Stable cyclosporine ophthalmic formulation and manufacturing process thereof

Abstract

A stable nanomicelle ophthalmic solution comprising cyclosporine and a method for preparing the nanomicelle solution. The present invention further relates to a stable nanomicelle solution comprising cyclosporine forms having characteristic XRD peaks at 6.9, 7.8, 9.4, and 15.9 2-theta (degrees), or amorphous cyclosporine. The present invention also relates to the use of the stable nanomicelle ophthalmic solution in the treatment of dry eye.

Description

[Technical field]

[0001] The present invention relates to a stable nanomicelle ophthalmic solution containing cyclosporine and a method for preparing the nanomicelle solution.The present invention further relates to a stable nanomicelle solution containing cyclosporine forms with characteristic XRD peaks at 6.9, 7.8, 9.4, and 15.9 2-theta (degrees), or amorphous cyclosporine.The present invention further relates to a stable nanomicelle solution containing cyclosporine forms with characteristic XRD peaks at 7.4, 8.7, 14.4, and 17.5 2-theta (degrees), or cyclosporine forms with characteristic XRD peaks at 8.5, 9.3, 11.6, and 20.3 2-theta (degrees).The present invention also relates to the use of this stable nanomicelle ophthalmic solution in dry eye. [Background technology]

[0002] A cyclosporine nanomicelle ophthalmic solution is generally disclosed in US Patent No. 10,918,694, which comprises 0.087-0.093% by weight of cyclosporine, polyoxyl lipid or fatty acid, and polyalkoxylated alcohol. Preferably, it comprises 0.087-0.093% by weight of cyclosporine, 0.5-5% of one or more selected from the group consisting of HCO-40, HCO-60, HCO-80, and HCO-100, and about 0.01-0.1% of octoxynol-40. Further, a method of preparing such a cyclosporine solution is disclosed. The method comprises dissolving cyclosporine in a polyoxyl castor oil, such as hydrogenated castor oil, and a polyalkoxylated alcohol, such as octoxynol, at 60° C. before adding to the aqueous phase. Specifically, the method of preparing the ophthalmic solution comprises the following steps: HCO-40 is melted in a flask heated to about 60°C with stirring. Once liquefied, the required amount of cyclosporine is added and mixed until dissolved and uniform. Octoxynol-40 is then heated to about 60°C and once liquefied, it is added to the cyclosporine HCO-40 mixture. Water for injection at about 25°C is charged to the flask containing the dissolved cyclosporine and stirred until dissolved. Other excipients such as sodium chloride and phosphate buffer are then added, followed by PVP-K90, mixed until dissolved, and made up to final volume with water for injection. However, current methods have the problem that when dissolving cyclosporine in polyoxyl castor oil such as HCO-40, there may be differences in the stability of different batches during production due to differences in the solubility and stability of different forms of cyclosporine. As a result, some batches are not stable and cyclosporine precipitates out of solution.

[0003] Furthermore, different forms of cyclosporine exhibit different solubilities and stabilities. Although cyclosporine with characteristic XRD peaks at 2-theta (degrees) 6.9, 7.8, 9.4, and 15.9 is the most soluble form of cyclosporine and is useful for preparing solution formulations, it is not the most stable form and may convert to a less soluble form of cyclosporine, thus affecting the stability of the solution. This form may convert to a more stable, more soluble cyclosporine with characteristic XRD peaks at 2-theta (degrees) 7.4, 8.7, 14.4, and 17.5 during dissolution of cyclosporine in a surfactant at 55-60°C. In turn, this may change to another more soluble form of cyclosporine with characteristic XRD peaks at 2-theta (degrees) 8.5, 9.3, 11.6, and 20.3. Furthermore, the amorphous form, which is also one of the more soluble forms and is useful for preparing solution formulations, may also recrystallize into these two less soluble forms. Such conversion is driven by several factors, including but not limited to moisture, water, solvent temperature, etc. This conversion is also dependent on temperature, stresses such as prolonged storage at elevated temperatures, etc. The process requires modification depending on the form of cyclosporine used, as the amorphous form, as well as forms with characteristic XRD peaks at 6.9, 7.8, 9.4, and 15.9 2-theta (degrees), require wetting followed by complete dissolution at temperatures up to 70°C. However, when less soluble forms are used, temperatures as high as 130°C are required. Furthermore, due to instability and conversion during the manufacturing steps, the solutions should be manufactured with strict control of API specifications, process temperatures, and times to avoid interconversion of the forms. It is somewhat difficult to determine the extent to which this conversion has occurred prior to completing the manufacturing process to obtain nanomicellar cyclosporine. This can result in the formation of seeds within the composition, either early in the process or during storage at higher temperatures, which subsequently crystallize out of the drug product, rendering the product useless. Soft mesophase or liquid crystalline forms of cyclosporine formed as non-aqueous phase intermediates can be a source of such instability.

[0004] Therefore, there is a need for a stable formulation and a method for preparing the same to prevent conversion to less soluble forms of cyclosporine during the manufacturing process and during storage. The present invention discloses a stable nanomicelle ophthalmic formulation and an improved method for making such a stable formulation. The method for making the formulation results in a stable formulation regardless of the form of cyclosporine used in the formulation. The method does not result in conversion of one form to another. More specifically, the method does not result in conversion of a dissolved form of cyclosporine to a less soluble form of cyclosporine, and further prevents precipitation of cyclosporine in the formulation during long-term stability. Summary of the Invention

[0005] According to some embodiments, one of the objects of the present invention is to provide a method for the preparation of a medicament for use in a pharmaceutical composition comprising: A method for making a stable nanomicellar ophthalmic formulation comprising cyclosporine, hydrogenated 40 polyoxyl castor oil, octoxynol-40, and an aqueous vehicle, comprising: The ophthalmic preparation is a) mixing cyclosporine with hydrogenated 40 polyoxyl castor oil at a temperature of 55° C.±2° C. or higher to form mixture A; b) prior to complete dissolution of the cyclosporine, reducing the temperature of mixture A to a temperature of 35° C.±2° C. and adding octoxynol-40; c) mixing the resulting mixture with an aqueous vehicle.

[0006] The inventors have surprisingly found that the solution stability of cyclosporine A at 35°C-40°C is greater than the solution stability at 55°C-60°C, and therefore, by reducing the temperature to 35°C-40°C, the above-mentioned stability concerns of the formulation are overcome and a more stable formulation is provided.

[0007] In another aspect, the present invention provides a method for producing a composition comprising: A stable nanomicelle ophthalmic formulation comprising cyclosporine, hydrogenated 40 polyoxyl castor oil, octoxynol-40, and an aqueous vehicle, The ophthalmic preparation is a) mixing cyclosporine with hydrogenated 40 polyoxyl castor oil at a temperature of 55° C.±2° C. or higher to form mixture A; b) prior to complete dissolution of the cyclosporine, reducing the temperature of mixture A to a temperature of 35° C.±2° C. and adding octoxynol-40; c) mixing the resulting mixture with an aqueous vehicle.

[0008] In one embodiment, the mixing of the aqueous vehicles occurs at a temperature of 35° C.±2° C. In another embodiment, the mixing of the aqueous vehicles occurs at a temperature of 55±2° C.

[0009] In one aspect, the present invention provides a stable nanomicellar ophthalmic formulation comprising cyclosporine forms having characteristic XRD peaks at 6.9, 7.8, 9.4, and 15.9 2-theta (degrees).

[0010] In another aspect, the present invention provides a stable nanomicellar ophthalmic formulation comprising an amorphous form of cyclosporine.

[0011] In another aspect, the present invention discloses a method for making a stable nanomicellar ophthalmic formulation comprising cyclosporine, hydrogenated 40 polyoxyl castor oil, octoxynol-40, and an aqueous vehicle, The ophthalmic preparation is a) mixing cyclosporine with hydrogenated 40 polyoxyl castor oil at a temperature of 55° C.±2° C. or higher to form mixture A; b) holding mixture A under vacuum to remove bubbles; c) optionally, prior to complete dissolution of cyclosporine, reducing the temperature of mixture A to a temperature of 35° C.±2° C.; d) adding octoxynol-40; e) mixing the resulting mixture with an aqueous vehicle.

[0012] In another embodiment, the present invention provides A stable nanomicelle ophthalmic formulation is provided, comprising cyclosporine, hydrogenated 40 polyoxyl castor oil, octoxynol-40, and an aqueous vehicle; The ophthalmic preparation is a) mixing cyclosporine with hydrogenated 40 polyoxyl castor oil at a temperature of 55° C.±2° C. or higher to form mixture A; b) holding mixture A under vacuum to remove bubbles; c) optionally, prior to complete dissolution of cyclosporine, reducing the temperature of mixture A to a temperature of 35° C.±2° C.; d) adding octoxynol-40; e) mixing the resulting mixture with an aqueous vehicle.

[0013] Surprisingly, it has been found that while the mixture is held under vacuum, the air bubbles are drawn out and accumulate on the surface, from which the air bubbles are gradually removed, resulting in a clean bottom.Therefore, dissolving under vacuum or intermittent removal of bubbles during dissolving may result in faster dissolution and may not require lowering the temperature of the mixture.This overcomes the above-mentioned stability concerns of the formulation and provides a more stable formulation.

[0014] In one embodiment, the invention discloses mixing the aqueous vehicles at a temperature of 35° C.±2° C. In another embodiment, the invention discloses mixing the aqueous vehicles at a temperature of 55±2° C.

[0015] In one aspect, the present invention provides a stable nanomicellar ophthalmic formulation comprising cyclosporine forms having characteristic XRD peaks at 6.9, 7.8, 9.4, and 15.9 2-theta (degrees).

[0016] In another aspect, the present invention provides a stable nanomicellar ophthalmic formulation comprising an amorphous form of cyclosporine.

[0017] In yet another aspect, the present invention provides a method for producing a composition comprising: A stable nanomicelle ophthalmic formulation is provided, comprising cyclosporine, hydrogenated 40 polyoxyl castor oil, octoxynol-40, and an aqueous vehicle; The ophthalmic preparation is a) mixing cyclosporin with hydrogenated 40 polyoxyl castor oil at a temperature of 127-130° C. until completely dissolved to form mixture A; b) adding Octoxynol-40 to mixture A at 127-130° C.; c) mixing the resulting mixture with an aqueous vehicle at a temperature of 127-130°C.

[0018] In one aspect, the present invention provides a stable nanomicellar ophthalmic formulation comprising cyclosporine forms having characteristic XRD peaks at 6.9, 7.8, 9.4, and 15.9 2-theta (degrees).

[0019] In another aspect, the present invention provides a stable nanomicellar ophthalmic formulation comprising an amorphous form of cyclosporine.

[0020] In yet another aspect, the present invention provides a method for producing a composition comprising: A stable nanomicelle ophthalmic formulation is provided, comprising cyclosporine, hydrogenated 40 polyoxyl castor oil, octoxynol-40, and an aqueous vehicle; The ophthalmic preparation is a) mixing cyclosporin with hydrogenated 40 polyoxyl castor oil at a temperature of 127-130° C. until completely dissolved to form mixture A; b) adding Octoxynol-40 to mixture A at 127-130° C.; c) mixing the resulting mixture with an aqueous vehicle at a temperature of 127-130°C; Cyclosporine exists in forms with characteristic XRD peaks at 7.4, 8.7, 14.4, and 17.5 2-theta (degrees).

[0021] In yet another aspect, the present invention provides a method for producing a composition comprising: A stable nanomicelle ophthalmic formulation is provided, comprising cyclosporine, hydrogenated 40 polyoxyl castor oil, octoxynol-40, and an aqueous vehicle; The ophthalmic preparation is a) mixing cyclosporin with hydrogenated 40 polyoxyl castor oil at a temperature of 127-130° C. until completely dissolved to form mixture A; b) adding Octoxynol-40 to mixture A at 127-130° C.; c) mixing the resulting mixture with an aqueous vehicle at a temperature of 127-130°C; Cyclosporine exists in forms with characteristic XRD peaks at 2-theta (degrees) 8.5, 9.3, 11.6, and 20.3.

[0022] In yet another aspect, the present invention provides a method for producing a composition comprising: A stable nanomicelle ophthalmic formulation is provided, comprising cyclosporine, hydrogenated 40 polyoxyl castor oil, octoxynol-40, and an aqueous vehicle; The ophthalmic preparation is a) mixing hydrogenated 40 polyoxyl castor oil and octoxynol-40 at a temperature of 127-130° C. to form a mixture A; b) adding cyclosporine to mixture A at 127-130°C; c) mixing the resulting mixture with an aqueous vehicle at a temperature of 127-130°C.

[0023] In one aspect, the present invention provides a stable nanomicellar ophthalmic formulation comprising cyclosporine forms having characteristic XRD peaks at 6.9, 7.8, 9.4, and 15.9 2-theta (degrees).

[0024] In another aspect, the present invention provides a stable nanomicellar ophthalmic formulation comprising an amorphous form of cyclosporine.

[0025] In another aspect, the present invention provides a stable nanomicellar ophthalmic formulation comprising cyclosporine in a form having characteristic XRD peaks at 7.4, 8.7, 14.4, and 17.5 2-theta (degrees).

[0026] In another aspect, the present invention provides a stable nanomicellar ophthalmic formulation comprising cyclosporine in a form having characteristic XRD peaks at 8.5, 9.3, 11.6, and 20.3 2-theta (degrees).

[0027] In a preferred embodiment, the present invention provides a method for producing a method for treating a cancer cell comprising the steps of: 0.09% by weight of cyclosporine, about 1.0% by weight of hydrogenated 40 polyoxyl castor oil, about 0.05% by weight of Octoxynol-40, and A method for making a stable nanomicellar ophthalmic formulation comprising an aqueous vehicle is provided, The method comprises: a) mixing cyclosporine with hydrogenated 40 polyoxyl castor oil at a temperature of 55° C.±2° C. or higher to form mixture A; b) prior to complete dissolution of the cyclosporine, reducing the temperature of mixture A to a temperature of 35° C.±2° C. and adding octoxynol-40; c) mixing the resulting mixture with an aqueous vehicle.

[0028] In one embodiment, the aqueous vehicle is mixed at a temperature of 35° C.±2° C. In another embodiment, the aqueous vehicle is mixed at a temperature of 55±2° C.

[0029] In a preferred embodiment, the present invention also provides a stable nanomicellar ophthalmic formulation prepared by the above method.

[0030] In a preferred embodiment, the present invention provides a method for producing a method for treating a cancer cell comprising the steps of: 0.09% by weight of cyclosporine, about 1.0% by weight of hydrogenated 40 polyoxyl castor oil, about 0.05% by weight of Octoxynol-40, and A method for making a stable nanomicellar ophthalmic formulation comprising an aqueous vehicle is provided, The ophthalmic preparation is a) mixing cyclosporine with hydrogenated 40 polyoxyl castor oil at a temperature of 55° C.±2° C. or higher to form mixture A; b) holding mixture A under vacuum to remove bubbles; c) optionally, prior to complete dissolution of cyclosporine, reducing the temperature of mixture A to a temperature of 35° C.±2° C.; d) adding octoxynol-40; e) mixing the resulting mixture with an aqueous vehicle.

[0031] In one embodiment, the aqueous vehicle is mixed at a temperature of 35° C.±2° C. In another embodiment, the aqueous vehicle is mixed at a temperature of 55±2° C.

[0032] In a preferred embodiment, the present invention also provides a stable nanomicellar ophthalmic formulation prepared by the above method.

[0033] In one aspect, the stable nanomicellar ophthalmic formulation comprises: About 0.20 to 0.550% by weight of sodium phosphate monobasic, about 0.23 to 0.465% by weight of dibasic sodium phosphate; about 0.05% by weight of sodium chloride, about 0.3% by weight povidone, Sodium hydroxide / hydrochloric acid to adjust the pH, and Further included is water for injection.

[0034] In one aspect, the present invention provides a stable nanomicellar ophthalmic formulation having a pH of about 5.0 to 8.0. More preferably, the pH of the formulation is about 6.5 to 7.2.

[0035] Furthermore, the present invention provides a stable nanomicellar ophthalmic formulation having an osmolality of about 150 to about 200 mOsmol / kg.

[0036] In another aspect, the present invention provides a stable nanomicellar ophthalmic formulation comprising cyclosporine forms having characteristic XRD peaks at 6.9, 7.8, 9.4, and 15.9 2-theta (degrees).

[0037] In yet another aspect, the present invention provides a stable nanomicellar ophthalmic formulation comprising an amorphous form of cyclosporine.

[0038] In another aspect, the present invention provides a stable nanomicellar ophthalmic formulation that is substantially free of cyclosporine forms having characteristic XRD peaks at 7.4, 8.7, 14.4, and 17.5 2-theta (degrees).

[0039] In another aspect, the present invention provides a stable nanomicellar ophthalmic formulation that is substantially free of cyclosporine forms having characteristic XRD peaks at 8.5, 9.3, 11.6, and 20.3 2-theta (degrees).

