High hyaluronate multi-use disinfectant solution for ophthalmic applications

Abstract

The present invention relates to an ophthalmic formulation that utilizes a synergistic combination of three different classes of disinfectants (a multimeric biguanide compound, a quaternary ammonium compound, and an antifungal / anti-acanthamoeba compound, specifically a combination of polyhexamethylene biguanide, polyquaternium-1, and myristamidopropyl dimethylamine) to allow for the use of higher concentrations of hyaluronate to provide better comfort and support for corneal health without compromising antimicrobial efficacy. The novel ophthalmic formulation supports the use of hyaluronate concentrations 7.5 times higher than those currently present in commercially available multi-use disinfecting solutions.

Description

[Technical Field]

[0001] FIELD OF THE INVENTION The present invention relates to a novel multi-use disinfecting solution for use in the eye. [Background technology]

[0002] background Multi-use solutions for cleaning and disinfecting contact lenses (called multi-use disinfecting solutions (MPDS)) are convenient because they allow lenses to be cleaned in one operation: neutralization of the cleaning solution is not required, and the lenses do not need to be rinsed, as must be done with, for example, hydrogen peroxide-based products. MPDSs must thoroughly clean and effectively disinfect contact lenses. Furthermore, they should provide comfort during contact lens wear. The aspect of comfort has received particular attention in recent years as changes in the working environment (e.g., prolonged use of screens and smartphones, exposure to air conditioning, etc.) have increasingly led to problems with dry and irritated eyes. These symptoms are often exacerbated in contact lens wearers. This condition, known as "Contact Lens-Associated Dry Eye," can cause patients to discontinue contact lens use (Brafman and Eiden. Review of Cornea & Contact Lenses, JAN / FEB 2012, pp. 18-21). Hyaluronic acid (HA) is frequently used as a rewetting and comfort agent in ophthalmic formulations and compositions. HA is a naturally occurring polymer composed of glucuronic acid and N-acetyl-D-glucosamine repeating units. HA typically has a high molecular weight (up to several million daltons) in mammals and is present in the extracellular matrix, epithelium, nerves, and connective tissues, as well as the vitreous of the eye (Stern et al. Eur. J. Cell Biol. (2006) 85: 699-715; Saranraj et al. Int. J. Pharma. Biol. Sci. Arch. (2013) 4: 853-859). Over the past few decades, several positive aspects of HA supplementation in cosmetics and especially ophthalmic products have been discovered.Thanks to its high capacity to absorb water (Holban & Grumezescu. Elsevier(2016). ISBN: 9780323477222), HA increases the wettability and lubrication of the corneal surface, positively influencing the stability of the tear film, resulting in a reduction in the rate of tear film evaporation and an increase in tear breakup time (Zheng et al. Cornea.(2013) 32:1260-1264; Invest. Ophthalmol. Vis. Sci.(2014) 55: 3454-60; Saeed et al. Pak. J. Med. Sci.(2013) 29: 1055-1058; You et al. J. Ocul. Pharmacol. Th.(2018) 34: 557-564; Oh et al. J. Ocul. Pharmacol. Th. (2014) 30: 533-542). These aspects are primarily responsible for the "sensation" of comfort and reduction of conditions such as dry eye. Furthermore, HA can function as a radical scavenger and prevent damage caused by reactive oxygen species (ROS). HA is known to promote corneal cell migration and corneal re-epithelialization and is active in various wound healing processes. Furthermore, the polymer has been reported to protect corneal epithelial cells from damage caused by detergents such as sodium lauryl sulfate and surfactants such as benzalkonium chloride (Wu et al. J. Ophthalmol. (2017) Article ID 3678586; Nishida et al. (1991) Exp. Eye Res. 53: 753-758; Saranraj et al. (2013) Int. J. Pharm. Biol. Sci. Arch. 4: 853-859; Carlson et al. (2018) J. Ocul. Pharmacol. Th. 34: 360-364). However, some of the physicochemical properties responsible for the latter positive effect also interfere with the use of HA in MPDS. While increased effective concentrations of HA can be used, for example, in preservative-free artificial tear solutions, the same concentrations inhibit the antimicrobial effects of disinfectants commonly used in MPDS (e.g., quaternary ammonium compounds).This inhibition occurs because HA is an anionic polymer that can complex with cationic antiseptics, causing their aggregation and inactivation (U.S. Patent Nos. 8,119,112, 5,858,346, and 5,559,104). In principle, the inhibitory effect of HA could be compensated for by increasing the concentration of the antiseptic. However, such compensation may not be easily achieved because antiseptics are cytotoxic at high concentrations (Pellinen et al. Curr. Eye Res. (2012) 37: 145-154; Paimela et al. Molecular Vision (2012) 18: 1189-1196). Therefore, it is not surprising that the concentration of HA in commercially available MPDS typically does not exceed about 0.01% (w / v). Because the comfort effect is predicted to increase with concentration, the development of ophthalmic solutions containing higher concentrations of HA should be an important goal (You et al. J. Ocul. Pharmacol. Th. (2018) 34: 557-564). The notion that higher HA concentrations are beneficial is further supported by studies by Torretta et al. (Int. J. Immunopathol. Pharmacol. (2016) 29: 438-442) and Romano et al. (J. Bone Jt. Infect. (2017) 2: 63-72). The latter study provided evidence that biofilm formation by certain bacterial strains does not occur at elevated HA concentrations. It is noted that U.S. Patent Nos. 8,119,112 and 8,664,180 suggest that HA concentrations greater than 0.01% w / v can be included in MPDS. However, U.S. Pat. No. 8,664,180 does not explicitly teach such compositions, and U.S. Pat. No. 8,119,112 describes MPDSs with elevated concentrations of HA, but only provides experimental evidence of adequate antimicrobial activity for compositions containing 0.01% w / v HA. The present disclosure relates to MPDSs that support higher concentrations of HA than those present in commercially available MPDSs, without compromising antimicrobial activity or cleaning effectiveness. [Prior art documents] [Patent documents]

[0003] [Patent Document 1] U.S. Patent No. 8,119,112 [Patent Document 2] U.S. Patent No. 8,664,180 [Non-patent literature]

[0004] [Non-Patent Document 1] Brafman and Eiden. Review of Cornea & Contact Lenses, JAN / FEB 2012, pp.18-21 [Non-patent document 2] Stern et al. Eur. J. Cell Biol. (2006) 85: 699-715 [Non-patent document 3] Saranraj et al. Int. J. Pharma. Biol. Sci. Arch. (2013) 4: 853-859 Summary of the Invention [Means for solving the problem]

[0005] Summary of the Invention The present invention relates to an ophthalmic formulation in which a synergistic combination of three different classes of antiseptics (multimeric biguanide compounds, quaternary ammonium compounds, and antifungal / anti-acanthamoeba compounds), specifically polyhexamethylene biguanide, polyquaternium-1, and myristamidopropyl dimethylamine, allows for the use of high concentrations of hyaluronic acid to provide better comfort and support for corneal health without compromising antimicrobial efficacy. The novel ophthalmic formulation of the present disclosure supports the use of hyaluronic acid at a concentration 7.5 times higher than that present in currently commercially available MPDSs. The solutions of the present invention may also preferably be referred to as ophthalmic preparations of the present invention or multi-use disinfecting solutions (MPDS) of the present invention.

[0006] Accordingly, the present disclosure relates to an ophthalmic MPDS comprising a multimeric biguanide compound, a quaternary ammonium compound, an antifungal / anti-acanthamoebic agent, and hyaluronic acid or a salt of hyaluronic acid. The multimeric biguanide compound may be present in an amount of about 0.00005 to about 0.0005% weight / volume (w / v), the quaternary ammonium compound in an amount of about 0.00005 to about 0.0025% w / v, and the antifungal / anti-acanthamoebic agent in an amount of about 0.00005 to about 0.0025% w / v. The antifungal / anti-acanthamoebic agent may be an amido-amine compound (e.g., myristamidopropyldimethylamine (MAPD) ​​or chlorhexidine sulfonate). MAPD is preferred. The hyaluronic acid or hyaluronic acid salt is present in an amount greater than 0.01% w / v but less than about 0.1% w / v. Preferred concentrations of the components of the MPDS of the present disclosure range from about 0.000075 to about 0.00025% for multimeric biguanide compounds, about 0.00005 to about 0.002% w / v for quaternary ammonium compounds, about 0.00005 to about 0.002% w / v for antifungal / anti-acanthamoebic agents, and about 0.0151% w / v to about 0.09% w / v for hyaluronic acid or hyaluronate salts. More preferred concentrations are about 0.0001 to about 0.0002% w / v, about 0.000075 to about 0.001% w / v, about 0.0001 to about 0.0018% w / v, and about 0.025 to about 0.085% w / v for the multimeric biguanide compound, quaternary ammonium compound, antifungal / anti-acanthamoebic agent, and HA, respectively. Most preferred concentrations are 0.00012 to 0.00016% w / v, 0.000085 to 0.00012% w / v, 0.0006 to 0.0014% w / v, and 0.04 to 0.08% w / v, respectively.

