Use of high molecular weight hyaluronic acid as an ophthalmic drug delivery vehicle.

Abstract

To provide a stable aqueous solution of a bioactive agent without the need for any additives not naturally occurring in the human eye. [Solution] The present invention relates to the use of ultra-high molecular weight hyaluronic acid (HMWHA) as a delivery vehicle for bioactive agents, such as ophthalmic drugs and other active pharmaceutical ingredients (APIs), with minimal side effects. One aspect of the invention is an ophthalmic drug delivery system (ODS) comprising an HMWHA fluid and a bioactive agent, wherein the hyaluronic acid is at least 2.5 ml. 3 / kg (i.e., 2.5m 3 / kg or more), wherein the HMWHA fluid is capable of transporting the bioactive agent to the eye; methods for delivering a bioactive agent to the eye; methods for treating, preventing, and / or delaying the onset or recurrence of an ocular disorder in a human or animal subject; and kits that can be used to practice the methods of the invention.

Description

[Technical Field]

[0001] The present invention relates to the use of ultra-high molecular weight hyaluronic acid (HMWHA) as a delivery vehicle for bioactive agents, such as ophthalmic drugs and other active pharmaceutical ingredients (APIs), with minimal side effects and no topical toxicity. [Background technology]

[0002] This application claims the benefit of U.S. Provisional Patent Application No. 63 / 041937, filed June 21, 2020, the entire disclosure of which, including any figures, tables, nucleic acid sequences, amino acid sequences, or drawings, is incorporated herein by reference.

[0003] Eye drops for the topical treatment of ocular diseases such as glaucoma, chronic inflammation, allergies, and atopy consist of an active pharmaceutical ingredient (API) with pharmacological, metabolic, or immunological activity dissolved or suspended in a vehicle. Potential functions of the vehicle include dissolving or suspending the API, stabilizing the solution during storage and patient use, extending the contact time between the API and the ocular surface, supporting the penetration of the API into the ocular surface, and enhancing the biocompatibility of the eye drop [1, 2].

[0004] Most eye drops are aqueous solutions that require surfactants to dissolve additives, especially lipophilic APIs. Eye drops must be sterile during patient use, which can be achieved by either disposable containers (single-dose), bottles with specific dispensers to prevent microbial contamination, or the addition of preservatives such as benzalkonium chloride. Contact time with the ocular surface can be extended by adding polymers that increase the solution's viscosity. Furthermore, mucoadhesive additives such as hyaluronic acid can adhere to the glycocalyx of apical epithelial cells, thereby facilitating contact between the API and the ocular surface. Penetration enhancers weaken the transcellular or paracellular epithelial barrier function, thereby facilitating the diffusion of APIs to the ocular surface. Salts are added to adjust osmolality, and buffers are added to adjust and stabilize the pH of eye drops at physiological levels and to stabilize the eye drops.

[0005] Surfactants can displace cell-associated mucins in the glycocalyx of apical epithelial cells and incorporate into the lipid bilayer that forms the cell membrane, thereby weakening the cell barrier function and supporting the transport of APIs through the membrane into the cells [3, 4]. Surfactants, such as benzalkonium chloride (BAK, cetalkonium chloride) and cationic polymers called polyquaterniums, remain widely used in ophthalmic medicines because of their combined effect of dissolving APIs in aqueous solutions, enhancing their penetration into the ocular surface, and simultaneously protecting the solution from microbial growth. These benefits come at the expense of local irritation and devastating long-term ocular surface disease [5, 6].

[0006] Additives such as ethylenediaminetetraacetic acid sodium salt (EDTA) remove Ca from the tight junctions between epithelial cells. 2+ It depletes ions, thereby weakening the paracellular barrier function of the epithelium.

[0007] Current eye drops for the treatment of glaucoma and chronic ocular inflammation cause serious side effects in a significant proportion of patients. These ocular side effects are caused not only by the API but, to a large extent, by the vehicle used. In clinical trials for regulatory approval and reimbursement, the safety and performance of new topical ophthalmic agents are often tested against the vehicle alone. This strategy can eliminate the adverse effects of the vehicle and enhance the efficacy of the product.

[0008] It would be advantageous to have available a vehicle that can transport an API across the ocular epithelial barrier without damaging the ocular surface or impairing its barrier function, allowing for the use of lower concentrations of the API to achieve the intended therapeutic effect, thereby reducing the inherent side effects that may be associated with the API. Eye drops containing such a vehicle could potentially serve as a platform for the development of next-generation topical ophthalmic drugs to treat sight-threatening diseases such as glaucoma and chronic ocular inflammation. [Prior art documents] [Non-patent literature]

[0009] [Non-Patent Document 1] Morrison, PW and VV Khutoryanskiy, Advances in ophthalmic drug delivery. Ther Deliv, 2014. 5(12): p. 1297-315. [Non-patent document 2] Moiseev, RV, et al., Penetration Enhancers in Ocular Drug Delivery. Pharmaceutics, 2019. 11(7). [Non-patent document 3] Kaur, IP and R. Smitha, Penetration enhancers and ocular bioadhesives: two new avenues for ophthalmic drug delivery. Drug Dev Ind Pharm, 2002. 28(4): p. 353-69. [Non-patent document 4] Patel, A., et al., Ocular drug delivery systems: An overview. World J Pharmacol, 2013. 2(2): p. 47-64. [Non-patent document 5] Patel, PB, et al., Ophthalmic Drug Delivery System: Challenges and Approaches. Systematic Reviews in Pharmacy, 2010. 1(2): p. 113-120. [Non-patent document 6] Burgalassi, S., et al., Cytotoxicity of potential ocular permeation enhancers evaluated on rabbit and human corneal epithelial cell lines. Toxicol Lett, 2001. 122(1): p. 1-8. [Non-Patent Document 7] Dogru, M., et al., Alterations of the ocular surface epithelial mucins 1, 2, 4 and the tear functions in patients with atopic keratoconjunctivitis. Clin Exp Allergy, 2006. 36(12): p. 1556-65. [Non-patent document 8] Dogru, M., et al., Alterations of ocular surface epithelial MUC16 and goblet cell MUC5AC in patients with atopic keratoconjunctivitis. Allergy, 2008. 63(10): p. 1324-3

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[0010] A major challenge in ocular delivery is bypassing the eye's protective barrier so that sufficient therapeutic molecules can penetrate the desired compartment or tissue to treat ocular diseases or exert their pharmacological effects. Traditional drug delivery systems, such as solutions, suspensions, gels, ointments, and inserts, have been investigated for controlled ocular delivery, but they suffer from problems such as poor drainage of eye drops, tear turnover, poor corneal permeability, nasolacrimal drainage, systemic absorption, and blurred vision. Traditional penetration enhancers for ophthalmic drugs enhance corneal absorption by altering the continuity of the corneal epithelial structure. Research has shown that such properties are exhibited by chelating agents, preservatives (e.g., benzalkonium chloride), surfactants, and bile salts. However, these substances exhibit local toxicity, limiting their use in ophthalmic drug formulations. [Means for solving the problem]

[0011] One aspect of the present invention is an ophthalmic drug delivery system (ODS) comprising a HMWHA fluid and a bioactive agent, wherein the hyaluronic acid is at least 2.5 ml 3 / kg (i.e., 2.5m 3 / kg or greater), and the HMWHA fluid comprises an ODS capable of transporting the bioactive agent to the eye. In some embodiments, the ODS is capable of transporting the bioactive agent across the epithelium of the ocular surface. In some embodiments, the ODS is capable of transporting the bioactive agent across the epithelium of the ocular surface and across the glycocalyx and lipid bilayer of the apical epithelial cells of the ocular surface. In some embodiments, the ODS is capable of transporting the bioactive agent across the paracellular barrier into the extracellular matrix (ECM). Any bioactive agent may be utilized. In some embodiments, the bioactive agent may be a small molecule. In other embodiments, the bioactive agent is a biologic such as a nucleic acid, peptide, or protein, or an antibody or antigen-binding fragment thereof. In some embodiments, the bioactive agent is a combination formulation (a combination of a small molecule and a biologic). In some embodiments, the ODS comprises two or more bioactive agents. In some embodiments, the bioactive agent is an anti-glaucoma agent, an anti-allergic agent, or an anti-inflammatory agent.

[0012] The inventors have determined that HA is not an inert thickening polymer, but rather actively contributes to the transport of bioactive agents, such as drugs, to their site of action. As shown in Example 1, 20 micrograms per milliliter of latanoprost dissolved in the HA vehicle of the present invention is more effective at reducing intraocular pressure than 50 micrograms per milliliter of latanoprost in the "gold standard" Xalatan® droplets, which contain benzalkonium chloride, known to transport latanoprost into the eye by weakening the ocular surface barrier. This enhanced efficacy with reduced side effects improves patient compliance.

[0013] The present invention provides a stable aqueous solution of bioactive agent without needing any additives that do not naturally occur in human eye.Therefore, this ophthalmic composition is completely biocompatible, non-sensitizing, and particularly useful for long-term use in delivering bioactive agent to eye.In addition, the HA of ODS can increase the solubility of the bioactive agent contained in this ODS, as demonstrated by latanoprost in Example 1.

[0014] Another aspect of the invention is a method for delivering a bioactive agent to the eye, comprising topically co-administering a HMWHA fluid and a bioactive agent to the ocular surface of a human or animal subject, wherein the hyaluronic acid is at least 2.5 ml 3 The HMWHA fluid has an intrinsic viscosity of 1000 u / kg, and the HMWHA fluid can transport the bioactive agent to the eye. The HMWHA fluid and the bioactive agent may be topically co-administered simultaneously or sequentially in any order as separate formulations or compositions, or may be administered together in the same formulation or composition as the ODS.

[0015] Another aspect of the present invention is a method for treating, preventing, or delaying the onset or recurrence of an ocular disorder in a human or animal subject, comprising topically co-administering to the ocular surface of the human or animal subject a HMWHA fluid and a bioactive agent, wherein the hyaluronic acid is at least 2.5 ml 3 The present invention relates to a method of treating an eye disorder, the method comprising: administering to the eye an HMWHA fluid having an intrinsic viscosity of 1000 u / kg, wherein the bioactive agent is capable of treating, preventing, or delaying the onset or recurrence of the eye disorder; and delivering the bioactive agent to the eye. The HMWHA fluid and the bioactive agent may be topically co-administered simultaneously or sequentially in any order as separate formulations or compositions, or may be administered together in the same formulation or composition as the ODS. DETAILED DESCRIPTION OF THE INVENTION

[0016] The use of hyaluronic acid (HA) in ophthalmic drug vehicles has been suggested in non-patent literature [3, 10-16]. HA has been shown to counteract the irritating effects of substances on the ocular epithelium [17-20]. When hyaluronic acid of three different molecular weights was examined using porcine buccal and vaginal tissues and cell monolayers (Caco-2 cell line), the hyaluronic acid with the lowest molecular weight showed improved mucoadhesion and the highest penetration enhancement on each substrate tested

[25] .

