Hemifluorinated compounds for ocular administration

Abstract

The present invention relates to semifluorinated compounds for ocular administration, in particular to ophthalmic compositions comprising semifluorinated compounds characterized by the general formula CF3-(CF2) n -CH(CH3)-(CH2) m -CH3wherein n is an integer selected from 3 to 5 and m is an integer selected from 1 to 5. The present invention also provides the use of said compositions as medicaments for topical administration to the eye, as well as a kit comprising said composition and a container comprising a dispensing device, such as a dropper.

Description

[0001] Related applications

[0002] This application is a divisional application of Chinese Patent Application No. 201680056178.5, filed on September 29, 2016, entitled "Semi-fluorinated compound for ocular drug delivery". Technical Field

[0003] This invention pertains to the field of semifluorinated compounds, compositions thereof, and their use as medicaments, particularly for ocular administration. Background Technology

[0004] Semi-fluorinated alkanes are compounds comprising at least one non-fluorinated hydrocarbon segment and at least one perfluorinated hydrocarbon segment. The general formula CF3(CF2) is described. n (CH2) m Straight-chain, unbranched, semi-fluorinated alkanes of CH, where n and m are integers representing the number of carbon atoms in each chain segment, are used for a variety of applications, such as commercially for unfolding and recoating the retina, as a long-term packing material as a substitute for vitreous fluid (H.Meinert et al., European Journal of Ophthalmology, Vol.10(3), pp.189-197, 2000), and as a washout of residual silicone oil after vitreoretinal surgery.

[0005] Formula CF3(CF2) n (CH2) m The semi-fluorinated alkanes of CH3 are described in other applications.

[0006] WO2011 / 073134 disclosed the CF3 (CF2) version. n (CH2) m A cyclosporine solution in a semi-fluorinated alkane of CH3, optionally with a co-solvent such as ethanol, wherein the semi-fluorinated alkane serves as a liquid drug delivery carrier for cyclosporine for the topical treatment of dry keratoconjunctivitis.

[0007] WO2014 / 041055 describes formula CF3(CF2). n (CH2) m CH3 (which can optionally be represented as F(CF2)) n (CH2) m A mixture of semi-fluorinated alkanes (H). These mixtures are described as ocular alternatives or for the treatment of patients with dry eye and / or meibomian gland dysfunction.

[0008] The naming convention commonly used for semi-fluorinated compounds with linear and unbranched segments is FnHm, where F represents a perfluorinated hydrocarbon segment, H represents a nonfluorinated segment, and n and m define the number of carbon atoms in the corresponding segment. For example, F3H3 is used for perfluoropropylpropane CF3(CF2)2(CH2)2CH3, which is 1-perfluoropropylpropane.

[0009] However, the general formula CF3-(CF2) n -CH(CH3)-(CH2) m -CH3 semifluorinated alkanes have not been described, and in particular, have not been described for ophthalmic applications. Therefore, one object of the present invention is to provide such compounds, particularly in relation to their use in compositions, especially in the field of ophthalmic applications. Summary of the Invention

[0010] In a first aspect, the present invention relates to a compound comprising the general formula CF3-(CF2). n -CH(CH3)-(CH2) m Compositions of semi-fluorinated compounds characterized by -CH3, wherein n is an integer selected from 3 to 5, and m is an integer selected from 1 to 5. Preferably, these compositions are in liquid form, wherein the compositions contain at least 1% by weight, particularly 1% to 5% by weight, of CF3-(CF2). n -CH(CH3)-(CH2) m -CH3 represents compounds, where n is an integer selected from 3 to 5 and m is an integer selected from 1 to 5.

[0011] On the other hand, the present invention relates to compositions comprising the compound in the form of a transparent liquid solution, and also to compositions substantially composed of the compound and optionally one or more lipophilic liquid components.

[0012] In another aspect, the present invention provides the use of compositions comprising the said compounds for treating dry eye disease and / or meibomian gland dysfunction and any symptoms or conditions associated therewith.

[0013] In another aspect, the present invention provides a method for treating dry eye disease and any related symptoms or conditions, including applying the composition topically to the lacrimal sac, the lower eyelid, the surface of the eye, or ocular tissue.

[0014] In another aspect, the present invention provides a kit comprising the composition of the present invention, the composition being contained in a container having a dispensing device adapted to apply the composition topically to the eye or eye tissue. Attached Figure Description

[0015] Figure 1It is a graph depicting the relative evaporation time of the composition composed of compounds CF3(CF2)5(CH2)7CH3 and CF3-(CF2)5-CH(CH3)-(CH2)5-CH3 as a function of the percentage of compound CF3-(CF2)5-CH(CH3)-(CH2)5-CH3 in the composition (represented by solid circles and dashed lines in the figure), and the relative evaporation time of the composition composed of compounds CF3(CF2)3(CH2)4CH3 and CF3-(CF2)3-CH(CH3)-(CH2)2-CH3 as a function of the percentage of compound CF3-(CF2)3-CH(CH3)-(CH2)2-CH3 in the composition (represented by hollow circles and dashed lines in the figure).