[0007] An amido-amine compound, as understood herein, is preferably defined as an amide of a fatty acid and a diamine compound formed from one fatty acid molecule and one diamine molecule. The amido-amine compound of the present invention preferably comprises an amide group -C(O)NH- and an amino group -NR2, where each R is independently hydrogen or an optionally substituted hydrocarbon. Fatty acids, as understood herein, preferably refer to alkanoic acids, optionally containing one or more C=C bonds and optionally substituted, in particular, with an -OH group. Preferably, the fatty acids have 2 to 30 carbon atoms. More preferably, the fatty acids have 4 to 28 carbon atoms. A diamine molecule, as defined herein, preferably comprises two amino groups, one of which may optionally be substituted with one or more C1-C4 alkyl and alkylene moieties having 1 to 10 carbon atoms.

[0008] The multimeric biguanide compound is selected from the group consisting of polyhexamethylene biguanide (PHMB), polyaminopropyl biguanide (PAPB), and 1,1'-hexamethylene-bis(5-[2-ethylhexyl]biguanide), and the quaternary ammonium compound is selected from the group consisting of polyquaternium-1 (PQ-1), polyquaternium-10 (PQ-10), polyquaternium-42, N-dodecyl{4,4'-(2,4,8,10-tetraoxaspiro[5.5]undecane-3,9-diyl)}dipyridinium dibromide, 3,3' The most preferred biguanide compound is PHMB, and the most preferred quaternary ammonium compound is PQ-1. Polyhexamethylene biguanide (PHMB) is preferably polyhexamethylene biguanide (PQ-1). The most preferred quaternary ammonium compound is PHMB, and the most preferred quaternary ammonium compound is PQ-1. Polyhexamethylene biguanide (PHMB) is preferably poly(hexamethylene biguanide). The most preferred quaternary ammonium compound is PHMB, and the most preferred quaternary ammonium compound is PQ-1. The most preferred quaternary ammonium compound is PHMB ... More preferably, each compound as described herein is understood to be that compound or a pharmaceutically acceptable salt thereof. Thus, for example, the term polyhexamethylene biguanide (PHMB) is preferably understood herein as polyhexamethylene biguanide (PHMB) or a pharmaceutical salt thereof.

[0009] The multimeric biguanide compounds, as understood herein, are preferably defined as compounds containing more than one biguanide moiety, -NH-C(=NH)-NH-C(=NH)-NH-, and more preferably, they are defined as compounds containing a repeating unit, which contains a biguanide moiety, -NH-C(=NH)-NH-C(=NH)-NH-. Multimeric biguanide compounds as described herein may also refer to pharmaceutically acceptable salts of the compounds (as defined herein).

[0010] Preferably, any compound described herein can also exist as a pharmaceutically acceptable salt. "Pharmaceutically acceptable salt" is preferably defined as a derivative of the described compound, wherein the parent compound is modified by making its acid salt or base salt. Examples of pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic residues (e.g., amines); alkali salts or organic salts of acidic residues (e.g., carboxylic acids); etc. The above-mentioned pharmaceutically acceptable salts include, for example, the conventional non-toxic salts or quaternary ammonium salts of the parent compound formed from non-toxic inorganic or organic acids. For example, such conventional non-toxic salts include those derived from inorganic acids (e.g., but not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, nitric acid, and the like); and salts prepared from organic acids (e.g., but not limited to, acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pamoic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, sulfanilic acid, 2-acetoxybenzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid, isethionic acid, and the like). The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound which contains a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or base form of these compounds with a stoichiometric amount of suitable base or acid in water or organic solvent, or in the mixture of the two.Organic solvents include but are not limited to non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile.A list of suitable salts can be found in Remington's Pharmaceutical Sciences, 18th edition, Mack Publishing Company, Easton, PA, 1990, p. 1445 (the disclosure of which is incorporated herein by reference).The MPDS of the present disclosure includes one or more, and more typically most or all, of the following additional components:

[0011] A chelating agent at a concentration of about 0.01-0.06% w / v, a buffer or buffering agent at a concentration of about 0.01-1% w / v, a viscosity adjusting agent at a concentration of about 0.01-0.25% w / v, a surface active compound at a concentration of about 0.01-1% w / v, a tonicity agent at a concentration of about 0.001-1% w / v, and a calcium and / or magnesium salt (e.g., MgCl2 and / or CaSO4) each at a concentration of about 0.001-0.015% w / v. Preferably, the MPDS of the present disclosure includes one or more, more typically most or all of the following additional components:

[0012] The composition contains a chelating agent at a concentration of about 0.01-0.06% w / v, a buffer or buffering agent at a concentration of about 0.01-1% w / v, a viscosity adjusting agent at a concentration of about 0.01-0.25% w / v, a surface-active compound at a concentration of about 0.01-1% w / v, an isotonicity adjusting agent at a concentration of about 0.001-1% w / v, and optionally a calcium and / or magnesium salt (e.g., MgCl2 and / or CaSO4) at a concentration of about 0.001-0.015% w / v, respectively. Most preferred is ethylenediaminetetraacetic acid (EDTA) as the chelating agent, borate buffer as the buffering agent, hydroxypropylmethylcellulose (HPMC) as the viscosity adjusting agent (more preferred compounds are carboxymethylcellulose (CMC) and hydroxyethylcellulose), poloxamine as the surfactant, and sodium chloride or a combination of sodium chloride and potassium chloride as the isotonicity adjusting agent.

[0013] Preferably, the multi-use disinfecting solutions of the present invention may further comprise one or more of a chelating agent in an amount ranging from about 0.01% w / v to about 0.06% w / v, a buffer in an amount ranging from about 0.01% w / v to about 1% w / v, a viscosity adjusting agent in an amount ranging from about 0.01% w / v to about 0.25% w / v, a surfactant compound in an amount ranging from about 0.01% w / v to about 1% w / v, a tonicity agent in an amount ranging from about 0.001% w / v to about 1% w / v, a calcium salt in an amount ranging from about 0.001% w / v to about 0.015% w / v, and a magnesium salt in an amount ranging from about 0.001% w / v to about 0.015% w / v. DETAILED DESCRIPTION OF THE INVENTION

[0014] Detailed Description of the Invention The present invention relates to a novel multi-use disinfecting solution (MPDS) that can be used for cleaning, disinfecting, and storing contact lenses. Applicant has discovered that in MPDSs containing disinfectants from two classes typically used in MPDSs (i.e., multimeric biguanide compounds and quaternary ammonium compounds), the concentration of hyaluronate (HA), included as a wetting, lubricating, and corneal support agent, cannot be significantly increased beyond 0.01% w / v without loss of effective antimicrobial activity. It is noted again that Bausch & Lomb's Biotrue (a representative MPDS of this type) contains 0.01% w / v HA. Presumably, higher concentrations of HA in such MPDSs are not tolerated due to concentration-dependent inactivation of cationic disinfectants by HA.

[0015] Surprisingly, it was found that the addition of a third class of disinfectant, namely antifungal / anti-acanthamoebic agents, effectively counteracted the negative effect of HA on the antimicrobial activity of MPDS: adequate antimicrobial activity was maintained at HA concentrations 7.5 times higher than those used in commercial MPDS.