[0017] The inventors have identified high molecular weight HA (HMWHA), i.e., at least 2.5 m 3 / kg(2.5m 3 We propose that HA with an intrinsic viscosity of 0.05% (0.05% saturates / kg or greater) can transport APIs across the ocular epithelial barrier without damaging the ocular surface or impairing its barrier function. The use of HMWHA in drug vehicles allows for the intended therapeutic effect to be achieved at lower concentrations of API, further reducing the inherent side effects of APIs. Eye drops containing HMWHA as a side-effect-free vehicle have the potential to become a platform for the development of next-generation topical ophthalmic drugs to treat sight-threatening diseases. In particular, by serving as a transport vehicle, HMWHA can replace current penetration enhancers in current eye drop formulations and significantly reduce side effects in the long-term topical treatment of ocular diseases such as ocular hypertension, glaucoma, allergy / atopy, and chronic ocular inflammation.

[0018] Healthy ocular surface epithelium is topographically smooth. The lipid bilayer plasma membrane of apical corneal epithelial cells is interwoven by microplicae lined with an anti-adhesive, water-binding, and protective glycocalyx (Wilcox MDP et al., [Non-Patent Document 23], especially Figure 3, originally published by Gipson IK and Argueso P. [Non-Patent Document 24]). The glycocalyx is covered by a mucous tear film composed primarily of membrane-bound mucins, which possess water-binding and lubricating properties due to the dissolved, gel-forming mucin MUC5AC, secreted primarily by conjunctival goblet cells (Non-Patent Documents 7-9). The largest membrane-bound mucin, MUC16, extends from the apex of the microvilli into the mucous tear film, preventing cell adhesion and bacterial attachment and invasion. MUC16 not only plays an important role in cell-epithelial barrier function, but also contributes to tight junctions between epithelial cells and, therefore, paracellular barrier function.

[0019] While not wishing to be bound by theory of its mechanism of action as a delivery vehicle, the inventors propose that, in addition to stabilizing epithelial barrier function, acting as an anti-inflammatory agent, and promoting intimate contact of bioactive agents (e.g., APIs) with the ocular surface, HMWHA may also perform one or more of the following functions: bind to MUC16 in the glycocalyx of apical epithelial cells; bind to the adhesion molecule CD44 on the apical surface of corneal and conjunctival epithelia; bind to the hyaluronan receptor for hyaluronan-mediated motility (RHAMM) on the apical surface of corneal and conjunctival epithelia; increase local tissue hydration, thereby allowing transient cell detachment, which may create a pathway or "highway" that allows cellular migration and transport of bioactive agents along the paracellular pathway; and bind to the HA receptor for endocytosis (HARE) on the apical surface of corneal and conjunctival epithelia, triggering HARE-mediated endocytosis of bioactive agents into the cytoplasm of epithelial cells. Regarding the latter proposed mechanism of action, the presence of HARE receptors on the surface of ocular epithelial cells allows them to internalize HA by endocytosis. This is a new option for transporting bioactive agents with HMWHA molecules as vehicles across epithelial cell membranes without damaging the membranes.

[0020] In subjects, a commercially available eye drop containing 50 μg / ml latanoprost induced a mean intraocular pressure (IOP) reduction of 3.24 mmHg, whereas a prototype eye drop containing 19 μg / ml latanoprost induced a mean IOP reduction of 5.87 mmHg. This finding suggests that the combination of latanoprost and high-molecular-weight hyaluronic acid is more effective at lowering IOP than latanoprost alone.

[0021] Combined with the ability of HA to mitigate the adverse effects of corneal toxicants [17-20], it is anticipated that patients, especially those requiring long-term topical treatment for diseases such as glaucoma, who currently suffer from the side effects of these treatments, will benefit from this new technology using HMWHA.

[0022] The properties of HA in eye drops depend on its chain length and concentration. While the concentration of HA is usually part of the labeling of the final product, it rarely includes information on chain length. This makes it very difficult to correlate the performance of different products reported in the literature. The average chain length or molecular weight of hyaluronic acid molecules is usually determined by gel electrophoresis, size exclusion chromatography, or small-angle light scattering, or calculated from the intrinsic viscosity [η]. Only methods for determining the intrinsic viscosity of hyaluronic acid have been standardized and published in the European and Japanese Pharmacopoeias [21, 22]. Furthermore, ultra-high molecular weight HA (i.e., 2.5 m 3 The clinical performance of eye drops containing low molecular weight HA (1.8m / kg or more) was 3 / kg) ~ Medium molecular weight HA (1.8m 3 / kg~2.5m 3 The clinical performance of eye drops containing HA (less than 1000 kJ / kg) is quite different from that of eye drops containing HA. Therefore, detailed description of the intrinsic viscosity of HA in future publications on HA-containing eye drops is highly recommended.

[0023] The high molecular weight hyaluronic acid or "HMWHA" used in the present invention is a hyaluronic acid having a molecular weight of at least 2.5 ml as measured by the method described in European Pharmacopoeia 9.0 "Sodium Hyaluronate", p. 3584 [Non-Patent Document 21]. 3 / kg (i.e., 2.5m 3 / kg or more). Briefly, the intrinsic viscosity [η] is calculated using the Martin formula: Log 10 (n r -1 / c)=log 10 [η] + κ[η]c, as calculated by linear least squares regression analysis. In some embodiments, the high molecular weight hyaluronic acid has a molecular weight of at least 2.9 m 3 / kg (i.e., 2.9m 3 / kg or more).

[0024] One aspect of the present invention is an ophthalmic drug delivery system (ODS) comprising a HMWHA fluid and a bioactive agent, wherein the hyaluronic acid is at least 2.5 ml 3 / kg, and the HMWHA fluid comprises an ODS capable of transporting the bioactive agent to the eye. In some embodiments, the ODS is capable of transporting the bioactive agent across the epithelium of the ocular surface. In some embodiments, the ODS is capable of transporting the bioactive agent across the epithelium of the ocular surface and across the glycocalyx and lipid bilayer of the apical epithelial cells of the ocular surface. In some embodiments, the ODS is capable of transporting the bioactive agent across the paracellular barrier into the extracellular matrix (ECM). Any bioactive agent may be utilized. In some embodiments, the bioactive agent may be a small molecule. In other embodiments, the bioactive agent is a biologic such as a nucleic acid, peptide, or protein, or an antibody or antigen-binding fragment thereof. In some embodiments, the bioactive agent is a combination formulation (a combination of a small molecule and a biologic). In some embodiments, the ODS comprises two or more bioactive agents. In some embodiments, the bioactive agent is an anti-glaucoma agent, an anti-allergic agent, or an anti-inflammatory agent.

[0025] In some embodiments, the ODS does not include a penetration enhancer.

[0026] In some embodiments, the ODS comprises one or more bioactive agents and a transport vehicle consisting of or consisting essentially of an HMWHA fluid, which can transport the one or more bioactive agents to the eye (e.g., through one or more of the ocular surface epithelium, the glycocalyx and lipid bilayer of the apical epithelial cells of the ocular surface, and the paracellular barrier to the extracellular matrix (ECM)).

[0027] Advantageously, the ODS is storage stable. In some embodiments, the ODS is an aqueous solution that is stable for at least 4 weeks, at least 3 months, or at least 6 months under one or more of the following conditions: (i) a temperature of 15-25°C, (ii) a temperature of 2-8°C, or (iii) a temperature of 25°C at 60% relative humidity.

[0028] Another aspect of the invention is a method for delivering a bioactive agent to the eye, comprising topically co-administering a HMWHA fluid and a bioactive agent to the ocular surface of a human or animal subject, wherein the hyaluronic acid is at least 2.5 ml 3 The HMWHA fluid has an intrinsic viscosity of 1000 u / kg, and the HMWHA fluid can transport the bioactive agent to the eye. The HMWHA fluid and the bioactive agent may be topically co-administered simultaneously or sequentially in any order as separate formulations or compositions, or may be administered together in the same formulation or composition as the ODS.

[0029] Another aspect of the present invention is a method for treating, preventing, or delaying the onset or recurrence of an ocular disorder in a human or animal subject, comprising topically co-administering a HMWHA fluid and a bioactive agent to the ocular surface of the human or animal subject, wherein the hyaluronic acid is at least 2.5 ml 3The present invention relates to a method of treating an eye disorder, the method comprising: administering to the eye an HMWHA fluid having an intrinsic viscosity of 1000 u / kg, wherein the bioactive agent is capable of treating, preventing, or delaying the onset or recurrence of the eye disorder; and delivering the bioactive agent to the eye. The HMWHA fluid and the bioactive agent may be topically co-administered simultaneously or sequentially in any order as separate formulations or compositions, or may be administered together in the same formulation or composition as the ODS.

[0030] In some embodiments, a penetration enhancer is not topically administered to the ocular surface before, during, and / or after topical administration of the bioactive agent and HMWHA.

[0031] In some embodiments, the hyaluronic acid has a concentration of less than 0.2% w / v, 0.1-0.19% w / v, or 0.15% w / v.

[0032] In some embodiments, the HMWHA fluid has the following composition / properties, which correspond to the composition / properties of COMFORT SHIELD® preservative-free sodium hyaluronate eye drops: a) pH 6.8-7.6; b) osmolality 240-330 mOsmol / kg; c) NaCl concentration of 7.6 to 10.5 g / L; and / or d) Phosphate concentration 1.0-1.4 mmol / l

[0033] In some embodiments, the fluid is a clear, colorless solution containing no visible impurities. The fluid is assumed to be sterile.

[0034] In some embodiments, a fluid according to the present invention is COMFORT SHIELD® preservative-free sodium hyaluronate eye drops.

[0035] In some embodiments, the HA has a molecular weight of at least 3 million tons as calculated by the Mark-Houwink formula, hi some embodiments, the HA has a molecular weight in the range of 3-4 million tons as calculated by the Mark-Houwink formula.

