HK40030606B - Ophthalmic compositions containing a nitric oxide releasing prostamide - Google Patents

Description

Ophthalmic composition comprising nitric oxide releasing prostamide

The present application is a divisional application of chinese patent application 201910622356.1 filed on 7/10/2019 entitled "ophthalmic composition comprising nitric oxide releasing prostamide".

The present invention relates to an aqueous ophthalmic composition in the form of a solution comprising hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester as active ingredient and polyethylene glycol 15 hydroxystearate.

Hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester has the following formula (I)

This compound has been shown to be an effective intraocular pressure (IOP) lowering drug (f.impagnatello, c.b.toris, m.batugo, g.prasanna, v.borghi, e.basita, e.ingini, a.h.p.krauss; Invest optohol Vis sci.2015; 56: 6558-64).

WO2009/136281 (Nicox SA) discloses the use of hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester for the treatment of glaucoma and ocular hypertension.

Impartiello francisco et al; british Journal of Pharmacology (2018), pages 1-11 disclose a non-clinical pharmacological study showing that the IOP-lowering potency of NCX470 is greater than that of an equimolar dose of bimatoprost. Both compounds were dissolved in vehicle containing 0.25% Tween 80, 0.02% BAK, 2% glycerol and 0.1% EDTA.

Topical instillation is the most widely preferred non-invasive route of drug administration for the treatment of elevated intraocular pressure. Most conventional dosage forms of the commercially available IOP lowering drugs are eye drops in the form of aqueous solutions, but the ocular bioavailability of the active ingredient upon topical drop administration is very low. Many anatomical and physiological factors limit ocular absorption of topically applied ophthalmic drugs, such as rapid corneal prodrug elimination due to solution drainage and systemic absorption from the conjunctival sac and corneal epithelial barrier. An important goal of pharmaceutical formulations is to enable IOP lowering drugs to penetrate into the eye after a unique or multiple topical instillation. It is known that the ocular residence time of the drug on the ocular surface after a single drop application is very short. As a result, several formulation approaches are commonly used to provide more opportunity for drug penetration into the eye following administration to the ocular surface, and it is desirable to use a minimum dosage regimen, such as once a day, to promote patient treatment compliance.

One approach is to increase the viscosity of the ophthalmic solution. As a result, the solution will adhere better to the ocular surface and correspondingly increase the ocular residence time of the drug on that side. As a result, an increased amount of the drug should penetrate into the interior of the eye. For example, WO 2012/001009 discloses a delivery system consisting of a gelling system based on the combination of two gelling agents to obtain a non-viscous polymer delivery system which ensures the desired viscosity level and enhances the solubility of the active ingredient prostaglandin.

Another approach is to utilize ocular penetration enhancers. One of the most well known penetration enhancers is benzalkonium chloride (BAK). In the field of prostaglandin ophthalmic formulations, Allergan utilizes its productsTo take advantage of this approach. The first generation products have been commercialized, wherein bimatoprost prostaglandins were formulated at 0.03% dosage. In this product, the benzalkonium chloride dose is 0.005%. This benzalkonium chloride dose is primarily used to ensure antimicrobial protection of the solution. Some years later, the second generationHas been commercialized with the same efficacy as the old product. The new formulation had a lower concentration of bimatoprost (0.01%), but the amount of benzalkonium chloride (0.02%) was increased by a factor of 4 compared to the original formulation (0.005%).

Another technical challenge of ophthalmic pharmaceutical formulations is to stabilize the active ingredient. It can be noted that some eye drop formulations require refrigeration to maintain their active ingredient levels, with the disadvantage of poor usability; for example, for multi-dose vials, if the formulation is not stored properly at low temperatures, the level of active ingredient may decrease during this period. Elderly people who are often difficult to apply "cold" drug storage requirements are prescribed eye drops for the treatment of glaucoma or ocular hypertension.

US 8,772,337(the Laboratories) is disclosed inProstaglandin-containing compounds stable at room temperature and method of makingAn ophthalmic solution of HS15 (polyethylene glycol 15hydroxystearate) which is free of antimicrobial preservatives.

US 8,772,337 disclosesHS15 is able to solubilize some prostaglandins such as latanoprost and impart stability to the solution at ambient temperature in the absence of quaternary ammonium agents such as benzalkonium chloride, which are used as common solubilizers for such prostaglandin analogs. More specifically, it confers stability to the packaging of prostaglandin analogue solutions, in particular LDPE type plastic packaging of European Pharmacopoeia (EP) quality. Therefore, the temperature of the molten metal is controlled,HS15 can be used as a substitute for polysorbate 80 as a solubilizer.

Test examples of US 8,772,337 disclose compositions containingHS15 (0.5%), phosphate buffer, sorbitol, and EDTA.

WO2013/003827(Allergan) discloses a formulation containing polyethylene glycol 15hydroxystearate and a preservative. This patent application discloses the use of polyethylene glycol 15 hydroxystearate(s) ((R))HS15) as a surfactant instead of polyethoxylated surfactant (polysorbate)80) There are several advantages, such as increased solubility of the API, improved stability of the API which tends to be susceptible to degradation by oxidative mechanisms, improved preservative effectiveness of benzalkonium chloride, improved tolerability for ophthalmic applications.

WO 2009/084021(Sun Pharmaceutical) discloses the addition of a stabilizing amount of a polyglycol ester of 12-hydroxystearic acid (polyglycol ester)(s) (glycol esters) to an ophthalmic composition containing one or more prostaglandin derivativesHS15) can reduce the adsorption of prostaglandin derivatives to polyethylene containers, and the addition of a small amount of oil further reduces the adsorption of prostaglandin derivatives to low density polyethylene containers.

US 8,795,634(Critical Pharma) disclosesThe use of HS15 as an absorption enhancer to improve systemic absorption of topically applied therapeutic agents through the mucosa of the nasal, oral and respiratory tracts. However, the adsorption mechanism of active ingredients applied topically to the mucosa differs from the route taken for topical application to therapeutic agents through the eye; in fact, the therapeutic agent must cross the intrinsic and unique anatomical barriers of the ocular anatomy (i.e., corneal epithelium, conjunctiva, and sclera) and bypass the eye's protective mechanisms (i.e., blinking, tear film renewal, and drainage).

Hydroxystearic acid polyethylene glycol 15 ester (C)HS15 from BASF, referred to above asHS15) is the main excipient for the formulation of the invention. As noted above, there are several prior art documents disclosing the use of polyethylene glycol 15hydroxystearate as an excipient for ophthalmic compositions.

The term "polyethylene glycol 15 hydroxystearate" refers to a mixture of monoesters and diesters of 12-hydroxystearic acid and polyethylene glycol obtained primarily by ethoxylation of 12-hydroxystearic acid. Polyethylene glycol 15hydroxystearate is also known as polyethylene glycol 12-hydroxystearate copolymer, polyethylene glycol-15-hydroxystearate and polyethylene glycol 66012-hydroxystearate. USP-NF also classified this compound as polyethylene glycol 15hydroxystearate (Polyoxyl 15 hydroxystearate).