[0016] Thus, the MPDS formulation of the present disclosure includes three types of disinfectants: a multimeric biguanide compound, a quaternary ammonium compound, and an antifungal / anti-acanthamoebic agent. The multimeric biguanide compound can be, but is not limited to, polyhexamethylene biguanide (PHMB), polyaminopropyl biguanide (PAPB), or 1,1'-hexamethylene-bis(5-[2-ethylhexyl]biguanide) (alexidine dihydrochloride). A preferred multimeric biguanide compound is PHMB. A typical concentration range for the multimeric biguanide compound (or a suitable salt thereof) is about 0.00005-0.0005% (w / v). Preferably, the compound is present at a concentration of about 0.000075-0.00025% (w / v). A more preferred range is about 0.0001-0.0002% (w / v), and most preferred is about 0.00012-0.00016% (w / v). Most preferably, the compound is present at a concentration of about 0.00015% (w / v).

[0017] Suitable quaternary ammonium compounds include, but are not limited to, polyquaternium-1 (PQ-1; also known as Polyquad (Alcon)), polyquaternium-10 (PQ-10) and polyquaternium-42 (PQ-42). These compounds may also be referred to as preferred quaternary ammonium compounds. Other suitable quaternary ammonium compounds include N-dodecyl{4,4'-(2,4,8,10-tetraoxaspiro[5.5]undecane-3,9-diyl)}dipyridinium dibromide (4TOSU-12), 3,3'-[1,4-phenylenebis(oxy)]bis(1-dodecylpyridinium)dibromide (3PHBO-12); These compounds are 3-(3-hydroxy-2-(hydroxymethyl)-2-{[(1-dodecylpyridinium-3-yl)oxy]methyl}propoxy)-1-dodecylpyridinium dibromide (3HHDMP-12), 5,50-[2,20-(tetramethylenedicarbonyldioxy)diethyl]bis(3-alkyl-4-methylthiazolium iodide) (5DEBT-4,8), and [4,40-(1,6-hexamethylenedithio)bis(1-octylpyridinium iodide)] (4DTBP-6,8) (Yamamoto et al. Biocontrol Science (2016) 21: 231-241; Ohkura et al. Bioorganic & Medicinal Chemistry (2005) 13: 2579-2587). Polyquaternium compounds are described, for example, in Iwata et al., Springer Science & Business Media, 2012-10-02, ISBN 9784431540618. For quaternary ammonium compounds (QACs) (or suitable salts thereof), a typical range is about 0.00005 to 0.0025% (w / v). Preferably, the concentration is about 0.00005 to about 0.0020% (w / v), more preferably about 0.000075 to about 0.001% (w / v), and most preferably about 0.000085 to about 0.00012% (w / v). Even more preferably, the concentration is about 0.00010% (w / v).Due to their low cytotoxicity and allergenic potential, multimeric QACs with several repeating units of quaternary ammonium groups are preferred over monomeric QACs. From this subclass, the most preferred multimeric QAC is polyquaternium-1 (PQ-1). In some embodiments, so-called bis-QACs (e.g., with two quaternary ammonium groups in the same entity (4TOSU-12)) are used because they have higher activity and better biocompatibility than conventional monomeric QACs. Mixtures of bis-QACs and PQ-1 may also be suitable for the ophthalmic formulations of the present disclosure.

[0018] Preferably, Polyquaternium-1 is understood herein as the compound having CAS number 75345-27-6. Polyquaternium-1 as defined herein is commercially available from Biosynth Carbosynth (FP163582).

[0019] Preferably, Polyquaternium-10 is understood herein as a compound or composition according to CAS number 68610-92-4. Polyquaternium-10 as defined herein is commercially available from Merck KGaA (525944).

[0020] Preferably, Polyquaternium 42 is understood herein as the compound or composition according to CAS number 31512-74-0. Polyquaternium-42 as defined herein is commercially available from Biosynth Carbosynth (FP59606).

[0021] Suitable antifungal / anti-acanthamoebic agents include myristamidopropyldimethylamine (MAPD; also known as Aldox (Alcon)) or other amido-amine compounds, or chlorhexidine gluconate. Most preferred is MAPD / Aldox. The antifungal / anti-acanthamoebic agent is typically included in the MPDS of the present disclosure at a concentration of about 0.00005 to about 0.0025% (w / v). Concentrations preferably range from about 0.00005 to about 0.0020% (w / v), more preferably from about 0.0001 to about 0.0018% (w / v), and most preferably from about 0.0006 to about 0.0014% (w / v). Even more preferably, the concentration is 0.00120% (w / v).

[0022] The HA (as the free acid or an acceptable salt thereof) utilized in the ophthalmic formulations of the present disclosure typically has a molecular weight of about 50,000 to about 2 million daltons (2 MDa), preferably about 0.4 to about 1.8 MDa, and most preferably about 1.2 to about 1.8 MDa. As understood herein, the molecular weight of HA preferably refers to the weight average molecular weight, determined as discussed below.

[0023] The molecular weight of HA was determined using gel permeation chromatography (GPC) with pullulan standards using the following protocol. A 3 mg / mL stock solution of hyaluronic acid was prepared in elution buffer (deionized water containing 0.05% (w / v) NaN3). 0.4 mL of the stock solution was then injected into the port of a tempered GPC device (1260 Infinity LC-System, Agilent, Santa Clara, CA). Chromatography was performed in elution buffer at a constant flow rate of 1.0 mL / min. Hyaluronic acid samples were separated on a Suprema 2 column system (precolumn, linear XL; 5 μm particle size; PSS, Mainz, Germany) placed in an external column oven at 55 °C. The copolymers were analyzed by RI (refractive index) and UV detection. A 10-point calibration curve was prepared for the following 10 polymers (showing Mw, Mn, and PDI): (1) Mw: 342 / Mn: 342, PDI 1.0; (2) Mw: 1320 / Mn: 1080, PDI 1.23; (3) Mw: 6200 / Mn: 5900, PDI 1.05; (4) Mw: 10000 / Mn: 9200, PDI 1.09; (5) Mw: 21700 / Mn: 20000, PDI 1.09; (6) Mw: 48800 / Mn: 45500, PDI 1.07; (7) Mw: 113000 / Mn: 100000, PDI 1.13; (8) Mw: 210000 / Mn The molecular weights of the characterized copolymers were estimated using pullulan standards obtained from PSS (Mainz, Germany), including (9) Mw: 366,000 / Mn: 318,000, PDI: 1.15; (10) Mw: 805,000 / Mn: 636,000, PDI: 1.27. For this purpose, the weight-average molecular weight (Mw), number-average molecular weight (Mn), and PDI of the polymer were determined based on GPC measurements using the software PSS WinGPC Unichrom V:8.1 Build 2827 (PSS; https: / / www.pss-polymer.com / ).

[0024] The concentration of HA is typically greater than 0.01% w / v and less than 0.1% w / v. Preferably, it is about 0.0151 to 0.09% (w / v), more preferably about 0.025 to 0.085% (w / v), and most preferably about 0.04 to about 0.08% (w / v). To maximize the safety of the ophthalmic formulation, the HA is preferably derived from a biotechnological process, for example, obtained by bacterial fermentation or in a cell-free system using appropriate enzymes.

[0025] HA is currently produced on an industrial scale either by extraction from animal tissues (mainly rooster combs) or via large-scale fermentation using genetically modified bacterial strains (Liu et al. Microb. Cell Fact. (2011) 10: 99). Both processes have their advantages, but from a safety standpoint, biotechnological methods are preferred for producing HA used in medical devices such as lens care solutions. Extraction from animal tissue requires harsh conditions (such as grinding, acid treatment, and the use of organic solvents), which results in partial degradation of HA and an increase in the polydispersity index (PDI) of the polymer. Consequently, batch-to-batch variability may increase (Boeriu et al. Int. J. Carbohydr. Chem. (2013) vol. 2013, Article ID 624967). Furthermore, HA derived from animal sources may still be bound to animal proteins and, depending on the source, may further contain nucleic acids, prions, and viruses, carrying the risk of immune reactions or disease transmission (Shiedlin et al. (2004) Biomacromolecules 5: 21222-2127). HA derived from biotechnological processes may be less likely to be contaminated. The presence of endotoxins can be avoided by using endotoxin-free strains (e.g., B. subtilis). Furthermore, biotechnologically produced HA has a lower PDI compared to tissue-extracted HA, provided that culture conditions are carefully monitored and controlled. To date, HA with molecular weights up to 2 MDa can be produced by fermentation. Cell-free systems using isolated enzymes produce HA with molecular weights of 1-2 MDa. This latter molecular weight range is well suited for ophthalmic formulations. This is because high molecular weight HA has a greater water absorption capacity and, due to their higher viscosity, remains on the contact lens / eye surface longer than HA with smaller molecular size (Sze et al. Biotech. (2016) 6: 67 and references cited therein).