[0036] In some embodiments, the HMWHA is hyaluronic acid. In some embodiments, the HMWHA is cross-linked. In some embodiments, the HMWHA is not cross-linked. In some embodiments, the HMWHA is linear. In some embodiments, the HMWHA is non-linear (e.g., branched). In some embodiments, the HMWHA is a derivative of hyaluronic acid, such as an ester derivative, an amide derivative, or a sulfated derivative, or a combination of two or more thereof.

[0037] As used herein in connection with the ODS, methods, and kits of the present invention, the term "bioactive agent" refers to any substance that, when administered in an amount effective to affect tissue, affects a human or non-human animal subject. Bioactive agents can be any type of substance, such as a drug molecule or a biological agent (e.g., polypeptide, carbohydrate, glycoprotein, immunoglobulin, nucleic acid), and can be natural or artificially produced. They can act by any mechanism, such as pharmacological, immunological, or metabolic mechanisms. Examples of types of bioactive agents include substances that regulate intraocular pressure (e.g., enzyme inhibitors) and antiangiogenic agents. Some specific examples of bioactive agents include steroids (e.g., corticosteroids), antibiotics, immunosuppressants, immunomodulators, tacrolimus, plasmin activators, antiplasmins, and cyclosporin A. In some embodiments, the bioactive agent is a steroid or antibiotic to treat, delay the onset of, or prevent an eye infection; a glaucoma or ocular hypertension medication such as a prostaglandin analog, beta-blocker, alpha-agonist, or carbonic anhydrase inhibitor; an allergy eye relief medication such as an antihistamine or nonsteroidal anti-inflammatory drug; or a mydriatic.

[0038] In some embodiments, the ocular disorder to be treated, prevented, or delayed is ocular hypertension or glaucoma, and the bioactive agent is a prostaglandin analog. In some embodiments, the prostaglandin analog is an F2a analog selected from latanoprost, travoprost, bimatoprost, tafluprost, prostaglandin F2a-ethanolamide, bimatoprost (free acid)-d4, bimatoprost-d4, latanoprost ethylamide, unoprostone, and unoprostone isopropyl ester, or a combination of two or more thereof.

[0039] In some embodiments, the bioactive agent is a therapeutic agent such as a small molecule drug or a biologic.

[0040] In some embodiments, the bioactive agent is an anti-glaucoma agent, an anti-allergic agent, or an anti-inflammatory agent.

[0041] In some embodiments, the bioactive agent is hydrophobic and / or poorly water soluble.

[0042] In some embodiments, the bioactive agent is encapsulated or attached to a lysosome or nanoparticle.

[0043] In some embodiments, two or more bioactive agents are used in the ODS, methods, and kits of the present invention, and the HMWHA fluid can deliver the two or more bioactive agents to the eye. For example, the first bioactive agent of the ODS, methods, and kits of the present invention can be a prostaglandin analog, and the second bioactive agent can be another drug that reduces intraocular pressure, for example, a drug that reduces intraocular pressure by a mechanism different from that of the prostaglandin analog. In some embodiments, the additional drug is a beta-adrenergic blocker (e.g., timolol), a cholinergic agonist, a carbonic anhydrase inhibitor (e.g., dorzolamide, brinzolamide), or an adrenergic receptor blocker (e.g., brimonidine). In some embodiments, the additional drug includes timolol (e.g., timolol maleate).

[0044] In some embodiments, the bioactive agent is associated with (e.g., bound to, encapsulated by, or loaded onto) a carrier or delivery system such as a nanoparticle (e.g., nanosphere or nanocapsule), liposome, niosome, discosome, micelle, dendrimer, or hydrogel [Non-Patent Documents 26-28].

[0045] Unfortunately, bioactive agent(s) co-administered with HMWHA fluids can be irritating or damaging to the eye (e.g., cyclosporine A). Advantageously, the HMWHA in the fluid, due to its rheological and other properties, can mitigate and / or protect the eye from the irritating and / or damaging effects of the bioactive agent(s) in the fluid (i.e., the bioactive agent would be more irritating or damaging to the eye if administered without the HMWHA).

[0046] In some embodiments, the ODS does not include any substances other than the bioactive agent(s) that are not naturally occurring in the human eye.

[0047] Preferably, the HMWHA fluid or ODS containing the HMWHA fluid does not contain preservatives or detergents (ie, the fluid is preservative-free and detergent-free).

[0048] In some embodiments, the HMWHA or HMWHA-containing ODS does not contain chemical or oxidative preservatives.

[0049] In some embodiments, the HMWHA or HMWHA-containing ODS does not contain preservatives or detergents that kill susceptible microbial cells by disrupting the lipid structure of the microbial cell membrane, thereby increasing the permeability of the microbial cell membrane.

[0050] In some embodiments, the HMWHA or HMWHA-containing ODS does not contain preservatives or detergents that typically cause damage to corneal tissues such as the corneal epithelium, endothelium, stroma, and membrane interfaces.

[0051] In some embodiments, the HMWHA or HMWHA-containing ODS does not contain one or more (or any) of the following preservatives or detergents: quaternary ammonium preservatives (e.g., benzalkonium chloride (BAK) or cetalkonium chloride), chlorobutanol, edetate disodium (EDTA), polyquaternium-1 (e.g., POLYQUAD™ preservative), stabilized oxidizing agents (e.g., stabilized oxychloro complexes (e.g., PURITE™ preservative)), ionic buffer system preservatives (e.g., SOFZIA™ preservative), polyhexamethylene biguanide (PHMB), sodium perborate (e.g., GENAQUA™ preservative), tyloxapol, and sorbate.

[0052] In some embodiments, the HMWHA fluid is at least essentially mucin-free, or in other words, has a mucin concentration of less than 0.3% w / v.

[0053] The fluid may be administered to the ocular surface of one or both eyes of a subject by any topical administration method. For example, the fluid may be administered as one or more drops from an eye dropper or other device for administering eye drops. The fluid may be self-administered or administered by a third party. The dose administered to the ocular surface as a single or multiple doses varies depending on various factors, including the patient's condition and characteristics, the severity of symptoms, concurrent treatments, frequency of treatment, and the desired effect. For example, one or more drops (e.g., about 30 microliters each) may be administered.

[0054] While administration of 1-3 drops 1-3 times daily may be sufficient for delivery of the bioactive agent in some circumstances, more frequent topical co-administration may be required, e.g., 1-3 drops 4, 5, 6, 7, 8, 9, 10, or more times daily, in some embodiments, 3 or more drops are administered one or more times daily.

[0055] A general aspect of the present invention provides a method for delivering a bioactive agent to the eye, comprising topically co-administering an HMWHA fluid and a bioactive agent to the ocular surface of a human or animal subject, wherein the HMWHA fluid transports the bioactive agent to the eye. A more specific aspect of the present invention provides a method for treating, preventing, or delaying the onset or recurrence of an ocular disorder in a human or animal subject, comprising topically co-administering an HMWHA fluid and a bioactive agent to the ocular surface of a human or animal subject, wherein the bioactive agent can treat, prevent, or delay the onset or recurrence of the ocular disorder, and the HMWHA fluid transports the bioactive agent to the eye.

[0056] In some embodiments of the aforementioned methods of the invention, a penetration enhancer is not topically administered to the ocular surface before, during, and / or after topical administration of the HMWHA and / or bioactive agent, whether in the same formulation as the HMWHA and / or bioactive agent or in a separate formulation.

[0057] In some embodiments of the aforementioned methods of the present invention, a transport vehicle consisting of or consisting essentially of an HMWHA fluid is topically administered to the ocular surface, wherein the HMWHA fluid is capable of transporting the one or more bioactive agents to the eye (e.g., via one or more of the ocular surface epithelium, the glycocalyx and lipid bilayer of the apical epithelial cells of the ocular surface, and the paracellular barrier to the extracellular matrix (ECM)).

[0058] In both of the above-mentioned methods of the present invention, the HMWHA fluid is locally co-administered with a bioactive agent to the ocular surface of a human or animal subject, and the HMWHA fluid transports the bioactive agent to the eye.The HMWHA fluid and the bioactive agent can be locally co-administered as separate formulations simultaneously or sequentially in any order.When co-administered simultaneously, the HMWHA fluid and the bioactive agent can be co-administered as separate formulations, or they can be co-administered together in a single formulation as the ODS of the present invention.

[0059] Whether HMWHA is co-administered with a bioactive agent in the same formulation, as an ODS, or in a separate formulation, the HMWHA fluid is co-administered locally to the ocular surface of the eye in an amount and duration sufficient to act as a transport vehicle for transporting the bioactive agent to the eye. In some embodiments of the method, the HMWHA fluid is co-administered locally to the ocular surface of the eye in an amount and duration sufficient to transport the bioactive agent through the epithelium of the ocular surface. In some embodiments of the method, the HMWHA fluid is co-administered locally to the ocular surface of the eye in an amount and duration sufficient to transport the bioactive agent through the epithelium of the ocular surface and through the glycocalyx and lipid bilayer of the apical epithelial cells of the ocular surface. In some embodiments of the method, the HMWHA fluid is co-administered locally to the ocular surface of the eye in an amount and duration sufficient to transport the bioactive agent through the paracellular barrier into the extracellular matrix (ECM).

[0060] In some embodiments, the HMWHA or ODS is formulated for topical administration to the ocular surface as eye drops, eye washes, or contact lenses (eg, corneal or scleral).

[0061] Another aspect of the present invention is a therapeutic method for treating, preventing, or delaying the onset or recurrence of an ocular disorder in a human or animal subject, comprising topically co-administering a high molecular weight hyaluronic acid (HMWHA) fluid and a bioactive agent to the ocular surface of said human or animal subject, wherein the hyaluronic acid is at least 2.5 ml 3 / kg, wherein the bioactive agent is capable of treating, preventing, or delaying the onset or recurrence of the ocular disorder, and the HMWHA fluid delivers the bioactive agent to the eye.