As mentioned above, some prior art documents disclose the use of polyethylene glycol 15hydroxystearate as an excipient for ophthalmic compositions, however, today, only one eye drop is approved in europe, which comprises polyethylene glycol 15hydroxystearate and prostaglandin analogues. The drug has been administered by Rafam S.AAnd which contains travoprost as a prostaglandin analogue and BAK as an antimicrobial preservative system.

Hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester is a viscous oil with a minimum water solubility of 0.02mg/ml, so hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, the aqueous pharmaceutical compositions of 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester must contain a solubilizing agent which increases the aqueous solubility of the compound and allows it to reach therapeutically active concentrations of the compound.

WO2009/136281 discloses a polysorbate 80 (C)80) 0.5%, benzalkonium chloride 0.02%, citrate buffer, water, hexanoic acid in a vehicle with pH 5.5, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl]-3, 5-dihydroxycyclopentyl]-1- (2-phenylethyl) -2-propen-1-yl ester.

Polysorbate 80 has been widely used as an excipient for ophthalmic compositions. For example, the product sold by NovartisOrnoprostone was combined with benzalkonium chloride and polysorbate 80 to a 0.015% by weight solution.

The present invention relates to an ophthalmic aqueous composition in the form of a solution comprising hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester and polyethylene glycol 15hydroxystearate, wherein polyethylene glycol 15hydroxystearate is preferably the only solubilizer.

The ophthalmic aqueous compositions of the present invention provide higher ocular penetration of the active ingredient hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester than the known formulations, thus allowing enhanced ocular absorption of the therapeutically active compound.

The present invention also provides an ophthalmic aqueous composition in the form of a solution comprising hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester, which is chemically and physically stable when stored at room temperature.

Summary of The Invention

The present invention provides an ophthalmic aqueous composition in the form of a solution comprising 0.005% to 0.18% w/w hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester, 0.5% w/w to 1.5% w/w polyethylene glycol 15hydroxystearate, wherein polyethylene glycol 15hydroxystearate is preferably the only solubilizer.

Another embodiment is an ophthalmic aqueous composition in the form of a solution comprising 0.005% to 0.10% w/w hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester, 0.5% w/w to 1.5% w/w polyethylene glycol 15hydroxystearate, wherein polyethylene glycol 15hydroxystearate is preferably the only solubilizer.

A preferred embodiment of the present invention provides an ophthalmic aqueous composition in the form of a solution comprising 0.005% w/w to 0.065% w/w hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester, 0.5-1.5% w/w of polyethylene glycol 15hydroxystearate and a pharmaceutically acceptable aqueous solvent, wherein the polyethylene glycol 15hydroxystearate is preferably the only solubilizer.

In the field of ophthalmology, and in particular in aqueous ophthalmic compositions, solubilizers are compounds that improve the dissolution of biologically active ingredients having relatively low water solubility.

Preferably, the amount of hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester in the ophthalmic solution is 0.01% w/w to 0.065% w/w, most preferably 0.021%, 0.042% w/w or 0.065% w/w.

The aqueous ophthalmic solution may contain benzalkonium chloride (BAK) as an antimicrobial preservative in an amount of 0.013% w/w to 0.02% w/w, Edetate (EDTA), such as edetate disodium salt, in an amount of 0.03% w/w to 0.07% w/w as an antimicrobial preservative adjuvant; preferably, the amount of benzalkonium chloride (BAK) is 0.013% w/w-0.02% w/w and the amount of Edetate (EDTA) is 0.05% w/w; most preferably, the amount of benzalkonium chloride (BAK) is 0.016% w/w and the amount of Edetate (EDTA) is 0.05% w/w.

The aqueous ophthalmic composition of the present invention further comprises a buffering agent selected from the group consisting of: sodium dihydrogen phosphate, disodium hydrogen phosphate heptahydrate, potassium dihydrogen phosphate or dipotassium hydrogen phosphate, boric acid and its salts, acetates such as sodium acetate and mixtures thereof. Preferably, the buffering agent of the aqueous ophthalmic composition of the present invention is a mixture of sodium phosphate dibasic heptahydrate and boric acid or a mixture of citric acid and sodium phosphate dibasic heptahydrate. The preferred pH of the formulation is 6.0. This pH has been found to be an optimal pH to allow suitable formulation stability under long term storage conditions and suitable ocular tolerance when the formulation is delivered on the ocular surface.

The pH of the aqueous ophthalmic composition is preferably adjusted to 5.5 to 6.5, more preferably to pH 6.0.

The aqueous ophthalmic composition of the present invention may comprise a pH adjusting agent selected from the group consisting of hydrochloric acid, citric acid, phosphoric acid, acetic acid, tartaric acid, sodium hydroxide, potassium hydroxide, sodium carbonate and sodium bicarbonate; preferably, the aqueous ophthalmic composition of the present invention contains hydrochloric acid and/or sodium hydroxide in an amount that can adjust the pH in the range of 5.5 to 6.5; more preferably pH 6.0.

Aqueous ophthalmic solutions may contain tonicity agents for adjusting the osmolality of the formulation and for targeting the desired isotonicity. Preferred tonicity agents are sodium chloride, sorbitol, glycerin and mannitol. Isotonicity was set at 300mOsm/kg, but a wider range of 260-340mOsm/kg is generally acceptable for use in formulating ophthalmic solutions. Preferably, the osmolality of the solution is 280-320 mOsm/kg.

The aqueous ophthalmic solutions of the present invention may contain viscosity modifiers to improve contact between the solution and the eye and to promote spreading of the product over the ocular surface. Preferred viscosity modifiers are cellulose derivative polymers such as carboxymethyl cellulose or hydroxypropyl methyl cellulose, hyaluronic acid, polyvinyl alcohol, carboxylic acid polymers such as carbomer or polycarbophil. Most preferably, the viscosity modifier is hydroxypropyl methylcellulose at a concentration of less than 0.5% w/w. When using cellulose derivatives, the viscosity of the aqueous ophthalmic solution is adjusted at 5-10m.pa.s for Newtonian solutions, but may be higher for high viscosity solutions with non-Newtonian/pseudoplastic rheology, typically obtained when using carboxylic acid polymers.

Another embodiment of the present invention provides an aqueous ophthalmic composition in the form of a solution comprising 0.005% w/w to 0.10% w/w hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester, 0.5% w/w to 1.5% w/w polyethylene glycol 15hydroxystearate, 0.013% w/w to 0.02% w/w benzalkonium chloride, 0.03% w/w to 0.07% w/w disodium edetate dihydrate, a pharmaceutically acceptable carrier, a pharmaceutically acceptable carrier, a pharmaceutically acceptable carrier, a pharmaceutically acceptable carrier, a pharmaceutically acceptable carrier, a pharmaceutically acceptable carrier, a pharmaceutically acceptable carrier, a pharmaceutically acceptable carrier, a pharmaceutically acceptable carrier, a, Sodium phosphate dibasic heptahydrate, boric acid and water, wherein the ophthalmic solution has a pH of 6, and wherein polyethylene glycol 15hydroxystearate is the sole solubilizer.