[0026] The MPDS of the present disclosure also typically includes a buffer component. The type and amount of buffer are selected so that the MPDS composition meets the required performance criteria (e.g., physicochemical attributes and shelf-life stability, antimicrobial effectiveness, buffering capacity, and similar factors). The buffer is also selected to provide a pH that is compatible with the target ocular tissue and any contact lenses for which the composition is intended to be used. Borate buffer, citrate buffer, histidine buffer, Tris buffer, Tris / glycine buffer, or Bis-Tris buffer, and combinations thereof may be suitable. Phosphate buffer may be even more suitable. Preferably, the buffer is borate buffer, phosphate buffer, citrate buffer, histidine buffer, Tris buffer, Tris / glycine buffer, Bis-Tris buffer, or combinations thereof. Generally, for ophthalmic formulations, a pH close to that of human tears (e.g., about pH 7.5) is very useful, although a broader pH range of about 6.0 to about 8.0, more preferably about 6.2 to about 7.8, and even more preferably about 7.0 to about 7.7, is also acceptable. Buffering substances are typically present at concentrations between about 0.01% w / v and about 1% w / v.

[0027] Ophthalmic formulations of the present disclosure intended to effectively clean contact lenses typically contain one or more surfactants. Useful surfactants include, but are not limited to, poloxamers (nonionic triblock copolymers composed of a central hydrophobic chain of polypropylene oxide (PPO) flanked by two hydrophilic chains of polyethylene oxide (PEO)), poloxamines (nonionic copolymers with an ethylenediamine core in which amine groups are substituted with PPO / PEO chains of various lengths and compositions), or polysorbate-type materials (oily liquids derived from ethoxylated sorbitan (a derivative of sorbitol) esterified with various fatty acids).

[0028] The inclusion of such surfactants results in effective lens cleaning during lens treatment without substantially affecting the antimicrobial activity of the MPDS composition. The concentration of surfactant in the formulation typically ranges from about 0.01% w / v to about 1% (w / v). Poloxamines are preferred. Examples of poloxamines are Tetronic 90R4, Tetronic 701, Tetronic 1304, and Tetronic 1107. The preferred poloxamines are Tetronic 90R4, Tetronic 1304, and Tetronic 1107. The cleaning properties of each poloxamine vary depending on the PEO / PPO ratio and molecular weight.

[0029] Tetronic 90R4 is preferably defined as the compound or composition according to CAS number 26316-40-5, which is available from Sigma Aldrich under catalog number 435546. Preferably, the number average molecular weight of Tetronic 90R4 is 7200 Da.

[0030] Tetronic 701 is preferably defined as the compound or composition according to CAS number 26316-40-5, which is available from Sigma Aldrich under catalog number 435511. Preferably, the number average molecular weight of Tetronic 701 is 3600 Da.

[0031] Tetronic 1304 is preferably defined as a compound or composition according to CAS number 26316-34-5. Preferably, Tetronic 1304 has a number average molecular weight of 10,500 Da.

[0032] Tetronic 1107 is preferably defined as a compound or composition according to CAS number 26316-40-5. Preferably, Tetronic 1107 has a number average molecular weight of 15,000 Da.

[0033] The MPDS formulations of the present disclosure typically contain a chelating agent or mixture of chelating agents. Such chelating agents should be capable of interacting with calcium and magnesium ions, as well as with residual heavy metal ions sometimes present in contact lenses. They should also be compatible with ophthalmic or medical applications. Suitable chelating agents include, but are not limited to, ethylenediaminetetraacetic acid (EDTA), N,N-bis(carboxymethyl)-L-glutamic acid tetrasodium salt (GLDA), iminodisuccinic acid tetrasodium salt, N-(1-carboxylatoethyl)iminodiacetate trisodium salt (MGDA) (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid) (DOTA), diethylenetriaminepentaacetic acid (DTPA), triethylenetetramine-N,N,N',N",N"',N"'-hexaacetic acid (TTHA), and citrate. EDTA and GLDA are preferred because they are excellent chelating agents and have good water solubility. GLDA is readily biodegradable. The concentration of the chelating agent in the formulation typically ranges from about 0.01% w / v to about 0.06% w / v. Commercially available MPDS, e.g., Bausch & Co., Inc., It is noted that Lomb's Biotrue contains higher chelating agent levels. The MPDS of the present disclosure may also contain low concentrations of calcium and / or magnesium salts at concentrations of about 0.001% w / v to about 0.015% w / v, whereby the combined molar amount of divalent cations added is less than that of the chelating agents used in the same formulation.

[0034] The osmolality of the ophthalmic formulations of the present disclosure can be adjusted with a tonicity agent. Examples of suitable tonicity agents include, but are not limited to, chloride salts of sodium, potassium, calcium, and magnesium, dextrose, glycerol, propylene glycol, sugars (e.g., sorbitol, mannitol), amino acids (e.g., glycine), and mixtures of these agents. A preferred tonicity agent is sodium chloride or a combination of sodium chloride and potassium chloride, which may be combined with mannitol, sorbitol, or glycine to increase buffering or radical scavenging capacity. Tonicity agents are typically used in amounts ranging from about 0.001% w / v to about 1% w / v. Preferably, the tonicity agent is used in an amount that results in a final osmolality of about 220-380 mOsm / kg, more preferably about 240-320 mOsm / kg, and most preferably about 280-310 mOsm / kg.

[0035] Viscosity modifiers that may be included in the formulations of the present disclosure include cellulose polymers (including hydroxypropylmethylcellulose, hydroxyethylcellulose, ethylhydroxyethylcellulose, hydroxypropylcellulose, methylcellulose, and carboxymethylcellulose), glycerol, carbomer, polyethylene glycol, polyvinyl alcohol, polyvinylpyrrolidone, poly(acryloyl-lysine) or copolymers containing amino acids with acrylated side chains, alginate, carrageenan, galactomannan polysaccharide (guar), karaya, agarose, locust bean gum, tragacanth gum, and xanthan gum. Viscosity modifiers are used in amounts effective to provide the desired lubricating effect; concentrations of these agents are typically between about 0.01 and 0.25% w / v. Preferred viscosity modifiers are hydroxypropylmethylcellulose (HPMC), carboxymethylcellulose (CMC), and hydroxyethylcellulose (HEC). HPMC, which is also an excellent wetting agent, is most preferred. HPMC is sold, for example, by The Dow Chemical Company (Dow) under the trademark METHOCEL. Numerous types are available, differing in their degree of functionalization with methoxyl / hydroxypropyl groups and molecular weight. The appropriate type of HPMC (or other viscosity modifier) ​​can be selected to achieve the desired viscosity, which is typically between about 0.5 and about 4 milliPascal-seconds (mPa·s), preferably between about 1.0 and about 3.5 mPa·s, more preferably between about 1.25 and about 3.25 mPa·s, and most preferably between about 2.0 and about 2.8 mPa·s.

[0036] Viscosity determinations as referred to herein were performed using a spindle viscosimeter. For this, 300 mL of sample solution was poured into a 500 mL glass beaker and conditioned in a water bath at 23° C. for 30 minutes. Viscosity was determined using a Brookfield Viscometer (DV-II, 60 rpm, spindle 1, 23° C.).