[0062] In some embodiments, the ocular disorder is glaucoma (including low-toned, normal-toned, and high-toned glaucoma), ocular hypertension, allergies, chronic inflammation, ocular surface disorders, age-related macular degeneration (AMD; atrophic (non-exudative or dry) or neovascular (exudative or wet)), early-onset macular degeneration (e.g., Stargardt's disease), macular telangiectasia, maculopathy (e.g., age-related maculopathy [ARM] and diabetic maculopathy [DMP] (including partial ischemic DMP), macular edema (e.g., diabetic macular edema [DME, including clinically significant DME, focal DME, and diffuse DME], Irvine-Gass syndrome [postoperative macular edema], and macular edema associated with retinal vein occlusion (RVO [including central RVO and branch RVO])), retinopathy (e.g., diabetic retinopathy [DR, including in patients with DME], proliferative vitreoretinopathy [PVR], perforation (Purtscher) retinopathy, and radiation retinopathy), retinal artery occlusion (RAO, e.g., central RAO ​​and branch RAO), retinal vein occlusion (RVO, e.g., central RVO [including central RVO with cystoid macular edema {CME}] and branch RVO [including branch RVO with CME]), ocular hypertension, retinitis (e.g., Coats' disease [exudative retinitis] and retinitis pigmentosa [RP]), chorioretinitis, choroiditis (e.g., creeping choroiditis), uveitis (including anterior uveitis with or without CME, intermediate uveitis, posterior uveitis, panuveitis, and non-infectious uveitis), retinal detachment (e.g., in von Hippel-Lindau disease), retinal pigment epithelium (RPE) detachment, rod and / or cone dystrophies, and diseases associated with increased intracellular or extracellular lipid storage or accumulation in addition to AMD.

[0063] In some embodiments, the ocular disorder is glaucoma or ocular hypertension, and the bioactive agent is a prostaglandin analog or other anti-glaucoma agent that reduces intraocular pressure (e.g., by reducing aqueous humor production or increasing aqueous humor outflow from the intraocular compartment). In some embodiments, the anti-glaucoma agent is a pharmacological agent selected from among miotics or cholinergic agonists (e.g., pilocarpine or eserine), beta-adrenergic antagonists or "beta-blockers" (e.g., timolol maleate or betaxolol), alpha-adrenergic agonists (e.g., epinephrine or dipivefrin), carbonic anhydrase inhibitors (e.g., dorzolamide), Rho-kinase inhibitors (e.g., netarsudil), and prodrugs of prostaglandin F2a (e.g., latanoprost).

[0064] In some embodiments, the bioactive agent is present at a concentration lower than that which would be effective to treat and / or prevent the ocular disorder without the HA (i.e., in the absence of the HA or the bioactive agent alone). For example, in embodiments in which the bioactive agent comprises a prostaglandin analog, such as latanoprost, the prostaglandin analog is present at a concentration lower than that which would be effective to treat and / or prevent ocular hypertension or glaucoma without the HA.

[0065] In some embodiments, the prostaglandin analog is latanoprost and is present at a concentration of less than 50 micrograms per milliliter (less than 50 μg / mL). In some embodiments, the latanoprost is present at a concentration of less than 0.005% by weight, based on the total volume (w / v) of the ophthalmic composition. In some embodiments, the latanoprost is present at a concentration of less than 30 micrograms per milliliter. In some embodiments, the latanoprost is present at a concentration within the range of about 2 micrograms per milliliter to about 45 micrograms per milliliter, about 10 micrograms per milliliter to about 40 micrograms per milliliter, about 15 micrograms per milliliter to about 25 micrograms per milliliter, or about 20 micrograms per milliliter to about 25 micrograms per milliliter. In some embodiments, the latanoprost is present at a concentration of about 20 micrograms per milliliter.

[0066] The prostaglandin analogue may be an F2a analogue such as latanoprost, travoprost, bimatoprost, tafluprost, prostaglandin F2a-ethanolamide, bimatoprost (free acid)-d4, bimatoprost-d4, latanoprost ethylamide, unoprostone, and unoprostone isopropyl ester, or a combination of two or more thereof. In some embodiments, the at least one prostaglandin comprises latanoprost.

[0067] In some embodiments, the at least one prostaglandin comprises latanoprost, and the latanoprost is present at a concentration ranging from about 20 micrograms per milliliter to about 25 micrograms per milliliter. In some embodiments, the at least one prostaglandin comprises latanoprost, and the latanoprost is present at a concentration of about 20 micrograms per milliliter.

[0068] In some embodiments, the prostaglandin analog is bimatoprost and is present at a concentration of less than 100 micrograms per milliliter (less than 100 μg / mL). In some embodiments, the bimatoprost is present at a concentration of less than 90 micrograms per milliliter. In some embodiments, the bimatoprost is present at a concentration within the range of about 5 micrograms per milliliter to about 90 micrograms per milliliter, about 10 micrograms per milliliter to about 80 micrograms per milliliter, about 20 micrograms per milliliter to about 70 micrograms per milliliter, or about 20 micrograms per milliliter to about 60 micrograms per milliliter. In some embodiments, the bimatoprost is present at a concentration of 50 micrograms per milliliter.

[0069] In some embodiments, the prostaglandin analog is travoprost and is present at a concentration of less than 30 micrograms per milliliter (less than 30 μg / mL). In some embodiments, the travoprost is present at a concentration of less than 25 micrograms per milliliter. In some embodiments, the travoprost is present at a concentration within the range of about 2 micrograms per milliliter to about 25 micrograms per milliliter, about 3 micrograms per milliliter to about 25 micrograms per milliliter, about 5 micrograms per milliliter to about 20 micrograms per milliliter, or about 10 micrograms per milliliter to about 15 micrograms per milliliter. In some embodiments, the travoprost is present at a concentration of about 15 micrograms per milliliter.

[0070] In some embodiments, the prostaglandin analog is tafluprost and is present at a concentration of less than 15 micrograms per milliliter (less than 15 μg / mL). In some embodiments, the tafluprost is present at a concentration of less than 12 micrograms per milliliter. In some embodiments, the tafluprost is present at a concentration within the range of about 1 microgram per milliliter to about 12 micrograms per milliliter, about 2 micrograms per milliliter to about 10 micrograms per milliliter, about 2 micrograms per milliliter to about 10 micrograms per milliliter, or about 3 micrograms per milliliter to about 9 micrograms per milliliter. In some embodiments, the tafluprost is present at a concentration of about 7.5 micrograms per milliliter.

[0071] In some embodiments, the prostaglandin analog is unoprostone and is present at a concentration of less than 1500 micrograms per milliliter (less than 1500 μg / mL). In some embodiments, the unoprostone is present at a concentration of less than 1350 micrograms per milliliter. In some embodiments, the unoprostone is present at a concentration within the range of about 50 micrograms per milliliter to about 1350 micrograms per milliliter, about 100 micrograms per milliliter to about 1200 micrograms per milliliter, about 200 micrograms per milliliter to about 1000 micrograms per milliliter, or about 250 micrograms per milliliter to about 900 micrograms per milliliter. In some embodiments, the unoprostone is present at a concentration of about 750 micrograms per milliliter.

[0072] In some embodiments, the eye disorder is an allergy and the bioactive agent is an anti-allergy agent, such as an antihistamine and / or a mast cell stabilizer (eg, ketotifen).

[0073] In some embodiments, the ocular disorder is chronic ocular inflammation and the bioactive agent is an immunosuppressant such as cyclosporin A.

[0074] Preferably, the HMWHA delivers the bioactive agent to an anatomical site within the eye where an ocular disorder is present or potentially may occur, and / or to tissues within the eye that are affected by or potentially may be affected by an ocular disorder.

[0075] In some embodiments, the ocular disorder is a disorder of the anterior segment of the eye, and the HMWHA delivers the bioactive agent to the anterior segment of the eye.

[0076] In some embodiments, the ocular disorder is a disorder of the posterior segment of the eye, and the HMWHA delivers the bioactive agent to the posterior segment of the eye.

[0077] The eye disorder may be at any stage, and may be acute or chronic.For example, HMWHA and bioactive agent may be co-administered at the early stage, intermediate stage, or advanced stage of the eye disorder.The eye disorder may be at any severity (for example, mild, moderate, or severe).

[0078] In some embodiments, the subject to whom the HMWHA fluid and bioactive agent are topically co-administered is a child (e.g., an infant, adolescent, or juvenile) under the age of 18. In other embodiments, the subject is an adult.

[0079] For therapeutic embodiments in which the subject has an ocular disorder at the time of co-administration, the treatment method may include identifying the subject as having the ocular disorder prior to topical co-administration of the HMWHA fluid and the bioactive agent. The subject may be identified by diagnosing the subject with an ocular disorder through one or more tests and / or diagnostic examinations. For example, glaucoma may be detected using one or more of tonometry (measuring intraocular pressure), ophthalmoscopy (examining the optic nerve), perimetry (a visual field test that creates a map of the subject's visual field to identify areas of vision loss), gonioscopy (determining whether the angle where the iris meets the cornea is open, wide, or narrow), and pachymetry (measuring the thickness of the cornea). Intraocular pressure is the best metric for assessing the state of ocular hypertension or glaucoma and its changes, such as progression, stabilization, or improvement (Konstas AG et al., Expert Opinion On Drug Safety, 2021 Apr;20(4):453-466; Kass MA et al., JAMA Ophthalmol, 2021;139(5):558-566; and Allis K et al., Cureus, 2020 Nov;12(11):e11686). Intraocular pressure may be measured using a Goldmann applanation tonometer, the gold standard instrument for measuring intraocular pressure.

[0080] Optionally, the subject is monitored one or more times during and / or after treatment, and the results can be compared with previous results to assess the status and progress of the treatment of the eye disorder.

[0081] Optionally, this method comprises, before administering the HMWHA fluid, identifying the subject as having one or more signs or symptoms of the eye disorder.For example, in the case of glaucoma, signs and symptoms vary according to the type and stage of the disorder.For example, in open-angle glaucoma, some signs and symptoms include: patchy blind spots in the subject's side (peripheral) or central vision, which often appear in both eyes; and tunnel vision in advanced stages.In acute angle-closure glaucoma, some signs and symptoms include: severe headache, eye pain, nausea and vomiting, blurred vision, halos around light, and redness of the eye.

[0082] Another aspect of the present invention relates to kits that can be used to carry out the methods of the present invention described herein, i.e., methods for delivering a bioactive agent to the eye and methods for treating, preventing, or delaying the onset or recurrence of eye disorders. The kits include the HMWHA fluid described herein and, optionally, one or more bioactive agents. If a bioactive agent is included, the bioactive agent may be packaged together with the HMWHA fluid in the same container or packaged separately from the HMWHA fluid in a separate container. Thus, the kit may contain one or more bioactive agents in a separate container from the HMWHA, or they may be contained together in the same container (e.g., "premixed"). Suitable containers include, for example, bottles, vials, syringes, blister packs, and the like. The containers may be formed from a variety of materials, such as glass or plastic.