[0016] Figure 2 It is a graph depicting the measured refractive index of a composition consisting of compounds CF3(CF2)5(CH2)7CH3 and CF3-(CF2)5-CH(CH3)-(CH2)5-CH3 as a function of the percentage of compound CF3-(CF2)5-CH(CH3)-(CH2)5-CH3 in the composition.

[0017] Figure 3 It is a graph depicting the measured refractive index of a composition consisting of compounds CF3(CF2)3(CH2)4CH3 and CF3-(CF2)3-CH(CH3)-(CH2)2-CH3 as a function of the percentage of compound CF3-(CF2)3-CH(CH3)-(CH2)2-CH3 in the composition.

[0018] Figure 4 This represents the corneal erosion size measurements (mm) over a three-day period in an in vitro eye stimulation test (EVEIT) comparing compositions containing CF3(CF2)5(CH2)7CH3 and CF3-(CF2)5-CH(CH3)-(CH2)5-CH3, a hyaluronic acid standard reference, and a 0.01% BAC positive control. 2 A schematic diagram of ). Detailed Implementation

[0019] In a first aspect, the present invention relates to general formula CF3-(CF2). n -CH(CH3)-(CH2) m -CH3 represents a semifluorinated compound, where n is an integer selected from 3 to 5, and m is an integer selected from 1 to 5.

[0020] The particularly preferred formula is CF3-(CF2). n -CH(CH3)-(CH2) mCompounds with -CH3 are those in which n is 5 and m is 5 (i.e., formula CF3-(CF2)5-CH(CH3)-(CH2)5-CH3) or those in which n is 3 or m is 2 (i.e., formula CF3-(CF2)3-CH(CH3)-(CH2)2-CH3). Other preferred compounds include CF3-(CF2)3-CH(CH3)-(CH2)3-CH3, CF3-(CF2)3-CH(CH3)-(CH2)5-CH3, CF3-(CF2)5-CH(CH3)-(CH2)1-CH3, and CF3-(CF2)5-CH(CH3)-(CH2)3-CH3.

[0021] Alternatively, the compounds of the present invention may also be commonly referred to as 2-perfluoroalkyl alkanes. For example, based on the root word alkyl, the compound CF3-(CF2)5-CH(CH3)-(CH2)5-CH3 may also be referred to as 2-perfluorohexyloctane.

[0022] These compounds have a stereocenter at the 2-alkyl position. As understood herein, the general formula includes two enantiomers, a rich mixture of the two enantiomers, and a racemic mixture.

[0023] The compounds defined above have been found to offer numerous unexpected advantages, as described below. They are particularly useful when provided as compositions, for example, for medical applications, especially for topical administration, such as topical ocular administration.

[0024] This invention relates to compounds comprising, as defined above, the general formula CF3-(CF2). n -CH(CH3)-(CH2) m Compositions of these semi-fluorinated compounds characterized by -CH3 are preferably in liquid form, for example, formulated as a liquid solution for application. In optional embodiments, the composition may be formulated as a gel, suspension, microemulsion, or spray for application. Preferably, the composition is provided in a sterile form.

[0025] In a particularly preferred embodiment, it includes the general formula CF3-(CF2). n -CH(CH3)-(CH2) m The composition of a semi-fluorinated compound characterized by -CH3 (where n is an integer selected from 3 to 5 and m is an integer selected from 1 to 5) is in liquid form and contains at least 1% by weight of the compound based on the total weight of the composition, particularly 1% to 5% by weight of the compound. In other embodiments, the composition may contain about 3% to 5% by weight, or about 5% to 10% by weight, or up to 25% by weight of the compound based on the total weight of the composition.

[0026] In a preferred embodiment, the composition as defined above is formulated into a transparent liquid solution. In this case, transparency means the absence of dispersed solid or liquid particles that would cause turbidity. In other words, this transparent solution is a pure single-phase liquid system, which may contain only small amounts of technically irrelevant particulate impurities.

[0027] Furthermore, the composition is preferably formulated as a liquid solution exhibiting a refractive index close to that of water (1.333) at room temperature (RT). In a particularly preferred embodiment, the refractive index of the liquid solution at 20°C is measured by a refractometer to be in the range of about 1.30 to about 1.35.