[0040] In another aspect, the present invention provides a stable nanomicelle ophthalmic formulation, comprising: a) mixing cyclosporine with hydrogenated 40 polyoxyl castor oil at a temperature of 55° C.±2° C. or higher to form mixture A; b) prior to complete dissolution of the cyclosporine, reducing the temperature of mixture A to a temperature of 35° C.±2° C. and adding octoxynol-40; c) then mixing the resulting mixture with an aqueous vehicle at 35° C.±2° C., wherein mixture A drops to a temperature of 35° C.±2° C. in less than 65 minutes. Preferably, mixture A drops to a temperature of 35° C.±2° C. in 40-50 minutes.

[0041] In one aspect, the present invention discloses mixing mixture A in step (a) for 20-30 minutes, preferably 20-25 minutes, more preferably 20±2 minutes.

[0042] In one embodiment, the invention discloses reducing the temperature to 35° C.±2° C. and stirring for 60-70 minutes. Preferably, the mixture is stirred at a temperature of 35° C.±2° C. for 60±5 minutes.

[0043] In yet another aspect, the present invention provides a method for producing a composition comprising: 0.09% by weight of cyclosporine, about 1.0% by weight of hydrogenated 40 polyoxyl castor oil, about 0.05% by weight of Octoxynol-40, and A stable nanomicellar ophthalmic formulation comprising an aqueous vehicle is provided, The ophthalmic preparation is a) mixing cyclosporin with hydrogenated 40 polyoxyl castor oil at a temperature of 127-130° C. until completely dissolved to form mixture A; b) adding Octoxynol-40 to mixture A at 127-130° C.; c) mixing the resulting mixture with an aqueous vehicle at a temperature of 127-130°C.

[0044] In one aspect, the present invention provides a stable nanomicellar ophthalmic formulation comprising cyclosporine forms having characteristic XRD peaks at 6.9, 7.8, 9.4, and 15.9 2-theta (degrees).

[0045] In another aspect, the present invention provides a stable nanomicellar ophthalmic formulation comprising an amorphous form of cyclosporine.

[0046] In yet another aspect, the present invention provides a method for producing a composition comprising: 0.09% by weight of cyclosporine, about 1.0% by weight of hydrogenated 40 polyoxyl castor oil, about 0.05% by weight of Octoxynol-40, and A stable nanomicellar ophthalmic formulation comprising an aqueous vehicle is provided, The ophthalmic preparation is a) mixing cyclosporin with hydrogenated 40 polyoxyl castor oil at a temperature of 127-130° C. until completely dissolved to form mixture A; b) adding Octoxynol-40 to mixture A at 127-130° C.; c) mixing the resulting mixture with an aqueous vehicle at a temperature of 127-130°C; Cyclosporine exists in forms with characteristic XRD peaks at 7.4, 8.7, 14.4, and 17.5 2-theta (degrees).

[0047] In yet another aspect, 0.09% by weight of cyclosporine, about 1.0% by weight of hydrogenated 40 polyoxyl castor oil, about 0.05% by weight of Octoxynol-40, and A stable nanomicellar ophthalmic formulation comprising an aqueous vehicle, The ophthalmic preparation is a) mixing cyclosporin with hydrogenated 40 polyoxyl castor oil at a temperature of 127-130° C. until completely dissolved to form mixture A; b) adding Octoxynol-40 to mixture A at 127-130° C.; c) mixing the resulting mixture with an aqueous vehicle at a temperature of 127-130°C; Cyclosporine exists in forms with characteristic XRD peaks at 2-theta (degrees) 8.5, 9.3, 11.6, and 20.3.

[0048] In yet another aspect, the present invention provides a method for producing a composition comprising: 0.09% by weight of cyclosporine, about 1.0% by weight of hydrogenated 40 polyoxyl castor oil, about 0.05% by weight of Octoxynol-40, and A stable nanomicellar ophthalmic formulation comprising an aqueous vehicle is provided, The ophthalmic preparation is a) mixing hydrogenated 40 polyoxyl castor oil and octoxynol-40 at a temperature of 127-130° C. to form a mixture A; b) adding cyclosporine to mixture A at 127-130°C; c) mixing the resulting mixture with an aqueous vehicle at a temperature of 127-130°C.

[0049] In one aspect, the present invention provides a stable nanomicellar ophthalmic formulation comprising cyclosporine forms having characteristic XRD peaks at 6.9, 7.8, 9.4, and 15.9 2-theta (degrees).

[0050] In another aspect, the present invention provides a stable nanomicellar ophthalmic formulation comprising an amorphous form of cyclosporine.

[0051] In another aspect, the present invention provides a stable nanomicellar ophthalmic formulation comprising cyclosporine in a form having characteristic XRD peaks at 7.4, 8.7, 14.4, and 17.5 2-theta (degrees).

[0052] In another aspect, the present invention provides a stable nanomicellar ophthalmic formulation comprising cyclosporine in a form having characteristic XRD peaks at 8.5, 9.3, 11.6, and 20.3 2-theta (degrees).

[0053] In yet another aspect, the present invention provides a method for producing a composition comprising: 0.09% by weight of cyclosporine, about 1.0% by weight of hydrogenated 40 polyoxyl castor oil, about 0.05% by weight of Octoxynol-40; About 0.20 to 0.550% by weight of sodium phosphate monobasic, about 0.23 to 0.465% by weight of dibasic sodium phosphate; about 0.05% by weight of sodium chloride, about 0.3% by weight povidone, Sodium hydroxide / hydrochloric acid to adjust the pH, and A method for making a stable nanomicellar ophthalmic formulation comprising water for injection is provided, The method comprises: a) mixing cyclosporine with hydrogenated 40 polyoxyl castor oil at a temperature of 55° C.±2° C. or higher to form mixture A; b) prior to complete dissolution of the cyclosporine, reducing the temperature of mixture A to a temperature of 35° C.±2° C. and adding octoxynol-40 to form an API mixture; c) adding the API mixture to water for injection (WFI); d) adding the remaining excipients to step (c) in the following order: sodium phosphate monobasic, then sodium phosphate dibasic, then sodium chloride, then polyvinylpyrrolidone; e) adjusting the pH to 6.5-7.2 and bringing to final volume with WFI.

[0054] In one aspect, the present invention discloses mixing mixture A in step (a) for 20-30 minutes, preferably 20-25 minutes, more preferably 20±2 minutes.

[0055] In one embodiment, the present invention discloses reducing the temperature of Mixture A to 35° C.±2° C. in less than 65 minutes. Preferably, the temperature of Mixture A is reduced to 35° C.±2° C. in 40-50 minutes.

[0056] In another aspect, the invention discloses stirring the mixture in step (b) for 60 to 70 minutes. Preferably, the mixture is stirred for 60±5 minutes at a temperature of 35° C.±2° C.

[0057] In one embodiment, the present invention discloses blending the API mixture into WFI at a temperature of 35° C.±2° C. In another embodiment, the present invention discloses blending the API mixture into WFI at a temperature of 55±2° C.

[0058] In yet another aspect, the present invention provides a method for producing a composition comprising: 0.09% by weight of cyclosporine, about 1.0% by weight of hydrogenated 40 polyoxyl castor oil, about 0.05% by weight of Octoxynol-40; About 0.20 to 0.550% by weight of sodium phosphate monobasic, about 0.23 to 0.465% by weight of dibasic sodium phosphate; about 0.05% by weight of sodium chloride, about 0.3% by weight povidone, Sodium hydroxide / hydrochloric acid to adjust the pH, and A method for making a stable nanomicellar ophthalmic formulation comprising water for injection is provided, The method comprises: a) mixing cyclosporine with hydrogenated 40 polyoxyl castor oil at a temperature of 55° C.±2° C. or higher to form mixture A; b) holding mixture A under vacuum to remove bubbles; c) optionally, prior to complete dissolution of cyclosporine, reducing the temperature of mixture A to a temperature of 35° C.±2° C.; d) adding octoxynol-40 to form an API mixture; e) adding the API blend to water for injection (WFI); f) adding the remaining excipients to step (c) in the following order: sodium phosphate monobasic, then sodium phosphate dibasic, then sodium chloride, then polyvinylpyrrolidone; g) adjusting the pH to 6.5-7.2 and bringing to final volume with WFI.

[0059] In one aspect, the present invention discloses mixing mixture A in step (a) for 20-30 minutes, preferably 20-25 minutes, more preferably 20±2 minutes.

[0060] In one embodiment, the present invention discloses reducing the temperature of Mixture A to 35° C.±2° C. in less than 65 minutes. Preferably, the temperature of Mixture A is reduced to 35° C.±2° C. in 40-50 minutes.

[0061] In one embodiment, the present invention discloses blending the API mixture into WFI at a temperature of 35° C.±2° C. In another embodiment, blending of the API mixture into WFI occurs at a temperature of 55±2° C.

[0062] In a more preferred aspect, the present invention provides a stable nanomicellar ophthalmic formulation prepared by any of the above methods. [Brief description of the drawings]

[0063] [Figure 1] Figures 1(a) and 1(b) show cyclosporine nanomicelle ophthalmic formulations prepared by the methods described in Example 1(a) and Example 1(b), respectively, where Figure 1(a) discloses that no particles were observed, while Figure 1(b) discloses that particles were observed at stability. [Diagram 2] 1 shows characteristic X-ray powder diffraction (XRPD) patterns of cyclosporine having characteristic XRD peaks at 6.9, 7.8, 9.4, and 15.9 2-theta (degrees) (A), cyclosporine having characteristic XRD peaks at 7.4, 8.7, 14.4, and 17.5 2-theta (degrees) (B), cyclosporine having characteristic XRD peaks at 8.5, 9.3, 11.6, and 20.3 2-theta (degrees) (C), and an amorphous form of cyclosporine. [Diagram 3] 3 shows the dissolution behavior of different cyclosporine forms at 55° C. FIG. 3(a) shows the dissolution behavior of CsA forms with characteristic XRD peaks at 6.9, 7.8, 9.4, and 15.9 2-theta (degrees). FIG. 3(b) shows the dissolution behavior of CsA forms with characteristic XRD peaks at 7.4, 8.7, 14.4, and 17.5 2-theta (degrees). FIG. 3(c) shows the dissolution behavior of CsA forms with characteristic XRD peaks at 8.5, 9.3, 11.6, and 20.3 2-theta (degrees), and FIG. 3(d) shows the dissolution behavior of amorphous CsA. [Figure 4] 1 shows X-ray powder diffraction (XRPD) patterns of cyclosporine precipitated from solutions prepared when cyclosporine was dissolved in Kolliphor® RH 40 at higher exposure and temperature. [Diagram 5] 1 shows X-ray powder diffraction (XRPD) patterns of precipitated cyclosporine when cyclosporine was dissolved in Kolliphor® RH 40 and held for longer periods until precipitation occurred. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0064] One embodiment of the present disclosure is a stable nanomicelle ophthalmic formulation comprising cyclosporine, a polyoxyl lipid or fatty acid, and a polyalkoxylated alcohol.

[0065] Another embodiment of the present disclosure is A method for making a stable nanomicelle ophthalmic formulation comprising cyclosporine, a polyoxyl lipid or fatty acid, and a polyalkoxylated alcohol, the ophthalmic formulation comprising: a) mixing cyclosporine with polyoxyl lipids or fatty acids at a temperature of 55° C. or higher to form mixture A; b) reducing the temperature of mixture A to a temperature below 40° C. prior to complete dissolution of cyclosporine.

[0066] Preferably, mixing the cyclosporine with the polyoxyl lipid or fatty acid is carried out at a temperature of 55°C to 60°C to form mixture A. Preferably, mixture A is cooled to a temperature of 35°C to 40°C before complete dissolution of the cyclosporine. More preferably, mixing the cyclosporine with the polyoxyl lipid or fatty acid is carried out at a temperature of 55°C ± 2°C to form mixture A, and mixture A is cooled to a temperature of 35°C ± 2°C or less before complete dissolution of the cyclosporine. Further, the method includes adding a polyalkoxylated alcohol and then mixing the resulting mixture with an aqueous vehicle at 35°C ± 2°C. In another embodiment, the resulting mixture is mixed with an aqueous vehicle at a temperature of 55°C ± 2°C.

[0067] Another embodiment of the present disclosure is A method for making a stable nanomicellar ophthalmic formulation comprising cyclosporine, a polyoxyl lipid or fatty acid, and a polyalkoxylated alcohol, and an aqueous vehicle, comprising: The ophthalmic preparation is a) mixing cyclosporine with polyoxyl lipids or fatty acids at a temperature of 55°C ± 2°C or higher to form mixture A; b) holding mixture A under vacuum to remove bubbles; c) optionally, prior to complete dissolution of the cyclosporine, reducing the temperature of mixture A to a temperature of 35° C.±2° C. and adding the polyalkoxylated alcohol; and d) mixing the resulting mixture with an aqueous vehicle.

[0068] In one embodiment, the aqueous vehicle is mixed at a temperature of 35° C.±2° C. In another embodiment, the aqueous vehicle is mixed at a temperature of 55±2° C.

[0069] It has been found that during dissolution of hydrophobic molecules such as cyclosporine A (CsA) in polyoxyl lipids such as polyoxyl hydrogenated castor oil, air is trapped in the bulk in the form of bubbles, creating foam. This occurs regardless of the temperature during the drug dissolution process, even when the temperature is reduced to 55°C or even 35°C. This may delay the wetting and dissolution of cyclosporine, as this trapped air may create an interface between the cyclosporine particles and the water. Surprisingly, it has been found that while the mixture is held under vacuum, the air bubbles are dragged out and accumulate on the surface, from which they are gradually removed, leaving a clean bottom. Thus, dissolution under vacuum or intermittent removal of bubbles during dissolution may result in faster dissolution and eliminate the need to reduce the temperature of the mixture.

[0070] In one aspect, the method is disclosed to make a stable nanomicellar ophthalmic formulation comprising cyclosporine forms having characteristic XRD peaks at 6.9, 7.8, 9.4, and 15.9 2-theta (degrees).

[0071] In another embodiment, the method of making a stable nanomicelle ophthalmic formulation comprises an amorphous form of cyclosporine.

[0072] Another embodiment of the present disclosure is a method of making a stable nanomicellar ophthalmic formulation comprising cyclosporine, a polyoxyl lipid or fatty acid, a polyalkoxylated alcohol, and an aqueous vehicle, the ophthalmic formulation comprising: a) mixing cyclosporine with polyoxyl lipid or fatty acid at a temperature of 127-130° C. until completely dissolved to form mixture A; b) adding a polyalkoxylated alcohol to mixture A at 127-130° C.; c) mixing the resulting mixture with an aqueous vehicle at a temperature of 127-130°C.

[0073] In one embodiment, the method of making a stable nanomicelle ophthalmic formulation comprises a cyclosporine form having characteristic XRD peaks at 6.9, 7.8, 9.4, and 15.9 2-theta (degrees).

[0074] In another embodiment, the method of making a stable nanomicelle ophthalmic formulation comprises an amorphous form of cyclosporine.

[0075] In another embodiment, the method of making a stable nanomicellar ophthalmic formulation comprises cyclosporine in a form having characteristic XRD peaks at 7.4, 8.7, 14.4, and 17.5 2-theta (degrees).

[0076] In yet another embodiment, the method of making a stable nanomicelle ophthalmic formulation comprises cyclosporine in a form having characteristic XRD peaks at 8.5, 9.3, 11.6, and 20.3 2-theta (degrees).

[0077] In yet another aspect, the present disclosure is a method of making a stable nanomicelle ophthalmic formulation comprising cyclosporine, a polyoxyl lipid or fatty acid, a polyalkoxylated alcohol, and an aqueous vehicle, the ophthalmic formulation comprising: a) mixing a polyoxyl lipid or a fatty acid with a polyalkoxylated alcohol at a temperature of 127 to 130° C. to form a mixture A; b) adding cyclosporine to mixture A at 127-130°C; c) mixing the resulting mixture with an aqueous vehicle at a temperature of 127-130°C.

[0078] In one embodiment, the method of making a stable nanomicelle ophthalmic formulation comprises a cyclosporine form having characteristic XRD peaks at 6.9, 7.8, 9.4, and 15.9 2-theta (degrees).

[0079] In another embodiment, the method of making a stable nanomicelle ophthalmic formulation comprises an amorphous form of cyclosporine.

[0080] In another embodiment, the method of making a stable nanomicellar ophthalmic formulation comprises cyclosporine in a form having characteristic XRD peaks at 7.4, 8.7, 14.4, and 17.5 2-theta (degrees).

[0081] In yet another embodiment, the method of making a stable nanomicelle ophthalmic formulation comprises cyclosporine in a form having characteristic XRD peaks at 8.5, 9.3, 11.6, and 20.3 2-theta (degrees).

[0082] Another embodiment of the present disclosure is a stable nanomicelle ophthalmic formulation comprising cyclosporine, a polyoxyl lipid or fatty acid, and a polyalkoxylated alcohol, the ophthalmic formulation comprising: a) mixing cyclosporine with polyoxyl lipids or fatty acids at a temperature of 55° C. or higher to form mixture A; b) reducing the temperature of mixture A to a temperature below 40° C. prior to complete dissolution of cyclosporine.

