surface.

11. The formulation according to claim 1, wherein the formulation is provided in single-dose units or multiple-dose units with respect to administration.

12. The formulation according to any one of claims 1 to 11, wherein the therapeutic agent, prophylactic agent, diagnostic agent, or functional nutritional factor is a protein or peptide, sugar or polysaccharide, lipid, glycolipid, glycoprotein, nucleic acid, its oligomer or polymer, or low molecular weight.

13. The preparation according to any one of claims 1 to 11, wherein the therapeutic agent, prophylactic agent, diagnostic agent, or functional nutritional factor is selected from the group consisting of steroids, glaucoma agents, tyrosine kinase inhibitors, immunosuppressants, antifibrotic agents, anti-infective agents, hormones, and chemotherapeutic agents.

Images