[0028] The compositions defined above may also contain other excipients, such as one or more acids, bases, electrolytes, buffers, solutes, antioxidants, stabilizers, and, if desired, preservatives. In a preferred embodiment, the compositions defined above are substantially free of water and / or substantially free of preservatives, such as benzalkonium chloride.

[0029] In another preferred embodiment, the compositions of the present invention are formulated as transparent liquid solutions substantially free of: (a) polymers, (b) perfluorinated compounds, and / or (c) dissolved pharmacologically active ingredients that are not semi-fluorinated alkanes. In another embodiment, the compositions as described herein may be substantially free of any form of pharmacologically active ingredient and are not semi-fluorinated alkanes.

[0030] As understood herein, the term “substantially free” regarding a component of a composition means that the component is present in no more than trace amounts, and if present in trace amounts, the component does not contribute technically to the composition.

[0031] Examples of polymers that are preferably absent in the compositions of the present invention include siloxane polymers (polymeric siloxanes), polyether polymers, and their fluorinated or perfluorinated derivatives.

[0032] Examples of perfluorinated compounds (i.e., compounds in which all hydrogen atoms are replaced by fluorine and are preferably absent in the compositions of the present invention) include perfluoroalkanes, such as perfluoronaphthenes, and haloperfluoroalkanes, such as perfluorooctyl bromide.

[0033] The compositions of the present invention, in the form of a transparent liquid solution, are also substantially free of dissolved pharmacologically active ingredients (which are not hemifluorinated alkanes). As used herein, the term "pharmacologically active ingredient" refers to any type of pharmaceutically active compound or drug that produces a pharmacological effect and is therefore suitable for prevention, diagnosis, stabilization, treatment, or generally for the treatment of a condition or disease.

[0034] However, the compounds of the present invention and the compositions comprising them have beneficial therapeutic effects at the site of administration, even if they do not contain other pharmacologically active ingredients.

[0035] In a further preferred embodiment, the composition of the present invention is essentially composed of the general formula CF3-(CF2). n -CH(CH3)-(CH2) m -CH3 (where n is an integer selected from 3 to 5 and m is an integer selected from 1 to 5) represents a compound and optionally one or more lipophilic liquid components. In one specific embodiment, the composition consists essentially of the compound CF3-(CF2)5-CH(CH3)-(CH2)5-CH3 or CF3-(CF2)3-CH(CH3)-(CH2)2-CH3, or a mixture thereof, and optionally one or more lipophilic liquid components.

[0036] As used herein, the term “consistently of…” is a so-called closed-ended language, meaning that only the mentioned components exist. Conversely, the terms “including,” “contains,” and “comprises” are used herein as so-called open-ended languages, meaning that other components may also exist.

[0037] The optional lipophilic liquid component is preferably a substantially non-aqueous and / or non-aqueous miscible excipient, such as an oily excipient, like lipids, triglyceride oils, and any other ocularly physiologically tolerant oil, or, for example, belonging to the general formula CF3 (CF2). n (CH2) m Other semi-fluorinated alkanes in the CH3 category, wherein n and m are independently selected from integers from 3 to 8, for example, CF3(CF2)3(CH2)4CH3 or CF3(CF2)5(CH2)7CH3.

[0038] This lipophilic liquid component may be present in the composition in an amount of up to about 25% by weight, up to about 50% by weight, or 75% by weight, or 90% by weight, or 95% by weight, or 97% by weight, based on the total weight of the composition.

[0039] In a further preferred embodiment, the composition according to the present invention comprises CF3-(CF2)5-CH(CH3)-(CH2)5-CH3 and CF3-(CF2)5-(CH2)7-CH3; or comprises CF3-(CF2)3-CH(CH3)-(CH2)2-CH3 and CF3-(CF2)3-(CH2)4-CH3, or is substantially composed of CF3-(CF2)5-CH(CH3)-(CH2)5-CH3 and CF3-(CF2)5-(CH2)7-CH3; or is substantially composed of (CF2)3-CH(CH3)-(CH2)2-CH3 and CF3-(CF2)3-(CH2)4-CH3.

[0040] The compositions defined above are preferably formulated to have a dynamic viscosity of not more than 10 mPa·s, and more preferably not more than 4 mPa·s, measured at standard ambient temperature and pressure (25°C, 1 atm). Preferably, the compositions have a dynamic viscosity of 1 mPa·s to 4 mPa·s. The viscosity of the compositions can be measured using any standard viscometer apparatus known in the art, such as a glass tube or capillary viscometer.

[0041] The general formula CF3-(CF2) described in this article n -CH(CH3)-(CH2) m Compounds and their compositions characterized by -CH3 (where n is an integer selected from 3 to 5 and m is an integer selected from 1 to 5) can be used for medical applications, particularly for ophthalmology, especially for topical application to the eye, such as to the lacrimal sac, to the inner lower eyelid, to the surface of the eye, or to any ocular tissue or anatomical structure associated with the eyeball that can be used for topical administration.