[0083] Preferably, mixing the cyclosporine with the polyoxyl lipid or fatty acid is performed at a temperature of 55°C to 60°C to form mixture A. Preferably, mixture A is cooled to a temperature of 35°C to 40°C before complete dissolution of the cyclosporine. More preferably, mixing the cyclosporine with the polyoxyl lipid is performed at a temperature of 55°C ± 2°C to form mixture A, and mixture A is cooled to a temperature of 35°C ± 2°C or less before complete dissolution of the cyclosporine. Further, the method includes adding a polyalkoxylated alcohol and then mixing the resulting mixture with an aqueous vehicle at 35°C ± 2°C. In another embodiment, the resulting mixture is mixed with an aqueous vehicle at a temperature of 55°C ± 2°C.

[0084] In one aspect, the present invention provides a stable nanomicellar ophthalmic formulation comprising cyclosporine forms having characteristic XRD peaks at 6.9, 7.8, 9.4, and 15.9 2-theta (degrees).

[0085] In another aspect, the present invention provides a stable nanomicellar ophthalmic formulation comprising an amorphous form of cyclosporine.

[0086] In another aspect, the present invention provides a stable nanomicellar ophthalmic formulation comprising cyclosporine, a polyoxyl lipid or fatty acid, and a polyalkoxylated alcohol, and an aqueous vehicle, The ophthalmic preparation is a) mixing cyclosporine with polyoxyl lipids or fatty acids at a temperature of 55°C ± 2°C or higher to form mixture A; b) holding mixture A under vacuum to remove bubbles; c) optionally, prior to complete dissolution of the cyclosporine, reducing the temperature of mixture A to a temperature of 35° C.±2° C. and adding the polyalkoxylated alcohol; and d) mixing the resulting mixture with an aqueous vehicle.

[0087] In one embodiment, the aqueous vehicle is mixed at a temperature of 35° C.±2° C. In another embodiment, the aqueous vehicle is mixed at a temperature of 55±2° C.

[0088] In one embodiment, the stable nanomicellar ophthalmic formulation comprises a cyclosporine form having characteristic XRD peaks at 6.9, 7.8, 9.4, and 15.9 2-theta (degrees).

[0089] In another embodiment, the stable nanomicellar ophthalmic formulation comprises an amorphous form of cyclosporine.

[0090] In yet another aspect, the present disclosure provides a method for producing a method for manufacturing a semiconductor device comprising: A stable nanomicelle ophthalmic formulation is provided, comprising cyclosporine, a polyoxyl lipid or fatty acid, a polyalkoxylated alcohol, and an aqueous vehicle, the ophthalmic formulation comprising: a) mixing cyclosporine with polyoxyl lipid or fatty acid at a temperature of 127-130° C. until completely dissolved to form mixture A; b) adding a polyalkoxylated alcohol to mixture A at 127-130° C.; c) mixing the resulting mixture with an aqueous vehicle at a temperature of 127-130°C.

[0091] In one embodiment, the stable nanomicellar ophthalmic formulation comprises a cyclosporine form having characteristic XRD peaks at 6.9, 7.8, 9.4, and 15.9 2-theta (degrees).

[0092] In another embodiment, the stable nanomicellar ophthalmic formulation comprises an amorphous form of cyclosporine.

[0093] In another embodiment, the stable nanomicellar ophthalmic formulation comprises cyclosporine in a form having characteristic XRD peaks at 7.4, 8.7, 14.4, and 17.5 2-theta (degrees).

[0094] In yet another embodiment, the stable nanomicellar ophthalmic formulation comprises cyclosporine in a form having characteristic XRD peaks at 8.5, 9.3, 11.6, and 20.3 2-theta (degrees).

[0095] In yet another aspect, the present disclosure provides a method for producing a method for manufacturing a semiconductor device comprising: A stable nanomicelle ophthalmic formulation is provided, comprising cyclosporine, a polyoxyl lipid or fatty acid, a polyalkoxylated alcohol, and an aqueous vehicle, the ophthalmic formulation comprising: a) mixing a polyoxyl lipid or a fatty acid with a polyalkoxylated alcohol at a temperature of 127 to 130° C. to form a mixture A; b) adding cyclosporine to mixture A at 127-130°C; c) mixing the resulting mixture with an aqueous vehicle at a temperature of 127-130°C.

[0096] In one embodiment, the stable nanomicellar ophthalmic formulation comprises a cyclosporine form having characteristic XRD peaks at 6.9, 7.8, 9.4, and 15.9 2-theta (degrees).

[0097] In another embodiment, the stable nanomicellar ophthalmic formulation comprises an amorphous form of cyclosporine.

[0098] In another embodiment, the stable nanomicellar ophthalmic formulation comprises cyclosporine in a form having characteristic XRD peaks at 7.4, 8.7, 14.4, and 17.5 2-theta (degrees).

[0099] In yet another embodiment, the stable nanomicellar ophthalmic formulation comprises cyclosporine in a form having characteristic XRD peaks at 8.5, 9.3, 11.6, and 20.3 2-theta (degrees).

[0100] Another embodiment of the present disclosure is a method of making a stable nanomicellar ophthalmic formulation comprising cyclosporine, polyoxyl lipid or fatty acid, and polyalkoxylated alcohol, the ophthalmic formulation being made by a method comprising the steps of: mixing cyclosporine with polyoxyl lipid or fatty acid at a temperature of 55° C. or higher to form mixture A; and, prior to complete dissolution of cyclosporine, changing the temperature of mixture A to prevent the formation of cyclosporine forms (B in FIG. 2) having characteristic XRD peaks at 7.4, 8.7, 14.4, and 17.5 2-theta (degrees).

[0101] Preferably, mixing cyclosporine with polyoxyl lipid or fatty acid is performed at a temperature of 55°C to 60°C to form mixture A. Preferably, mixture A is cooled to a temperature of 35°C to 40°C before complete dissolution of cyclosporine. More preferably, mixing cyclosporine with polyoxyl lipid is performed at a temperature of 55°C ± 2°C to form mixture A, and mixture A is cooled to a temperature of 35°C ± 2°C or less before complete dissolution of cyclosporine. In one embodiment, the method of making a stable nanomicellar ophthalmic formulation comprises cyclosporine (A in Figure 2) having characteristic XRD peaks at 6.9, 7.8, 9.4, and 15.9 2-theta (degrees). In another embodiment, the method of making a stable nanomicellar ophthalmic formulation comprises an amorphous form of cyclosporine. Further, the method comprises adding a polyalkoxylated alcohol and then mixing the resulting mixture with an aqueous vehicle at 35°C ± 2°C. In another embodiment, the resulting mixture is mixed with an aqueous vehicle at a temperature of 55±2°C.

[0102] Another embodiment of the present disclosure is a method of making a stable nanomicellar ophthalmic formulation comprising cyclosporine, polyoxyl lipid or fatty acid, and polyalkoxylated alcohol, the ophthalmic formulation being made by a method comprising the steps of: mixing cyclosporine with polyoxyl lipid or fatty acid at a temperature of 55° C. or greater to form mixture A; and applying a vacuum to the mixture to prevent the formation of cyclosporine forms (B in FIG. 2) having characteristic XRD peaks at 7.4, 8.7, 14.4, and 17.5 2-theta (degrees).

[0103] Preferably, mixing the cyclosporine with the polyoxyl lipid or fatty acid is performed at a temperature of 55°C to 60°C to form mixture A. Optionally, mixture A is lowered to a temperature of 35°C to 40°C before complete dissolution of the cyclosporine. More preferably, mixing the cyclosporine with the polyoxyl lipid is performed at a temperature of 55°C ± 2°C to form mixture A, and optionally lowered to a temperature of 35°C ± 2°C or less before complete dissolution of the cyclosporine. Further, the method includes adding a polyalkoxylated alcohol and then mixing the resulting mixture with an aqueous vehicle at 35°C ± 2°C. In another embodiment, the resulting mixture is mixed with an aqueous vehicle at a temperature of 55°C ± 2°C.

[0104] Another embodiment of the present disclosure is a method of making a stable nanomicellar ophthalmic formulation comprising cyclosporine, polyoxyl lipid or fatty acid, and polyalkoxylated alcohol, the ophthalmic formulation being made by a method comprising the steps of: mixing cyclosporine with polyoxyl lipid or fatty acid at a temperature of 55° C. or higher to form mixture A; and, prior to complete dissolution of cyclosporine, changing the temperature of mixture A to prevent the formation of cyclosporine forms (C in FIG. 2) having characteristic XRD peaks at 8.5, 9.3, 11.6, and 20.3 2-theta (degrees).

[0105] Preferably, mixing cyclosporine with polyoxyl lipid or fatty acid is performed at a temperature of 55°C to 60°C to form mixture A. Furthermore, mixture A is cooled to a temperature of 35°C to 40°C before complete dissolution of cyclosporine. More preferably, mixing cyclosporine with polyoxyl lipid is performed at a temperature of 55°C ± 2°C to form mixture A, and then mixture A is cooled to a temperature of 35°C ± 2°C or less before complete dissolution of cyclosporine. In one embodiment, the method of making a stable nanomicellar ophthalmic formulation comprises cyclosporine (A in FIG. 2) having characteristic XRD peaks at 6.9, 7.8, 9.4, and 15.9 2-theta (degrees). In another embodiment, the method of making a stable nanomicellar ophthalmic formulation comprises an amorphous form of cyclosporine. Further, the method comprises adding a polyalkoxylated alcohol and then mixing the resulting mixture with an aqueous vehicle at 35°C ± 2°C. In another embodiment, the resulting mixture is mixed with an aqueous vehicle at a temperature of 55±2°C.

[0106] Another embodiment of the present disclosure is a method of making a stable nanomicellar ophthalmic formulation comprising cyclosporine, polyoxyl lipid or fatty acid, and polyalkoxylated alcohol, the ophthalmic formulation being made by a method comprising the steps of mixing cyclosporine with polyoxyl lipid or fatty acid at a temperature of 55° C. or greater to form mixture A, and applying a vacuum to the mixture to prevent the formation of cyclosporine forms (C in FIG. 2) having characteristic XRD peaks at 8.5, 9.3, 11.6, and 20.3 2-theta (degrees).

[0107] Preferably, mixing the cyclosporine with the polyoxyl lipid or fatty acid is performed at a temperature of 55°C to 60°C to form mixture A. Optionally, mixture A is lowered to a temperature of 35°C to 40°C before complete dissolution of the cyclosporine. More preferably, mixing the cyclosporine with the polyoxyl lipid is performed at a temperature of 55°C ± 2°C to form mixture A, and optionally lowered to a temperature of 35°C ± 2°C or less before complete dissolution of the cyclosporine. Further, the method includes adding a polyalkoxylated alcohol and then mixing the resulting mixture with an aqueous vehicle at 35°C ± 2°C. In another embodiment, the resulting mixture is mixed with an aqueous vehicle at a temperature of 55°C ± 2°C.

[0108] Another embodiment of the present disclosure is a stable nanomicelle ophthalmic formulation comprising cyclosporine, a polyoxyl lipid or fatty acid, and a polyalkoxylated alcohol, the formulation being a solution, the formulation exhibiting stability at room temperature (20-25° C.) for 6 months to at least 24 months. Typically, the ophthalmic formulation is stable for at least 6 months, at least 8 months, at least 10 months, at least 12 months, at least 14 months, at least 16 months, at least 18 months, at least 20 months, and at least 24 months when maintained at room temperature.

[0109] In another embodiment, the present disclosure provides stable nanomicellar ophthalmic formulations exhibiting stability for 6 months to at least 24 months at 2° C. to 8° C. Typically, the ophthalmic formulations are stable for at least 6 months, at least 8 months, at least 10 months, at least 12 months, at least 14 months, at least 16 months, at least 18 months, at least 20 months, and at least 24 months when maintained at 2° C. to 8° C.

[0110] Another embodiment of the present disclosure is a method of treating or preventing an ophthalmic disease or condition, the method comprising administering to a patient in need thereof a formulation described in any of the preceding embodiments, from 6 months to at least 24 months after preparation of the formulation.

[0111] In another embodiment, the present disclosure is a stable nanomicellar ophthalmic formulation according to any of the previous embodiments for use in treating an ophthalmic disease or condition. Preferably, the ophthalmic disease or condition is dry eye syndrome.

[0112] Materials useful in the formulations of the present disclosure include, but are not limited to, those disclosed in US Pat. No. 10,918,694.

[0113] As used herein, the terms "cyclosporin," "cyclosporine," "cyclosporine A," or "CsA" may be used interchangeably and include pharma- ceutically acceptable salts thereof.

[0114] As used herein in connection with numerical values, the terms "approximately" and "about" mean ±10% of the stated value, inclusive of the stated value.

[0115] As used herein, the term "substantially free" refers to an amount of a reference material that is 10% or less, e.g., in another form, preferably 8%, 5%, 4%, 3%, 2%, 1%, 0.5% or less of another form.

[0116] As used herein, the term "polyoxyl lipid or fatty acid" refers to mono- and diesters of lipid or fatty acid and polyoxyethylene diol. Polyoxyl lipid or fatty acid may be numbered ("n") according to the average polymer length of oxyethylene units (e.g., 40, 60, 80, 100), as is well understood in the art. The term "n≧40 polyoxyl lipid" means that the polyoxyl lipid or fatty acid has an average oxyethylene polymer length of 40 units or more. Hydrogenated castor oil stearate and castor oil are common lipids / fatty acids commercially available as polyoxyl lipids or fatty acids, but it is understood that any lipid or fatty acid can be polyoxylated to become a polyoxyl lipid or fatty acid contemplated herein. Examples of polyoxyl lipids or fatty acids include, but are not limited to, hydrogenated polyoxyl castor oil, e.g., HCO-40, HCO-60, HCO-80, HCO-100, polyoxyl 40 stearate, polyoxyl 35 castor oil.

[0117] As used herein, the term "micelle" or "nanomicelles" refers to aggregates (or clusters) of surfactant molecules. Micelles form only when the concentration of surfactant is higher than the critical micelle concentration (CMC). Surfactants are chemicals that are amphiphilic, meaning that they contain both hydrophobic and hydrophilic groups. Micelles can exist in different shapes, including spherical, cylindrical, and discoidal. Micelles that contain at least two different molecular species are mixed micelles. In some embodiments, the ophthalmic compositions of the present disclosure comprise an aqueous clear mixed micellar solution.

[0118] In some embodiments, the formulation includes, but is not limited to, nanomicelles as disclosed in U.S. Patent No. 10,918,694. For example, the ophthalmic composition can be administered topically to the eye as a biocompatible aqueous transparent mixed micellar solution. The composition has the drug incorporated and / or encapsulated in the micelles dispersed in the aqueous medium.

[0119] In some aspects of the embodiment, the polyoxyl lipid or fatty acid is polyoxyl castor oil. In some embodiments, the polyoxyl lipid or fatty acid is one or more selected from hydrogenated polyoxyl castor oil, such as HCO-40, HCO-60, HCO-80, or HCO-100. In some embodiments, the polyoxyl lipid or fatty acid (e.g., polyoxyl castor oil such as HCO-60, HCO-80, or HCO-100) is present at 0.5-2%, or 0.7-2%, or 1-6%, or 2-6%, or 2-6%, or 3-6%, or 4-6%, or 2-5%, or 3-5%, or 3-5%, or 2-6%, or about 4%, or greater than 0.7%, or greater than 1%, or greater than 1.5%, or greater than 2%, or greater than 3%, or greater than 4% by weight of the formulation. In some embodiments, the polyoxyl lipid is HCO-40. In some embodiments, the polyoxyl lipid is HCO-60. In some embodiments, the polyoxyl lipid is HCO-80. In some embodiments, the polyoxyl lipid is HCO-100.

[0120] In some aspects of the embodiment, the formulation comprises a polyalkoxylated alcohol. In some embodiments, the polyalkoxylated alcohol is octoxynol-40. In some aspects of the embodiment, the formulation comprises a polyalkoxylated alcohol (such as octoxynol-40) present at 0.002-4%, or 0.005-3%, or 0.005-2%, or 0.005-1%, or 0.005-0.5%, or 0.005-0.1%, or 0.005-0.05%, or 0.008-0.02%, or 0.01-0.1%, or 0.02-0.08%, or 0.005-0.08%, or about 0.05%, or about 0.01% by weight of the formulation.

[0121] One embodiment of the present disclosure is a stable nanomicelle ophthalmic formulation. The ophthalmic formulation comprises cyclosporine, hydrogenated polyoxyl castor oil, and octoxynol-40. Preferably, the hydrogenated polyoxyl castor oil is hydrogenated 40 polyoxyl castor oil (HCO-40). Preferably, the ophthalmic formulation comprises 0.09% by weight cyclosporine, 1.0% by weight hydrogenated 40 polyoxyl castor oil, and 0.05% by weight octoxynol-40.

