Silica-Based Formulations of Therapeutic Oligopeptides and Peptidomimetics

Abstract

The present invention discloses a depot formulation comprising a biodegradable silica hydrogel composite, the hydrogel composite comprising: (i) a silica hydrogel; and (ii) a plurality of microparticles comprising silica and an oligopeptide or peptidomimetic compound or a pharmaceutically acceptable salt thereof. The depot formulation described herein provides a sustained long-term release of an active pharmaceutical compound. The depot formulation can be implanted in the eye and is useful for treating ocular diseases. The plurality of microparticles comprising silica and an oligopeptide or peptidomimetic compound or a pharmaceutically acceptable salt thereof can be prepared using a continuous flow process.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 171,723, filed April 7, 2021, and Finnish Patent Application No. 20215537, filed May 6, 2021; both of which are incorporated herein by reference. [Background technology]

[0002] Oligopeptide and peptidomimetic compounds are useful for treating various diseases. For example, oligopeptide and peptidomimetic compounds have been used to treat diabetes, skin diseases, heart diseases, and obesity. Oligopeptide and peptidomimetic compounds have also demonstrated utility in treating various ophthalmic indications, such as Leber's Hereditary Optic Neuropathy and Fuchs' Corneal Endothelial Dystrophy. However, the method of administering these compounds to the eye remains an unsolved problem. Thus, there is a need to develop formulations and methods for administering oligopeptide and peptidomimetic compounds for ophthalmic indications. Summary of the Invention [Means for solving the problem]

[0003] The present application provides a depot formulation comprising a biodegradable silica hydrogel composite, the hydrogel composite comprising: a silica hydrogel; and a plurality of microparticles comprising silica and an oligopeptide or peptidomimetic compound or a pharma- ceutically acceptable salt thereof; the oligopeptide or peptidomimetic compound has a free base equivalent (defined below) of less than 1,200 amu, the hydrogel composite being non-flowable and structurally stable during static storage and shear-thinning when subjected to shear stress. The depot formulation is, for example, a controlled release depot, such as a zero-order release depot, a first-order release depot, a second-order release depot, a delayed release depot, a sustained release depot, an immediate release depot, or any combination thereof.

[0004] In certain embodiments, a plurality of microparticles comprising silica and an oligopeptide or peptidomimetic compound, or a pharma- ceutically acceptable salt thereof, are formed using a continuous flow process.

[0005] In certain embodiments, the formulation is pre-packaged in a syringe and needle configuration for single-use intravitreal injection.

[0006] In certain embodiments, the formulation is sterilized, for example, by gamma irradiation.

[0007] In another aspect, the present application provides a method of treating, inhibiting, ameliorating or delaying the onset of a disease, disorder or condition, comprising administering to a subject in need thereof an effective amount of a depot formulation disclosed herein.

[0008] In certain embodiments, the condition is Leber's hereditary optic neuropathy. In another aspect, the present application provides a method for treating Fuchs' endothelial corneal dystrophy (FECD), age-related macular degeneration (AMD), glaucoma, optic neuropathy, retinopathy, diabetic macular edema, diabetic retinopathy, retinal telangiectasia, or retinitis pigmentosa, comprising administering to a subject in need thereof an effective amount of the depot formulation disclosed herein. Age-related macular degeneration (AMD) can be wet AMD or dry AMD, or a combination thereof.

[0009] In another aspect, the present application provides a prefilled syringe comprising the depot formulation disclosed herein. Optionally, the prefilled syringe is sterilized by exposure to gamma radiation.

[0010] The present application also relates to a method of making the depot formulation disclosed herein, comprising: a) silica particles comprising an oligopeptide or peptidomimetic compound or a pharma- ceutically acceptable salt thereof, optionally in suspension, having a maximum diameter of 1000 μm or less and a molecular weight of the oligopeptide or peptidomimetic compound free base equivalent of less than 1,200 amu; and b) Silica hydrogel

[0033] i) the silica hydrogel has a solids content of 50% by weight or less; ii) the silica particles comprise a polymerized alkoxysilane; iii) the depot formulation comprises up to 85% by weight of said silica particles; iv) The hydrogel composite is non-flowable and structurally stable upon static storage, and shear thinning when subjected to shear stress by injection.

[0011] The method of making the depot formulation may comprise one or more steps carried out in a continuous flow process.