[0037] Representative MPDS formulations according to the present disclosure are set forth in Tables 1 and 2. As known to those skilled in the art, the formulations of the present invention can be obtained by mixing and diluting aqueous stock solutions of each of the above components. Those skilled in the art will recognize that, for example, in the case of a hydrophobic agent such as Aldox, an ethanol stock solution can be prepared for adding the hydrophobic agent to an aqueous formulation. [Table 1]

[0038] [Table 2]

[0039] The invention thus generally described will be more readily understood by reference to the following examples, which are offered by way of illustration and are not intended to be limitations of the invention. [Example]

[0040] Example Example 1: Antimicrobial preservative effectiveness according to ISO 14729 In an attempt to obtain an MPDS that offers greater patient comfort, Formulations A and B were prepared. These formulations are based on the same multimeric biguanides and quaternary ammonium compounds used in commercially available MPDSs, but contain HA at a 50% higher concentration (Table 3). These formulations were tested for their ability to inactivate microbial growth according to the ISO 14729 test protocol, using MPDS Biotrue containing 0.01% (w / v) HA as a positive control for the test procedure. Note that the spiral plate method was used for cell counting. [Table 3]

[0041] *Formulations A-E further contain boric acid (0.1% w / v), disodium tetraborate decahydrate (0.3% w / v), HPMC (0.08% w / v), Tetronic 90R4 (0.1% w / v), EDTA (sodium salt dihydrate - 0.04% w / v), potassium chloride (0.04% w / v), sodium chloride (0.5% w / v), calcium chloride dihydrate (0.002% w / v) and magnesium sulfate heptahydrate (0.002% w / v); pH 7.5 (adjusted by adding a 32% w / w aqueous solution of HCl). **Formulation F further contains sodium phosphate monobasic (monohydrate, final concentration 0.1% w / v), sodium phosphate dibasic (heptahydrate, final concentration 0.4% w / v), HPMC (0.08% w / v), Tetronic 90R4 (0.1% w / v), EDTA (sodium salt dihydrate - 0.04% w / v), potassium chloride (0.04% w / v), sodium chloride (0.5% w / v); pH 7.5 (adjusted by adding a 20% w / w aqueous solution of NaOH).

[0042] The latter component is present in concentrations as indicated in parentheses within the ranges described above.

[0043] 1.0 x 10 5 ~1.0×10 6 An appropriate number of challenge organisms sufficient to provide a final count between cfu / ml was inoculated and then incubated at 20-25°C for up to 32 hours. At 2, 4, 6, and 8 hours, 1 ml aliquots of the solution were withdrawn and analyzed for residual viable microbial content using the spiral plate method. This method is used to measure the residual viable microbial content of 10 2 have a limit of detection of microorganisms (i.e., at the time of inoculation, the microorganism count is 10 6(The maximum detectable log reduction is 4.0 if the serotonin concentration is 0.05). In the case of yeast and fungal challenges (C. albicans and F. solani), additional samples were removed after 32 hours. Representative test results are shown in Table 4. The symbol ">" in Table 4 indicates that no colony forming units (CFU) were observed.

[0044] Viable organism count determinations and log reduction determinations were performed according to ISO 14729 (1st Edition 2001-04-15), except that the spiral plate method (Gilchrist et al. Appl Microbiol. 1973 Feb;25(2):244-52) was used to recover the challenge organisms. Colony-forming units (CFU) were recorded only for enumerable plates. As defined herein, enumerable plates refer to plates with 30 to 300 CFU / plate for bacteria and yeast, and 8 to 80 CFU / plate for filamentous fungi. The average number of CFU for the enumerable plates (triplicates) was recorded, and the microbial reduction at specific time points [2, 4, 6, and 8 hours (bacteria) or 2, 4, 6, 8, and 32 hours (yeast and filamentous fungi)] was calculated by allocating the inoculum value for each challenge organism. [Table 4]

[0045] The data in this table represent the average from three independent experiments, which were carried out in the same laboratory and by the same examiner.

[0046] Neither Formulation A nor Formulation B met the primary criteria of the ISO test procedure (a 3.0-log reduction of bacteria and a 1.0-log reduction of yeast and mold within the recommended soak time) (Table 4). Thus, the combination of PHMB and PQ-1, even when used at higher concentrations than the positive control formulation, Biotrue, fails to provide adequate antimicrobial activity when HA concentrations are significantly increased. Antifungal activity is particularly affected. Replacing PQ-1 with a second biguanide compound resulted in even lower antimicrobial activity. Both antibacterial and antifungal activity were low. Collectively, the test results indicated that the presence of only two disinfectants (either from two different classes or from the same class) is insufficient to provide adequate antimicrobial activity in MDSs with significantly increased concentrations of HA compared to commercially available MPDSs.

[0047] Applicant then conducted experiments to determine whether the addition of a third disinfectant could remedy the deficiencies of Formulations A and B. Excellent results were obtained with MAPD / Aldox as the third disinfectant (see Table 3 for a description of Formulation C and Table 4 for data). The effect of MAPD was strong enough to allow the reduction of PQ-1 levels to those present in the positive control formulation (Biotrue, a commercially available MPDS (see Table 3 for a description of Formulation D and Table 4 for data)). Assuming comfort increases with concentration, the concentration of HA was further increased in Formulation E (see Table 2) to levels 7.5-fold higher than those present in commercially available MPDSs (e.g., Biotrue). As the data in Table 4 reveal, the bactericidal and fungicidal activity of Formulation E readily met the primary criteria of the ISO test procedure at 4 hours and all subsequent time points. Thus, formulation E, which contains disinfectants from three classes, i.e., biguanides, quaternary ammonium compounds, and amido-amine compounds, appears to offer the optimal combination of patient comfort (due to the presence of high levels of HA) and antimicrobial activity. It is noted that the slightly lower antimicrobial activity at 2 hours of formulation E compared to that of formulations containing lower concentrations of HA (e.g., formulations C and D) is not of direct practical relevance, since lens wearers typically incubate their contact lenses in MPDS overnight (6-8 hours).

[0048] Additionally, Formulation F, based on a phosphate buffer system, was tested in the same regimen according to ISO 14729. The primary criteria were: a >3.0 log reduction of the three test organisms (bacteria) should be achieved after 4 hours, and a >1.0 log reduction of the two fungal strains should be achieved after 6-8 hours. These results demonstrate that the disclosed synergistic effect of the three classes of disinfectants works in a variety of buffer systems.

[0049] Example 2: Long-term effectiveness of antimicrobial preservation according to ISO 14730 To evaluate the long-term effectiveness of the MPDS presented in this disclosure, Formulation E was inoculated with five standard test organisms and then stored at 20-25°C for up to 28 days. Aliquots from the test solution were removed and assayed for surviving organisms immediately after addition of the microorganisms (T 0 time), and after 7, 14, 21, and 28 days of storage according to the ISO 14730 test procedure. Immediately after the 14-day aliquot was withdrawn, the test solution was diluted with 1 x 10 4 ~1×10 5 The same microorganisms were re-inoculated at CFU / ml. The Log reduction in the levels of the test organisms was calculated for each time point. The results are shown in Table 6: [Table 6]

[0050] The "greater than" symbol (>) is used when no colonies were observed on the plate, taking into account the sensitivity of the assay (see Example 1 for details).

[0051] Test method pass criteria: Positive controls are positive for growth of indicator organisms. Negative controls are negative for growth of indicator organisms. Suitability controls must demonstrate ≥ 50% organism recovery in the reported test formulation solution.

[0052] Testing criteria according to the United States Pharmacopeia (USP) for Category 1 products (injectables, emulsions made with an aqueous base or vehicle, and other parenterals including otic products, sterile nasal products, and ophthalmic products):

[0053] Bacteria: ≥ 1.0 log reduction from initial calculated count at 7 days, ≥ 3.0 log reduction from initial count at 14 days, and no increase from 14 day count at 28 days. Yeast and mold: No increase from initial calculated counts at 7, 14, and 28 days.

[0054] Formulation E met all test criteria. In particular, with respect to the difficult-to-treat fungus Aspergillus brasiliensis, Formulation E's performance was excellent, with a log reduction of 2.35 at 14 days and >2 at 21 and 28 days, i.e., after reinoculation of the fungus at 14 days. This indicates that the synergistic effect of the three classes of disinfectants used in this MPDS is strong and long-lasting, even though it contains a 7.5-fold higher concentration of HA than the commercial product.

[0055] Example 3: Biocompatibility of MPDS Formulation E according to ISO 10993-5 The biocompatibility of Formulation E, which has a high content of hyaluronic acid (0.075% w / v), was evaluated for potential cytotoxic effects using an in vitro mammalian cell culture model. Biotrue MPDS served as a comparison product. The test was performed in accordance with the guidelines of ISO 10993-5, Biological Evaluation of Medical Devices - Part 5: Tests for In Vitro Cytotoxicity.