[0122] Another embodiment of the present disclosure is a method of making a stable nanomicelle ophthalmic formulation comprising cyclosporine, hydrogenated polyoxyl castor oil, and octoxynol-40, the ophthalmic formulation comprising: a) mixing cyclosporine with hydrogenated polyoxyl castor oil at a temperature of 55° C. or higher to form mixture A; b) reducing the temperature of mixture A to a temperature below 40° C. prior to complete dissolution of cyclosporine.

[0123] Preferably, mixing the cyclosporine with the hydrogenated polyoxyl castor oil is carried out at a temperature of 55°C to 60°C to form mixture A. Preferably, mixture A is reduced to a temperature of 35°C to 40°C prior to complete dissolution of the cyclosporine. More preferably, mixing the cyclosporine with the hydrogenated polyoxyl castor oil is carried out at a temperature of 55°C ± 2°C to form mixture A, and then mixture A is reduced to a temperature of 35°C ± 2°C or less prior to complete dissolution of the cyclosporine. Further, the method includes adding octoxynol-40 and then mixing the resulting mixture with an aqueous vehicle at 35°C ± 2°C. In another embodiment, the aqueous vehicle is mixed at a temperature of 55±2°C.

[0124] Preferably, the hydrogenated polyoxyl castor oil is hydrogenated 40 polyoxyl castor oil (HCO-40). More preferably, the ophthalmic formulation comprises 0.09% by weight cyclosporine, 1.0% by weight hydrogenated 40 polyoxyl castor oil, and 0.05% by weight octoxynol-40.

[0125] Another embodiment of the present disclosure is a method of making a stable nanomicelle ophthalmic formulation comprising cyclosporine, hydrogenated polyoxyl castor oil, and octoxynol-40, the ophthalmic formulation comprising: a) mixing cyclosporine with hydrogenated polyoxyl castor oil at a temperature of 55° C. or higher to form mixture A; b) holding mixture A under vacuum to remove bubbles; c) optionally, prior to complete dissolution of cyclosporine, reducing the temperature of mixture A to a temperature of 35° C.±2° C. and adding octoxynol-40; and d) mixing the resulting mixture with an aqueous vehicle.

[0126] Preferably, mixing the cyclosporine with the hydrogenated polyoxyl castor oil is carried out at a temperature of 55° C. to 60° C. to form mixture A. Optionally, mixture A is lowered to a temperature of 35° C. to 40° C. prior to complete dissolution of the cyclosporine. More preferably, mixing the cyclosporine with the hydrogenated polyoxyl castor oil is carried out at a temperature of 55° C.±2° C. to form mixture A, and optionally lowered to a temperature of 35° C.±2° C. or less prior to complete dissolution of the cyclosporine. Further, the method includes adding octoxynol-40 and then mixing the resulting mixture with an aqueous vehicle at 35° C.±2° C. In another embodiment, the aqueous vehicle is mixed at a temperature of 55° C.±2° C.

[0127] Preferably, the hydrogenated polyoxyl castor oil is hydrogenated 40 polyoxyl castor oil (HCO-40). More preferably, the ophthalmic formulation comprises 0.09% by weight cyclosporine, 1.0% by weight hydrogenated 40 polyoxyl castor oil, and 0.05% by weight octoxynol-40.

[0128] Another embodiment of the present disclosure is a stable nanomicelle ophthalmic formulation comprising cyclosporine, hydrogenated polyoxyl castor oil, and octoxynol-40, the ophthalmic formulation comprising: a) mixing cyclosporine with hydrogenated polyoxyl castor oil at a temperature of 55° C. or higher to form mixture A; b) reducing the temperature of mixture A to a temperature below 40° C. prior to complete dissolution of cyclosporine.

[0129] Preferably, mixing the cyclosporine with the hydrogenated polyoxyl castor oil is carried out at a temperature of 55°C to 60°C to form mixture A. Preferably, mixture A is reduced to a temperature of 35°C to 40°C prior to complete dissolution of the cyclosporine. More preferably, mixing the cyclosporine with the hydrogenated polyoxyl castor oil is carried out at a temperature of 55°C ± 2°C to form mixture A, and then mixture A is reduced to a temperature of 35°C ± 2°C or less prior to complete dissolution of the cyclosporine. Further, the method includes adding octoxynol-40 and then mixing the resulting mixture with an aqueous vehicle at 35°C ± 2°C. In another embodiment, the aqueous vehicle is mixed at a temperature of 55±2°C.

[0130] Preferably, the hydrogenated polyoxyl castor oil is hydrogenated 40 polyoxyl castor oil (HCO-40). More preferably, the ophthalmic formulation comprises 0.09% by weight cyclosporine, 1.0% by weight hydrogenated 40 polyoxyl castor oil, and 0.05% by weight octoxynol-40.

[0131] Another embodiment of the present disclosure is A method for making a stable nanomicelle ophthalmic formulation comprising cyclosporine, a polyoxyl lipid or fatty acid, and a polyalkoxylated alcohol, the ophthalmic formulation comprising: a) mixing cyclosporine with hydrogenated 40 polyoxyl castor oil at a temperature of 55° C.±2° C. or higher to form mixture A; b) reducing the temperature of mixture A to a temperature of 35° C.±2° C. prior to complete dissolution of cyclosporine; c) adding Octoxynol-40; and d) mixing the resulting mixture with an aqueous vehicle.

[0132] Another embodiment of the present disclosure is a stable nanomicelle ophthalmic formulation comprising cyclosporine, hydrogenated polyoxyl castor oil, and octoxynol-40, the ophthalmic formulation comprising: a) mixing cyclosporine with hydrogenated polyoxyl castor oil at a temperature of 55° C. or higher to form mixture A; b) holding mixture A under vacuum to remove bubbles; c) optionally, prior to complete dissolution of cyclosporine, reducing the temperature of mixture A to a temperature of 35° C.±2° C.; d) adding octoxynol-40; e) mixing the resulting mixture with an aqueous vehicle.

[0133] Preferably, mixing the cyclosporine with the hydrogenated polyoxyl castor oil is carried out at a temperature of 55° C. to 60° C. to form mixture A. Optionally, mixture A is lowered to a temperature of 35° C. to 40° C. prior to complete dissolution of the cyclosporine. More preferably, mixing the cyclosporine with the hydrogenated polyoxyl castor oil is carried out at a temperature of 55° C.±2° C. to form mixture A, and optionally lowered to a temperature of 35° C.±2° C. or less prior to complete dissolution of the cyclosporine. Further, the method includes adding octoxynol-40 and then mixing the resulting mixture with an aqueous vehicle at 35° C.±2° C. In another embodiment, the aqueous vehicle is mixed at a temperature of 55° C.±2° C.

[0134] Preferably, the hydrogenated polyoxyl castor oil is hydrogenated 40 polyoxyl castor oil (HCO-40). More preferably, the ophthalmic formulation comprises 0.09% by weight cyclosporine, 1.0% by weight hydrogenated 40 polyoxyl castor oil, and 0.05% by weight octoxynol-40.

[0135] Another embodiment of the present disclosure is a stable nanomicellar ophthalmic formulation comprising cyclosporine, hydrogenated 40 polyoxyl castor oil, octoxynol-40, and an aqueous vehicle; The ophthalmic preparation is a) mixing cyclosporine with hydrogenated 40 polyoxyl castor oil at a temperature of 55° C.±2° C. or higher to form mixture A; b) reducing the temperature of mixture A to a temperature of 35° C.±2° C. prior to complete dissolution of cyclosporine; c) adding Octoxynol-40; and d) mixing the resulting mixture with an aqueous vehicle.

[0136] In one embodiment, the aqueous vehicle is mixed at a temperature of 35° C.±2° C. In another embodiment, the aqueous vehicle is mixed at a temperature of 55±2° C.

[0137] In one embodiment, the present invention provides a stable nanomicellar ophthalmic formulation comprising cyclosporine forms having characteristic XRD peaks at 6.9, 7.8, 9.4, and 15.9 2-theta (degrees).

[0138] In another embodiment, the present invention provides a stable nanomicellar ophthalmic formulation comprising an amorphous form of cyclosporine.

[0139] Another embodiment of the present disclosure is a method of making a stable nanomicellar ophthalmic formulation comprising cyclosporine, hydrogenated polyoxyl castor oil, and octoxynol-40, the ophthalmic formulation being made by a method comprising the steps of: mixing cyclosporine with hydrogenated polyoxyl castor oil at a temperature of 55° C. or higher to form mixture A; and, prior to complete dissolution of cyclosporine, changing the temperature of mixture A to prevent the formation of cyclosporine forms (FIG. 2B) having characteristic XRD peaks at 7.4, 8.7, 14.4, and 17.5 2-theta (degrees).

[0140] Preferably, mixing cyclosporine with hydrogenated polyoxyl castor oil is performed at a temperature of 55°C to 60°C to form mixture A. Furthermore, mixture A is cooled to a temperature of 35°C to 40°C before complete dissolution of cyclosporine. More preferably, mixing cyclosporine with hydrogenated polyoxyl castor oil is performed at a temperature of 55°C ± 2°C to form mixture A, and then mixture A is cooled to a temperature of 35°C ± 2°C or less before complete dissolution of cyclosporine. In one embodiment, the stable nanomicelle ophthalmic formulation comprises cyclosporine (A in Figure 2) having characteristic XRD peaks at 2-theta (degrees) 6.9, 7.8, 9.4, and 15.9. In another embodiment, the stable nanomicelle ophthalmic formulation comprises an amorphous form of cyclosporine. Further, the method comprises adding octoxynol-40 and then mixing the resulting mixture with an aqueous vehicle at 35°C ± 2°C. In another embodiment, the resulting mixture is mixed with an aqueous vehicle at a temperature of 55±2° C. Preferably, the hydrogenated polyoxyl castor oil is hydrogenated 40 polyoxyl castor oil (HCO-40). More preferably, the ophthalmic formulation comprises 0.09% by weight cyclosporine, 1.0% by weight hydrogenated 40 polyoxyl castor oil, and 0.05% by weight octoxynol-40.

[0141] Another embodiment of the present disclosure is a method of making a stable nanomicellar ophthalmic formulation comprising cyclosporine, hydrogenated polyoxyl castor oil, and octoxynol-40, the ophthalmic formulation being made by a method comprising the steps of: mixing cyclosporine with hydrogenated polyoxyl castor oil at a temperature of 55° C. or higher to form mixture A; and, prior to complete dissolution of cyclosporine, changing the temperature of mixture A to prevent the formation of cyclosporine forms (C in FIG. 2) having characteristic XRD peaks at 8.5, 9.3, 11.6, and 20.3 2-theta (degrees).

[0142] Preferably, mixing cyclosporine with hydrogenated polyoxyl castor oil is performed at a temperature of 55°C to 60°C to form mixture A. Furthermore, mixture A is then lowered to a temperature of 35°C to 40°C prior to complete dissolution of cyclosporine. More preferably, mixing cyclosporine with hydrogenated polyoxyl castor oil is performed at a temperature of 55°C ± 2°C to form mixture A, and then the temperature of mixture A is lowered to a temperature of 35°C ± 2°C or lower prior to complete dissolution of cyclosporine. In one embodiment, the method of making a stable nanomicellar ophthalmic formulation comprises cyclosporine (A in FIG. 2) having characteristic XRD peaks at 6.9, 7.8, 9.4, and 15.9 2-theta (degrees). In another embodiment, the method of making a stable nanomicellar ophthalmic formulation comprises an amorphous form of cyclosporine. Further, the method comprises adding octoxynol-40 and then mixing the resulting mixture with an aqueous vehicle at 35°C ± 2°C. In another embodiment, the aqueous vehicle is mixed at a temperature of 55±2° C. Preferably, the hydrogenated polyoxyl castor oil is hydrogenated 40 polyoxyl castor oil (HCO-40). More preferably, the ophthalmic formulation comprises 0.09% by weight cyclosporine, 1.0% by weight hydrogenated 40 polyoxyl castor oil, and 0.05% by weight octoxynol-40.

[0143] Another embodiment of the present disclosure is a method of making a stable nanomicellar ophthalmic formulation comprising cyclosporine, hydrogenated polyoxyl castor oil, and octoxynol-40, the ophthalmic formulation being made by a method comprising the steps of mixing cyclosporine with hydrogenated polyoxyl castor oil at a temperature of 55° C. or greater to form mixture A, and applying a vacuum to the mixture to prevent the formation of cyclosporine forms (FIG. 2B) having characteristic XRD peaks at 7.4, 8.7, 14.4, and 17.5 2-theta (degrees).

[0144] Preferably, mixing the cyclosporine with the hydrogenated polyoxyl castor oil is carried out at a temperature of 55° C. to 60° C. to form mixture A. Optionally, mixture A is lowered to a temperature of 35° C. to 40° C. prior to complete dissolution of the cyclosporine. More preferably, mixing the cyclosporine with the hydrogenated polyoxyl castor oil is carried out at a temperature of 55° C.±2° C. to form mixture A, and optionally lowered to a temperature of 35° C.±2° C. or less prior to complete dissolution of the cyclosporine. Further, the method includes adding octoxynol-40 and then mixing the resulting mixture with an aqueous vehicle at 35° C.±2° C. In another embodiment, the resulting mixture is mixed with an aqueous vehicle at a temperature of 55° C.±2° C. Preferably, the cyclosporine is present in a cyclosporine form (A in FIG. 2) having characteristic XRD peaks at 6.9, 7.8, 9.4, and 15.9 2-theta (degrees). In another embodiment, preferably, cyclosporine is present in amorphous form. Preferably, the hydrogenated polyoxyl castor oil is hydrogenated 40 polyoxyl castor oil (HCO-40). More preferably, the ophthalmic formulation comprises 0.09% by weight cyclosporine, 1.0% by weight hydrogenated 40 polyoxyl castor oil, and 0.05% by weight octoxynol-40.

[0145] Another embodiment of the present disclosure is a method of making a stable nanomicellar ophthalmic formulation comprising cyclosporine, hydrogenated polyoxyl castor oil, and octoxynol-40, the ophthalmic formulation being made by a method comprising the steps of mixing cyclosporine with hydrogenated polyoxyl castor oil at a temperature of 55° C. or greater to form mixture A, and applying a vacuum to the mixture to prevent the formation of cyclosporine forms (C in FIG. 2) having characteristic XRD peaks at 8.5, 9.3, 11.6, and 20.3 2-theta (degrees).

[0146] Preferably, mixing the cyclosporine with the hydrogenated polyoxyl castor oil is carried out at a temperature of 55° C. to 60° C. to form mixture A. Optionally, mixture A is lowered to a temperature of 35° C. to 40° C. prior to complete dissolution of the cyclosporine. More preferably, mixing the cyclosporine with the hydrogenated polyoxyl castor oil is carried out at a temperature of 55° C.±2° C. to form mixture A, and optionally lowered to a temperature of 35° C.±2° C. or less prior to complete dissolution of the cyclosporine. Further, the method includes adding octoxynol-40 and then mixing the resulting mixture with an aqueous vehicle at 35° C.±2° C. In another embodiment, the resulting mixture is mixed with an aqueous vehicle at a temperature of 55° C.±2° C. Preferably, the cyclosporine is present in a cyclosporine form (A in FIG. 2) having characteristic XRD peaks at 6.9, 7.8, 9.4, and 15.9 2-theta (degrees). In another embodiment, preferably, cyclosporine is present in amorphous form. Preferably, the hydrogenated polyoxyl castor oil is hydrogenated 40 polyoxyl castor oil (HCO-40). More preferably, the ophthalmic formulation comprises 0.09% by weight cyclosporine, 1.0% by weight hydrogenated 40 polyoxyl castor oil, and 0.05% by weight octoxynol-40.

[0147] In yet another aspect, the present disclosure provides a method for producing a method for manufacturing a semiconductor device comprising: A method for making a stable nanomicellar ophthalmic formulation comprising cyclosporine, hydrogenated 40 polyoxyl castor oil, octoxynol-40, and an aqueous vehicle, comprising: The ophthalmic preparation is a) mixing cyclosporin with hydrogenated 40 polyoxyl castor oil at a temperature of 127-130° C. until completely dissolved to form mixture A; b) adding Octoxynol-40 to mixture A at 127-130° C.; c) mixing the resulting mixture with an aqueous vehicle at a temperature of 127-130°C.

[0148] In one aspect, the present disclosure is a method of making a stable nanomicellar ophthalmic formulation comprising cyclosporine forms having characteristic XRD peaks at 6.9, 7.8, 9.4, and 15.9 2-theta (degrees).

[0149] In another aspect, the present disclosure is a method of making a stable nanomicellar ophthalmic formulation comprising an amorphous form of cyclosporine.

[0150] Another embodiment of the present disclosure is A method for making a stable nanomicellar ophthalmic formulation comprising cyclosporine, hydrogenated polyoxyl castor oil, octoxynol-40, and an aqueous vehicle, comprising: The method comprises: a) mixing cyclosporin with hydrogenated polyoxyl castor oil at a temperature of 127-130° C. until completely dissolved to form mixture A; b) adding Octoxynol-40 to mixture A at 127-130° C.; c) mixing the resulting mixture with an aqueous vehicle at a temperature of 127-130°C; Cyclosporine exists in a form (FIG. 2B) with characteristic XRD peaks at 7.4, 8.7, 14.4, and 17.5 2-theta (degrees).