[0083] The kit may include a delivery agent (separately or together with a fluid) that is contacted with the ocular surface or other portion of the eye. For example, the kit may include particles (e.g., microparticles or nanoparticles) that are coated with the fluid and / or release the fluid onto the ocular surface.

[0084] Optionally, the kit may include a device for dispensing the eye drops (e.g., an eye dropper), which may or may not function as a container for the HMWHA fluid in the kit before the outer packaging of the kit is accessed (e.g., opened). That is, the eye drop dispensing device may function to contain the fluid provided in the unaccessed (unopened) kit, or may be empty and receive fluid after the kit is accessed. Optionally, the kit may include a label or package insert with printed or digital instructions for using the kit, e.g., to practice the methods of the invention.

[0085] The kit can include a packaging material compartmentalized to accommodate one or more containers, such as vials, tubes, etc., each containing one of the separate components used in the methods described herein. Packaging materials for use in packaging pharmaceuticals include, by way of example only, U.S. Patent Nos. 5,323,907, 5,052,558, and 5,033,252. Examples of pharmaceutical packaging materials include, but are not limited to, blister packs, bottles, tubes, pumps, bags, vials, light-tight seals, syringes, bottles, and any packaging material suitable for the selected formulation and intended mode of administration and treatment.

[0086] The kit may contain one or more additional containers, each containing one or more of a variety of materials desirable from a commercial and user standpoint for using the compositions described herein. Non-limiting examples of such materials include, but are not limited to, buffers, diluents, carriers, packages, containers, vials, and / or tube labels that describe the contents and / or instructions for use, and package inserts containing instructions for use.

[0087] A label can be present on or associated with a container. A label can be present on a container when letters, numbers, or other characters forming the label are attached, molded, or etched onto the container itself. A label can be associated with a container when it is present in a receptacle or carrier that also holds the container, for example, as a package insert. A label can be used to indicate that the contents should be used for a specific therapeutic application. A label can also indicate instructions for using the contents, such as in the methods described herein.

[0088] In some embodiments of the kit, the HMWHA fluid can be present in a pack or dispenser device that can contain one or more unit dose forms containing the compositions disclosed herein.The pack can include, for example, metal or plastic foil, such as a blister pack.The pack or dispenser device can be accompanied by instructions for administration.

[0089] Preparation of HMWHA fluid The hyaluronic acid of the fluid is at least 2.5 ml 3 / kg (i.e., 2.5m 3 / kg or more), and preferably has an intrinsic viscosity of less than 0.2% w / v. In some embodiments, the hyaluronic acid has an intrinsic viscosity of at least 2.9 m 3 / kg(2.9m 3 / kg or more).

[0090] Viscoelasticity is defined as the property of a fluid that has both viscous and elastic properties. Zero shear viscosity is determined as the steady shear plateau viscosity at the vanishing shear rate. For high viscosity formulations, measurement using a controlled stress rheometer is preferred.

[0091] m 3The relationship between molecular weight in units of mol / kg and intrinsic viscosity [η] is given by the Mark-Houwink equation: [η]=k·(Mrm) a (Mrm is the molecular weight in MDa, Coefficient k=1.3327·10 -4 and The coefficient a=0.6691, The values ​​of k and a were found to be most predictive.

[0092] HMWHA fluids may be manufactured by sterilizing a filling line; adding purified water or water for injection (WFI) to a stainless steel mixing tank; adding salt while mixing; slowly adding HA and mixing until a homogenous solution / fluid is obtained; optionally, adding one or more bioactive agents; adjusting the pH value, if necessary, by adding NaOH or HCl while continuing the mixing process; transferring the solution to a sterile holding tank through a 1 μm pore size filter cartridge; and aseptically filling the solution into sterile primary packages (single doses or vials) by sterile filtration. For single doses, this may be done by a blow-fill-seal (BFS) process.

[0093] Preferably, the HMWHA fluid is at least essentially mucin-free, or in other words, has a mucin concentration of less than 0.3% w / v, meaning that the flow behavior or flow properties are essentially provided or regulated by hyaluronic acid, rather than by mucins that are naturally present in the subject's tears and are primarily responsible for their flow behavior.

[0094] When viscosity-increasing substances are added, they are preferably added toward, during, or as the final step. Mixing is performed to obtain a uniform mixture. Alternatively, or in addition, it is preferred to first provide purified water or water for injection as a base, and then optionally add electrolytes, buffers, and non-viscosity-increasing substances to the purified water or water for injection first.

[0095] HA is further described in the monograph of the European Pharmacopoeia 9.0, page 3583 ("Sodium Hyaluronate"), which is incorporated herein by reference in its entirety.

[0096] In one embodiment, the fluids used in the ophthalmic drug delivery systems (ODS), methods, and kits of the present invention have the properties listed in Table 1.

[0097] Table 1 [Table 1]

[0098] definition The terms "a," "an," "the," and similar terms used in the context of the present invention (particularly in the context of the claims) should be interpreted as including both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. Thus, for example, a reference to a "cell" or a "bioactive agent" should be interpreted as including both a single cell or a single bioactive agent and a plurality of cells or a plurality of bioactive agents, unless otherwise indicated or clearly contradicted by context. Similarly, the term "or" is intended to include "and" unless the context clearly indicates otherwise. The abbreviation "eg" is derived from the Latin "exempli gratia" and is used herein to indicate a non-limiting example. Thus, the abbreviation "eg" is synonymous with the term "for example."

[0099] The terms "comprising," "including," "having," and "containing" (and grammatical variations thereof) are interchangeable and are open (inclusive) terms that do not exclude the presence of one or more additional elements, components, or process steps not expressly recited. On the other hand, the term "consisting of" and its grammatical variations are closed terms that exclude the presence of other additional elements, steps, or components not expressly recited. The term "consisting essentially of" and its grammatical variations are partially open terms that do not exclude the presence of one or more additional elements, components, or steps, as long as these do not essentially affect the basic and novel characteristics of the invention. Thus, the transitional term "comprising" (or grammatical variations such as "comprises / comprise") includes the term "consisting of" as well as the term "essentially consisting of" and its grammatical variations. The transitional terms / phrases (and any grammatical variations thereof) "comprising," "comprises," "comprise," "consisting essentially of," "consists essentially of," "consisting of," and "consists of" may be used interchangeably to attach the specific meaning associated with each term.

[0100] The term "co-administration," in the context of co-administration of HMWHA and one or more bioactive agents, refers to topical administration of the HMWHA fluid and one or more bioactive agents to the ocular surface, either in the same composition or in separate compositions, simultaneously or sequentially in any order. When administered sequentially, the HMWHA fluid and one or more bioactive agents are administered sufficiently close in time that the HMWHA contributes to the transport of the one or more bioactive agents to the eye.

[0101] The term "effective amount" in the context of the administration fluids of the present invention means the amount of fluid needed to achieve the desired result, eg, the amount needed to deliver a bioactive agent to the eye.

[0102] The term "isolated," when used as a modifier of a composition, means that the composition has been produced by human intervention or has been separated from its naturally occurring in vivo environment. Generally, such isolated compositions are substantially free of one or more materials with which they are normally associated in nature, such as one or more proteins, nucleic acids, lipids, carbohydrates, and cell membranes. "Substantially pure" molecules can be combined with one or more other molecules. Thus, the term "substantially pure" does not exclude combinations of compositions. Substantial purity may be at least about 60% by weight or more of the molecule. Purity may also be about 70% or 80% or more, or even greater, e.g., 90% or more. Purity can be determined by any appropriate method, including, for example, UV spectroscopy, chromatography (e.g., high-performance liquid chromatography (HPLC), gas phase), gel electrophoresis (e.g., silver or Coomassie staining), and sequence analysis (for nucleic acids and peptides).

[0103] As used herein, the term "hyaluronic acid" (HA) refers to the naturally occurring glycosaminoglycan, also known as hyaluronan, composed of disaccharide repeats of N-acetylglucosamine and glucuronic acid (e.g., a linear glycosaminoglycan polymer formed from repeating units of the disaccharide [-D-glucuronic acid-β1,3-N-acetyl-D-glucosamine-β1,4-]n), as well as derivatives of hyaluronic acid with chemical modifications such as esters, amide derivatives, alkylamine derivatives, low- and high-molecular-weight forms of hyaluronic acid, and crosslinked forms such as hylan. Thus, the disaccharide chains may be linear or nonlinear. Hyaluronic acid can be crosslinked by attaching crosslinkers such as thiols, methacrylates, hexadecylamides, and tyramine. Hyaluronic acid can also be directly crosslinked with formaldehyde and divinyl sulfone.Examples of hyaluronans include hyaluronan A, hyaluronan B, and hyaluronan G-F20 (Hargittai M and I Hargittai, "More Conversations with Hyaluronan Scientists," from Hyaluronan - From Basic Science to Clinical Applications, Balazs EA, Ed., Vol. 3, 2011, PubMatrix, Edgewater, NJ; Cowman MK et al., Carbohydrate Polymers 2000, 41:229-235; Takigami S et al., Carbohydrate Polymers 1993, 22:153-160; Balazs EA et al., "Hyaluronan, its cross-linked derivative - Hylan - and their medical applications," in Cellulosics Utilization: Research and Rewards in Cellulosics, Proceedings of Nisshinbo International Conference on Cellulosics Utilization in the Near Future (Eds. Inagaki, H and Phillips GO), Elsevier Applied Science (1989), NY, pp. 233-241; Koehler L et al., Scientific Reports, 2017, 7, article no. 1210; and Pavan M et al., Carbohydr Polym, 2013, 97(2): 321-326, each of which is incorporated herein by reference in its entirety.

[0104] The term "hyaluronic acid" or HA includes HA itself and its pharmaceutically acceptable salts, such as sodium hyaluronate. HA can be formulated into a pharmaceutically acceptable salt form. Pharmaceutically acceptable salts of HA can be prepared using conventional techniques.

[0105] The term "high molecular weight" or "HMW" in the context of the hyaluronic acid of the present invention means a hyaluronic acid having a molecular weight of at least 2.5 mg / L as determined by the method of the European Pharmacopoeia 9.0 ("Sodium Hyaluronate"), page 3584, which is incorporated herein by reference in its entirety. 3 / kg (i.e., 2.5m 3 / kg or more). Briefly, the intrinsic viscosity [η] is calculated using the Martin formula: Log 10 (n r -1 / c)=log 10 [η] + κ[η]c, as calculated by linear least squares regression analysis. In some embodiments, the high molecular weight hyaluronic acid has a molecular weight of at least 2.9 m 3 / kg (i.e., 2.9m 3 / kg or more).