[0042] In particular, the compounds of the present invention are beneficial for use in the treatment of diseases and conditions, as they will benefit from the stabilization of the tear film and the tear film lipid layer and the lubrication of the ocular surface. Therefore, the compositions of the present invention are particularly suitable for treating dry eye disease (keratoconjunctivitis sicca) and / or meibomian gland dysfunction (MGD) and any symptoms or any symptoms associated therewith.

[0043] Dry eye syndrome, also known as keratoconjunctivitis sicca, can be divided into two categories: dehydrating dry eye and evaporative dry eye. These conditions are not necessarily mutually exclusive. Dehydrating dry eye is commonly observed in patients with Sjögren's syndrome or those with lacrimal gland insufficiency, lacrimal duct obstruction, or reflex hypotonia. On the other hand, evaporative dry eye has different underlying causes and is associated with increased / abnormal evaporation loss of the tear film, such as due to meibomian gland disorders, eyelid aperture disorders, blink disorders, or ocular surface diseases.

[0044] Symptoms of dry eye include dry, itchy, gritty, sandy, or foreign body sensation in the eyes; pain, soreness, stinging, or burning; itching, increased need to blink, eye fatigue, photophobia, blurred vision, redness and inflammation of the eye tissues, excessive mucus discharge and crusting / clumping, contact lens intolerance, and excessive reflex tearing.

[0045] Meibomian gland insufficiency (MGD) refers to a condition where the meibomian glands cannot produce enough oil, or the oily secretion is of poor quality or abnormal. Normally, the openings of the oil glands may become blocked or clogged, resulting in less oil secretion. The oil secreted from the glands may be granular (skinny) or otherwise abnormal and may irritate the eyes. In the early stages, patients are usually asymptomatic, but if left untreated, MGD can cause or worsen dry eye symptoms and eyelid inflammation. The oil glands become clogged due to thickened secretions. Long-term blockage of the glands eventually prevents them from producing any more oil, which can lead to permanent changes in the tear film and dry eyes.

[0046] Symptoms of meibomian gland dysfunction include dryness, burning, itching, stickiness / crusting, watering, photosensitivity, red eyes, foreign body sensation, chalazion / stye, or intermittent blurred vision.

[0047] In a preferred embodiment of the invention, the compounds and compositions thereof described above are used for topical ocular treatment of evaporative dry eye disease and / or meibomian gland dysfunction, and to alleviate any of the associated symptoms.

[0048] In one embodiment of the invention, the ophthalmic composition comprises the general formula CF3-(CF2). n -CH(CH3)-(CH2) m A semi-fluorinated compound characterized by -CH3, wherein n is an integer selected from 3 to 5, and m is an integer selected from 1 to 5; or preferably, wherein n is 5 and m is 5 (i.e., formula CF3-(CF2)5-CH(CH3)-(CH2)5-CH3), or wherein n is 3 and m is 2 (i.e., formula CF3-(CF2)3-CH(CH3)-(CH2)2-CH3), which is used to treat dry eye disease (keratoconjunctivitis sicca) or related symptoms. In another embodiment, this composition can be used to treat meibomian gland dysfunction or related symptoms.

[0049] In a further embodiment, treatment of these conditions is preferably carried out by administering an effective amount of the composition to a patient in need, the composition consisting essentially of the semi-fluorinated alkane and optionally one or more lipophilic liquid components, the lipophilic liquid components preferably being substantially non-water-soluble and / or non-water-soluble excipients, such as oily excipients, like lipids, triglyceride oils and any other ocularly physiologically tolerant oils, or other semi-fluorinated alkane, such as those belonging to the general formula CF3 (CF2). n (CH2) m The type of CH3, where n and m are independently selected from integers from 3 to 8, such as CF3(CF2)3(CH2)4CH3 or CF3(CF2)5(CH2)7CH3.

[0050] In a further preferred embodiment of the invention, the compounds and compositions thereof described above are used for topical ocular treatment of corneal injuries. Therefore, the compounds and compositions are actively supporting the corneal healing process for corneal injuries (e.g., corneal erosion).

[0051] The advantages of the above-mentioned compounds in the context of their use according to the present invention are considered to relate to their properties particularly suitable for ophthalmic applications. The refractive index of the compounds of the invention is very close to that of water, meaning that there will be little or no effect on the patient's vision after administration, unlike ophthalmic compositions based on oily carriers that may cause blurred vision upon administration. The generally low viscosity and low surface tension of these compounds, and especially their high wetting and spreading abilities, also ensure that they are rapidly contained and adapted for administration onto the ocular surface.