[0151] Preferably, the hydrogenated polyoxyl castor oil is hydrogenated 40 polyoxyl castor oil (HCO-40). More preferably, the ophthalmic formulation comprises 0.09% by weight cyclosporine, 1.0% by weight hydrogenated 40 polyoxyl castor oil, and 0.05% by weight octoxynol-40.

[0152] Another embodiment of the present disclosure is A method for making a stable nanomicellar ophthalmic formulation comprising cyclosporine, hydrogenated polyoxyl castor oil, octoxynol-40, and an aqueous vehicle, comprising: The method comprises: a) mixing cyclosporin with hydrogenated polyoxyl castor oil at a temperature of 127-130° C. until completely dissolved to form mixture A; b) adding Octoxynol-40 to mixture A at 127-130° C.; c) mixing the resulting mixture with an aqueous vehicle at a temperature of 127-130°C; Cyclosporine exists in a form (FIG. 2C) with characteristic XRD peaks at 8.5, 9.3, 11.6, and 20.3 2-theta (degrees).

[0153] Preferably, the hydrogenated polyoxyl castor oil is hydrogenated 40 polyoxyl castor oil (HCO-40). More preferably, the ophthalmic formulation comprises 0.09% by weight cyclosporine, 1.0% by weight hydrogenated 40 polyoxyl castor oil, and 0.05% by weight octoxynol-40.

[0154] In yet another aspect, the present disclosure provides a method for producing a method for manufacturing a semiconductor device comprising: A method for making a stable nanomicellar ophthalmic formulation comprising cyclosporine, hydrogenated polyoxyl castor oil, octoxynol-40, and an aqueous vehicle, comprising: The method comprises: a) mixing hydrogenated 40 polyoxyl castor oil and octoxynol-40 at a temperature of 127-130° C. to form a mixture A; b) adding cyclosporine to mixture A at 127-130°C; c) mixing the resulting mixture with an aqueous vehicle at a temperature of 127-130°C.

[0155] In one aspect, the present disclosure is a method of making a stable nanomicellar ophthalmic formulation comprising cyclosporine forms having characteristic XRD peaks at 6.9, 7.8, 9.4, and 15.9 2-theta (degrees).

[0156] In another aspect, the present disclosure is a method of making a stable nanomicellar ophthalmic formulation comprising an amorphous form of cyclosporine.

[0157] In another aspect, the present disclosure is a method of making a stable nanomicellar ophthalmic formulation comprising cyclosporine in a form having characteristic XRD peaks at 7.4, 8.7, 14.4, and 17.5 2-theta (degrees).

[0158] In another aspect, the present disclosure is a method of making a stable nanomicellar ophthalmic formulation comprising cyclosporine in a form having characteristic XRD peaks at 8.5, 9.3, 11.6, and 20.3 2-theta (degrees).

[0159] Preferably, the hydrogenated polyoxyl castor oil is hydrogenated 40 polyoxyl castor oil (HCO-40). More preferably, the ophthalmic formulation comprises 0.09% by weight cyclosporine, 1.0% by weight hydrogenated 40 polyoxyl castor oil, and 0.05% by weight octoxynol-40.

[0160] In yet another aspect, the present disclosure provides a method for producing a method for manufacturing a semiconductor device comprising: A stable nanomicelle ophthalmic formulation comprising cyclosporine, hydrogenated 40 polyoxyl castor oil, octoxynol-40, and an aqueous vehicle, The ophthalmic preparation is a) mixing cyclosporin with hydrogenated 40 polyoxyl castor oil at a temperature of 127-130° C. until completely dissolved to form mixture A; b) adding Octoxynol-40 to mixture A at 127-130° C.; c) mixing the resulting mixture with an aqueous vehicle at a temperature of 127-130°C.

[0161] In one aspect, the present disclosure is a stable nanomicellar ophthalmic formulation comprising cyclosporine forms having characteristic XRD peaks at 6.9, 7.8, 9.4, and 15.9 2-theta (degrees).

[0162] In another aspect, the present disclosure is a stable nanomicellar ophthalmic formulation comprising an amorphous form of cyclosporine.

[0163] Preferably, the hydrogenated polyoxyl castor oil is hydrogenated 40 polyoxyl castor oil (HCO-40). More preferably, the ophthalmic formulation comprises 0.09% by weight cyclosporine, 1.0% by weight hydrogenated 40 polyoxyl castor oil, and 0.05% by weight octoxynol-40.

[0164] In yet another embodiment of the present disclosure, A stable nanomicelle ophthalmic formulation comprising cyclosporine, hydrogenated polyoxyl castor oil, octoxynol-40, and an aqueous vehicle, The ophthalmic preparation is a) mixing cyclosporin with hydrogenated polyoxyl castor oil at a temperature of 127-130° C. until completely dissolved to form mixture A; b) adding Octoxynol-40 to mixture A at 127-130° C.; c) mixing the resulting mixture with an aqueous vehicle at a temperature of 127-130°C; Cyclosporine exists in a form (FIG. 2B) with characteristic XRD peaks at 7.4, 8.7, 14.4, and 17.5 2-theta (degrees).

[0165] Preferably, the hydrogenated polyoxyl castor oil is hydrogenated 40 polyoxyl castor oil (HCO-40). More preferably, the ophthalmic formulation comprises 0.09% by weight cyclosporine, 1.0% by weight hydrogenated 40 polyoxyl castor oil, and 0.05% by weight octoxynol-40.

[0166] Yet another embodiment of the present disclosure comprises: A stable nanomicelle ophthalmic formulation comprising cyclosporine, hydrogenated polyoxyl castor oil, octoxynol-40, and an aqueous vehicle, The ophthalmic preparation is a) mixing cyclosporin with hydrogenated polyoxyl castor oil at a temperature of 127-130° C. until completely dissolved to form mixture A; b) adding Octoxynol-40 to mixture A at 127-130° C.; c) mixing the resulting mixture with an aqueous vehicle at a temperature of 127-130°C; Cyclosporine exists in a form (FIG. 2C) with characteristic XRD peaks at 8.5, 9.3, 11.6, and 20.3 2-theta (degrees).

[0167] Preferably, the hydrogenated polyoxyl castor oil is hydrogenated 40 polyoxyl castor oil (HCO-40). More preferably, the ophthalmic formulation comprises 0.09% by weight cyclosporine, 1.0% by weight hydrogenated 40 polyoxyl castor oil, and 0.05% by weight octoxynol-40.

[0168] In yet another aspect, the present disclosure provides a method for producing a method for manufacturing a semiconductor device comprising: A stable nanomicelle ophthalmic formulation comprising cyclosporine, hydrogenated 40 polyoxyl castor oil, octoxynol-40, and an aqueous vehicle, The ophthalmic preparation is a) mixing hydrogenated 40 polyoxyl castor oil and octoxynol-40 at a temperature of 127-130° C. to form a mixture A; b) adding cyclosporine to mixture A at 127-130°C; c) mixing the resulting mixture with an aqueous vehicle at a temperature of 127-130°C.

[0169] In one aspect, the present disclosure is a stable nanomicellar ophthalmic formulation comprising cyclosporine forms having characteristic XRD peaks at 6.9, 7.8, 9.4, and 15.9 2-theta (degrees).

[0170] In another aspect, the present disclosure is a stable nanomicellar ophthalmic formulation comprising an amorphous form of cyclosporine.

[0171] In another aspect, the present disclosure is a stable nanomicellar ophthalmic formulation comprising cyclosporine in a form having characteristic XRD peaks at 7.4, 8.7, 14.4, and 17.5 2-theta (degrees).

[0172] In another aspect, the present disclosure is a stable nanomicellar ophthalmic formulation comprising cyclosporine in a form having characteristic XRD peaks at 8.5, 9.3, 11.6, and 20.3 2-theta (degrees).

[0173] Another embodiment of the present disclosure is a stable nanomicelle ophthalmic formulation comprising cyclosporine, hydrogenated 40 polyoxyl castor oil (HCO-40), and octoxynol-40, the formulation being a solution, the formulation exhibiting stability at room temperature (20-25° C.) for 6 months to at least 24 months. Typically, the ophthalmic formulation is stable for at least 6 months, at least 8 months, at least 10 months, at least 12 months, at least 14 months, at least 16 months, at least 18 months, at least 20 months, and at least 24 months when maintained at room temperature. More preferably, the stable nanomicelle ophthalmic formulation is a solution and comprises 0.09% by weight of cyclosporine, 1.0% by weight of hydrogenated 40 polyoxyl castor oil, and 0.05% by weight of octoxynol-40.

[0174] Another embodiment of the present disclosure is a method of treating or preventing an ocular disease or condition, e.g., dry eye, comprising administering a stable nanomicellar ophthalmic formulation comprising 0.09% by weight cyclosporine, 1.0% by weight hydrogenated 40 polyoxyl castor oil, and 0.05% by weight octoxynol-40 to a patient in need thereof, 6 months to at least 24 months after preparation of the formulation.

[0175] The cyclosporine present in certain formulations according to embodiments of the present disclosure is preferably amorphous when in solution. Alternatively, the cyclosporine may be present in solution as a cyclosporine form having characteristic XRD peaks at 6.9, 7.8, 9.4, and 15.9 2-theta (degrees). In certain embodiments, the solution according to the present disclosure is free of crystalline cyclosporine. In certain embodiments, the formulation is substantially free of cyclosporine forms having characteristic XRD peaks at 7.4, 8.7, 14.4, and 17.5 2-theta (degrees). In alternative embodiments, the formulation according to the present disclosure is substantially free of cyclosporine forms having characteristic XRD peaks at 8.5, 9.3, 11.6, and 20.3 2-theta (degrees).

[0176] Additional formulation ingredients The compositions of the present disclosure may also contain other components, such as, but not limited to, additives, adjuvants, buffers, tonicity agents, bioadhesive polymers, and preservatives. In any of the compositions of the present disclosure for topical administration to the eye, the mixture is preferably formulated at about pH 5 to about pH 8. This pH range may be achieved by adding a buffer to the composition, as described in the Examples. In one embodiment, the pH range in the composition in the formulation is about pH 6.5 to about pH 7.2. The compositions of the present disclosure may be buffered with any common buffer system, such as phosphate, borate, acetate, citrate, carbonate, and borate-polyol complexes, and the pH and osmolality are adjusted according to well-known techniques to the appropriate physiological values. The mixed micelle compositions of the present disclosure are stable in aqueous buffer solutions. That is, there is no deleterious interaction between the buffer and any other components that may destabilize the composition.

[0177] Examples of isotonicity agents include mannitol, sodium chloride, sodium nitrate, sodium sulfate, dextrose, xylitol, or combinations thereof. These isotonicity agents may be used to adjust the osmolarity of the composition. In one embodiment, the osmolarity of the formulation is adjusted to within the range of about 150 to about 200 mOsmol / kg. In a preferred embodiment, the osmolarity of the formulation is adjusted to between about 160 to about 190 mOsmol / kg.

[0178] Additives such as sugars, glycerol, and other sugar alcohols may be included in the compositions of the present disclosure. Pharmaceutical additives may be added to increase the effectiveness or potency of other components in the compositions. Pharmaceutical additives may be added to the compositions of the present disclosure to improve the stability of calcineurin inhibitors, to adjust the osmolality of the composition, to adjust the viscosity of the composition, or for other reasons such as effecting drug delivery. Non-limiting examples of pharmaceutical additives of the present disclosure include sugars such as trehalose, mannose, D-galactose, and lactose. In one embodiment, sugars may be incorporated into the composition prior to hydrating the thin film (i.e., internally). In another embodiment, sugars may be incorporated into the composition during the hydration step (i.e., externally). In one embodiment, the aqueous clear mixed micellar solution of the present disclosure includes an additive such as a sugar.

[0179] In one embodiment, the composition of the present disclosure further comprises one or more bioadhesive polymers. Bioadhesion refers to the ability of certain synthetic and biological macromolecules, as well as hydrocolloids, to adhere to biological tissues. Bioadhesion is a complex phenomenon that depends in part on the properties of the polymer, the biological tissue, and the surrounding environment. Several factors have been found to contribute to the bioadhesive ability of a polymer: the presence of functional groups capable of forming hydrogen bridges (--OH, COOH), the presence and strength of anionic charges, sufficient elasticity for the polymer chain to penetrate the mucosal layer, and high molecular weight. Bioadhesive systems have been used in dental, orthopedic, ophthalmic, and surgical applications. However, recently, significant interest has emerged in the use of bioadhesive materials in other areas, such as soft tissue-based artificial replacements and controlled release systems for localized release of bioactive agents. Such applications include systems for the release of drugs in the oral or nasal cavity, and systems for intestinal or rectal administration.

[0180] In one embodiment, the composition of the present disclosure includes at least one bioadhesive polymer. The bioadhesive polymer can increase the viscosity of the composition, thereby increasing its residence time in the eye. Bioadhesive polymers of the present disclosure include, for example, carboxylic acid polymers such as Carbopol® (carbomer), Noveon® (polycarbophil), cellulose derivatives including alkyl and hydroxyalkyl celluloses such as methylcellulose, hydroxypropylcellulose, carboxymethylcellulose, gums such as locust bean, xanthan, agarose, karaya, guar, and other polymers such as, but not limited to, polyvinyl alcohol, povidone, polyethylene glycol, Pluronic® (poloxamer), tragacanth, and hyaluronic acid; phase transition polymers for providing sustained and controlled delivery of encapsulated agents to the eye (e.g., alginate, carrageenan (e.g., Eucheuma), xanthan, and locust bean gum mixtures, pectin, cellulose acetate phthalate, alkyl hydroxyalkyl cellulose ... cellulose acetate phthalate, alkyl hydroxyalkyl cellulose, cellulose acetate phthalate, cellulose acetate phthalate, cellulose acetate phthalate, cellulose acetate phthalate, cellulose acetate phthalate, cellulose acetate phthalate, cellulose acetate phthalate Examples of bioadhesive polymers include polycellulose and its derivatives, hydroxyalkylated polyacrylic acid and its derivatives, poloxamers and its derivatives, etc. The physical properties of these polymers may be mediated by changing environmental factors such as ionic strength, pH, or temperature, alone or in combination with other factors. In one embodiment, the optional one or more bioadhesive polymers are present in the composition at about 0.01% to about 10% by weight / volume, preferably about 0.1 to about 5% by weight / volume. In one embodiment, the composition of the present disclosure further comprises at least one hydrophilic polymeric excipient, e.g., selected from PVP-K-30, PVP-K-90, HPMC, HEC, and polycarbophil. In one embodiment, the polymeric excipient is selected from PVP-K-90, PVP-K-30, or HPMC. In one embodiment, the polymeric excipient is selected from PVP-K-90 or PVP-K-30.

[0181] In one embodiment, if a preservative is desired, the composition may be optionally preserved with any of a number of well-known preservatives, including benzyl alcohol with or without EDTA, benzalkonium chloride, chlorhexidine, Cosmocil® CQ, or Dowicil® 200. In certain embodiments, it may be desirable for the formulations described herein to be free of any preservatives. In this regard, a preservative may not be necessary or desirable in some embodiments in formulations contained in single-use containers. In other embodiments, it may be advantageous to include a preservative, such as in certain embodiments in which the formulation is contained in a multi-use container.

[0182] In a preferred embodiment, the present disclosure provides 0.09% by weight of cyclosporine, about 1.0% by weight of hydrogenated 40 polyoxyl castor oil, about 0.05% by weight of Octoxynol-40, and A method for making a stable nanomicellar ophthalmic formulation comprising an aqueous vehicle, comprising: The ophthalmic preparation is a) mixing cyclosporine with hydrogenated 40 polyoxyl castor oil at a temperature of 55° C.±2° C. or higher to form mixture A; b) prior to complete dissolution of the cyclosporine, reducing the temperature of mixture A to a temperature of 35° C.±2° C. and adding octoxynol-40; c) mixing the resulting mixture with an aqueous vehicle.

[0183] In another preferred embodiment, the present disclosure provides: 0.09% by weight of cyclosporine, about 1.0% by weight of hydrogenated 40 polyoxyl castor oil, about 0.05% by weight of Octoxynol-40, and A method for making a stable nanomicellar ophthalmic formulation comprising an aqueous vehicle, comprising: The ophthalmic preparation is a) mixing cyclosporine with hydrogenated 40 polyoxyl castor oil at a temperature of 55° C.±2° C. or higher to form mixture A; b) holding mixture A under vacuum to remove bubbles; c) optionally, prior to complete dissolution of cyclosporine, reducing the temperature of mixture A to a temperature of 35° C.±2° C.; d) adding octoxynol-40; e) mixing the resulting mixture with an aqueous vehicle.

[0184] In one embodiment, the mixing of the aqueous vehicles occurs at a temperature of 35° C.±2° C. In another embodiment, the mixing of the aqueous vehicles is performed at a temperature of 55±2° C.