[0106] As used herein, the term "ocular disorder" is intended to broadly include any abnormality (e.g., disease, illness, trauma) of the eye that may benefit from a co-administered bioactive agent (therapeutically or prophylactically). The disorder may be at any stage, and may be an acute or chronic disorder. For example, the HMWHA and bioactive agent may be co-administered at an early, intermediate, or advanced stage of the ocular disorder. The disorder may be of any severity (e.g., mild, moderate, or severe). In some embodiments, the ocular disorder is a disorder of the anterior segment, posterior segment, or both.

[0107] As used herein, the term "ocular surface" refers to the cornea and conjunctiva, and portions thereof, including the conjunctiva overlying the upper and lower eyelids. The HMWHA fluid and one or more bioactive agents may be co-administered topically to one or more portions of the ocular surface (e.g., including the entire ocular surface).

[0108] As used herein, the term "penetration enhancer" refers to an agent that can enhance the delivery of a bioactive agent, such as a drug, across an otherwise impermeable or permeability-limited membrane, such as the cornea, by any mechanism of action, such as acting on the epithelium. A penetration enhancer may be any type of substance that functions as a penetration enhancer. For example, a penetration enhancer may be a drug molecule or a biological agent, may be natural or artificially produced, and may act by any mechanism to enhance penetration, either by itself or in conjunction with another agent. Specific examples of penetration enhancers and classes of penetration enhancers are identified in Moiseev, RV et al., "Penetration Enhancers in Ocular Drug Delivery," Pharmaceutics, 2019. 11(7), incorporated herein by reference in its entirety. Examples include, but are not limited to, cyclodextrins, chelating agents, preservatives (such as benzalkonium chloride), surfactants, crown ethers, bile acids, bile salts, cell-penetrating peptides, and other amphiphilic compounds.

[0109] " Pharmaceutically acceptable salt " includes both acid addition salt and base addition salt. Any one of the pharmaceutically acceptable salts of HA or other compounds described herein is intended to include any and all pharmaceutically suitable salt forms.Preferred pharmaceutically acceptable salts described herein are pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts.

[0110] "Pharmaceutically acceptable acid addition salts" refer to salts formed with inorganic acids, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, hydroiodic acid, hydrofluoric acid, phosphorous acid, and the like, which retain the biological effectiveness and properties of the free base and are not biologically or otherwise undesirable. Also included are salts formed with organic acids, such as aliphatic mono- and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxyalkanoic acids, alkanedioic acids, aromatic acids, aliphatic and aromatic sulfonic acids, and the like, including, for example, acetic acid, trifluoroacetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and the like. Thus, exemplary salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate, trifluoroacetate, propionate, caprylate, isobutyrate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, mandelate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, phthalate, benzenesulfonate, toluenesulfonate, phenylacetate, citrate, lactate, malate, tartrate, methanesulfonate, etc. Also contemplated are salts of amino acids such as alginate, gluconate, and galacturonate (see, e.g., Berge SM et al., [Non-Patent Document 29], incorporated herein by reference in its entirety). Acid addition salts of basic compounds may be prepared by contacting the free base form with a sufficient amount of the desired acid to produce the salt according to methods and techniques familiar to those skilled in the art.

[0111] "Pharmaceutically acceptable base addition salts" refer to salts that retain the biological effectiveness and properties of the free acids and are not biologically or otherwise undesirable. These salts are prepared by adding an inorganic or organic base to the free acid. Pharmaceutically acceptable base addition salts may be formed with metals or amines, such as alkali and alkaline earth metals or organic amines. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts, and the like. Salts derived from organic bases include, but are not limited to, salts of primary, secondary, and tertiary amines, substituted amines such as naturally occurring substituted amines, cyclic amines, and basic ion exchange resins, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, N,N-dibenzylethylenediamine, chloroprocaine, hydrabamine, choline, betaine, ethylenediamine, ethylenedianiline, N-methylglucamine, glucosamine, methylglucamine, theobromine, purines, piperazine, piperidine, N-ethylpiperidine, polyamine resins, and the like (see Berge et al., supra). In some embodiments, the pharmaceutically acceptable salt is a sodium salt (see European Pharmacopoeia 9.0, p. 3583, "Sodium Hyaluronate," incorporated herein by reference).

[0112] As used herein, the terms "subject," "patient," and "individual" refer to a human or non-human animal. A subject also refers to, for example, a primate (e.g., a human), cow, sheep, goat, horse, dog, cat, rabbit, rat, mouse, fish, bird, etc. In some embodiments, the subject is a mammal. In some embodiments, the subject is a human. In some embodiments, the subject is a bird or fish. Thus, the method may be practiced in medical and veterinary settings. The non-human animal subject may be, for example, a pet or animal model of an ocular or non-ocular disease. In some embodiments, the subject is an adult. In other embodiments, the subject is a child (e.g., an infant, adolescent, or juvenile) under the age of 18.

[0113] The term "topical administration" is used herein in its conventional sense to mean local delivery to a desired anatomical site, such as the ocular surface. A fluid containing high molecular weight hyaluronic acid may be applied directly or indirectly to the ocular surface by any method that allows an effective amount of the fluid to contact the ocular surface. For example, the fluid may be applied directly to the ocular surface via eye drops or irrigation solution, or indirectly via a delivery agent (i.e., a fluid delivery agent) that is brought into contact with the ocular surface or other parts of the eye. An example of a delivery agent is a particle (e.g., a microparticle or nanoparticle) that is coated with the fluid and / or releases the fluid onto the ocular surface. Such particles may be composed of various materials, such as natural or synthetic polymers. In some embodiments, the delivery agent itself may be administered as drops.

[0114] The terms "treat," "treating," and "treatment" include alleviating, ameliorating, slowing the progression of, reversing, or arresting a medical condition, such as an eye disorder, or one or more symptoms or complications associated with the condition, and alleviating, ameliorating, or eradicating one or more causes of the condition.

[0115] The present invention is described by way of example only by the embodiments in the specification and drawings, and is not limited thereto, but includes all variations, modifications, substitutions and combinations that a specialist may take from the full text of this application in view of and / or in combination with his particular knowledge.

[0116] All patents, patent applications, provisional applications, and publications mentioned or cited herein are hereby incorporated by reference in their entirety, including any drawings and tables, to the extent they do not contradict the explicit teachings of this specification.

[0117] The following are examples illustrating procedures for carrying out the present invention. These examples should not be construed as limiting. Unless otherwise noted, all percentages are by weight and all solvent mixture proportions are by volume.

[0118] ( Example 1 - Comparison of the combination of latanoprost and high molecular weight hyaluronic acid with latanoprost alone in reducing intraocular pressure ) Materials and Methods The sterile bulk solution for manufacturing COMFORT SHIELD® MDS eye drops (i.com medical GmbH, Munich, Germany) was used as a vehicle for the preparation of a prototype latanoprost bulk solution (PLBS). The vehicle consisted of 0.15% w / v hylan A (2.9 mmol / L) dissolved in phosphate-buffered saline (8.035 g / L NaCl; 1.2 mmol / L NaHPO / NaHPO; pH 7.4). 3 The PLBS contained HA (HA with an intrinsic viscosity of 100000000 / kg). Latanoprost was obtained from Yonsung Fine Chemicals Co. Ltd. (Gyeonggi-do, Korea). PLBS was prepared by medi-pharm Laboratorium GmbH (Falkensee, Germany) by dissolving 20 ± 1 μg / ml of latanoprost in the vehicle.

[0119] Sterile 10 ml bottles with ophthalmic squeeze dispensers (OSDs) were obtained from Aptar Radolfzell GmbH (Radolfzell, Germany). Medi-pharm Laboratorium GmbH prepared two batches of prototype latanoprost test samples (PLTS-A) for stability screening by aseptically filling 9 ml of PLBS into the Aptar bottles and closing them with the OSDs. Sterile Novelia® 11 ml soft bottles with a 1.6 mm valve diameter and PureFlow® 1500 droppers were obtained from Nemera (La Verpilliere, France). Pharmpur GmbH (Königsbrunn, Germany) prepared prototype latanoprost test samples (PLTS-N) for IOP self-testing by aseptically filling 10 ml of PLBS into sterile Novelia bottles and closing them with the droppers. Because latanoprost tends to adsorb to the silicone contained in the dropper, these test samples had to be held upright to minimize contact between the solution and the dropper. The latanoprost concentration in the PLTS-N bottle used for self-testing was 19 μg / ml.

[0120] Xalatan® eye drops (PFIZER OFG Germany GmbH, Berlin, Germany) containing 50 μg / ml latanoprost, 0.2 mg / ml benzalkonium chloride, and 6.3 mg / ml phosphate were used as a control sample in the IOP self-test.

[0121] COMFORT SHIELD® MDS 0.15% Hylan A eye drops (i.com medical GmbH, Munich, Germany), consisting of the vehicle of the prototype latanoprost test sample, were used as a control during the washout period of the IOP self-test.

[0122] subject The subject (TS) was a 71-year-old male with a healthy ocular surface, no history of ocular trauma or ocular surgery, or use of stored eye drops, and untreated ocular hypertension not related to glaucoma.

[0123] method In preliminary studies, 25 μg / ml and 50 μg / ml of latanoprost were added to the vehicle, and the solution was stirred at 40° C. for 18 hours. The latanoprost content was determined by HPLC using a 5 μm, 150.0 × 4.0 mm Hypersil BDS C18 column (VDS optilab) and a UV detector (200 nm). Regardless of the starting amount, 20.8 μg / ml of latanoprost was found to dissolve in the vehicle, compared with a water solubility of 12.9 μg / ml (PubChem Compound Summary for CID 5311221, Latanoprost, National Center for Biotechnology Information).

[0124] Samples from two batches of PLTS-A were stored for 6 months at room temperature (15-25°C), 2-8°C, 25°C / 60% relative humidity (RH), and 40°C / 75% RH. Initially, after 4 weeks, 3 months, and 6 months, samples were visually inspected for appearance (clarity) and the presence of particles, and tested for pH, latanoprost content, and weight loss.

[0125] Throughout the self-examination, IOP was measured using a handheld icare HOME Model rebound tonometer TA022 (Icare Finland Oy, Vantaa, Finland) (Liu, J. et al., Icare Home Tonometer: A Review of Characteristics and Clinical Utility, Clin Ophthalmol, 2020. 14: pp. 4031-4045). Three measurements were taken, and the average value was recorded.