Another embodiment of the present invention provides an aqueous ophthalmic composition in the form of a solution comprising 0.005% w/w to 0.065% w/w hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester, 0.5% w/w to 1.5% w/w polyethylene glycol 15hydroxystearate, 0.013% w/w to 0.02% w/w benzalkonium chloride, 0.03% w/w to 0.07% w/w disodium edetate dihydrate, a pharmaceutically acceptable carrier, a pharmaceutically acceptable carrier, a pharmaceutically acceptable carrier, a pharmaceutically acceptable carrier, a pharmaceutically acceptable carrier, a pharmaceutically acceptable carrier, a pharmaceutically acceptable carrier, a, Sodium phosphate dibasic heptahydrate and boric acid and water, wherein the ophthalmic solution has a pH of 6, and wherein polyethylene glycol 15hydroxystearate is the only solubilizer; preferably the amount of hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester in the ophthalmic solution is 0.01% w/w to 0.065% w/w, most preferably 0.042% w/w, 0.021% w/w or 0.065% w/w; optionally, the ophthalmic solution further comprises HCl 1.2M/NaOH 1M to adjust the pH to pH 6.0.

Another embodiment of the present invention provides an aqueous ophthalmic composition in the form of a solution comprising 0.005% w/w to 0.10% w/w hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester, 1.0% w/w polyethylene glycol 15hydroxystearate, 0.016% w/w benzalkonium chloride, 0.05% w/w disodium edetate dihydrate, disodium hydrogen phosphate heptahydrate and boric acid and water, wherein the ophthalmic solution has a pH of 6 and wherein polyethylene glycol 15hydroxystearate is the sole solubilizer.

Another embodiment of the present invention provides an aqueous ophthalmic composition in the form of a solution comprising 0.005% w/w to 0.065% w/w hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester, 1.0% w/w polyethylene glycol 15hydroxystearate, 0.016% w/w benzalkonium chloride, 0.05% w/w edetate disodium dihydrate, disodium hydrogen phosphate heptahydrate and boric acid and water, wherein the ophthalmic solution has a pH of 6 and wherein polyethylene glycol 15hydroxystearate is the only solubilizer; preferably the amount of hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester in the ophthalmic solution is 0.01% w/w to 0.065% w/w, most preferably 0.042% w/w, 0.021% w/w or 0.065% w/w; optionally, the ophthalmic solution further comprises HCl 1.2M/NaOH 1M to adjust the pH to pH 6.0.

Another embodiment of the present invention provides an aqueous ophthalmic composition in the form of a solution, consisting of: 0.005% w/w to 0.10% w/w hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl]-3, 5-dihydroxycyclopentyl]-1- (2-phenylethyl) -2-propen-1-yl ester, 1.0% -1.5% w/w polyethylene glycol 15hydroxystearate, 0.016% w/w benzalkonium chloride, 0.05% w/w disodium edetate dihydrate, 2.76% w/w sorbitol, 1.33% w/w disodium hydrogen phosphate heptahydrate (Na)₂HPO₄ 7H₂O), 0.5% w/w boric acid and water, wherein the ophthalmic solution has a pH of 6 and wherein polyethylene glycol 15hydroxystearate is the only solubilizer.

Another embodiment of the present invention provides an aqueous ophthalmic composition in the form of a solution, consisting of: 0.005% w/w-0.065% w/w hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl]-3, 5-dihydroxycyclopentyl]-1- (2-phenylethyl) -2-propen-1-yl ester, 1.0% w/w polyethylene glycol 15hydroxystearate, 0.016% w/w benzalkonium chloride, 0.05% w/w disodium edetate dihydrate, 2.76% w/w sorbitol, 1.33% w/w disodium hydrogen phosphate heptahydrate (Na)₂HPO₄ 7H₂O), 0.5% w/w boric acid and water, wherein the ophthalmic solution has a pH of 6, and wherein polyethylene glycol 15hydroxystearate is the only solubilizer; preferably hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl]-3, 5-dihydroxycyclopentyl](iii) the amount of (E) -1- (2-phenylethyl) -2-propen-1-yl ester is from 0.01% w/w to 0.065% w/w, most preferablyPreferably 0.021%, 0.042% or 0.065% w/w; optionally, the ophthalmic solution further comprises HCI 1.2M/NaOH 1M to adjust the pH to pH 6.0.

Specific examples of aqueous ophthalmic compositions in the form of solutions according to the invention are:

-0.042% w/w hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester, 1.0% w/w polyethylene glycol 15hydroxystearate, 0.016% w/w benzalkonium chloride, 0.05% w/w disodium edetate dihydrate, 2.76% w/w sorbitol, 1.33% w/w disodium hydrogenphosphate heptahydrate, 0.5% w/w boric acid and water and having a pH of 6;

-0.021% w/w hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester, 1.0% w/w polyethylene glycol 15hydroxystearate, 0.016% w/w benzalkonium chloride, 0.05% w/w disodium edetate dihydrate, 2.76% w/w sorbitol, 1.33% w/w disodium hydrogenphosphate heptahydrate, 0.5% w/w boric acid and water and having a pH of 6;

0.065% w/w hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester, 1.0% w/w polyethylene glycol 15hydroxystearate, 0.016% w/w benzalkonium chloride, 0.05% w/w disodium edetate dihydrate, 2.76% w/w sorbitol, 1.33% w/w disodium hydrogen phosphate heptahydrate, 0.5% w/w boric acid and water and having a pH of 6;

-0.10% w/w hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester, 1.5% w/w polyethylene glycol 15hydroxystearate, 0.016% w/w benzalkonium chloride, 0.05% w/w disodium edetate dihydrate, 0.52% w/w glycerol, 2.33% w/w disodium hydrogen phosphate heptahydrate, 0.36% w/w citric acid and water and having a pH of 6.

In the above aqueous ophthalmic composition, hydrochloric acid or sodium hydroxide may be used as a pH adjuster.

The ophthalmic aqueous compositions reported above can be packaged in Low Density Polyethylene (LDPE) primary containers, typically multi-dose ophthalmic bottles with a cap, i.e. dropper tip, capable of delivering calibrated drops of pressurized controlled drop size. Examples of such primary containers may be those from Berry-PlasticsBottle, Boston from Amcor3-piece vials from Gerresheimer, Philips-Medisize, Bormioli, or equivalents.

The aqueous ophthalmic composition in solution form may be provided as an antimicrobial aqueous ophthalmic solution without preservatives.

One embodiment of the present invention provides a preservative-free antimicrobial aqueous ophthalmic composition in the form of a solution comprising 0.005% w/w to 0.10% w/w hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester, 0.5% w/w to 1.5% w/w polyethylene glycol 15hydroxystearate, disodium hydrogen phosphate heptahydrate and boric acid and water, wherein the ophthalmic solution has a pH of 6, also comprises a tonicity agent and optionally another pH adjusting agent, wherein the polyethylene glycol 15hydroxystearate is the only solubilizer.

Specific examples of preservative-free aqueous ophthalmic compositions in the form of solutions are the following compositions consisting of: 0.042% w/w hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester, 1.0% w/w polyethylene glycol 15hydroxystearate, 2.76% w/w sorbitol, 1.33% w/w disodium hydrogenphosphate heptahydrate, 0.5% w/w boric acid and water and has a pH of 6.