[0056] Briefly, sample solutions of Formulation E or Biotrue were mixed with double-strength minimum essential medium (2x MEM) to a 50% concentration for testing (test solution). Controls (negative control, reagent control, and positive control) were extracted in 1x MEM for 24 hours at 37°C. Triplicate monolayers of L929 mouse fibroblast cells were overlaid with the test solution or control extract and incubated for 48 hours at 37°C in the presence of 5% CO2. After incubation, the monolayers were examined microscopically for abnormal cell morphology and cell degeneration using the scoring system in Table 5. No pH shift (change in medium color) was observed.

[0057] Negative control: High density polyethylene (HDPE), extraction ratio 3 cm 2 : 1mL; total extraction area 31.5cm 2 Extraction with 10 mL extraction vehicle at 37°C for 24 hours

[0058] Reagent control: One-strength minimal essential medium (92% Gibco MEM with Earle's salts) supplemented with 5% fetal bovine serum, 2% antibiotics (100 units / mL penicillin, 100 μg / mL streptomycin, and 2.5 μg / mL amphotericin B), and 1% (2 mM) L-glutamine.

[0059] Positive control: Powder-free latex glove (composition: natural rubber latex, zinc carbamate accelerator, zinc oxide, and titanium dioxide), extraction ratio 6 cm 2 : 1mL; total extraction area 60cm 2 Extraction with 10 mL extraction vehicle at 37°C for 24 hours.

[0060] Dilution vehicle: double strength minimal essential medium (2xMEM) supplemented with 10% fetal bovine serum, 4% antibiotics (200 units / mL penicillin, 200 μg / mL streptomycin, and 5.0 μg / mL amphotericin B) and 2% (4 mM) L-glutamine.

[0061] Control extraction vehicle: 1x MEM (minimum essential medium).

[0062] The results of the experiment are shown in Table 8. Both Formulation E and Biotrue showed slight cytotoxicity (Grade 1), which was less than or equal to Grade 2 (mild cytotoxicity), thus meeting the ISO 10993-5 acceptance criteria for medical devices. Reagent, negative, and positive controls were performed as expected. Formulation E demonstrated better biocompatibility than Biotrue: cell lysis was 10% for Formulation E and 20% for Biotrue. The results indicate that Formulation E, which contains a 7.5-fold higher concentration of HA than Biotrue, has significantly reduced (50%) cytotoxic potential compared to Biotrue. [Table 7]

[0063] [Table 8]

[0064] A larger-scale batch (100 L) of Formulation E was further evaluated in more detail with an XTT test according to ISO 10993, "Biological Evaluation of Medical Devices," Part 1: "Evaluation and Testing within the Risk Management Process," 2018, Part 5: "Testing for In Vitro Cytotoxicity," published by the International Organization for Standardization (ISO 10993), using validated methods and Xenometrix test kits from ICCR-Rossdorf GmbH (Germany). The XTT assay is used to calorimetrically determine cell proliferation and viability as well as mitochondrial metabolic capacity of cells after treatment with the test article. The XTT test is based on the cleavage of XTT [=(3'-(1-phenylaminocarbonyl)-(3,4-tetrazolium)-bis-(4-methoxy-6-nitro)-benzenesulfonate sodium hydrate)], a yellow tetrazolium salt, which forms an orange, water-soluble formazan dye via dehydrogenase activity in active mitochondria. This method was first described by Scudiero et al. in 1988.

[0065] Briefly, the potential cytotoxic effect was evaluated by serial dilutions of Formulation E in complete medium (RPMI 1640 medium (with glutamax) supplemented with 10% FCS, 1 mM sodium pyruvate, 100 units / mL penicillin, and 100 μg / mL streptomycin) resulting in the following final concentrations: 39.1 μg / mL, 78.1 μg / mL, 156 μg / mL, 313 μg / mL, 625 μg / mL, 1250 μg / mL; 2500 μg / mL, 5000 μg / mL. The effect of these solutions was then tested after a 24±1 hour treatment period on previously prepared mouse L929 cell lines (provided by DSMZ, 38124 Braunschweig, Germany). In parallel, several controls were also tested:

[0066] Medium control = complete medium; solvent control = complete medium; positive control = sodium dodecyl sulfate (SDS; purity: ≥ 99%); solvent = deionized water (diluted with complete medium at a ratio of 1:10 (v / v)). The positive control was tested at eight concentrations ranging from 3.1 to 250 μg / mL. Solvent control for the positive control = complete medium and 10% (v / v) deionized water.

[0067] At the end of the incubation period, 50 μL of XTT labeling mixture (Xenometrix, Allschwil, Switzerland) was added to each well. The cells were incubated and then transferred to a microplate reader, and their absorbance values ​​were determined at 450 nm (reference wavelength 690 nm) (Versamax® Molecular Devices, software SoftMax Pro Enterprise (version 4.7.1)). A decrease in the number of viable cells results in a decrease in the overall activity of mitochondrial dehydrogenase in the sample. This decrease directly correlates with the amount of orange formazan formed, as monitored by absorbance. The relative absorbance (= viability) compared to the solvent control is calculated using the formula:

number

number

[0068] The lower the XTT50 value, the greater the cytotoxic potential of the test article. A test article is considered to have a cytotoxic effect if at least one concentration results in cell viability <70% compared to the solvent control. [Table 9]

[0069] Test groups shaded in grey showed cytotoxic effects in photometric evaluation. *Average absolute absorbance of 7 wells **Relative absorbance [rounded value] Preparation E XTT 50 -values ​​could not be determined if viability was not reduced below 50%; Positive control: XTT 50 -Value: 72.09μg / mL

[0070] Vehicle control (positive control) / medium control ratio: 90.44%.

[0071] In this XTT assay, Formulation E showed no negative effect on cell proliferation of mouse L929 fibroblasts, a result consistent with the above morphological evaluation and verification of the high biocompatibility of the solution.

[0072] Example 4: Long-term elution of HA from silicone hydrogel contact lenses It is known from the literature that silicone hydrogel contact lenses can absorb HA from lens care solutions during incubation, thereby forming a depot of HA within the contact lens. From this depot, HA is slowly released during lens wear, improving surface wettability and comfort (Scheuer et al., Clin. Ophthalmol., 2016, pp. 1945-1952). Because the ophthalmic formulations of the present disclosure contain HA at concentrations 1.5 to 7.5 times higher than commercially available formulations (e.g., Biotrue), a better depot effect of the silicone hydrogel lenses is expected with the formulations of the present disclosure. To verify this hypothesis, a side-by-side comparison of Formulation E and Biotrue was performed.

[0073] In the experiment, HA release was evaluated using four commercially available lens types: Comfilcon A (Biofinity), fanfilcon A (Avaira Vitality), lotrafilcon B (Air Optix Aqua), and senofilcon A (Acuvue Oasys). Multiple lenses per lens type were removed from their packaging and equilibrated in phosphate-buffered saline (PBS) for 7 hours with agitation at ambient temperature (in a 24-well plate; two lenses per well). The lenses were then immersed in 3 mL of either Formulation E or Biotrue for 16 hours under overnight storage conditions (ambient temperature, no agitation). After lens care storage, the lenses were gently blotted on laboratory tissue to remove carryover solution and transferred to a 24-well plate containing 500 μL PBS per well. The plate was then incubated at ambient temperature for 24 hours. Aliquots of 100 μL were removed from the wells at 1, 3, 5, 7, and 24 hours and stored at 40°C until analysis. Fresh PBS (100 μL) was added to the wells after each withdrawal. Aliquots were diluted 1:200, and HA was quantified by ELISA according to the manufacturer's instructions (ELISA Hyaluronan kit, R&D Systems, Minneapolis, MN).

[0074] The results are shown in Table 10. The data show that greater amounts of HA were released at each time point from lenses soaked in Formulation E than from lenses incubated with Biotrue (except for the Avira Vitality lenses at the 1 hour time point). [Table 10]

[0075] Example 5: Long-term elution of HA from silicone hydrogel contact lenses into synthetic tears Experiments similar to those discussed in Example 4 can be performed to evaluate the release of HA into tears from lenses incubated in different MPDSs. Because human tears vary in composition (e.g., in pH, salt content, and osmolality) between individuals and results can be influenced by the behavior of study participants, standardized in vitro test procedures are used for such comparisons.