[0126] IOP is known to fluctuate significantly over a 24-hour period (circadian period), and the time of peak IOP varies among patients (Barkana, Y. et al., "Clinical utility of intraocular pressure monitoring outside of normal office hours in patients with glaucoma," Arch Ophthalmol, 2006. 124(6): pp. 793-7; Mansouri, K. et al., "Review of the measurement and management of 24-hour intraocular pressure in patients with glaucoma," Surv Ophthalmol, 2020. 65(2): pp. 171-186). Little is known about daily IOP fluctuations. Therefore, TS measured IOP in both eyes for 7 consecutive days at 08:00 (8:00 AM), 11:00 (11:00 AM), 15:00 (3:00 PM), 19:00 (7:00 PM), and 22:00 (10:00 PM). The individual time of peak IOP (11:00) of the TS was selected for IOP monitoring throughout the screening test.

[0127] Eight weeks before the self-study, TSs applied one drop of COMFORT SHIELD eye drops (vehicle) to each eye in the morning and evening. The self-study lasted for 5 weeks, with IOP measurements in both eyes taken daily at 11:00 AM. During weeks 1, 3, and 4, vehicle was applied in the morning and evening. During weeks 2 and 5, vehicle was applied only in the morning (7:00 AM–8:00 AM), and one drop of latanoprost eye drops was applied to each eye in the evening between 7:00 PM and 8:00 PM. During week 2, Xalatan® eye drops (50 μg / ml latanoprost) were applied, and during week 5, PLTS-N eye drops (19 μg / ml latanoprost) were applied.

[0128] result Stability Screening The results of the stability screening for two batches of prototype latanoprost test sample (PLTS-A) are summarized in Tables 2 and 3.

[0129] Table 2. Stability test results for PLTS-A, batch E030219 [Table 2]

[0130] Table 3. Stability test results for PLTS-A, batch E040219 [Table 3] *nd=not determined

[0131] IOP self-test To study individual circadian rhythms and diurnal variations, TS underwent IOP measurements for 7 consecutive days. IOP in TS peaked in the late morning and then decreased continuously until the evening (see Table 4).

[0132] Table 4. Circadian and diurnal variations of IOP in the right eye (OD) and left eye (OS) of TS. [Table 4]

[0133] Therefore, we performed IOP measurements at 11:00 AM to compare the effectiveness of latanoprost eye drops in eyes with TS. Because significant differences were observed between days, IOP measurements were performed with and without latanoprost eye drops for 7 consecutive days. The results of the self-test are summarized in Table 5.

[0134] Table 5. IOP values ​​before (week 1) and during (week 2) application of commercially available latanoprost eye drops 50 μg / mL, and before (week 4) and during (week 5) application of PLTS-N 19 μg / mL latanoprost eye drops. [Table 5]

[0135] Application of commercially available eye drops containing 50 μg / ml latanoprost reduced intraocular pressure by 3.24 mmHg from a mean baseline value of 27.62 mmHg to a mean value of 24.38 mmHg, whereas application of the prototype latanoprost test sample PLTS-N, containing only 19 μg / ml latanoprost, reduced IOP by 5.87 mmHg from a mean baseline value of 27.30 mmHg to a mean value of 21.43 mmHg.

[0136] conclusion In human subjects' (TS) eyes, a commercially available eye drop containing 50 μg / ml latanoprost reduced IOP by a mean of 3.24 mmHg, whereas the prototype eye drop PL20 containing 19 μg / ml latanoprost reduced IOP by a mean of 5.87 mmHg. These findings suggest that the combination of latanoprost and high molecular weight hyaluronic acid is more effective at lowering IOP than latanoprost alone.

[0137] Example 2 - Cyclosporine and Ketotifen as Active Ingredients in HMWHA Fluid as Matrix Solution The immunosuppressant cyclosporine is used in ophthalmology for the treatment of severe keratitis in adults with dry eye that has not improved despite treatment with tear substitutes, for example, marketed by Santen GmbH under the trade name IKERVIS® 1 mg / ml eye drops.

[0138] Cyclosporine (C 62 H 111 N 11 O 12Cyclosporine (Cyclosporin, molecular weight 1202.62) is an 11-amino acid, pH-neutral cyclic peptide with strong hydrophobic / lipophilic properties. It is practically insoluble in water. These properties make it extremely difficult to formulate an effective and ophthalmically acceptable preparation. In principle, it is possible to prepare cyclosporine for ocular application in pharmaceutical dosage forms: suspension, emulsion, or solution. The literature contains various suggestions for the formulation of cyclosporine eye drops. Numerous patent applications have also proposed formulations and claimed specific compositions as inventions.

[0139] When formulating cyclosporine eye drops, the following problems are encountered:

[0140] 1. Emulsion Preparation Cyclosporine is dissolved in a lipophilic solvent, such as a medium-chain triglyceride, and emulsified in water using a surface-active substance (surfactant). Generally, a relatively high proportion of surfactant is required to obtain a stable emulsion. Surfactants act like soap and are poorly tolerated by the eye. Redness, burning, itching, and a foreign body sensation often occur. They also adversely affect the stability of the tear film. Santen's Ikervis® product is an emulsion eye drop.

[0141] 2. Solution Preparation Due to its chemical nature, cyclosporine is practically insoluble in water. Therefore, the active ingredient can only be dissolved in lipophilic solvents. Medium-chain triglycerides (neutral oils) or vegetable oils such as castor oil are suitable. Oil-based eye drops are poorly tolerated by the eye and cause significant visual disturbances after application. Therefore, they are usually rejected by patients. In recent years, the complexation of water-insoluble substances with so-called cyclodextrins has also been discussed extensively. The active substance cyclodextrin complexes obtained in this way can exhibit improved water solubility. However, many questions remain to be clarified for widespread ocular application, such as long-term stability, ocular compatibility during prolonged use, and release of the active ingredient from the complex, which are prerequisites for efficacy (bioavailability).

[0142] 3. Formulation as a suspension Water-insoluble active ingredients can be micronized to achieve a fine dispersion. However, without additional additives, the micronized active ingredient particles settle virtually immediately and become stable as aggregates that are difficult or impossible to shake. These formulation problems are usually alleviated by adding thickening additives and surfactants. Cellulose derivatives, such as methylcellulose (MC) or methylhydroxypropylcellulose (MHPC), are often used as thickening additives. However, to reduce the tendency to settle, these additives must be used at relatively high concentrations. This can lead to adhesion and incompatibility when applied to the eye.

[0143] The development of a new formulation for cyclosporine eye drops led to the idea of ​​using COMFORT SHIELD® eye drops (i.com medical GmbH) as the base (matrix solution). COMFORT SHIELD® is a hyaluronic acid-containing eye drop with a relatively low viscosity. These have been used very successfully in the treatment of dry eye and are very well tolerated.

[0144] Cyclosporine was introduced as a fine powder into the COMFORT SHIELD® matrix solution at a concentration of 0.05% and uniformly dispersed by stirring. A small amount of polysorbate 80 (a well-tolerated surfactant) was added to improve the wettability of the active ingredient particles. Surprisingly, the active ingredient particles were rapidly and uniformly dispersed in the matrix solution. Only after prolonged standing, up to several hours, did slight settling occur. A brief shaking was sufficient to re-establish a uniform suspension. The dreaded "caking," a solid, barely agitated aggregate precipitate, could not be observed even after several weeks of standing.

[0145] Recipe: Cyclosporine: 500mg Tween 80: 2.5ml Matrix solution: 1000 ml

[0146] manufacturing First, 2.5 ml of Tween 80 was added to a total of 700 ml of matrix solution. The mixture was stirred until the Tween 80 was completely dissolved. Subsequently, a total of 500 mg of cyclosporin was transferred to the remaining approximately 300 ml of matrix solution. The entire preparation was then stirred with a magnetic stirrer until the active substance cyclosporin was completely and uniformly dispersed.

[0147] The cyclosporine suspension was filled into special containers for unpreserved eye drops. These containers consisted of conventional plastic bottles for eye drops with a maximum volume of 10 ml and a top that prevented the entry of bacteria during drop withdrawal. By utilizing this innovative concept, it was possible to avoid the addition of preservatives.

[0148] Two batches of cyclosporine eye drops were manufactured and filled in this manner (Batch E281019-1 and Batch E281019-2).

[0149] Samples of cyclosporine eye drops were stored under different climatic conditions to test their shelf life. The conditions for these stability tests were as follows: Storage conditions Test points 2°C~8°CT=0, T=4 weeks, T=3 months 15°C~25°CT=0, T=4 weeks, T=3 months 25°C / 60%rH T=0, T=4 weeks, T=3 months 40°C / 75%rH T=0, T=4 weeks, T=3 months rH = relative humidity T = test time (immediately after production, T = 0)

[0150] Discussion of stability results The determination of the content of the active substance cyclosporine was carried out using an HPLC method against a cyclosporine standard. Further test parameters were the appearance and pH value of the suspension.

[0151] Batch E281019-1 The pH value of the suspension immediately after production at test time T=0 was 7.21. The cyclosporine content was 100.89%. The appearance of the suspension was white to creamy, and the suspension could be shaken without any problems.

[0152] The pH value was 6.23 when stored at 2°C to 8°C for 4 weeks. The cyclosporine content was 100.31%. The appearance of the suspension remained white to cream-colored, and the suspension could be shaken without any problems.

[0153] The pH value was 6.93 when stored at 2°C to 8°C for 3 months. The cyclosporine content was 96.71%. The appearance of the suspension remained white to cream-colored, and the suspension could be shaken without any problems.

[0154] The pH was 6.27 when stored at 15°C to 25°C for 4 weeks. The cyclosporine content was 99.63%. The appearance of the suspension remained white to cream-colored, and the suspension could be shaken without any problems.

[0155] The pH value was 6.93 when stored at 15°C to 25°C for 3 months. The cyclosporine content was 94.04%. The appearance of the suspension remained white to cream-colored, and the suspension could be shaken without any problems.

[0156] The pH value was 6.29 when stored at 25°C / 60% rH for a test time of T = 4 weeks. The cyclosporine content was 98.62%. The appearance of the suspension remained white to cream-colored, and the suspension could be shaken without any problems.

[0157] The pH value was 6.94 when stored at 25°C / 60% rH at test time T = 3 months. The cyclosporine content was 97.88%. The appearance of the suspension remained white to cream-colored, and the suspension could be shaken without any problems.