[0052] As will be more clearly shown in the examples below, these compounds and their compositions have been found to be biocompatible and do not exhibit significant cytotoxic effects. Furthermore, it has been demonstrated that these compounds and their compositions not only exhibit good tolerability in the eye but also provide beneficial effects in ocular lubrication and tear film stabilization, in the form of relieving symptoms in patients with mild to moderate symptoms associated with dry eye disease and / or meibomian gland dysfunction. Patients with dry eye disease and / or dysfunctional meibomian glands often express opaque and thicker tarsal plates, which can lead to an abnormal lipid layer in the tear film. Not wishing to be bound by theory, it is believed that the physicochemical properties of the compounds of the present invention can play a role in stabilizing the lipid layer of the tear film, for example, by dissolving certain lipid components or improving the fluidity of the lipid layer.

[0053] On the other hand, the present invention provides a method for treating dry eye disease and any related symptoms or conditions, comprising applying the composition of the invention topically to the lacrimal sac, to the inner lower eyelid, to the surface of the eye, or to ocular tissues. Preferably, the composition may be applied to the eye or ocular tissues up to four times daily.

[0054] Furthermore, the present invention provides a kit comprising any of the compositions described above and a container for containing the compositions. The container preferably includes a dispensing device, such as an eye dropper, suitable for topical application of the composition to the ocular sac, lower eyelid, or ocular tissue.

[0055] In a further preferred embodiment, the dispensing device includes a dropper of a specified size to dispense droplets with a volume of about 8 μL to 15 μL, preferably about 8 μL to 12 μL, and more preferably about 10 μL. With small droplet volumes, precise dosing to the eye can be achieved, and excessive excretion of most of the composition from the eye after administration can be avoided.

[0056] Item 1. An ophthalmic composition comprising a compound characterized by the following general formula:

[0057] CF3-(CF2) n -CH(CH3)-(CH2) m -CH3

[0058] Where n is an integer selected from 3 to 5, and m is an integer selected from 1 to 5.

[0059] Item 2. The composition according to Item 1, wherein n is 5 and m is 5, or wherein n is 3 and m is 2.

[0060] Item 3. The composition according to any one of items 1 or 2, which is in liquid form and contains at least 1% by weight of the compound based on the total weight of the composition, particularly from 1% to 5% by weight of the compound.

[0061] Item 4. The composition according to any one of the preceding items is formulated as a transparent liquid solution.

[0062] Item 5. The composition according to any one of the preceding items, which substantially does not contain:

[0063] (a) Polymer,

[0064] (b) perfluorinated compounds, and / or

[0065] (c) Not a dissolved pharmacologically active ingredient of a semi-fluorinated alkane.

[0066] Item 6. The composition according to any one of the preceding items has a dynamic viscosity of not more than 10 mPa·s at ambient temperature and pressure, and preferably not more than 4 mPa·s.

[0067] Item 7. The composition according to any one of the preceding items, which is substantially composed of the said compound and optionally one or more lipophilic liquid components.

[0068] Item 8. The composition according to any one of the preceding items, used as a medicine.

[0069] Item 9. The composition according to Item 8, wherein the composition is applied topically to the lacrimal sac, the lower eyelid, the surface of the eye, or the ocular tissue.

[0070] Item 10. The composition according to Item 8 or 9, for the treatment of dry eye disease (keratoconjunctivitis sicca) and / or meibomian gland dysfunction or related symptoms or conditions.

[0071] Item 11. The composition according to any one of items 8 to 10, used for treating corneal damage.

[0072] Item 12. A kit comprising an ophthalmic composition according to any one of items 1 to 7 and a container for containing the composition, wherein the container includes a dispensing device adapted to apply the composition topically to the lacrimal sac, the inner lower eyelid, the surface of the eye, or ocular tissue.

[0073] Item 13. The kit according to Item 12, wherein the dispensing device includes a dropper size for dispensing droplets with a volume of 8 μL to 15 μL.

[0074] Example

[0075] CF3-(CF2)5-CH(CH3)-(CH2)5-CH3(2-perfluorohexyl-octane, C 14 F 13 H 17 Preparation of )

[0076] The compound CF3-(CF2)5-CH(CH3)-(CH2)5-CH3 can be prepared as follows: In the presence of a radical initiator, perfluorohexyl iodine is radically added to 1-octene (in this paper, perfluorohexyl iodine is mixed with 1-octene and a radical initiator of AIBN, and the resulting solution is maintained at 80°C for 30 minutes and then cooled). The resulting iodine adduct is then reduced with a hydride (i.e., LiAlH4) or by hydrogenation (i.e., catalytic hydrogenation in the presence of a catalyst such as Pd / C) to form 2-perfluorohexyl-octane, which is then purified by fractional distillation. The formula CF3-(CF2) is as defined above. n -CH(CH3)-(CH2) m Other compounds of -CH3 can be prepared similarly using this general method.