[0076] In the experiment, two types of commercially available silicone hydrogel contact lenses (Biofinity, Comfilcon A Cooper Vision, and Acuvue Oasys, Senofilcon A J&J) were placed in the wells of a 24-well cell culture plate (a plate without surface modification; Eppendorf AG, Hamburg, Germany). A 1.8 ml aliquot of the MPDS formulation to be tested (Formulation E; Biotrue by B&L; Hycare by Cooper Vision) was added to the wells, and the plate was incubated for 8 hours at room temperature (4 contact lenses per MPDS formulation). The soaked contact lenses were then gently rinsed with 0.9% NaCl solution to remove residual MPDS on the surface, and carefully transferred to the wells of a new 24-well plate containing 1.8 ml / well of complex synthetic tear solution (prepared according to Lorentz et al. Mol. Vis. (2011) 17: 3392-3405). To assess HA release from contact lenses into synthetic tears, 40 μL aliquots were withdrawn at 0, 0.5, 1, 2, 4, 8, 12, and 24 h (the withdrawn volume was replaced with fresh synthetic tears to simulate tear turnover (approximately 1.2 μL / min according to Wilson, Cambridge University Press, 2004. ISBN: 9780521841580)). Aliquots were stored at 4°C until analysis of HA concentration by Hyaluronan Enzyme-Linked Immunosorbent Assay (HA ELISA, Echelon Biosciences, USA).

[0077] This experiment is expected to reveal that greater amounts of HA are released into the tear fluid at each time point from lenses incubated with Formulation E compared to lenses incubated with the other formulations.

[0078] The recitation of ranges of values ​​herein is merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated herein as if individually set forth herein. Unless otherwise stated, all exact values ​​provided herein represent the equivalent approximation, and vice versa (e.g., all exact exemplary values ​​provided with respect to a particular factor or measurement may also be considered to provide the equivalent approximation of the measurement, and may be modified by "about" where appropriate, and vice versa).

[0079] The terms "a," "an," "the," "the," and similar terms are to be construed to cover both the singular and the plural unless otherwise indicated or clearly contradicted by context.

[0080] A description herein of any aspect or embodiment of the invention using such language with reference to an element is intended to provide support for similar aspects or embodiments of the invention that "consist of," "consist essentially of," or "substantially comprise" that particular element, unless stated otherwise or clearly contradicted by context (e.g., a composition described herein as comprising a particular element should also be understood to describe a composition consisting of that element, unless stated otherwise or clearly contradicted by context).

[0081] Preferably, as understood herein, the ratio term weight to volume, also referred to herein as w / v, or (w / v), which may also be expressed as a percentage (%) value, refers to the ratio of the weight of the dissolved substance as expressed in grams (g) to the volume of the solution as expressed in milliliters (mL).

[0082] This invention includes all modifications and equivalents of the subject matter recited in the aspects or claims presented herein to the maximum extent permitted by applicable law.

[0083] All references cited in this application, including publications, patents, and patent applications, shall be deemed to be incorporated by reference in their entirety.

[0084] Further embodiments of the present invention are disclosed in the following numbered paragraphs: 1. A multi-use disinfecting solution for application to the eye, comprising a multimeric biguanide compound in an amount ranging from about 0.00005 to about 0.0005% w / v, a quaternary ammonium compound in an amount ranging from about 0.00005 to about 0.0025% w / v, an antifungal / anti-acanthamoebic agent in an amount ranging from about 0.00005 to about 0.0025% w / v, and hyaluronic acid or a salt thereof in an amount ranging from about 0.015 to about 0.1% w / v.

[0085] 2. The multi-use disinfectant solution of item 1, wherein the antifungal / anti-acanthamoebic agent is an amido-amine compound.

[0086] 3. The multi-use disinfectant solution of item 2, wherein the multimeric biguanide compound is present in an amount ranging from about 0.0001 to about 0.0002% w / v, the quaternary ammonium compound is present in an amount ranging from about 0.000075 to about 0.001% w / v, and the amido-amine compound is present in an amount ranging from about 0.0001 to about 0.0018% w / v.

[0087] 4. The multi-use disinfectant solution of item 2, wherein the multimeric biguanide compound is present in an amount ranging from about 0.00012 to about 0.00016% w / v, the quaternary ammonium compound is present in an amount ranging from about 0.000085 to about 0.00012% w / v, and the amido-amine compound is present in an amount ranging from about 0.0006 to about 0.0014% w / v.

[0088] 5. The multi-use disinfectant solution of any of items 1 to 4, wherein the hyaluronic acid or salt thereof is present in an amount ranging from about 0.025% w / v to about 0.085% w / v.

[0089] 6. The multi-use disinfectant solution of any one of items 1 to 4, wherein the hyaluronic acid or salt thereof is present in an amount ranging from about 0.04% w / v to about 0.08% w / v.

[0090] 7. The multimeric biguanide compound is selected from the group consisting of polyhexamethylene biguanide, polyaminopropyl biguanide, and 1,1'-hexamethylene-bis(5-[2-ethylhexyl]biguanide), and the quaternary ammonium compound is polyquaternium-1, polyquaternium-10, polyquaternium-42, N-dodecyl{4,4'-(2,4,8,10-tetraoxaspiro[5.5]undecane-3,9-diyl)}dipyridinium dibromide, 3,3'-[1,4-phenylenebis(oxy)]bis(1-dodecylpyridinium) dibromide; 7. The multi-use disinfectant solution according to any one of items 1 to 6, wherein the amido-amine compound is selected from the group consisting of 3-(3-hydroxy-2-(hydroxymethyl)-2-{[(1-dodecylpyridinium-3-yl)oxy]methyl}propoxy)-1-dodecylpyridinium dibromide, 5,50-[2,20-(tetramethylenedicarbonyldioxy)diethyl]bis(3-alkyl-4-methylthiazolium iodide), and [4,40-(1,6-hexamethylenedithio)bis(1-octylpyridinium iodide)], and the amido-amine compound is myristamidopropyl dimethylamine.

[0091] 8. The multi-use disinfectant solution of item 7, wherein the multimeric biguanide compound is polyhexamethylene biguanide and the quaternary ammonium compound is polyquaternium-1.

[0092] 9. The multi-use disinfectant solution of any of items 1 to 8, further comprising one or more of a chelating agent in an amount ranging from about 0.01% w / v to about 0.06% w / v, a buffer in an amount ranging from about 0.01% w / v to about 1% w / v, a viscosity modifier in an amount ranging from about 0.01% w / v to about 0.25% w / v, a surfactant compound in an amount ranging from about 0.01% w / v to about 1% w / v, an isotonicity agent in an amount ranging from about 0.001% w / v to about 1% w / v, a calcium salt in an amount ranging from about 0.001% w / v to about 0.015% w / v, and a magnesium salt in an amount ranging from about 0.001% w / v to about 0.015% w / v.

[0093] 10. The multi-use disinfectant solution of item 9, wherein the chelating agent is ethylenediaminetetraacetic acid.

[0094] 11. The multi-use disinfectant solution of item 9, wherein the buffer is a borate buffer.

[0095] 12. The multi-use disinfectant solution of item 9, wherein the viscosity modifier is selected from the group consisting of hydroxypropyl methylcellulose, carboxymethylcellulose, and hydroxyethylcellulose.

[0096] 13. The multi-use disinfectant solution of item 12, wherein the viscosity modifier is hydroxypropyl methylcellulose.

[0097] 14. The multi-use disinfectant solution of item 9, wherein the surfactant compound is a poloxamine.