[0012] In certain embodiments, the method further comprises sterilizing the depot formulation. [Brief description of the drawings]

[0013] [Figure 1] FIG. 1 is a graph showing the cumulative in vitro dissolution of elamipretide depot formulation #05D under sink conditions. [Diagram 2] 1 is a graph showing the cumulative in vitro dissolution of elamipretide depot formulation #08D under sink conditions. [Diagram 3] 1 is a graph showing the cumulative in vitro dissolution of Compound I (defined below) depot formulation #05D under sink conditions. [Figure 4] 1 is a graph showing the cumulative in vitro dissolution of Compound I (defined below) depot formulation #08D under sink conditions. [Diagram 5] FIG. 1 is a flow diagram for the manufacture of a pre-filled syringe-packaged depot formulation as disclosed herein, using a continuous flow process to produce a plurality of microparticles comprising silica and an oligopeptide or peptidomimetic compound or a pharma-ceutically acceptable salt thereof. [Figure 6] FIG. 1 is a diagram of the chemical polymerization of hydrolyzed silica. [Figure 7] FIG. 1 shows a schematic diagram of a continuous flow reactor system. [Figure 8] Comparison of cumulative in vitro dissolution of silica matrix (left) and release of Compound I (right) from microparticle formulations #01-#03 (2% API load, pH 5.9) with different first R values ​​in 50 mM TRIS buffer (pH 7.4, 37 °C) supplemented with 0.01% (v / v) TWEEN® 80 under sink conditions. Mean values ​​of triplicate analyses at each time point. [Figure 9]Comparison of cumulative in vitro dissolution of silica matrix (left) and release of Compound I (right) from microparticle formulations #03-#06 (R3-100, loaded with 2% API) with different pH in 50 mM TRIS buffer (pH 7.4, 37° C.) supplemented with 0.01% (v / v) TWEEN® 80 under sink conditions. Mean values ​​of triplicate analyses at each time point. [Figure 10] Comparison of cumulative in vitro dissolution of silica matrix (left) and release of Compound I (right) from microparticle formulations #03, #05, #07 and #08 (R3-100 based formulations) with different API loading and pH in 50 mM TRIS buffer (pH 7.4, 37° C.) supplemented with 0.01% (v / v) TWEEN® 80 under sink conditions. Mean values ​​of triplicate analyses at each time point. [Figure 11] Figure 1 shows the cumulative in vitro dissolution of silica matrix and release of Compound I from depot formulation #05D in 50 mM TRIS buffer (pH 7.4, 37°C) supplemented with 0.01% (v / v) TWEEN® 80 under sink conditions. The dissolution profile of the corresponding microparticle formulation #05 (R3-100, 2% API loading) is shown for comparison. Mean values ​​of triplicate analyses at each time point. [Figure 12] Figure 1 shows the cumulative in vitro dissolution of silica matrix and release of Compound I from depot formulation #08D in 50 mM TRIS buffer (pH 7.4, 37°C) supplemented with 0.01% (v / v) TWEEN® 80 under sink conditions. The dissolution profile of the corresponding microparticle formulation #08 (R3-100, 5% API loading) is shown for comparison. Mean values ​​of triplicate analyses at each time point. [Figure 13] Storage modulus (G') and tan δ (loss factor) values ​​for depot formulations #05D and #08D are shown. Measurements were performed under controlled deformation (γ<0.002) within the linear viscoelastic range of the material. Values ​​reported represent a single measurement. [Figure 14] Figure 1 shows dynamic viscosity as a function of shear rate for depot formulations #05D and #08D. Values ​​reported represent a single measurement. [Figure 15] Figure 1 shows a comparison of the cumulative in vitro dissolution and release of elamipretide from the silica matrix from microparticle formulations #01-#03 (2% API loading, pH 5.9) with different first R values ​​in 50 mM TRIS buffer (pH 7.4, 37 °C) containing 0.01% (v / v) TWEEN® 80 under sink conditions. Mean values ​​of triplicate analyses at each time point. [Figure 16] Figure 1 shows a comparison of the cumulative in vitro dissolution of silica matrix and release of elamipretide from microparticle formulations #03-#06 (R3-100, loaded with 2% API) with different pH in 50 mM TRIS buffer (pH 7.4, 37°C) supplemented with 0.01% (v / v) TWEEN® 80 under sink conditions. Mean values ​​of triplicate analyses at each time point. [Figure 17] Comparison of the cumulative in vitro dissolution of silica matrix (left) and release of elamipretide (right) from microparticle formulations #03, #05, #07 and #08 (R3-100 based formulations) with different API loading and pH in 50 mM TRIS buffer (pH 7.4, 37° C.) supplemented with 0.01% (v / v) TWEEN® 80 under sink conditions. Mean values ​​of triplicate analyses at each time point. [Figure 18] Figure 1 shows the cumulative in vitro dissolution of silica matrix and release of elamipretide from depot formulation #05D in 50 mM TRIS buffer (pH 7.4, 37°C) supplemented with 0.01% (v / v) TWEEN® 80 under sink conditions. The dissolution profile of the corresponding microparticle formulation #05 (R3-100, 2% API loading) is shown for comparison. Mean values ​​of triplicate analyses at each time point. [Figure 19] Figure 1 shows the cumulative in vitro dissolution of silica matrix and release of elamipretide from depot formulation #08D in 50 mM TRIS buffer (pH 7.4, 37°C) supplemented with 0.01% (v / v) TWEEN® 80 under sink conditions. The dissolution profile of the corresponding microparticle formulation #08 (R3-100, 5% API loaded) is shown for comparison. Mean values ​​of triplicate analyses at each time point. [Figure 20] Storage modulus (G') and tan δ (loss factor) values ​​for depot formulations #05D and #08D are shown. Measurements were performed under controlled deformation (γ<0.002) within the linear viscoelastic range of the material. Values ​​reported represent a single measurement. [Figure 21] Figure 1 shows dynamic viscosity as a function of shear rate for depot formulations #05D and #08D. Values ​​reported represent a single measurement. [Figure 22] 1 shows a graph of particle size distribution for consistency batches of formulations #09 and #10-5. [Diagram 23] 1 shows a graph of cumulative in vitro dissolution under sink conditions for consistency batches of elamipretide microparticle formulations #09 and #10-5. [Figure 24] 1 shows a graph of particle size distribution of continuous flow 10x batches of formulations #09 and #10-5. [Figure 25-1] 1 shows a graph of cumulative in vitro dissolution under sink conditions for continuous flow 10x batches of elamipretide microparticle formulations #09 and #10-5. [Figure 25-2] Continued from Figure 25-1. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0014] The present application relates to a depot formulation comprising a biodegradable silica hydrogel composite. The hydrogel composite comprises a silica hydrogel and a plurality of microparticles comprising silica and an oligopeptide or peptidomimetic compound or a pharma- ceutically acceptable salt thereof, wherein the free base equivalent of the oligopeptide or peptidomimetic compound has a molecular weight of less than 1,200 amu. The hydrogel composite is non-flowable and structurally stable when stored at rest, and shear-thinning when subjected to shear stress (e.g., force applied by injection through a syringe).