[0185] The stable nanomicellar ophthalmic formulation of the preceding embodiment comprises: About 0.20 to 0.550% by weight of sodium phosphate monobasic, about 0.23 to 0.465% by weight of dibasic sodium phosphate; about 0.05% by weight of sodium chloride, about 0.3% by weight povidone, Sodium hydroxide / hydrochloric acid to adjust the pH, and Further included is water for injection.

[0186] In another preferred embodiment, the present disclosure is a stable nanomicellar ophthalmic formulation having a pH of about 5.0 to 8.0. More preferably, the pH of the formulation is about 6.5 to 7.2.

[0187] Further disclosed herein is a stable nanomicellar ophthalmic formulation having an osmolality of about 150 to about 200 mOsmol / kg.

[0188] Further disclosed herein are stable nanomicelle ophthalmic formulations in which the mixed nanomicelle size and polydispersity index were determined with a Zetasizer (Malvern Instruments, NJ). Briefly, approximately 1 ml of each formulation was transferred to a cuvette and placed in the instrument. A laser beam was used to determine the mixed nanomicelle size. Nanomicelles contemplated by the present disclosure typically range in size from about 1-100 nm, in some embodiments the size ranges from about 5-50 nm, in some embodiments the size ranges from about 10-40 nm, and in some embodiments the size ranges from about 13-16 nm.

[0189] In another embodiment, the present disclosure relates to cyclosporine forms having characteristic XRD peaks at 6.9, 7.8, 9.4, and 15.9 2-theta (degrees).

[0190] In yet another embodiment, the present disclosure relates to a stable nanomicellar ophthalmic formulation comprising an amorphous form of cyclosporine.

[0191] In yet another embodiment, the present disclosure relates to a stable nanomicellar ophthalmic formulation substantially free of cyclosporine forms having characteristic XRD peaks at 7.4, 8.7, 14.4, and 17.5 2-theta (degrees).

[0192] In yet another embodiment, the present disclosure is a stable nanomicellar ophthalmic formulation that is substantially free of cyclosporine forms having characteristic XRD peaks at 8.5, 9.3, 11.6, and 20.3 2-theta (degrees). The XRD data for these cyclosporine forms is presented in FIG. 2.

[0193] The competitive dissolution time of cyclosporine depends on the amount of cyclosporine A dissolved based on the batch size. In general, the dissolution time of cyclosporine in hydrogenated 40 polyoxyl castor oil (Kolliphor® RH 40) was found to be 130 minutes or more at a ratio of 9:10 (cyclosporine:Kolliphor® RH 40). Cyclosporine A slowly dissolved over a period of time with stirring, and during this complete dissolution period the solution became clear. If cyclosporine A reprecipitates during the dissolution period, it is possible that a slightly cloudy solution migrated to the aqueous phase during batch manufacture. This may initiate a seeding effect on crystal growth in the final formulation during storage, leading to batch failure. Based on these observations, the dissolution behavior of cyclosporine was tested in hydrogenated 40 polyoxyl castor oil (Kolliphor® RH 40) at 55° C. and 35° C. It was found that the solution stability at 35° C. was relatively higher than that at 55° C. (see Example 2 in Table 4). Based on the above observations, if cyclosporine was not completely dissolved in Kolliphor® RH 40 at 55° C., it is believed that the temperature can be reduced to 35° C. This is because the solution stability of cyclosporine at 35° C. is higher than that at 55° C.

[0194] In another embodiment, when cyclosporine is present in a form having characteristic XRD peaks at 7.4, 8.7, 14.4, and 17.5 2-theta (degrees) (FIG. 2B), cyclosporine is mixed with hydrogenated polyoxyl castor oil at a temperature of 127-130° C. until completely dissolved, and octoxynol-40 is added to the mixture at 127-130° C.

[0195] In another embodiment, when cyclosporine is present in a form having characteristic XRD peaks at 8.5, 9.3, 11.6, and 20.3 2-theta (degrees) (FIG. 2C), cyclosporine is mixed with hydrogenated polyoxyl castor oil at a temperature of 127-130° C. until completely dissolved, and octoxynol-40 is added to the mixture at 127-130° C.

[0196] In certain embodiments of the present invention, in the step of reducing the temperature of mixture A, mixture A is reduced to a temperature of 35° C.±2° C. The step of reducing the temperature of mixture A may occur in less than 65 minutes, preferably about 60 minutes. In certain embodiments, water may be added after the step of reducing the temperature of mixture A.

[0197] In another aspect, the present disclosure provides a stable nanomicelle ophthalmic formulation, comprising: a) mixing cyclosporine with hydrogenated 40 polyoxyl castor oil at a temperature of 55° C.±2° C. or higher to form mixture A; b) reducing the temperature of mixture A to a temperature of 35° C.±2° C. prior to complete dissolution of cyclosporine; c) adding Octoxynol-40; d) then mixing the resulting mixture with an aqueous vehicle at 35° C.±2° C., wherein mixture A drops to a temperature of 35° C.±2° C. in less than 65 minutes. Preferably, mixture A drops to a temperature of 35° C.±2° C. in 60 minutes.

[0198] In one embodiment, mixture A in step (a) is mixed for 20 to 30 minutes, preferably for 20 to 25 minutes, more preferably for 20±2 minutes.

[0199] In one embodiment, the temperature is reduced to 35° C.±2° C. and stirred for 60 to 70 minutes. Preferably, the mixture is stirred for 60±5 minutes at a temperature of 35° C.±2° C.

[0200] In yet another aspect, the present invention provides a method for producing a composition comprising: 0.09% by weight of cyclosporine, about 1.0% by weight of hydrogenated 40 polyoxyl castor oil, about 0.05% by weight of Octoxynol-40; About 0.20 to 0.550% by weight of sodium phosphate monobasic, about 0.23 to 0.465% by weight of dibasic sodium phosphate; about 0.05% by weight of sodium chloride, about 0.3% by weight povidone, Sodium hydroxide / hydrochloric acid to adjust the pH, and A method for making a stable nanomicelle ophthalmic formulation with water for injection, comprising: The method comprises: a) mixing cyclosporine with hydrogenated 40 polyoxyl castor oil at a temperature of 55° C.±2° C. or higher to form mixture A; b) prior to complete dissolution of the cyclosporine, reducing the temperature of mixture A to a temperature of 35° C.±2° C. and adding octoxynol-40 to form an API mixture; c) adding the API mixture to water for injection (WFI); d) adding the remaining excipients to step (c) in the following order: sodium phosphate monobasic, then sodium phosphate dibasic, then sodium chloride, then polyvinylpyrrolidone; e) adjusting the pH to 6.5-7.2 and bringing to final volume with WFI.

[0201] In one embodiment, the mixing of the cyclosporine in step (a) is performed at 200-300 RPM.

[0202] In one embodiment, the mixing of mixture A in step (a) is carried out for 20 to 30 minutes, preferably 20 to 25 minutes, more preferably 20±2 minutes.

[0203] In one embodiment, the temperature of mixture A is decreased to 35° C.±2° C. in less than 65 minutes. Preferably, the temperature of mixture A is decreased to 35° C.±2° C. in 40 to 50 minutes.

[0204] In another embodiment, the mixture of step (b) is stirred for 60 to 70 minutes. Preferably, the mixture is stirred for 60±5 minutes at a temperature of 35° C.±2° C.

[0205] In one embodiment, blending the API mixture with the WFI is performed at a temperature of 35° C.±2° C. In another embodiment, blending the API mixture with the WFI is performed at a temperature of 55±2° C.

[0206] In yet another aspect, the present invention provides a method for producing a composition comprising: 0.09% by weight of cyclosporine, about 1.0% by weight of hydrogenated 40 polyoxyl castor oil, about 0.05% by weight of Octoxynol-40; About 0.20 to 0.550% by weight of sodium phosphate monobasic, about 0.23 to 0.465% by weight of dibasic sodium phosphate; about 0.05% by weight of sodium chloride, about 0.3% by weight povidone, Sodium hydroxide / hydrochloric acid to adjust the pH, and A method for making a stable nanomicelle ophthalmic formulation with water for injection, comprising: The method comprises: a) mixing cyclosporine with hydrogenated 40 polyoxyl castor oil at a temperature of 55° C.±2° C. or higher to form mixture A; b) holding mixture A under vacuum to remove bubbles; c) optionally, prior to complete dissolution of cyclosporine, reducing the temperature of mixture A to a temperature of 35° C.±2° C.; d) adding octoxynol-40; e) adding the API blend to water for injection (WFI); f) adding the remaining excipients to step (c) in the following order: sodium phosphate monobasic, then sodium phosphate dibasic, then sodium chloride, then polyvinylpyrrolidone; g) adjusting the pH to 6.5-7.2 and bringing to final volume with WFI.

[0207] In one embodiment, the mixing of mixture A in step (a) is carried out for 20 to 30 minutes, preferably 20 to 25 minutes, more preferably 20±2 minutes.

[0208] In one embodiment, the temperature of mixture A is decreased to 35° C.±2° C. in less than 65 minutes. Preferably, the temperature of mixture A is decreased to 35° C.±2° C. in 40 to 50 minutes.

[0209] In one embodiment, blending the API mixture with the WFI is performed at a temperature of 35° C.±2° C. In another embodiment, blending the API mixture with the WFI is performed at a temperature of 55±2° C.

[0210] In a more preferred aspect, the present invention provides a stable nanomicellar ophthalmic formulation prepared by any of the above methods.

[0211] In yet another aspect, the present disclosure provides a method for producing a method for manufacturing a semiconductor device comprising: 0.09% by weight of cyclosporine, about 1.0% by weight of hydrogenated 40 polyoxyl castor oil, about 0.05% by weight of Octoxynol-40, and A stable nanomicellar ophthalmic formulation comprising an aqueous vehicle, The ophthalmic preparation is a) mixing cyclosporin with hydrogenated 40 polyoxyl castor oil at a temperature of 127-130° C. until completely dissolved to form mixture A; b) adding Octoxynol-40 to mixture A at 127-130° C.; c) mixing the resulting mixture with an aqueous vehicle at a temperature of 127-130°C.

[0212] In one aspect, the present disclosure is a stable nanomicellar ophthalmic formulation comprising cyclosporine forms having characteristic XRD peaks at 6.9, 7.8, 9.4, and 15.9 2-theta (degrees).

[0213] In another aspect, the present disclosure is a stable nanomicellar ophthalmic formulation comprising an amorphous form of cyclosporine.

[0214] In yet another aspect, the present disclosure provides a method for producing a method for manufacturing a semiconductor device comprising: 0.09% by weight of cyclosporine, about 1.0% by weight of hydrogenated 40 polyoxyl castor oil, about 0.05% by weight of Octoxynol-40, and A stable nanomicellar ophthalmic formulation comprising an aqueous vehicle, The ophthalmic preparation is a) mixing cyclosporin with hydrogenated 40 polyoxyl castor oil at a temperature of 127-130° C. until completely dissolved to form mixture A; b) adding Octoxynol-40 to mixture A at 127-130° C.; c) mixing the resulting mixture with an aqueous vehicle at a temperature of 127-130°C; Cyclosporine exists in forms with characteristic XRD peaks at 7.4, 8.7, 14.4, and 17.5 2-theta (degrees).

[0215] In yet another aspect, the present disclosure provides a method for producing a method for manufacturing a semiconductor device comprising: 0.09% by weight of cyclosporine, about 1.0% by weight of hydrogenated 40 polyoxyl castor oil, about 0.05% by weight of Octoxynol-40, and A stable nanomicellar ophthalmic formulation comprising an aqueous vehicle, The ophthalmic preparation is a) mixing cyclosporin with hydrogenated 40 polyoxyl castor oil at a temperature of 127-130° C. until completely dissolved to form mixture A; b) adding Octoxynol-40 to mixture A at 127-130° C.; c) mixing the resulting mixture with an aqueous vehicle at a temperature of 127-130°C; Cyclosporine exists in forms with characteristic XRD peaks at 2-theta (degrees) 8.5, 9.3, 11.6, and 20.3.

[0216] In yet another aspect, the present disclosure provides a method for producing a method for manufacturing a semiconductor device comprising: 0.09% by weight of cyclosporine, about 1.0% by weight of hydrogenated 40 polyoxyl castor oil, about 0.05% by weight of Octoxynol-40, and A stable nanomicellar ophthalmic formulation comprising an aqueous vehicle, The ophthalmic preparation is a) mixing hydrogenated 40 polyoxyl castor oil and octoxynol-40 at a temperature of 127-130° C. to form a mixture A; b) adding cyclosporine to mixture A at 127-130°C; c) mixing the resulting mixture with an aqueous vehicle at a temperature of 127-130°C.

[0217] In one aspect, the present disclosure is a stable nanomicellar ophthalmic formulation comprising cyclosporine forms having characteristic XRD peaks at 6.9, 7.8, 9.4, and 15.9 2-theta (degrees).

[0218] In another aspect, the present disclosure is a stable nanomicellar ophthalmic formulation comprising an amorphous form of cyclosporine.

[0219] In another aspect, the present disclosure is a stable nanomicellar ophthalmic formulation comprising cyclosporine in a form having characteristic XRD peaks at 7.4, 8.7, 14.4, and 17.5 2-theta (degrees).

[0220] In another aspect, the present disclosure is a stable nanomicelle ophthalmic formulation comprising cyclosporine.

[0221] In yet another aspect, the present disclosure provides a method for producing a method for manufacturing a semiconductor device comprising: 0.09% by weight of cyclosporine, about 1.0% by weight of hydrogenated 40 polyoxyl castor oil, about 0.05% by weight of Octoxynol-40; About 0.20 to 0.550% by weight of sodium phosphate monobasic, about 0.23 to 0.465% by weight of dibasic sodium phosphate; about 0.05% by weight of sodium chloride, about 0.3% by weight povidone, Sodium hydroxide / hydrochloric acid to adjust the pH, and A stable nanomicelle ophthalmic formulation comprising water for injection, The ophthalmic preparation is a) mixing cyclosporin with hydrogenated 40 polyoxyl castor oil at a temperature of 127-130° C. until completely dissolved to form mixture A; b) adding Octoxynol-40 to mixture A at 127-130° C. to form an API mixture; c) adding the API mixture to water for injection (WFI); d) adding the remaining excipients to step (c) in the following order: sodium phosphate monobasic, then sodium phosphate dibasic, then sodium chloride, then polyvinylpyrrolidone; e) adjusting the pH to 6.5-7.2 and bringing to final volume with WFI.

[0222] In one aspect, the present disclosure is a stable nanomicellar ophthalmic formulation comprising cyclosporine forms having characteristic XRD peaks at 6.9, 7.8, 9.4, and 15.9 2-theta (degrees).

[0223] In another aspect, the present disclosure is a stable nanomicellar ophthalmic formulation comprising an amorphous form of cyclosporine.

[0224] In another aspect, the present disclosure is a stable nanomicellar ophthalmic formulation comprising cyclosporine in a form having characteristic XRD peaks at 7.4, 8.7, 14.4, and 17.5 2-theta (degrees).

[0225] In another aspect, the present disclosure is a stable nanomicelle ophthalmic formulation comprising cyclosporine.

[0226] In yet another aspect, the present invention provides a method for producing a composition comprising: 0.09% by weight of cyclosporine, about 1.0% by weight of hydrogenated 40 polyoxyl castor oil, about 0.05% by weight of Octoxynol-40; About 0.20 to 0.550% by weight of sodium phosphate monobasic, about 0.23 to 0.465% by weight of dibasic sodium phosphate; about 0.05% by weight of sodium chloride, about 0.3% by weight povidone, Sodium hydroxide / hydrochloric acid to adjust the pH, and A method for making a stable nanomicelle ophthalmic formulation with water for injection, comprising: The method comprises: a) mixing cyclosporin with hydrogenated 40 polyoxyl castor oil at a temperature of 127-130° C. until completely dissolved to form mixture A; b) adding Octoxynol-40 to mixture A at 127-130° C. to form an API mixture; c) adding the API mixture to water for injection (WFI); d) adding the remaining excipients to step (c) in the following order: sodium phosphate monobasic, then sodium phosphate dibasic, then sodium chloride, then polyvinylpyrrolidone; e) adjusting the pH to 6.5-7.2 and bringing to final volume with WFI.

[0227] In one aspect, the present disclosure is a method of making a stable nanomicellar ophthalmic formulation comprising cyclosporine forms having characteristic XRD peaks at 6.9, 7.8, 9.4, and 15.9 2-theta (degrees).

[0228] In another aspect, the present disclosure is a method of making a stable nanomicellar ophthalmic formulation comprising an amorphous form of cyclosporine.

[0229] In another aspect, the present disclosure is a method of making a stable nanomicellar ophthalmic formulation comprising cyclosporine in a form having characteristic XRD peaks at 7.4, 8.7, 14.4, and 17.5 2-theta (degrees).

[0230] In another aspect, the present disclosure is a method of making a stable nanomicellar ophthalmic formulation comprising cyclosporine.