Further specific examples of preservative-free ophthalmic aqueous compositions in the form of solutions are the following compositions:

-0.042% w/w hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester, 1.0% w/w polyethylene glycol 15hydroxystearate, 2.76% w/w sorbitol, 1.33% w/w disodium hydrogenphosphate heptahydrate, 0.5% w/w boric acid and water and having a pH of 6.

-0.021% w/w hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester, 1.0% w/w polyethylene glycol 15hydroxystearate, 2.76% w/w sorbitol, 1.33% w/w disodium hydrogen phosphate heptahydrate, 0.5% w/w boric acid and water and having a pH of 6.

-0.065% w/w hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester, 1.0% w/w polyethylene glycol 15hydroxystearate, 2.76% w/w sorbitol, 1.33% w/w disodium hydrogenphosphate heptahydrate, 0.5% w/w boric acid and water and having a pH of 6.

-0.10% w/w hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester, 1.5% w/w polyethylene glycol 15hydroxystearate, 0.52% w/w glycerol, 2.33% w/w disodium hydrogenphosphate heptahydrate, 0.36% w/w citric acid and water and having a pH of 6.

In the above aqueous ophthalmic composition, hydrochloric acid or sodium hydroxide may be used as a pH adjuster.

The antimicrobial preservative-free ophthalmic aqueous composition of the present invention can be packaged in a container that prevents microbial contamination of the formulation even after multiple uses; or the antimicrobial preservative-free ophthalmic aqueous composition is packaged in a unit dose container that is sterile in unopened form.

Such antimicrobial preservative-free formulations can be packaged in single or unit dose LDPE primary containers or inside preservative-free multi-dose container systems.

Typically, the preservative-free ophthalmic aqueous compositions of the present invention may be packaged in single dose containers intended for single use by a patient. In another embodiment, the antimicrobial preservative-free ophthalmic aqueous compositions of the present invention may be packaged in multi-dose containers, which enable the formulation to remain sterile even after multiple uses by a patient. Examples of preservative-free multi-dose containers are: from AptarFrom NemeraFrom AerocumpOr an equivalent device.

The polyethylene glycol 15hydroxystearate used in the present invention is commercially availableHS15, which is referred to in advance asHS15 and also listed in USP as polyethylene glycol-15 hydroxystearate; it consists of polyethylene glycol mono-and diesters of 12-hydroxystearic acid and about 30% free polyethylene glycol.

The aqueous ophthalmic compositions of the present invention in the form of solutions exhibit efficacy in lowering intraocular pressure and are therefore useful in the treatment of ocular hypertension, glaucoma or in methods of lowering intraocular pressure.

Another object of the present invention relates to an ophthalmic aqueous composition in the form of a solution according to the invention for use in the treatment of ocular hypertension, glaucoma or in a method of lowering intraocular pressure.

Another object of the present invention relates to an ophthalmic aqueous composition as defined above, in the form of a solution, for the treatment of ocular hypertension, glaucoma or reduction of intraocular pressure.

Another embodiment of the present invention relates to a method of treating ocular hypertension or glaucoma or a method of lowering intraocular pressure, which method comprises administering to a patient in need thereof a therapeutically effective amount of an ophthalmic aqueous composition in the form of a solution as defined above.

Another embodiment of the present invention relates to a method of treating ocular hypertension or glaucoma or a method of lowering intraocular pressure comprising administering to a patient in need thereof a therapeutically effective amount of an ophthalmic aqueous composition comprising 0.005% w/w to 0.065% w/w hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester, 0.5% w/w to 1.5% w/w polyethylene glycol 15hydroxystearate and a pharmaceutically acceptable aqueous soluble coal, wherein the hydroxystearic acid polyethylene glycol 15 ester is the only solubilizer.

Another embodiment of the present invention relates to a method of treating ocular hypertension or glaucoma or a method of lowering intraocular pressure comprising administering to a patient in need thereof a therapeutically effective amount of an ophthalmic aqueous composition comprising 0.005% w/w to 0.065% w/w hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl 1-1- (2-phenylethyl) -2-propen-1-yl ester, 1.0% w/w polyethylene glycol hydroxystearate 15 ester, 0.016% w/w benzalkonium chloride, a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable carrier, or a pharmaceutically acceptable salt thereof, 0.05% w/w disodium edetate dihydrate, disodium hydrogen phosphate heptahydrate, and boric acid and water, wherein the ophthalmic solution has a pH of 6, and wherein polyethylene glycol 15hydroxystearate is the only solubilizer; preferably the amount of hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester in the ophthalmic solution is 0.01% w/w to 0.065% w/w, most preferably 0.042% w/w, 0.021% w/w or 0.065% w/w; optionally the ophthalmic solution further comprises HCl 1.2M/NaOH 1M to adjust the pH to pH 6.0.

Another embodiment of the present invention relates to a method of treating ocular hypertension or glaucoma or a method of reducing intraocular pressure comprising administering to a patient in need thereof a therapeutically effective amount of an ophthalmic aqueous composition in the form of a solution consisting of: 0.005% w/w-0.065% w/w hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester, 1.0% w/w polyethylene glycol 15hydroxystearate, 0.016% w/w benzalkonium chloride, 0.05% w/w disodium edetate dihydrate, 2.76% w/w sorbitol, 1.33% w/w sodium phosphate dibasic heptahydrate, 0.5% w/w boric acid and water, and having a pH of 6; preferably in the ophthalmic aqueous composition in the form of a solution, the amount of hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester is 0.01% w/w to 0.065% w/w, most preferably 0.042% w/w, 0.021% w/w or 0.065% w/w; optionally the above mentioned ophthalmic solution in the form of a solution further comprises HCl 1.2M/NaOH 1M in order to adjust the pH to pH 6.0.

The ophthalmic composition of the present invention may be administered as eye drops for the treatment of chronic ophthalmic diseases, such as glaucoma or ocular hypertension. It is contemplated that the ophthalmic solution is typically administered once daily in each eye at a frequency of once daily.

Another embodiment of the present invention relates to a method for preparing an ophthalmic aqueous solution of the present invention, the method comprising the steps of:

step 1) preparation of a concentrated solution of hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester (API concentrated solution) comprising:

1a) heating a mixture of water for injection and polyethylene glycol 15hydroxystearate at 32 ℃ until the polyethylene glycol 15hydroxystearate melts;

1b) adding the molten polyethylene glycol 15 hydroxystearate/water mixture to a pre-weighed amount of hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester (API);

1c) mixing the resulting mixture to complete dissolution of hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester while maintaining the mixture at a temperature of 32 ℃; the amount of water for injection used in step 1a) is about 1.5% of the total weight of water used in the preparation; the amount of polyethylene glycol 15hydroxystearate and hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester is their total weight corresponding to its percentage in the final solution.

Step 2) an aqueous solution of the remaining vehicle components was prepared by adding excipients in the following specific order in a manufacturing tank containing water for injection: edetate disodium dihydrate selected from boric acid and sodium phosphate dibasic heptahydrate or citric acid, sodium phosphate dibasic heptahydrate buffer, sorbitol or glycerol and benzalkonium chloride; each excipient is completely dissolved before the next excipient is added, and the preparation of the solution is carried out at a temperature of 25-30 ℃; the amount of each excipient is its total weight corresponding to its percentage in the final ophthalmic solution; the water for injection used in step 2) is 80% to 90% of the total weight in the final ophthalmic solution.