[0077] CF3-(CF2)5-CH(CH3)-(CH2)5-CH3: 1H-NMR (CDCl3, 400MHz): 2.17-2.33(m,1H,CH), 1.67-1.77(m,2H,CH2), 1.25-1.40(m,8H,CH2), 1.15(d,3H,CH3), 0.90(t,3H,CH3).

[0078] CF3-(CF2)3-CH(CH3)-(CH2)2-CH3(2-perfluorobutylpentane, C9F9H) 11 Preparation of )

[0079] The compound CF3-(CF2)3-CH(CH3)-(CH2)2-CH3 can be prepared using perfluorobutyl iodine and 1-pentene as starting materials according to the general method described above.

[0080] CF3-(CF2)3-CH(CH3)-(CH2)2-CH3: 1 H-NMR (CDCl3, 400MHz): 2.21-2.32(m,1H,CH), 1.68-1.74(m,2H,CH2), 1.45-1.55(m,2H,CH2), 1.12(d,3H,CH3), 0.92(t,3H,CH3).

[0081] In vitro cytotoxicity assay

[0082] The cytotoxicity of a composition comprising 1.3 wt% CF3-(CF2)5-CH(CH3)-(CH2)5-CH3 and 95.8 wt% CF3(CF2)5(CH2)7CH3 was evaluated by a cell growth inhibition assay, which predicted cytotoxic or necrotic effects that correlated well with animal experiments and high sensitivity.

[0083] The composition was extracted with cell culture medium (DMEM supplemented with 10% FBS) for approximately 24 hours under stirring. The resulting extract was then incubated with mouse cell line L929 for 68–72 hours, and protein content was analyzed as a measure of cytotoxicity using a BCA (dicaprinic acid) assay. No inhibition of cell growth or cell lysis was observed.

[0084] Similar in vitro cytotoxicity assays were performed on compositions containing about 23.7% by weight of CF3-(CF2)5-CH(CH3)-(CH2)5-CH3 and about 75.6% by weight of F6H8.

[0085] Tear film analysis study

[0086] A composition comprising 98.3% by weight CF3(CF2)5(CH2)7CH3 and 1.2% by weight CF3-(CF2)5-CH(CH3)-(CH2)5-CH3 was tested in an observational study of patients with mild to moderate evaporative dry eye syndrome. The clear, colorless liquid composition was provided in 5 ml vials fitted with droppers sized to dispense approximately 10 μl of the liquid into the ocular sac per drop. Patients wearing contact lenses were excluded from the study. With informed consent, patients were advised to instill 3–4 drops into both eyes daily, transitioning to a daily dose of 30–40 μl. Patients returned after a 5–7 week follow-up. Clinical data were collected from 29 patients at baseline and during the 5–7 week follow-up.

[0087] a) Tear film analysis

[0088] Improvements in tear film volume and tear film stability were observed during the study, as evidenced by increases in Schirmer I and TFBUT. Retrospective statistical analysis reinforced this observation, as the differences in TFBUT between baseline and follow-up were highly significant (paired two-sided t-test: p = 0.0026 (right eye) and p = 0.0006 (left eye)). No changes were detected in tear osmolality.

[0089] Subjective dry eye questionnaire (ocular surface disease index, OSDI) showed a reduction in the severity of patients’ subjective symptoms after using a composition containing 98.3% by weight CF3(CF2)5(CH2)7CH3 and 1.2% by weight CF3-(CF2)5-CH(CH3)-(CH2)5-CH3 for 5–7 weeks, which was reflected in the lower scores in follow-up and retrospective statistical analysis (paired two-sided t-test: p<0.0001).

[0090]

[0091] b) Corneal staining (Oxford classification system)

[0092] Corneal fluorescein staining is an indicator of corneal damage (loss of cell-to-cell connections). Data showed a reduction in corneal damage after 5–7 weeks of treatment, which can be seen in the change in the number of patients diagnosed as grade 1 or 2 at baseline to grade 0 at follow-up. This difference from the initial degree of damage was statistically significant, as shown by the Wilcoxon signed-rank test: p = 0.0013 (right eye) and p = 0.0041 (left eye).

[0093]

[0094] c) Doctor's symptom assessment

[0095] At baseline and follow-up, physicians inquired whether patients currently experienced typical dry eye symptoms. As shown in the table below, a small number of DED-related symptoms were reported 5–7 weeks after treatment.