[0158] The pH value was 6.24 when stored at 40°C / 75% rH for a test time of T = 4 weeks. The cyclosporine content was 93.47%. The appearance of the suspension remained white to cream-colored, and the suspension could be shaken without any problems.

[0159] The pH value was 6.71 when stored at 40°C / 75% rH at test time T = 3 months. The cyclosporine content was 98.37%. The appearance of the suspension remained white to cream-colored, and the suspension could be shaken without any problems.

[0160] The results indicate a good shelf life of the preparation.

[0161] The pH value of approximately 7.2 initially measured after manufacture dropped to approximately 6.3 within four weeks and rose again to approximately 6.9 after three months of storage. The pH value measured after four weeks of storage was clearly incorrect; most likely, the pH meter used was not properly calibrated. The cyclosporine content did not decrease significantly under any storage conditions up to three months of storage. Storage at 40°C / 75% rH (stress test) was unremarkable. The somewhat lower content value after four weeks of storage under these stress conditions is still within the approximate accuracy of the measurement.

[0162] Batch E281019-2 The pH value of the suspension immediately after production at test time T=0 was 7.36. The cyclosporine content was 100.43%. The appearance of the suspension was white to creamy, and the suspension could be shaken without any problems.

[0163] The pH value was 6.26 when stored at 2°C to 8°C for 4 weeks. The cyclosporine content was 99.17%. The appearance of the suspension remained white to cream-colored, and the suspension could be shaken without any problems.

[0164] The pH value was 6.93 when stored at 2°C to 8°C for 3 months. The cyclosporine content was 96.87%. The appearance of the suspension remained white to cream-colored, and the suspension could be shaken without any problems.

[0165] The pH value was 6.29 when stored at 15°C to 25°C for 4 weeks. The cyclosporine content was 78.86%. The appearance of the suspension remained white to cream-colored, and the suspension could be shaken without any problems.

[0166] The pH value was 6.93 when stored at 15°C to 25°C for 3 months. The cyclosporine content was 93.65%. The appearance of the suspension remained white to cream-colored, and the suspension could be shaken without any problems.

[0167] The pH value was 6.26 when stored at 25°C / 60% rH for a test time of T = 4 weeks. The cyclosporine content was 57.12%. The appearance of the suspension remained white to cream-colored, and the suspension could be shaken without any problems.

[0168] The pH value was 6.90 when stored at 25°C / 60% rH at test time T = 3 months. The cyclosporine content was 73.56%. The appearance of the suspension remained white to cream-colored, and the suspension could be shaken without any problems.

[0169] The pH value was 6.23 when stored at 40°C / 75% rH for a test time of T = 4 weeks. The cyclosporine content was 92.65%. The appearance of the suspension remained white to cream-colored, and the suspension could be shaken without any problems.

[0170] The pH value was 6.80 when stored at 40°C / 75% rH at test time T = 3 months. The cyclosporine content was 97.12%. The appearance of the suspension remained white to cream-colored, and the suspension could be shaken without any problems.

[0171] The results appear to be partially contradictory. However, the measured pH values ​​support the observations made for batch E281019-1 that the pH meter was not properly calibrated.

[0172] The pH value of approximately 7.2 initially measured after production dropped to approximately 6.3 within four weeks and rose again to approximately 6.9 after three months of storage. The pH value measured after four weeks of storage was clearly incorrect. Most likely, the pH meter used was not properly calibrated (see batch E281019-1). The cyclosporine content did not decrease significantly up to three months of storage under any of the storage conditions. Storage at 40°C / 75% rH (stress test) showed no significant results. The somewhat lower content values ​​after four weeks of storage under these stress conditions are still roughly within the accuracy of the measurements.

[0173] Table 6. Cyclosporine A eye drops, 10 ml, batch number E281019-1, store at 15°C to 25°C [Table 6]

[0174] Table 7. Cyclosporine A eye drops, 10 ml, batch number E281019-1, store at 2°C to 8°C [Table 7]

[0175] Table 8. Cyclosporine A eye drops, 10 ml, batch number 281019-1, stored at 25°C / 60% RH [Table 8]

[0176] Table 9. Cyclosporine A eye drops, 10 ml, batch number E281019-1, stored at 40°C / 75% RH [Table 9]

[0177] Table 10. Cyclosporine A eye drops, 10 ml, batch number E281019-2, store at 15°C to 25°C [Table 10]

[0178] Table 11. Cyclosporine A eye drops, 10 ml, batch number E281019-2, store at 2°C to 8°C [Table 11]

[0179] Table 12. Cyclosporine A eye drops, 10 ml, batch number 281019-2, stored at 25°C / 60% RH [Table 12]

[0180] Table 13. Cyclosporine A eye drops, 10 ml, batch number E281019-2, stored at 40°C / 75% RH [Table 13]

[0181] summary Two batches of cyclosporine eye drops (Batch E281019-1 and E281019-2) were prepared, filled into special eye drop bottles for unpreserved eye drops, and stored under different climatic conditions for 3 months. The stability of the preserved samples was tested for shelf life immediately after manufacture and after 4 weeks and 3 months of storage. Both batches showed good stability up to 3 months of storage, even when stored under stress conditions (40°C / 75%RH).

[0182] The present formulation is an improvement over formulations described in the literature and over the commercially available formulation (Ikervis® eye drops) since, with the exception of 2.5 ml of Tween 80, it is possible to omit the adjuvants normally required for suspension eye drops, such as cellulose derivatives to increase viscosity or surfactants to wet the suspended active ingredient particles.

[0183] Two batches of ketotifen (Batch E180419-1 and E180419-2) were prepared, filled identically into special eye drop bottles for unpreserved eye drops, and stored under different climatic conditions for 3 to 6 months.

[0184] Table 14. Ketotifen eye drops, 9 ml, batch number E180419-1, store at 15°C to 25°C [Table 14]

[0185] Table 15. Ketotifen eye drops, 9 ml, batch number E180419-1, store at 2°C to 8°C [Table 15]

[0186] Table 16. Ketotifen eye drops, 9 ml, batch number E180419-1, stored at 25°C / 60% RH [Table 16]

[0187] Table 17. Ketotifen eye drops, 9 ml, batch number E180419-1, stored at 40°C / 75% RH [Table 17]

[0188] Table 18. Ketotifen eye drops, 9 ml, batch number E180419-2, store at 15°C to 25°C [Table 18]

[0189] Table 19. Ketotifen eye drops, 9 ml, batch number E180419-2, store at 2°C to 8°C [Table 19]

[0190] Table 20. Ketotifen eye drops, 9 ml, batch number E180419-2, stored at 25°C / 60% RH [Table 20]

[0191] Table 21 Ketotifen eye drops, 9 ml, batch number E180419-2, stored at 40°C / 75% RH [Table 21]

[0192] It should be understood that the examples and embodiments described herein are for illustrative purposes only, and that various modifications or changes in light thereof will be suggested to those skilled in the art, and are within the spirit and scope of this application and the appended claims. Furthermore, any element or limitation of any invention or embodiment thereof disclosed herein may be combined (individually or in any combination) with any and / or all other elements or limitations of any other invention or embodiment thereof disclosed herein, and all such combinations are intended to be within the scope of the present invention, without limitation thereto.

Claims

1. 1. An ophthalmic drug delivery system (ODS) comprising a fluid comprising high molecular weight hyaluronic acid and a bioactive agent, The high molecular weight hyaluronic acid has a molecular weight of at least 2.5 m ３ / kg and a molecular weight in the range of 3 million to 4 million tonnes; the bioactive agent is latanoprost and is present in the ODS at a concentration of 2 micrograms per milliliter to 45 micrograms per milliliter, or at a concentration of less than 30 micrograms per milliliter; The fluid is capable of transporting the bioactive agent into the eye. ODS.

2. The ODS according to claim 1, The latanoprost is present in the ODS at a concentration of less than 30 micrograms per milliliter. ODS.

3. The ODS according to claim 1, The latanoprost is present in the ODS at a concentration of 2 micrograms per milliliter to 45 micrograms per milliliter. ODS.

4. An ODS according to any one of claims 1 or 3, The latanoprost is present in the ODS at a concentration of 10 micrograms per milliliter to 40 micrograms per milliliter. ODS.

5. An ODS according to any one of claims 1 to 4, The latanoprost is present in the ODS at a concentration of 15 micrograms per milliliter to 25 micrograms per milliliter. ODS.

6. An ODS according to any one of claims 1 to 4, The latanoprost is present in the ODS at a concentration ranging from 20 micrograms per milliliter to 25 micrograms per milliliter. ODS.

7. An ODS according to any one of claims 1 to 6, The latanoprost is present in the ODS at a concentration of 20±1 μg / ml. ODS.

8. An ODS according to any one of claims 1 to 7, the fluid or an ODS containing the fluid does not contain preservatives or detergents that would normally cause damage to the corneal tissue; The preservative or detergent is selected from quaternary ammonium preservatives, chlorobutanol, edetate disodium (EDTA), polyquaternium-1, stabilizing oxidizing agents, ionic buffer system preservatives, polyhexamethylene biguanide (PHMB), sodium perborate, tyloxapol, and sorbate. ODS.

9. The ODS according to any one of claims 1 to 8, Formulated for topical administration to the ocular surface as eye drops, eye washes, or contact lenses. ODS.

10. An ODS according to any one of claims 1 to 9 for use in a method for treating, preventing, and / or delaying the onset or recurrence of an ocular disorder in a human or animal subject, comprising topically co-administering a fluid comprising high molecular weight hyaluronic acid and a bioactive agent to the ocular surface of said human or animal subject, the fluid transports the bioactive agent into the eye; The eye disorder is ocular hypertension or glaucoma. ODS.

11. The ODS of claim 10, The eye disorder is glaucoma ODS.

12. The ODS according to claim 10 or 11, The glaucoma is open-angle glaucoma, acute angle-closure glaucoma, low-tension glaucoma, normal-tension glaucoma, or high-tension glaucoma. ODS.

13. The ODS of claim 10, The eye disorder is ocular hypertension. ODS.

14. The ODS according to any one of claims 10 to 13, The subject is a child under the age of 18. ODS.

15. The ODS according to any one of claims 10 to 13, The subject is an adult. ODS.

16. The ODS of claim 14, The child is an infant, adolescent, or juvenile. ODS.

17. The ODS of claim 5, The latanoprost is present in the ODS at a concentration of 19 μg / ml. ODS.

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