The order of excipient addition reported above must be observed to meet pH and tonicity requirements of pH6.0 and 300 mOsm/kg.

Step 3) the whole ophthalmic solution is prepared by adding the API concentrated solution of step 1 to the manufacturing tank containing the aqueous solution of step 2 and water for injection until the target final weight.

Optionally, the pH of the bulk ophthalmic solution is adjusted to pH6.0 with 1M sodium hydroxide or 1.2M hydrochloric acid.

Step 4) the whole ophthalmic solution of step 3 was filtered through a Polyethersulfone (PES) filter having a pore size of about 0.2 μm and sterilized.

Step 5) optionally, filling the bulk ophthalmic solution in a Low Density Polyethylene (LDPE) ophthalmic primary container.

Optionally, when the aqueous ophthalmic solution further comprises a viscosity modifier, in step 2), the viscosity modifier is added to the manufacturing tank containing water for injection as a first ingredient, and other excipients are added once it is completely dissolved.

Another embodiment of the present invention relates to a method of preparing the antimicrobial preservative-free aqueous ophthalmic solution of the present invention, the method comprising the steps of:

step 1) preparation of a concentrated solution of hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester (API concentrated solution) comprising:

1a) heating a mixture of water for injection and polyethylene glycol 15hydroxystearate at 32 ℃ until the polyethylene glycol 15hydroxystearate melts;

1b) adding the molten polyethylene glycol 15 hydroxystearate/water mixture to a pre-weighed amount of hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester (API);

1c) mixing the resulting mixture to complete dissolution of hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester while maintaining the mixture at a temperature of 32 ℃; the amount of water for injection used in step 1a) is about 1.5% of the total weight of water used in the preparation; the amount of polyethylene glycol 15hydroxystearate and hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester is their total weight corresponding to its percentage in the final solution.

Step 2) an aqueous solution of the remaining coal-dissolving component was prepared by adding excipients in the following specific order in a manufacturing tank containing water for injection: a buffering agent and a tonicity agent; each excipient is completely dissolved before the next excipient is added, and the preparation of the solution is carried out at a temperature of 25-30 ℃; the amount of each excipient is its total weight corresponding to its percentage in the final ophthalmic solution; the water for injection used in step 2) is 80% to 90% of the total weight in the final ophthalmic solution.

The order of excipient addition reported above must be observed to meet pH and tonicity requirements of pH6.0 and 300 mOsm/kg.

Step 3) the whole ophthalmic solution is prepared by adding the API concentrated solution of step 1 to the manufacturing tank containing the aqueous solution of step 2 and water for injection until the target final weight.

Optionally, the pH of the bulk ophthalmic solution is adjusted to pH6.0 with 1M sodium hydroxide or 1.2M hydrochloric acid.

Step 4) the whole ophthalmic solution of step 3 was filtered through a Polyethersulfone (PES) filter having a pore size of about 0.2 μm and sterilized.

Step 5) optionally, filling the bulk ophthalmic solution in a Low Density Polyethylene (LDPE) ophthalmic primary container.

When the aqueous ophthalmic solution further comprises a viscosity modifier, in step 2), the viscosity modifier is added to the manufacturing tank containing water for injection as a first ingredient, and other excipients are added once it is completely dissolved.

Example 1

Preparation of an ophthalmic composition comprising hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester (NCX470) 0.042% w/w (100 liter batch)

The components of the ophthalmic composition are listed as follows:

step 1) preparation of concentrated API solution

1.5L of water for injection is added to a pre-weighed amount of 1,000g of polyethylene glycol 15hydroxystearate at about 32 ℃ in a container, which is placed in a hot water bath until the polyethylene glycol 15hydroxystearate is completely melted. The molten polyethylene glycol 15 hydroxystearate/water mixture was added to a 4L batch tank containing 42g NCX-470.

A 4L batch tank was thermostated with a water bath at 32 ℃, and the API solution was stirred until all ingredients were completely dissolved and held under continuous stirring until it was added to the solution of the remaining excipient in coal.

Step 2) preparation of solution of remaining excipient solvent

100L of water for injection was poured into a manufacturing tank made of stainless steel (316L grade) and cooled between 25 deg.C and 30 deg.C. About 12 liters of this water for injection is taken out of the tank and stored in another container for use at the time of preparation.

The following compounds were added to a manufacturing tank containing water, with continuous stirring, in exactly the following order; each compound was completely dissolved before the next compound was added:

50g disodium ethylenediaminetetraacetate dihydrate;

500g of boric acid;

1326g disodium phosphate heptahydrate (Na)₂HPO₄ 7H₂O)；

2760g sorbitol;

32g of a 50% benzalkonium chloride solution;

the container containing the benzalkonium chloride solution is rinsed several times with a sufficient amount of water for injection and the rinse is added to the manufacturing tank to complete the transfer.

Step 3) preparation of Whole ophthalmic solution

Transferring the API concentrated solution into a manufacturing tank; the 4L batch tank can be rinsed with water for injection and the rinse added to the bulk solution to complete the transfer.

Water for injection was added to the manufacturing tank to adjust the final target weight of 100 kg.

4) Sterilization of whole eye solutions

Passing through a 0.2 μm Polyethersulfone (PES) filterRedundant filtration sterilizes the ophthalmic solution. 5) Integral ophthalmic solution filled with LDPE ophthalmic bottleLiquid for treating urinary tract infection

After the filtration step, the ophthalmic solution is filled into LDPE multi-dose containers of appropriate volume, under a grade a environment, according to conventional aseptic process practice.

Example 2

Stability study

The stability of the ophthalmic formulations of the present invention stored in multi-dose Low Density Polyethylene (LDPE) containers sterilized by different sterilization methods was tested.

The stability of the ophthalmic formulations containing NCX 4700.042% w/w was evaluated at 25 ℃, initially, 3.5 months and 6 months, 9 months (long term storage conditions), and at 40 ℃, Relative Humidity (RH) not exceeding 25%, initially, at 3.5 months and 6 months, at accelerated stability evaluations.

The ophthalmic preparation comprises the following components:

the results are shown in tables 1 to 4.

The results of the stability tests at 25 ℃ (tables 1 and 3) and 40 ℃ (tables 2 and 4) demonstrate the good stability of the ophthalmic solutions of the present invention, and therefore the shelf life of the ophthalmic solutions filled in LDPE primary containers is expected to be at least 24 months for the product under storage conditions at ambient temperature.

Example 2A:

primary container: transparent LDPE/previously sterilized by irradiation (gamma ray/25 kGy).

Primary container construction: 2.5mL of fill in a 7.5mL vial.

Example 2B:

primary container: LDPE/previously sterilized with ethylene oxide.

Primary container construction: 2.5mL of fill in a 7.5mL vial.

As a conclusion, the ophthalmic solution from the present invention can be packaged in LDPE primary containers, which in any case are pre-sterilized versions of such containers. The multi-dose primary container may be sterilized with gamma radiation or ethylene oxide gas. BySingle-use or unit dose containers produced by (blow-fill-seal) technology provide natural sterilized LDPE containers, and such LDPE material articles are also suitable for packaging preservative-free ophthalmic solutions from the present invention.