[0096]

[0097] d) Meibomian secretion analysis

[0098] In a healthy eye, meibum is a clear fluid secreted by the meibomian glands. More opaque and thicker meibum is an indicator of meibomian gland dysfunction. A descriptive examination of the patients' meibum was performed at baseline and during follow-up. Based on the data obtained, meibum quality improved in many cases. In seven cases, treatment induced a reduction in expressible meibum (from clear meibum to none).

[0099]

[0100]

[0101] e) Safety parameters

[0102] No changes in visual acuity or intraocular pressure were observed, indicating that the use of the composition containing 98.3% by weight of CF3(CF2)5(CH2)7CH3 and 1.2% by weight of CF3-(CF2)5-CH(CH3)-(CH2)5-CH3 is safe and does not interfere with these ophthalmic parameters.

[0103]

[0104] Differential scanning calorimetry

[0105] Differential scanning calorimetry (DSC 1, Mettler Toledo, Greifensee, Switzerland) was used to characterize the structure and phase properties of mixtures of CF3(CF2)5-CH(CH3)-(CH2)5-CH3 and CF3(CF2)5(CH2)7CH3. Data on the transformations were obtained using DSC through heating scans at a rate of 1 °C / min. A sealed standard aluminum crucible (40 μl, Mettler Toledo) was used.

[0106]

[0107]

[0108] The presence of CF3(CF2)5-(CH2)7-CH3(F6H8) in the mixture results in a significant decrease in melting temperature. The enthalpy of fusion is also reduced, indicating that the semi-fluorinated alkane does not crystallize. This difference will have beneficial effects on the application of this compound to the eye as a tear substitute or lubricant; for example, in terms of its ability to mix with and modulate the tear film lipid layer. Furthermore, this effect can be advantageously modulated by varying the amount of the compound added to the ophthalmic composition.

[0109] DSC measurements were also performed on a series of mixtures of 23.74 wt% F3(CF2)5-CH(CH3)-(CH2)5-CH3 and CF3(CF2)5(CH2)7CH3 with tetradecane (C14). Conversion data were obtained using temperature sweeps (heating rates of 0.2 °C / min, 0.5 °C / min, and 1 °C / min). The ending temperature was determined using a heating rate extrapolated to 0 °C / min, while the starting temperature was determined using the average of the three measurements. A decrease in enthalpy of fusion was observed compared to the mixture of tetradecane and pure CF3(CF2)5(CH2)7CH3, indicating that some tetradecane dissolves in the liquid portion of CF3(CF2)5-CH(CH3)-(CH2)5-CH3, and that this compound has a stronger solubilizing ability compared to CF3(CF2)5(CH2)7CH3.

[0110] Refractive index and evaporation time .

[0111] The evaporation time of mixtures of semi-fluorinated alkanes CF3(CF2)3(CH2)4CH3(F4H5) and CF3-(CF2)3-CH(CH3)-(CH2)2-CH3 was evaluated. 10 μL droplets of each mixture were placed on a glass surface at room temperature. The time until evaporation was recorded via video monitoring.

[0112]

[0113] It was observed that increasing the amount of compound CF3-(CF2)3-CH(CH3)-(CH2)2-CH3 appeared to increase the evaporation time of the mixture (see [link]). Figure 1 ).

[0114] Similarly, the evaporation time of mixtures of semi-fluorinated alkanes CF3(CF2)5(CH2)7CH3(F6H8) and CF3(CF2)5-CH(CH3)-(CH2)5-CH3 was evaluated.

[0115]

[0116] In contrast, it was unexpectedly observed that the increase in the percentage of semi-fluorinated alkanes CF3(CF2)5-CH(CH3)-(CH2)5-CH3 significantly reduced the evaporation time of the mixture (see [link]). Figure 1 ).

[0117] Therefore, it depends on the general formula CF3-(CF2). n -CH(CH3)-(CH2) m The compound characterized by -CH3, wherein n is an integer selected from 3 to 5, m is an integer selected from 1 to 5, and the amount present in the mixture, can adjust and fine-tune the composition to the requirements of the intended ocular use, such as prolonging or reducing the time the composition remains on the ocular surface.

[0118] The refractive index of the mixture was also determined. For topical ophthalmic compositions, the refractive index of the composition should preferably be similar to or adjusted to the refractive index of the eye or lens, for example, as close as possible to the refractive index of physiological tears. If the refractive index of the composition is not similar, patients may experience blurred vision or impaired vision when applied to the ocular surface. The amount of compound CF3(CF2)5-CH(CH3)-(CH2)5-CH3 was observed to have an effect on the refractive index. Figure 2 The study depicts the increase in refractive index value as the content of 2-perfluorohexyloctane in a mixture with 1-perfluorohexyloctane increases. Figure 3 Similarly, it was shown that the refractive index value increased with the increase of the content of 2-perfluorobutylpentane in the mixture with 1-perfluorobutylpentane.