[0231] In yet another aspect, the present disclosure provides a method for producing a method for manufacturing a semiconductor device comprising: 0.09% by weight of cyclosporine, about 1.0% by weight of hydrogenated 40 polyoxyl castor oil, about 0.05% by weight of Octoxynol-40; About 0.20 to 0.550% by weight of sodium phosphate monobasic, about 0.23 to 0.465% by weight of dibasic sodium phosphate; about 0.05% by weight of sodium chloride, about 0.3% by weight povidone, Sodium hydroxide / hydrochloric acid to adjust the pH, and A stable nanomicelle ophthalmic formulation comprising water for injection, The ophthalmic preparation is a) mixing hydrogenated 40 polyoxyl castor oil and octoxynol-40 at a temperature of 127-130° C. to form a mixture A; b) adding cyclosporine to mixture A at 127-130° C. and stirring until completely dissolved to form an API mixture; c) adding the API mixture to water for injection (WFI); d) adding the remaining excipients to step (c) in the following order: sodium phosphate monobasic, then sodium phosphate dibasic, then sodium chloride, then polyvinylpyrrolidone; e) adjusting the pH to 6.5-7.2 and bringing to final volume with WFI.

[0232] In one aspect, the present disclosure is a stable nanomicellar ophthalmic formulation comprising cyclosporine forms having characteristic XRD peaks at 6.9, 7.8, 9.4, and 15.9 2-theta (degrees).

[0233] In another aspect, the present disclosure is a stable nanomicellar ophthalmic formulation comprising an amorphous form of cyclosporine.

[0234] In another aspect, the present disclosure is a stable nanomicellar ophthalmic formulation comprising cyclosporine in a form having characteristic XRD peaks at 7.4, 8.7, 14.4, and 17.5 2-theta (degrees).

[0235] In another aspect, the present disclosure is a stable nanomicelle ophthalmic formulation comprising cyclosporine.

[0236] In yet another aspect, the present invention provides a method for producing a composition comprising: 0.09% by weight of cyclosporine, about 1.0% by weight of hydrogenated 40 polyoxyl castor oil, about 0.05% by weight of Octoxynol-40; About 0.20 to 0.550% by weight of sodium phosphate monobasic, about 0.23 to 0.465% by weight of dibasic sodium phosphate; about 0.05% by weight of sodium chloride, about 0.3% by weight povidone, Sodium hydroxide / hydrochloric acid to adjust the pH, and A method for making a stable nanomicelle ophthalmic formulation with water for injection, comprising: The method comprises: a) mixing hydrogenated 40 polyoxyl castor oil and octoxynol-40 at a temperature of 127-130° C. to form a mixture A; b) adding cyclosporine to mixture A at 127-130° C. and stirring until completely dissolved to form an API mixture; c) adding the API mixture to water for injection (WFI); d) adding the remaining excipients to step (c) in the following order: sodium phosphate monobasic, then sodium phosphate dibasic, then sodium chloride, then polyvinylpyrrolidone; e) adjusting the pH to 6.5-7.2 and bringing to final volume with WFI.

[0237] In one aspect, the present disclosure is a method of making a stable nanomicellar ophthalmic formulation comprising cyclosporine forms having characteristic XRD peaks at 6.9, 7.8, 9.4, and 15.9 2-theta (degrees).

[0238] In another aspect, the present disclosure is a method of making a stable nanomicellar ophthalmic formulation comprising an amorphous form of cyclosporine.

[0239] In another aspect, the present disclosure is a method of making a stable nanomicellar ophthalmic formulation comprising cyclosporine in a form having characteristic XRD peaks at 7.4, 8.7, 14.4, and 17.5 2-theta (degrees).

[0240] In another aspect, the present disclosure is a method of making a stable nanomicellar ophthalmic formulation comprising cyclosporine.

[0241] In one embodiment, mixing speed, time, and energy input play a role in the complete dissolution of cyclosporine in hydrogenated polyoxyl castor oil. When a lower speed is used, the time for dissolution increases. In one aspect of this embodiment, typically, cyclosporine is dissolved in hydrogenated polyoxyl castor oil by stirring at approximately 100-200 RPM for 90 minutes, 85 minutes, 80 minutes, 75 minutes, 70 minutes, 65 minutes, 60 minutes, 55 minutes, 50 minutes, 45 minutes, 40 minutes, 35 minutes, 30 minutes, 25 minutes, 20 minutes, 15 minutes, 10 minutes. At 200-300 RPM, 75 minutes, 70 minutes, 65 minutes, 60 minutes, 55 minutes, 50 minutes, 45 minutes, 40 minutes, 35 minutes, 30 minutes, 25 minutes, 20 minutes, 15 minutes, 10 minutes. In another embodiment, cyclosporine is typically dissolved in hydrogenated polyoxyl castor oil by stirring at approximately 300-400 RPM for 65 minutes, 60 minutes, 55 minutes, 50 minutes, 45 minutes, 40 minutes, 35 minutes, 30 minutes, 25 minutes, 20 minutes, 15 minutes, 10 minutes. In another embodiment, cyclosporine is typically dissolved in hydrogenated polyoxyl castor oil by stirring at approximately 350-400 RPM for 60 minutes, 55 minutes, 50 minutes, 45 minutes, 40 minutes, 35 minutes, 30 minutes, 25 minutes, 20 minutes, 15 minutes, 10 minutes. In another embodiment, cyclosporine is typically dissolved in hydrogenated polyoxyl castor oil by stirring at approximately 400-450 RPM for 50 minutes, 45 minutes, 40 minutes, 35 minutes, 30 minutes, 25 minutes, 20 minutes, 15 minutes, 10 minutes. In another embodiment, the cyclosporine is typically dissolved in the hydrogenated polyoxyl castor oil by stirring at greater than approximately 450 RPM for 40 minutes, 35 minutes, 30 minutes, 25 minutes, 20 minutes, 15 minutes, 10 minutes, 5 minutes, etc. In yet another embodiment, the cyclosporine is dissolved in the hydrogenated polyoxyl castor oil by stirring at approximately 200-300 RPM until complete dissolution.

[0242] The energy input into the mixture is defined in Equation 1 as follows: E / V=n 3 D 5 t / V(1) where E is the theoretical energy input, n is the rpm of the shear plate, D is the shear plate diameter, t is time, and V is the solution volume. The energy input per volume is scale independent. (Diaz, M., et al., "Mixing Power, External Convection, and Effectiveness in Bioreactors," Biotechnology and Bioengineering, Vol. 51, 1996, pp. 131-140). This is a simple, fast and reliable method for scaling up the preparation of nanomicelle formulations.

[0243] Alternative methods of mixing, such as the use of sonicators or the use of solvents, water, or pressure, may be used to affect the temperature or time of each process step, for example, if a solvent is used, it may be possible to utilize a lower temperature in the process step.

[0244] In one embodiment, the mixture is mixed using a homogenizer or sonicator or mixer equipped with an agitator. The agitator may be located at the overhead or bottom or both the overhead and bottom of the homogenizer or sonicator or mixer. Preferably, the agitator is present at both the overhead and bottom of the homogenizer or sonicator or mixer.

[0245] The present disclosure further relates to the treatment or prevention of ocular diseases or disorders, for example, by topical administration of the formulations described herein.

[0246] The term "treatment" refers to preventing a disease, disorder, or condition from occurring in a cell, tissue, system, animal, or human that may be predisposed to, but has not yet been diagnosed as having, the disease, disorder, and / or condition; stabilizing the disease, disorder, or condition, i.e., halting its development; and / or alleviating one or more symptoms of the disease, disorder, or condition, i.e., causing regression of the disease, disorder, and / or condition.

[0247] As used herein, a composition that "prevents" a disorder or condition refers to a compound that, in a statistical sample, reduces the occurrence of the disorder or condition in a treated sample compared to an untreated control sample, or delays the onset of or reduces the severity of one or more symptoms of the disorder or condition compared to an untreated control sample.

[0248] As used herein, the term "ocular disease" refers to a disease / condition that can threaten vision, lead to ocular discomfort, and may be a precursor to systemic health problems.

[0249] The patient or subject treated by any of the compositions or methods of the present disclosure may refer to either a human or a non-human animal. In one embodiment, the present disclosure provides a method for treating an ocular disease in a human patient in need thereof. In one embodiment, the present disclosure provides a method for treating an inflammatory ocular disease in a human patient in need thereof. In another embodiment, the present disclosure provides a method for treating an ocular disease in a veterinary patient in need thereof, including but not limited to dogs, horses, cats, rabbits, gerbils, hamsters, rodents, birds, aquatic mammals, cows, pigs, camelids, and other zoological animals.

[0250] In some embodiments of the compositions and methods disclosed herein, the cyclosporine further comprises one or more additional active ingredients, e.g., an active agent selected from the group consisting of a resolvin or resolvin-like compound, a steroid (such as a corticosteroid), and the like. In some embodiments, the additional active agent comprises a resolvin. In some embodiments, the additional active agent comprises a corticosteroid. In some embodiments, the additional active agent comprises a resolvin and a corticosteroid. In some embodiments, the additional active agent comprises an antibiotic, e.g., one or more antibiotics selected from the group consisting of azithromycin, ciprofloxacin, ofloxacin, gatifloxacin, levofloxacin, moxifloxacin, besifloxacin, and levofloxacin. In some embodiments, the additional active agent comprises an antibiotic, e.g., one or more antibiotics selected from the group consisting of azithromycin, ciprofloxacin, ofloxacin, gatifloxacin, levofloxacin, moxifloxacin, besifloxacin, and levofloxacin, and a second such active agent is a resolvin as described herein (including, but not limited to, compound 1001). In some embodiments, the active agent comprises two or more active agents, and one of the active agents is an antiviral agent, e.g., one or more antiviral agents selected from the group consisting of ganciclovir, trifluridine, acyclovir, famciclovir, valacyclovir, penciclovir, and cidofovir. In some embodiments, the active agent comprises two or more active agents, one of which is an antibiotic, e.g., one or more antiviral agents selected from the group consisting of ganciclovir, trifluridine, acyclovir, famciclovir, valacyclovir, penciclovir, and cidofovir, and the second of which is a resolvin as described herein (including, but not limited to, compound 1001).

[0251] Thus, in another aspect, there is provided a method of treating or preventing an ocular disease or condition comprising topically administering a formulation of any of the aspects or embodiments disclosed herein. In some embodiments, the ocular disease is an anterior ocular disease. In some embodiments, the ocular disease is a posterior ocular disease. In some embodiments, the ocular disease is one or more selected from the group consisting of dry eye syndrome, Sjogren's syndrome, uveitis, anterior uveitis (iritis), chorioretinitis, posterior uveitis, conjunctivitis, allergic conjunctivitis, keratitis, keratoconjunctivitis, vernal keratoconjunctivitis (VKC), atopic keratoconjunctivitis, cicatricial conjunctivitis and other autoimmune disorders of the ocular surface, blepharitis, scleritis, age-related macular degeneration (AMD), diabetic retinopathy (DR), diabetic macular edema (DME), ocular neovascularization, age-related macular degeneration (ARMD), proliferative vitreoretinopathy (PVR), cytomegalovirus (CMV) retinitis, optic neuritis, retrobulbar optic neuritis, and macular pucker. In one embodiment, the ocular disease is dry eye. In one embodiment, the ocular disease is allergic conjunctivitis. In one embodiment, the eye disease is age-related macular degeneration (AMD). In one embodiment, the eye disease is diabetic retinopathy.

[0252] The daily dose of the ophthalmic formulation effective for relieving dry eye symptoms and / or improving tear film can be divided into one or several unit dose administrations. The subject can use the product as needed, generally not more than twice a day, and in many cases the product is used only once a day. The preferred regimen of the nanomicelle ophthalmic formulation of the present invention is one drop of 0.09% (w / w) solution per eye, twice a day (approximately 12 hours apart). EXAMPLES

[0253] The following examples have been prepared to illustrate non-limiting embodiments of the present disclosure.

[0254] Example 1 [Table 1]

[0255] Cyclosporine nanomicelle ophthalmic solution was prepared as follows: In Example 1(a), polyoxyl 40 hydrogenated castor oil (Kolliphor® RH 40) was melted at 55-60°C with stirring at about 200 rpm. Cyclosporine A was added to the melted Kolliphor® RH 40 at 55-60°C and the reaction mixture was mixed at the same temperature range until completely dissolved. After Cyclosporine A was dissolved, a surfactant (Octoxynol-40) was added under stirring and after stirring for 10 minutes, this non-aqueous solution was delivered to 90% water for injection at 55-60°C. The temperature of the water for injection was maintained below 22°C. Sodium phosphate monobasic, sodium phosphate dibasic, sodium chloride, and povidone were added sequentially to the bulk solution under stirring until completely dissolved. When all components were completely dissolved in the bulk solution, the volume was made up to 100% with water for injection to 1 L. In Example 1(b), the procedure of Example 1(a) was followed except that the solution of cyclosporine A in Kolliphor® RH 40 at 55-60° C. was added to the water immediately after it began to become cloudy.

[0256] Both batches of Example 1(a) and Example 1(b) were filled into three 5 mL low density polyethylene (LDPE) vials in a sterile area. The vials were exposed to accelerated temperatures of 40° C. and 30° C. in a chamber (to accelerate particle formation). Samples were visually inspected daily for any signs of turbidity and / or visible particle formation. The two batches were analyzed for critical quality parameters such as assays for cyclosporine, pH, osmolality, and micelle size, all of which were found to be well within specifications. The results are provided in Table 2 below. Figure 1 shows photographs of a stable batch (Example 1a) and an unstable batch (Example 1b). [Table 2]

[0257] The data in Table 2 show that when batches are made using a clear CsA non-aqueous phase (cyclosporine in Kolliphor® RH 40 and Octoxynol-40) and then the mixture is added to the aqueous phase, the batches remain stable for a longer period of time. However, when batches are made using a cloudy cyclosporine non-aqueous phase and then the mixture is added to the aqueous phase, the batches show less stability. This indicates that non-uniform distribution of cyclosporine within the micelles may promote nucleation and particle formation in the final product upon storage.

[0258] Example 2 Solution stability of cyclosporine A lots in polyoxyl 40 hydrogenated castor oil (Kolliphor® RH 40) at 55-60 °C is a critical process parameter for a stable formulation. Surprisingly, it was also found that the dissolution behavior changed over storage time, as shown in Table 3. [Table 3]

[0259] Example 3 Similarly, the dissolution behavior test of Cyclosporine A in polyoxyl 40 hydrogenated castor oil (Kolliphor® RH 40) was examined at 35° C. Table 4 shows the results of the dissolution behavior test of Cyclosporine A in Kolliphor® RH 40 at 35° C. As can be seen from Table 4, the solution stability at 35° C. was found to be relatively higher than that at 55° C. [Table 4]

[0260] As can be seen from Table 4, if Cyclosporine A is not completely dissolved in Kolliphor® RH 40 at 55° C., the temperature can be reduced to 35° C. Based on this data, the solution stability of Cyclosporine A at 35° C. is believed to be greater than that at 55° C. The complete dissolution step of Cyclosporine A in Kolliphor® RH 40 was modified as follows: (i) Dispersing the API for 20 minutes at 55°C ± 2°C and stirring for an additional 15 minutes; (ii) The temperature is then reduced from 55° C.±2° C. to 35° C.±2° C. in less than 55 minutes (approximately 40 minutes) and the solution is stirred at 35° C.±2° C. until complete dissolution of the API is achieved; (iii) Octoxynol-40 was added with stirring and the non-aqueous phase was delivered to water for injection at 35° C.±2° C.

[0261] This change in the dissolution process accommodates lot-to-lot variability in solution stability of the API in Kolliphor® RH-40 at 55° C. and can also accommodate variability during storage.

[0262] Example 4 The dissolution behavior of the three cyclosporine forms disclosed in Figure 2 was tested in polyoxyl 40 hydrogenated castor oil (Kolliphor® RH 40) at 55°C. The results are shown in Table 5. [Table 5]

[0263] 3(a)-3(d) are photographs of the results of the tests reported in Table 5.

[0264] As can be seen from Table 5 above and the associated figures, the CsA forms having characteristic XRD peaks at 7.4, 8.7, 14.4, and 17.5 2-theta (degrees), as well as the CsA forms having characteristic XRD peaks at 8.5, 9.3, 11.6, and 20.3 2-theta (degrees), are much less soluble compared to the CsA forms having characteristic XRD peaks at 6.9, 7.8, 9.4, and 15.9 2-theta (degrees). During dissolution of cyclosporine in Kolliphor® RH 40 at 55° C., the CsA forms having characteristic XRD peaks at 6.9, 7.8, 9.4, and 15.9 2-theta (degrees) may change to a less soluble form, or the amorphous cyclosporine may recrystallize to a relatively less soluble form. This transformation may be dependent on stresses on the system, including temperature, extended storage periods at elevated temperatures, etc. For this purpose, cyclosporine ophthalmic formulations were exposed to higher temperatures and times and PXRD data was obtained from the precipitated portion. As shown in Figure 4, the precipitate was found to be close to the CsA form with characteristic XRD peaks at 6.9, 7.8, 9.4 and 15.9 2-theta (degrees). Since this CsA form has low solubility in water, its use may result in a seeding effect either at the initial stage or during storage at higher temperatures, thus leading to batch failure.