Example 3

Antimicrobial effectiveness test

Example 3A

Antimicrobial effectiveness tests were conducted to evaluate the ability of the ophthalmic solutions of the present invention to meet antimicrobial preservation efficacy standards.

Ophthalmic solutions containing disodium edetate 0.05% (w/w) (see table 5) and varying concentrations of benzalkonium chloride (see table 6) were tested. The ophthalmic composition was prepared by applying the method disclosed in example 1.

The test was carried out according to the method for Testing performance disclosed in the United states pharmacopoeia monograph <51>, Antimicrobial Effectiveness Testing (AET). The success of this USP test is equivalent to matching european pharmacopoeia B.

The results reported in table 6 show that ophthalmic compositions comprising benzalkonium chloride from 0.012% (w/w) to 0.02% (w/w) meet the criteria for microbial effectiveness and pass the Antimicrobial Effectiveness Test (AET).

NCX470 ═ hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester

Example 3B

To evaluate the effect of EDTA as an antimicrobial preservative aid, the efficacy of ophthalmic compositions comprising benzalkonium chloride (0.016% w/w) and disodium edetate (0.05% w/w) (formulation a-table 7) and ophthalmic compositions comprising benzalkonium chloride (0.016% w/w) but no disodium edetate (formulation B-table 7) in meeting preservative criteria was evaluated.

The results show that disodium edetate is required to meet the preservative criteria defined in the united states pharmacopeia monograph <51> or european pharmacopeia standard B.

WO2013/003827(Allergan) discloses that polyethylene glycol 15hydroxystearate improves the effectiveness of antimicrobial preservatives, allowing a reduction in the dose of the antimicrobial preservative benzalkonium chloride matching USP <51> or EP-Standard B. Preservative studies conducted with the ophthalmic solutions of the present invention show that polyethylene glycol 15hydroxystearate does not improve the antimicrobial preservative efficacy of benzalkonium chloride and, to ensure the antimicrobial preservative efficacy of the ophthalmic solutions, benzalkonium chloride must be present in an amount which must be higher than 0.12% w/w in the presence of 0.5% w/w EDTA. The most suitable antimicrobial preservative dosage target was achieved using a mixture of 0.16% w/w benzalkonium chloride and 0.05% w/w EDTA.

Example 4

Pharmacokinetic evaluation in Dutch black-banded rabbits following single ocular instillation

Study 1

Hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester is a dual action prostaglandin analogue derivative that combines the pharmacological activity of bimatoprost with nitric oxide. Bimatoprost free acid is one of the active metabolites of hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester; in this study, bimatoprost free acid levels were used as markers to compare aqueous ophthalmic solutions of the present invention to a commercially available ophthalmic solution containing bimatoprost and a reference preparation containing hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester, hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-propen-1-yl ester following topical instillation Oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl hexanoic acid.

The objective of this study was to comparatively assess the amount of bimatoprost free acid in aqueous humor samples taken after instillation of the tested aqueous ophthalmic solution.

Two different aqueous ophthalmic solutions (formulations 1 and 2) comprising hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester and a commercial eye drop comprising bimatoprost (formulation 3) were evaluated in a rabbit ocular pharmacokinetic study.

Two different aqueous solutions comprising hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester are an ophthalmic solution according to the invention (formulation 1) and a formulation disclosed in WO2009/136281 (formulation 2).

Test solution

Table 8 reports the vehicle for two aqueous ophthalmic solutions comprising hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester 0.042% w/w.

Preparation 3 (commercially available eye drops- Allergan)

Active ingredients: bimatoprost 0.03%;

hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester, at a concentration of 0.042% w/w was equimolar compared to the bimatoprost moles of the commercial eye drops (formulation 3).

Material

Polyethylene glycol 15hydroxystearate is commercially availableHS15 (BASF). Polysorbate 80 is a commercially available polysorbate 80Super(Croda)。

Experimental methods

Groups of 10 male naive Dutch rabbits were assigned to the study and formulations 1 and 2 or 3 were administered to each eye by instillation at a nominal target dose of 12.6 μ g/eye for formulations 1 and 2 and 9 μ g/eye for bimatoprost. A further group of 5 male dutch black-banded rabbits was used to provide a blank control matrix (aqueous humor) for bioanalytical purposes.

During the study, body weight measurements and clinical observations were made and at predetermined time points, animals were sacrificed for the purpose of harvesting Aqueous Humor (AH) at 1h, 2h, 4h and 8 h.

There were no clinical signs associated with the dosing observed during the study.

Results

The results reported in table 9 show that the ophthalmic aqueous solution of the invention (formulation 1) exhibits a greater bimatoprost acid exposure (in terms of C) than that obtained after administration of the reference formulation (formulation 2) and of the commercially available eye drops (formulation 3)_maxAnd AUC values).

C_maxIs the maximum concentration of bimatoprost free acid achieved in the aqueous humor.

AUC (area under the curve) represents the total amount of bimatoprost free acid in the aqueous humor as a function of time available to produce a biological effect.

Study 2

The objective of this study was to evaluate the aqueous humor content of bimatoprost free acid following topical ocular administration of three different aqueous ophthalmic solutions containing hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester (0.042% w/w) in three different vehicles reported in table 10.

Vehicles 1 and 2 contain polysorbate 80, which is a solubilizer of the formulation disclosed in prior art comparison document WO 2009/136281; vehicle 1 differs from vehicle 2 in that it contains benzalkonium chloride, whereas vehicle 2 does not.

Vehicle 3 contains a mixture of polysorbate 80 and polyethylene glycol 15 hydroxystearate.

Test solution

Table 10 reports containing hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxy cyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester 0.042% w/w aqueous ophthalmic solution of coal.

Additional reference formulations were tested; this formulation is a commercially available eye drop containing 0.03% bimatoprost.

Material

Polyethylene glycol 15hydroxystearate is commercially availableHS15 (BASF). Polysorbate 80 is a commercially available polysorbate 80Super(Croda)。

Experimental methods

The study included 40 animals. They were divided into 4 groups of 10 animals each and the test aqueous ophthalmic solutions reported above were administered.

All animals included in this study were administered by eye instillation of both eyes with a graduated pipette of volume 30 μ L/eye. 2 animals in each formulation were sacrificed at 0h (pre-dose), 1h, 2h, 4h and 8h and aqueous humor samples were obtained immediately from both eyes.

Results

The pharmacokinetic data reported in table 11 show the amount of bimatoprost acid exposed to aqueous ophthalmic solutions containing vehicle 1 or vehicle 2 (in terms of C)_maxAnd AUC values) were equivalent, whereas bimatoprost free acid in aqueous ophthalmic solutions containing vehicle 3 showed higher exposureAmount (higher concentration of bimatoprost free acid in aqueous humor).

The lowest exposure parameter for bimatoprost acid was obtained in the reference formulation group.

Furthermore, the results show that benzalkonium chloride does not exert any effect on ocular penetration of hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester hexanoic acid salt at concentrations up to 0.02% w/w, indeed the pharmacokinetic data are considered to be equivalent for both aqueous ophthalmic solutions containing vehicle 1 or 2.