[0119] By varying the amount of these compounds in the mixture, for example in the general formula CF3-(CF2) n -CH(CH3)-(CH2) m The amount of the compound characterized by -CH3, where n is an integer selected from 3 to 5 and m is selected from 1 to 5, is also feasible to adapt the composition to the requirements of the intended ocular use, such as to patients with altered tear composition and refractive index due to ocular condition and / or age.

[0120] Ex vivo eye stimulation test (EVEIT)

[0121] For two compositions containing CF3(CF2)5(CH2)7CH3, namely compositions consisting of a mixture of semi-fluorinated alkanes CF3(CF2)5(CH2)7CH3 and CF3(CF2)5-CH(CH3)-(CH2)5-CH3 (composition A having 0.17 wt% CF3(CF2)5-CH(CH3)-(CH2)5-CH3 and composition B having 64 wt% CF3(CF2)5-CH(CH3)-(CH2)5-CH3), hyaluronic acid is used. As a reference, 0.01% BAC (benzalkonium chloride) was used as a positive control, and the corneal healing process was compared with a similar ex vivo eye stimulation test (EVEIT) described in M. Frentz et al, Altern. to Lab. Anim., 2008(36)p 25-32; and N. Schrage et al, Graefes Arch Clin Exp Ophthalmol 2012(250),1330-1340).

[0122] Methods. Rabbit corneas were obtained and placed in an artificial anterior chamber, which was gently filled with serum-free minimum essential culture medium (Eagle's MEM) containing Earle's salts and HEPES buffer for nutrition. This medium was continuously replenished by a micropump to mimic the physiological conditions of the eye. The culture chamber was maintained at 32°C in normal air, without supplemental CO2 and >95% relative humidity. Five corneas (n=5) / test material were used, except for a positive control of two corneas (n=2).

[0123] After stabilizing in the culture chamber for 12 hours, the corneas were evaluated under a microscope, and corneas with intact epithelium and no opacity were selected. Four small abrasions (2.3–4.3 mm) were made on the selected corneal surface using a corneal drill. 2 All defects were monitored by fluorescein staining (0.17% aqueous solution) and microscopy.

[0124] The test substance was applied 1 hour after induced corneal erosion and then applied six times daily to the apex of the cornea (30-50 μL every four hours). A soft-top cannula for continuous aspiration was placed at the lowest point of the corneal scleral region in the culture chamber to remove any excess fluid. The experiment was terminated after 3 days of application. Biomicroscopic images of the cornea were taken daily using a phase-contrast microscope integrated camera (KY-F1030U, JVC, (BadVilbel, DE) mounted on a Z16 APO microscope (Wetzlar, DE)) to document the corneal healing process. All defects were monitored by staining with sodium fluorescein (0.17% aqueous solution), with yellow-green fluorescence indicating areas of epithelial defects. Erosion size was determined using the software tools of the microscope (DISKUS). At the end of 3 days, the experiment was terminated and all corneas were fixed in 3.7% formaldehyde and stained with hematoxylin-eosin for microscopic evaluation. To monitor corneal metabolic activity, glucose and lactate concentrations were spectrophotometrically quantified in the effluent medium from the artificial anterior chamber.

[0125] Results. The composition compared to the standard reference hyaluronic acid. In contrast, after inducing corneal erosion, the two mixtures of semi-fluorinated alkane (compositions A and B as described above) were observed to have similar positive effects on the corneal treatment process.

[0126] Corneal erosion size measurement / average mm 2 (SD)

[0127]

[0128] Day 3 Histological observation

[0129]

[0130]

[0131] No significant difference in positive corneal healing was observed between composition B and composition A, which contain 64% by weight of the semi-fluorinated alkane CF3(CF2)5-CH(CH3)-(CH2)5-CH3 in the total weight of the compositions. For both compositions, as with the reference composition, mechanically induced epithelial erosion was significantly reduced and essentially nonexistent after day 2 of treatment. Figure 3 The results of corneal erosion size measurements for the test composition, reference, and positive control during the 0-3 day EVEIT experiment are described.

[0132] As shown in the table above, microscopic histological examination of the cornea in cross-section after the experiment on day 3 showed that the results were obtained using compositions A, B, and the reference. No significant residual defects or differences were found in the treated corneas.

[0133] Furthermore, no corneal toxicity was observed for these compositions based on metabolic activity indicated by glucose / lactate measurements.

[0134] In stark contrast, during the three-day experiment, the positive control containing 0.01% preservative BAC showed a gradual increase in induced epithelial damage.

Claims

1. A compound, characterized in that... The compound has the structural formula CF3-(CF2)5-CH(CH3)-(CH2)5-CH3.

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