[0265] In another attempt, the behavior of CsA in Kolliphor® RH 40 alone in the absence of water is determined. CsA was dissolved in Kolliphor® RH 40 at 55° C. and held for a longer time until precipitation occurred. The precipitated portion was separated and PXRD was performed. As shown in FIG. 5, cyclosporine was found to have characteristic XRD peaks at 8.5, 9.3, 11.6, and 20.3 2-theta (degrees). As a result, the process disclosed in the present disclosure, which reduces the exposure of cyclosporine to Kolliphor® RH 40 at, for example, 55° C., can minimize this precipitation phenomenon, which may be due to the possible conversion of cyclosporine to the less soluble CsA form. Thus, the present disclosure provides, among other things, improved storage stability.

[0266] Example 5 Table 1. Preparation method of nanomicelle solution Polyoxyl 40 hydrogenated castor oil (Kolliphor® RH40) was heated to approximately 50-60°C until liquefied and then introduced into a 10 L glass vessel. Cyclosporine (CsA) was added while maintaining the vessel temperature at 55±2°C, dissolved by stirring at approximately 200-300 RPM for 75 minutes, and visually inspected to confirm a clear solution without visible particles. The temperature was gradually reduced to 35°C. Once completely dissolved, the temperature was increased to 55±2°C. Octoxynol-40 was then added. If Octoxynol-40 solidified, it was added after heating at approximately 50-60°C until liquefied.

[0267] A portion (approximately 90%) of water for injection (WFI) was charged to a stainless steel mixing tank and the temperature was maintained at 20-30° C. throughout the process. With stirring, the API blend was added to the mixing tank at 55±2° C. and the remaining excipients were added in the following order with stirring: sodium phosphate monobasic, then sodium phosphate dibasic, then sodium chloride, then polyvinylpyrrolidone, while stirring for approximately 15 minutes.

[0268] After mixing for 15 minutes, the pH was checked and adjusted to 6.8±0.2 using hydrochloric acid (1N) or sodium hydroxide (1N) if necessary. The solution was adjusted to final volume with WFI and filtered through a 0.2 μm filter.

[0269] Example 6 Table 1. Preparation method of nanomicelle solution Polyoxyl 40 hydrogenated castor oil (Kolliphor® RH 40) was heated to approximately 50-60°C until liquefied and then introduced into a 10 L glass vessel. Cyclosporine (CsA) was added while maintaining the vessel temperature at 55±2°C for 20±2 minutes, followed by stirring at approximately 200-300 RPM for 15 minutes. The temperature was gradually reduced to 35°C under stirring, and once the temperature reached 35°C, stirring was continued for 60±5 minutes. Octoxynol-40 was then added. If Octoxynol-40 solidified, it was heated to approximately 50-60°C until liquefied and then added.

[0270] A portion (approximately 90%) of water for injection (WFI) was charged to a stainless steel mixing tank and the temperature was maintained at 20-30° C. throughout the process. The CsA mixture was added to the mixing tank at 35±2° C. with stirring, and the remaining excipients were added in the following order: sodium phosphate monobasic, then sodium phosphate dibasic, then sodium chloride, then polyvinylpyrrolidone while stirring for approximately 15 minutes.

[0271] After mixing for 15 minutes, the pH was checked and adjusted to 6.8±0.2 using hydrochloric acid (1N) or sodium hydroxide (1N) if necessary. The solution was adjusted to final volume with WFI and filtered through a 0.2 μm filter.

[0272] Example 7 Table 1 Preparation method of nanomicelle solution - Process under high speed Polyoxyl 40 hydrogenated castor oil (Kolliphor® RH 40) was heated to approximately 50-60° C. until liquefied and then introduced into a 10 L glass vessel. Cyclosporine (CsA) was added and stirred at approximately >450 RPM for 15 minutes while maintaining the vessel temperature at 55±2° C. for 20±2 minutes. The temperature was gradually reduced to 35° C. under stirring and stirred for 60±5 minutes. Octoxynol-40 was then added. If Octoxynol-40 solidified, it was added after heating at approximately 50-60° C. until liquefied.

[0273] A portion (approximately 90%) of water for injection (WFI) was charged to a stainless steel mixing tank and the temperature was maintained at 20-30° C. throughout the process. The CsA mixture was added to the mixing tank at 35±2° C. with stirring, and the remaining excipients were added in the following order: sodium phosphate monobasic, then sodium phosphate dibasic, then sodium chloride, then polyvinylpyrrolidone while stirring for approximately 15 minutes.

[0274] After mixing for 15 minutes, the pH was checked and adjusted to 6.8±0.2 using hydrochloric acid (1N) or sodium hydroxide (1N) if necessary. The solution was adjusted to final volume with WFI and filtered through a 0.2 μm filter.

[0275] Example 8 Table 1 Preparation method of nanomicelle solution - Process at high temperature Polyoxyl 40 hydrogenated castor oil (Kolliphor® RH 40) was heated to approximately 50-60°C until liquefied and then introduced into a 10 L glass vessel. The temperature was increased to 127-130°C. While maintaining the vessel temperature at 127-130°C, cyclosporine (CsA) was added and stirred at approximately 200-300 RPM to completely dissolve. Octoxynol-40 was then added. If the Octoxynol-40 had solidified, it was heated at approximately 50-60°C until liquefied and then added.

[0276] A portion (approximately 90%) of water for injection (WFI) was charged to a stainless steel mixing tank and the temperature was maintained at 20-30° C. throughout the process. With stirring, the CsA mixture was added to the mixing tank at 127-130° C., and the remaining excipients were added in the following order: sodium phosphate monobasic, then sodium phosphate dibasic, then sodium chloride, then polyvinylpyrrolidone, while stirring for approximately 15 minutes.

[0277] After mixing for 15 minutes, the pH was checked and adjusted to 6.8±0.2 using hydrochloric acid (1N) or sodium hydroxide (1N) if necessary. The solution was adjusted to final volume with WFI and filtered through a 0.2 μm filter.

[0278] Example 9 Polyoxyl 40 hydrogenated castor oil (Kolliphor® RH 40) was heated to approximately 50-60°C until liquefied and then introduced into a 10 L glass vessel. The temperature was increased to 127-130°C. Octoxynol-40 was then added. If the Octoxynol-40 solidified, it was added after heating at approximately 50-60°C until liquefied. The mixture was stirred at 127-130°C for 20 minutes. While maintaining the vessel temperature at 127-130°C, cyclosporine (CsA) was added and stirred using a stirrer at approximately 200-300 RPM to ensure complete dissolution.

[0279] A portion (approximately 90%) of water for injection (WFI) was charged to a stainless steel mixing tank and the temperature was maintained at 20-30° C. throughout the process. With stirring, the CsA mixture was added to the mixing tank at 127-130° C., and the remaining excipients were added in the following order: sodium phosphate monobasic, then sodium phosphate dibasic, then sodium chloride, then polyvinylpyrrolidone while stirring for approximately 15 minutes using both overhead and bottom stirrers.

[0280] After mixing for 15 minutes, the pH was checked and adjusted to 6.8±0.2 using hydrochloric acid (1N) or sodium hydroxide (1N) if necessary. The solution was adjusted to final volume with WFI and filtered through a 0.2 μm filter.

[0281] Example 10 Stability testing The nanomicellar ophthalmic formulation of Example 6 was examined after storage at 25° C. / 40% RH for 6 months.

[0282] The formulations were examined for appearance, pH, osmolality, viscosity, cyclosporine assay by HPLC Method II, micelle size determination by laser light scattering, and the presence of particulate matter. [Table 6]

[0283] As can be seen from the data in Table 6, the formulation was found to be both chemically and physically stable.

[0284] Example 11 Stability testing The nanomicellar ophthalmic formulation of Example 8 was examined after storage at 25° C. / 40% RH for 24 months, at 30° C. / 35% RH for 6 months, and at 40° C. / 25% RH for 6 months. [Table 7] [Table 8]

[0285] Unless otherwise indicated, documents referred to herein are incorporated by reference in their entirety.

[0286] Although certain specific embodiments are fully described in this application, it is to be understood that the same concepts disclosed with respect to those specific embodiments are also applicable to other embodiments. Moreover, the individual elements of the formulations and methods disclosed herein are described with reference to specific embodiments for convenience only. It is to be understood that the individual elements of the formulations and methods disclosed herein are applicable to embodiments other than the specific embodiment for which they are described.

[0287] In addition, the scope of the present disclosure is not limited to the above embodiments, and those skilled in the art will understand that various modifications and changes can be made without departing from the scope of the present disclosure. For example, the batch size may be changed by those skilled in the art while remaining within the present disclosure.

Claims

1. A stable nanomicelle ophthalmic formulation comprising cyclosporine, hydrogenated 40 polyoxyl castor oil, octoxynol-40, and an aqueous vehicle, the ophthalmic formulation comprising: a) mixing said cyclosporine with said hydrogenated 40 polyoxyl castor oil at a temperature of not less than 55°C ± 2°C to form mixture A; b) reducing the temperature of Mixture A to a temperature of 35°C ± 2°C prior to complete dissolution of the cyclosporine; c) adding the octoxynol-40; and d) mixing the resulting mixture with said aqueous vehicle.

2. A stable nanomicellar ophthalmic formulation as described in claim 1, wherein the cyclosporine in the formulation comprises a cyclosporine form having characteristic XRD peaks at 2-theta (degrees) 6.9, 7.8, 9.4, and 15.

9.

3. The stable nanomicellar ophthalmic formulation of claim 1, wherein the cyclosporine in the formulation comprises an amorphous form of cyclosporine.

4. 2. The stable nanomicellar ophthalmic formulation of claim 1, wherein the formulation is substantially free of (i) cyclosporine forms having characteristic XRD peaks at 7.4, 8.7, 14.4, and 17.5 2-theta (degrees), and / or (ii) cyclosporine forms having characteristic XRD peaks at 8.5, 9.3, 11.6, and 20.3 2-theta (degrees).

5. A stable nanomicelle ophthalmic formulation described in any one of claims 1 to 4, wherein the temperature of mixture A is reduced to 35°C ± 2°C in less than 65 minutes and the mixing in d) is carried out at 35°C ± 2°C.

6. The stable nanomicelle ophthalmic formulation of claim 5, wherein the temperature of the mixture A decreases to 35°C ± 2°C in about 40 to about 50 minutes.

7. A stable nanomicelle ophthalmic formulation described in any one of claims 1 to 4, wherein the method further comprises stirring the mixture A at 35°C ± 2°C for 60 to 70 minutes following the reduction of b).

8. 1. A method of making a stable nanomicelle ophthalmic formulation, the ophthalmic formulation comprising cyclosporine, hydrogenated 40 polyoxyl castor oil, octoxynol-40, and an aqueous vehicle, the method comprising: a) mixing said cyclosporine with said hydrogenated 40 polyoxyl castor oil at a temperature of not less than 55°C ± 2°C to form mixture A; b) reducing the temperature of Mixture A to a temperature of 35°C ± 2°C prior to complete dissolution of the cyclosporine; c) adding the octoxynol-40; and d) mixing the resulting mixture with said aqueous vehicle.

9. The method described in claim 8, wherein the temperature of mixture A is reduced to 35°C ± 2°C in less than 65 minutes and the mixing in d) is carried out at 35°C ± 2°C.

10. The method of claim 9, wherein the temperature of mixture A is reduced to 35°C ± 2°C in about 40 to about 50 minutes.

11. The method of claim 8, further comprising stirring the mixture A at 35°C ± 2°C for 60 to 70 minutes following the reduction in b).

12. A stable nanomicelle ophthalmic formulation comprising cyclosporine, hydrogenated 40 polyoxyl castor oil, octoxynol-40, and an aqueous vehicle, the ophthalmic formulation comprising: a) mixing said cyclosporine with said hydrogenated 40 polyoxyl castor oil at a temperature of not less than 55°C ± 2°C to form mixture A; b) holding said mixture A under vacuum to remove bubbles; c) optionally, reducing the temperature of Mixture A to a temperature of 35°C ± 2°C prior to complete dissolution of the cyclosporine; d) adding the octoxynol-40; and e) mixing the resulting mixture with said aqueous vehicle.

13. 1. A method of making a stable nanomicelle ophthalmic formulation, the ophthalmic formulation comprising cyclosporine, hydrogenated 40 polyoxyl castor oil, octoxynol-40, and an aqueous vehicle, the method comprising: a) mixing said cyclosporine with said hydrogenated 40 polyoxyl castor oil at a temperature of not less than 55°C ± 2°C to form mixture A; b) holding said mixture A under vacuum to remove bubbles; c) optionally, reducing the temperature of Mixture A to a temperature of 35°C ± 2°C prior to complete dissolution of the cyclosporine; d) adding the octoxynol-40; e) mixing the resulting mixture with the aqueous vehicle.

14. A stable nanomicelle ophthalmic formulation comprising cyclosporine, hydrogenated 40 polyoxyl castor oil, octoxynol-40, and an aqueous vehicle, the ophthalmic formulation comprising: a) mixing the cyclosporin with the hydrogenated 40 polyoxyl castor oil at a temperature of 127-130°C until completely dissolved to form mixture A; b) adding the Octoxynol-40 to the mixture A at 127-130°C; and c) mixing the resulting mixture with said aqueous vehicle at a temperature of 127-130°C.

15. A method for making a stable nanomicelle ophthalmic formulation, wherein the ophthalmic formulation comprises cyclosporine, hydrogenated 40 polyoxyl castor oil, octoxynol-40, and an aqueous vehicle, the method comprising: a) mixing the cyclosporin with the hydrogenated 40 polyoxyl castor oil at a temperature of 127-130°C until completely dissolved to form mixture A; b) adding the Octoxynol-40 to the mixture A at 127-130°C; and c) mixing the resulting mixture with said aqueous vehicle at a temperature of 127-130°C.

16. A stable nanomicellar ophthalmic formulation comprising: comprising cyclosporine, hydrogenated 40 polyoxyl castor oil, octoxynol-40, and an aqueous vehicle; The ophthalmic preparation comprises: a) mixing the hydrogenated 40 polyoxyl castor oil and the octoxynol-40 at a temperature of 127-130°C to form mixture A; b) adding said cyclosporine to said mixture A at 127-130°C; and c) mixing the resulting mixture with said aqueous vehicle at a temperature of 127-130°C.

17. A method for making a stable nanomicellar ophthalmic formulation, comprising: the ophthalmic formulation comprising cyclosporine, hydrogenated 40 polyoxyl castor oil, octoxynol-40, and an aqueous vehicle; The method comprises: a) mixing the hydrogenated 40 polyoxyl castor oil and the octoxynol-40 at a temperature of 127-130°C to form mixture A; b) adding said cyclosporine to said mixture A at 127-130°C; and c) mixing the resulting mixture with said aqueous vehicle at a temperature of 127-130°C.

18. 17. The stable nanomicellar ophthalmic formulation of claim 14 or claim 16, wherein the cyclosporine in the formulation comprises a cyclosporine form having characteristic XRD peaks at 2-theta (degrees) 6.9, 7.8, 9.4, and 15.

9.

19. 17. The stable nanomicellar ophthalmic formulation of claim 14 or claim 16, wherein the cyclosporine in the formulation comprises an amorphous form of cyclosporine.

20. 17. The stable nanomicellar ophthalmic formulation of claim 14 or claim 16, wherein the cyclosporine in the formulation comprises cyclosporine in a form having characteristic XRD peaks at 2-theta (degrees) 7.4, 8.7, 14.4, and 17.

5.

21. 17. The stable nanomicellar ophthalmic formulation of claim 14 or claim 16, wherein the cyclosporine in the formulation comprises cyclosporine in a form having characteristic XRD peaks at 2-theta (degrees) 8.5, 9.3, 11.6, and 20.

3.

22. 0.09% by weight of said cyclosporine. about 1.0% by weight of said hydrogenated 40 polyoxyl castor oil, and about 0.05% by weight of said Octoxynol-40; 17. The stable nanomicellar ophthalmic formulation of any one of claims 1 to 4, claim 12, claim 14 or claim 16, comprising:

23. about 0.20 to 0.550% by weight of monobasic sodium phosphate; about 0.23 to 0.465% by weight of dibasic sodium phosphate; about 0.05% by weight of sodium chloride; about 0.3% by weight of povidone, and 23. The stable nanomicellar ophthalmic formulation of claim 22, further comprising sodium hydroxide / hydrochloric acid.

24. The stable nanomicellar ophthalmic formulation of claim 22 or claim 23, wherein the osmolality of the formulation is about 150 to about 200 mOsmol / kg.

25. The stable nanomicellar ophthalmic formulation of claim 24, wherein the osmolality of the formulation is about 160 to about 190 mOsmol / kg.

Images