In summary, the results of the studies reported above indicate that polyethylene glycol 15hydroxystearate enhances ocular penetration of hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester. Enhanced absorption is a surprising effect due to its independence from the known solubilizing activity of polyethylene glycol 15hydroxystearate, since the therapeutically active compound was solubilized in all aqueous ophthalmic solutions tested.

In fact, hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl]-3, 5-dihydroxycyclopentyl]-1- (2-phenylethyl) -2-propen-1-yl ester comprising 0.5% (w/w) polyethylene glycol hydroxystearate 15 ester(s) ((w/w))HS15) or 0.5% (w/w) polysorbate 80 (C: (A)80) Is 0.070% (w/w) and 0.074% (w/w), respectively, and is thus considered to be substantially equivalent, so in all tested vehicles, hexanoic acid, 6- (nitroxide), isBase) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl]-3, 5-dihydroxycyclopentyl]-1- (2-phenylethyl) -2-propen-1-yl ester was solubilised.

Furthermore, the results of study 2 show that benzalkonium chloride has no effect on the ocular bioavailability of the active compound; in fact, vehicle 1, containing polysorbate 80 and benzalkonium chloride, and vehicle 2, containing polysorbate 80 but no benzalkonium chloride, showed comparable concentrations of bimatoprost free acid in aqueous humor.

Example 5

Intraocular pressure in dogs with normal ocular blood pressure

In this study, the efficacy of the ophthalmic aqueous composition of the present invention in lowering intraocular pressure (IOP) in a bigote with normotensive intraocular pressure was evaluated.

Compositions tested

Preparation 1

Formulation 1 comprises:

0.042% w/w hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester;

polyethylene glycol 15 hydroxystearate: 1.0% w/w

Benzalkonium chloride: 0.016% w/w

Boric acid: 0.5% w/w

Disodium ethylene diamine tetraacetate: 0.05% w/w

Sorbitol: 2.76% w/w

Disodium hydrogen phosphate heptahydrate (Na)₂HPO₄ 7H₂O)：1.326％w/w

Proper amount of water for injection to 100g

The composition had a final pH of 6.0 and an osmolality of about 300 mOsm/kg.

Experimental methods

The aqueous composition tested was administered to the conjunctival sac of both eyes of a beagle by a topical route, 2 times daily, spaced about 4 hours apart, for 28 days. Groups were included, each group containing 3 males and 3 females compared to the brother:

group 1 (control animals): solvent

And 2, group: preparation 1

Each animal received 30 μ L/eye of the indicator substance at each treatment.

The first dose of the day is administered between 7 and 9 am, approximately simultaneously each day.

After the lower eyelid was gently pulled away from the eyeball, the test article was placed in the conjunctiva of both eyes of each animal by an automatic pipette. The eyelids were then held together gently for about 1 second to prevent loss of the test article.

Intraocular pressure was measured by electronic tonometer 1-1.5 hours before application and after daily dose at treatment days 2, 4, 8, 20 and 27.

Instilling oxybuprocaine hydrochloride into pupils before measuring intraocular pressure0.4%) or oxybuprocaine hydrochloride, or tetracaine hydrochloride (Colircusi)Anestesico) eye drops.

Results

The results are reported in table 12 as changes in IOP from baseline.

The aqueous composition of the present invention is effective in lowering IOP in dogs with normal ocular blood pressure. Furthermore, repeated daily administration of the aqueous composition of the present invention results in a sustained IOP lowering activity over time.

Example 6

Stability study

The ophthalmic formulations of the present invention containing 0.065% w/w NCX470 were evaluated for stability at 25 ℃, initially, for 3 months and for 6 months, and accelerated stability at 40 ℃, not exceeding 25% Relative Humidity (RH), initially, for 3 months and for 6 months, stored in 7.5mL gamma-ray sterilized LDPE bottles.

Ophthalmic formulations were prepared according to the method described in example 1.

The ophthalmic preparation comprises the following components:

0.065% w/w hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester (NCX 470);

1.0% w/w polyethylene glycol 15 hydroxystearate;

0.016% w/w benzalkonium chloride;

0.05% w/w disodium edetate dehydrate;

2.76% w/w sorbitol;

1.326% w/w sodium phosphate dibasic heptahydrate;

0.5% w/w boric acid; and

the proper amount of water for injection is 100% w/w;

primary container structure: 2.5mL of fill in a 7.5mL vial.

The results of the stability tests at 25 ℃ (table 13) and 40 ℃ (table 14) indicate that the ophthalmic formulations of the present invention are stable, and thus the ophthalmic solutions are expected to have a product shelf life of at least 24 months at ambient temperature storage conditions when storing LDPE containers.

Example 7

Preparation of a preservative-free ophthalmic composition comprising 0.065% w/w hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester (NCX470)

Ophthalmic compositions were as follows:

step 1) preparation of concentrated API solution

1.5L of water for injection is added at about 32 ℃ to 1000g of pre-weighed polyethylene glycol 15hydroxystearate in a container, which is placed in a hot water bath until the polyethylene glycol 15hydroxystearate is completely melted. The molten polyethylene glycol 15 hydroxystearate/water mixture was added to a 4L batch tank containing 65g NCX-470.

A 4L batch tank was thermostated with a water bath at 32 ℃, and the API solution was stirred until all ingredients were completely dissolved and maintained under continuous stirring until it was added to the solution of the remaining excipient carrier vehicle.

Step 2) preparation of residual excipient solvent solution

100L of water for injection was poured into a manufacturing tank made of stainless steel (316L grade) and cooled between 25 ℃ and 30 ℃. About 12 liters of this water for injection is taken out of the tank and stored in another container for use at the time of preparation.

The following compounds were added in the following order to a manufacturing tank containing water under continuous stirring; each compound was completely dissolved before the next compound was added:

500g of boric acid;

1300g sodium phosphate dibasic heptahydrate (Na)₂HPO₄ 7H₂O)；

2760g sorbitol;

the container containing the solution is rinsed multiple times with enough water for injection and the rinse is added to the manufacturing tank to complete the transfer.

Step 3) preparation of Whole ophthalmic solution

Transferring the concentrated API solution to a manufacturing tank; the 4L batch tank was rinsed several times with water for injection and the rinse was added to the whole to complete the transfer.

Water for injection was added to the manufacturing tank to adjust the final target weight to 100 kg.

The pH can be more precisely fine-tuned by using sodium hydroxide and/or hydrochloric acid.

4) Sterilization of whole eye solutions

Passing through a 0.2 μm Polyethersulfone (PES) filter (from Pall)) Redundant filtration of (a) sterilizes the ophthalmic solution.

5) Integral ophthalmic solution fill in LDPE ophthalmic primary container

Following the filtration step, the ophthalmic solution is filled in a suitable volume of preservative-free LDPE multi-dose primary container system (e.g., Aptar) in a class A environment according to conventional aseptic process practiceA system). As an alternative to preservative-free primary container systems, the entire sterile solution can be filled into single-dose primary containers, for example by the BFS technique (/Rommelag) single-dose primary container.