[0098] 15. The multi-use disinfectant solution of item 9, wherein the tonicity agent is sodium chloride or a combination of sodium chloride and potassium chloride. (Item 1) 1. A solution comprising a multimeric biguanide compound in an amount ranging from about 0.00005 to about 0.0005% w / v, a quaternary ammonium compound in an amount ranging from about 0.00005 to about 0.0025% w / v, an antifungal / anti-acanthamoebic agent in an amount ranging from about 0.00005 to about 0.0025% w / v, and hyaluronic acid or a salt thereof in an amount ranging from about 0.015 to about 0.1% w / v. (Item 2) 2. The solution of item 1, wherein the antifungal / anti-acanthamoebic agent is an amido-amine compound. (Item 3) 3. The solution of item 2, wherein the multimeric biguanide compound is present in an amount ranging from about 0.0001 to about 0.0002% w / v, the quaternary ammonium compound is present in an amount ranging from about 0.000075 to about 0.001% w / v, and the amido-amine compound is present in an amount ranging from about 0.0001 to about 0.0018% w / v. (Item 4) 3. The solution of item 2, wherein the multimeric biguanide compound is present in an amount ranging from about 0.00012 to about 0.00016% w / v, the quaternary ammonium compound is present in an amount ranging from about 0.000085 to about 0.00012% w / v, and the amido-amine compound is present in an amount ranging from about 0.0006 to about 0.0014% w / v. (Item 5) 5. The solution according to any one of items 1 to 4, wherein the hyaluronic acid or a salt thereof is present in an amount ranging from about 0.025% w / v to about 0.085% w / v. (Item 6) 5. The solution according to any one of items 1 to 4, wherein the hyaluronic acid or a salt thereof is present in an amount ranging from about 0.04% w / v to about 0.08% w / v. (Item 7) the multimeric biguanide compound is selected from the group consisting of polyhexamethylene biguanide, polyaminopropyl biguanide, and 1,1'-hexamethylene-bis(5-[2-ethylhexyl]biguanide), and the quaternary ammonium compound is selected from the group consisting of polyquaternium-1, polyquaternium-10, polyquaternium-42, N-dodecyl{4,4'-(2,4,8,10-tetraoxaspiro[5.5]undecane-3,9-diyl)}dipyridinium dibromide, 3,3'-[1,4-phenylenebis(oxy)]bis(1-dodecylpyridinium) dibromide; 7. The solution according to any one of items 1 to 6, wherein the amido-amine compound is selected from the group consisting of 3-(3-hydroxy-2-(hydroxymethyl)-2-{[(1-dodecylpyridinium-3-yl)oxy]methyl}propoxy)-1-dodecylpyridinium dibromide, 5,50-[2,20-(tetramethylenedicarbonyldioxy)diethyl]bis(3-alkyl-4-methylthiazolium iodide), and [4,40-(1,6-hexamethylenedithio)bis(1-octylpyridinium iodide)], and the amido-amine compound is myristamidopropyldimethylamine. (Item 8) 8. The solution of claim 7, wherein the multimeric biguanide compound is poly(hexamethylene biguanide) and the quaternary ammonium compound is polyquaternium-1. (Item 9) 9. The solution of any one of items 1 to 8, further comprising one or more of: a chelating agent in an amount ranging from about 0.01% w / v to about 0.06% w / v, and / or a buffer in an amount ranging from about 0.01% w / v to about 1% w / v, and / or a viscosity adjusting agent in an amount ranging from about 0.01% w / v to about 0.25% w / v, and / or a surfactant compound in an amount ranging from about 0.01% w / v to about 1% w / v, and / or a tonicity adjusting agent in an amount ranging from about 0.001% w / v to about 1% w / v, and / or optionally a calcium salt in an amount ranging from about 0.001% w / v to about 0.015% w / v, and / or optionally a magnesium salt in an amount ranging from about 0.001% w / v to about 0.015% w / v. (Item 10) 10. The solution of claim 9, wherein the chelating agent is ethylenediaminetetraacetic acid. (Item 11) 10. The solution according to item 9, wherein the buffer is a borate buffer or a phosphate buffer. (Item 12) 10. The solution of claim 9, wherein the viscosity modifier is selected from the group consisting of hydroxypropyl methylcellulose, carboxymethyl cellulose, and hydroxyethyl cellulose. (Item 13) Item 13. The solution of item 12, wherein the viscosity modifier is hydroxypropyl methylcellulose. (Item 14) 10. The solution of claim 9, wherein the surface-active compound is a poloxamine. (Item 15) 10. The solution of item 9, wherein the tonicity agent is sodium chloride or a combination of sodium chloride and potassium chloride. (Item 16) 16. The solution according to any one of items 1 to 15, wherein the solution is a multi-use disinfecting solution. (Item 17) 17. A solution according to any one of items 1 to 16 for use in therapy. (Item 18) 17. A solution according to any one of items 1 to 16 for use in disinfection, in particular in ocular application. (Item 19) 17. Non-therapeutic use of a solution according to any of items 1 to 16 in disinfection, in particular in disinfecting contact lenses.

Claims

1. A solution comprising a multimeric biguanide compound in an amount ranging from 0.00012 to 0.00016% w / v, a quaternary ammonium compound in an amount ranging from 0.000085 to 0.00012% w / v, an amido-amine compound in an amount ranging from 0.0006 to 0.0014% w / v, and hyaluronic acid or a salt thereof in an amount ranging from 0.04 to 0.08% w / v, wherein the solution is a multi-use disinfectant solution.

2. The multimeric biguanide compound is selected from the group consisting of polyhexamethylene biguanide, polyaminopropyl biguanide, and 1,1'-hexamethylene-bis(5-[2-ethylhexyl]biguanide), and the quaternary ammonium compound is selected from the group consisting of polyquaternium-1, polyquaternium-10, polyquaternium-42, N-dodecyl{4,4'-(2,4,8,10-tetraoxaspiro[5.5]undecane-3,9-diyl)}dipyridinium dibromide, 3,3'-[1,4-phenylenebis(oxy)]bis(1-dodecylpyridinium)dibromide; 2. The solution of claim 1, wherein the amido-amine compound is selected from the group consisting of 3-(3-hydroxy-2-(hydroxymethyl)-2-{[(1-dodecylpyridinium-3-yl)oxy]methyl}propoxy)-1-dodecylpyridinium dibromide, 5,50-[2,20-(tetramethylenedicarbonyldioxy)diethyl]bis(3-alkyl-4-methylthiazolium iodide), and [4,40-(1,6-hexamethylenedithio)bis(1-octylpyridinium iodide)], and the amido-amine compound is myristamidopropyldimethylamine.

3. 3. The solution of claim 2, wherein the quaternary ammonium compound is 3,3'-[1,4-phenylenebis(oxy)]bis(1-dodecylpyridinium) dibromide.

4. 3. The solution according to claim 1, wherein the quaternary ammonium compound is N-dodecyl{4,4'-(2,4,8,10-tetraoxaspiro[5.5]undecane-3,9-diyl)}dipyridinium dibromide.

5. 3. The solution of claim 2, wherein the multimeric biguanide compound is poly(hexamethylene biguanide) and the quaternary ammonium compound is polyquaternium-1.

6. A solution according to any one of claims 1 to 5, further comprising one or more of: a chelating agent in an amount ranging from 0.01% w / v to 0.06% w / v, and / or a buffer in an amount ranging from 0.01% w / v to 1% w / v, and / or a viscosity modifier in an amount ranging from 0.01% w / v to 0.25% w / v, and / or a surfactant compound in an amount ranging from 0.01% w / v to 1% w / v, and / or an isotonicity agent in an amount ranging from 0.001% w / v to 1% w / v, and / or optionally a calcium salt in an amount ranging from 0.001% w / v to 0.015% w / v, and / or optionally a magnesium salt in an amount ranging from 0.001% w / v to 0.015% w / v.

7. 7. The solution of claim 6, wherein the chelating agent is ethylenediaminetetraacetic acid or tetrasodium N,N-bis(carboxymethyl)-L-glutamate.

8. 7. The solution of claim 6, wherein the buffer is a borate buffer or a phosphate buffer.

9. 7. The solution of claim 6, wherein the viscosity modifier is selected from the group consisting of hydroxypropyl methylcellulose, carboxymethyl cellulose, and hydroxyethyl cellulose.

10. 10. The solution of claim 9, wherein the viscosity modifier is hydroxypropyl methylcellulose.

11. 7. The solution of claim 6, wherein the viscosity modifier is poly(acryloyl-lysine) or a copolymer containing amino acids whose side chains are acrylated.

12. The solution of claim 6 , wherein the surfactant compound is a poloxamine.

13. 7. The solution of claim 6, wherein the tonicity agent is sodium chloride or a combination of sodium chloride and potassium chloride.

14. The solution of claim 13, wherein the solution further contains mannitol.

15. The solution of claim 6, wherein the isotonic agent is mannitol.

16. A solution according to any one of claims 1 to 15 for use in therapy.

17. A solution according to any one of claims 1 to 15 for use in disinfection, in particular in ocular application.

18. Non-therapeutic use of a solution according to any one of claims 1 to 15 in disinfection, in particular in disinfecting contact lenses.

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