Example 8

Stability study

Stability of ophthalmic formulations of the present invention comprising 0.01% w/w NCX470 and 0.18% NCX470 stored in 5mL glass vials was evaluated at 25 ℃ and 40 ℃ at various time points.

Ophthalmic formulations were prepared according to the method described in example 7. In these formulations, the buffer system is a mixture of sodium phosphate dibasic heptahydrate and citric acid monohydrate, and the tonicity agent is glycerin.

0.01% w/w NCX470 ophthalmic formulation composition:

0.01% w/w hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester (NCX 470);

1.5% w/w polyethylene glycol hydroxystearate 15 ester(s) ((R))HS15)；

0.52% w/w glycerol;

2.33% w/w sodium phosphate dibasic heptahydrate;

0.36% w/w citric acid monohydrate; and

the proper amount of water for injection is 100% w/w.

Primary container structure: 3mL of filling in a 5mL glass vial.

0.18% w/w NCX470 ophthalmic formulation composition:

0.18% w/w hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester (NCX 470);

1.5% w/w polyethylene glycol hydroxystearate 15 ester (C:)HS15)；

0.52% w/w glycerol;

2.33% w/w sodium phosphate dibasic heptahydrate;

0.36% w/w citric acid monohydrate; and

the proper amount of water for injection is 100% w/w.

Primary container structure: 3mL of filling in a 5mL bottle.

The stability results for both ophthalmic formulations are reported in tables 15-18 below; the results show that the preservative-free ophthalmic formulations of the present invention are stable.

Example 9 (viscous solution)

Preparation of an ophthalmic composition comprising hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester (NCX470) 0.065% w/w and a viscosity modifier.

Ophthalmic compositions were as follows:

step 1) preparation of concentrated API solution

1.5L of water for injection is added to 1000g of pre-weighed polyethylene glycol 15hydroxystearate at about 32 ℃ in a vessel, which is placed in a hot water bath until the polyethylene glycol 15hydroxystearate is completely melted. The molten polyethylene glycol 15 hydroxystearate/water mixture was added to a 4L batch tank containing 65g NCX-470.

The 4L batch was thermostated with a water bath at 32 ℃, and the API solution was stirred until all ingredients were completely dissolved and maintained under continuous stirring until it was added to the solution of the remaining excipient vehicle.

Step 2) preparation of remaining excipient vehicle solution (viscous solution example)

100L of water for injection is poured into a manufacturing tank made of stainless steel (316L grade) at a temperature higher than 85 ℃. About 12 liters of this water for injection is taken out of the tank and stored in another container for use at the time of preparation.

200g of HPMC (/Shin-Etsu) was slowly introduced into the interior of the tank while mixing. The cellulose was slowly introduced within 15 min. Once the polymer was completely dispersed into the tank, mixing was maintained at a temperature of at least 85 ℃ for an additional 15 min. After the hold time, the bulk solution is cooled to a temperature of 25 ℃ to 30 ℃.

The following compounds were then added to the tank containing water, thus under continuous stirring, in exactly the following order; each compound was completely dissolved before the next compound was added:

50g disodium ethylenediaminetetraacetate dihydrate;

500g of boric acid;

1326g disodium phosphate heptahydrate (Na)₂HPO₄ 7H₂O)；

2760g sorbitol;

32g of benzalkonium chloride solution 50%;

the container containing the benzalkonium chloride solution is rinsed multiple times with enough water for injection and the rinse is added to the manufacturing tank to complete the transfer.

Step 3) preparation of Whole ophthalmic solution

Transferring the API concentrated solution into a manufacturing tank; the 4L batch tank was rinsed with water for injection and the rinse was added to the bulk solution to complete the transfer.

Water for injection was added to the manufacturing tank to adjust the final target weight to 100 kg.

4) Sterilization of whole eye solutions

By passing through a 0.2 μm Polyethersulfone (PES) filter (Pall) to sterilize the ophthalmic solution;

step 5) optionally, filling the bulk ophthalmic solution at a low densityPolyethylene (LDPE) ophthalmic primary containers. Typically, the formulation can be packaged in a multi-dose ophthalmic vial with a cap and dropper tip, enabling the delivery of calibrated droplets with controlled droplet size. An example of such a primary container may be from Berry-PlasticsBottle, Boston from Amcor3-piece flasks from Gerresheimer, Philips-Medisize, Bormioli or equivalent.

Claims (7)

1. An ophthalmic aqueous composition in the form of a solution comprising 0.005% to 0.10% w/w hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester, 0.5% w/w to 1.5% w/w polyethylene glycol 15hydroxystearate, 0.013% w/w to 0.02% w/w benzalkonium chloride, 0.03% w/w to 0.07% w/w disodium edetate dihydrate, a pharmaceutically acceptable carrier, a pharmaceutically acceptable carrier, a pharmaceutically acceptable carrier, a pharmaceutically acceptable carrier, a pharmaceutically acceptable carrier, a pharmaceutically acceptable carrier, a pharmaceutically acceptable carrier, a carrier, sodium phosphate dibasic heptahydrate, boric acid and water, wherein the ophthalmic aqueous composition has a pH of 5.5 to 6.5 and wherein polyethylene glycol 15hydroxystearate is the only solubilizer.

2. The aqueous ophthalmic composition of claim 1, further comprising a tonicity agent selected from sorbitol or glycerin.

3. The aqueous ophthalmic composition of claim 1, further comprising hydrochloric acid and sodium hydroxide as pH adjusting agents.

4. The aqueous ophthalmic composition according to any one of claims 1 to 3, which has an osmolality of 260-340 mOsm/kg.

5. The aqueous ophthalmic composition of any one of claims 1 to 3, wherein the amount of hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl ] -3, 5-dihydroxycyclopentyl ] -1- (2-phenylethyl) -2-propen-1-yl ester is 0.021% w/w, 0.042% w/w, 0.065% w/w or 0.10 w/w.

6. The aqueous ophthalmic composition of claim 5, consisting of: 0.065% w/w hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl]-3, 5-dihydroxycyclopentyl]-1- (2-phenylethyl) -2-propen-1-yl ester, 1.0% w/w polyethylene glycol 15hydroxystearate, 0.016% w/w benzalkonium chloride, 0.05% w/w disodium edetate dihydrate, 2.76% w/w sorbitol, 1.33% w/w disodium hydrogen phosphate heptahydrate (Na)₂HPO₄ 7H₂O), 0.5% w/w boric acid and water, wherein the pH is 6.

7. The aqueous ophthalmic composition of claim 5, consisting of: 0.10% w/w hexanoic acid, 6- (nitrooxy) -, (1S,2E) -3- [ (1R,2R,3S,5R) -2- [ (2Z) -7- (ethylamino) -7-oxo-2-hepten-1-yl]-3, 5-dihydroxycyclopentyl]-1- (2-phenylethyl) -2-propen-1-yl ester, 1.0% polyethyleneglycol 15hydroxystearate, 0.016% w/w benzalkonium chloride, 0.05% w/w disodium edetate dihydrate, 2.76% w/w sorbitol, 1.33% w/w disodium hydrogen phosphate heptahydrate (Na)₂HPO₄ 7H₂O), 0.5% w/w boric acid and water, wherein the pH is 6.