[0015] In some embodiments, the oligopeptide or peptidomimetic compound of the depot formulation is positively charged (eg, has a net charge of +2 or greater) in an aqueous buffer solution at neutral pH.

[0016] In some embodiments, the oligopeptide or peptidomimetic compound of the depot formulation exhibits good water solubility (eg, has a water solubility of >50 mg / mL, >75 mg / mL, >100 mg / mL, or >150 mg / mL).

[0017] In some embodiments, the plurality of microparticles of the hydrogel composite are present in various amounts. In some embodiments, the microparticles are present in the depot formulation in an amount of about 30 to about 95% by weight, or about 45 to about 90% by weight, or about 50 to about 85% by weight, or about 75 to 90% by weight. In some embodiments, the microparticles are present in the depot formulation in an amount of about 80% by weight. In some embodiments, the microparticles are present in the depot formulation in an amount of about 30 to about 70% by weight, or about 40 to about 60% by weight, or about 20 to about 40% by weight. In some embodiments, the microparticles are present in the depot formulation in an amount of about 50% by weight. In some embodiments, the microparticles are present in the depot formulation in an amount of about 20 to about 50% by weight, or about 20 to about 40% by weight.

[0018] The depot formulation may include an oligopeptide, which may be present in a range of amounts. In some embodiments, the free base equivalent of the oligopeptide in the depot formulation is present in an amount of about 0.1% to about 10% by weight, or about 0.3% to about 5% by weight, or about 1% to about 3% by weight, or about 2% to about 4% by weight. In some embodiments, the free base equivalent of the oligopeptide in the depot formulation is present in an amount of about 2% to about 5% by weight, or about 0.25% to about 2.5% by weight, or 0.45% to 2% by weight.

[0019] The depot formulation may include a peptidomimetic compound, which may be present in a range of amounts. In some embodiments, the free base equivalent of the peptidomimetic compound in the depot formulation is present in an amount of about 0.1% to about 10% by weight, or about 0.3% to about 5% by weight, or about 1% to about 3% by weight, or about 2% to about 4% by weight. In some embodiments, the free base equivalent of the peptidomimetic compound in the depot formulation is present in an amount of about 2% to about 5% by weight, or about 0.25% to about 2.5% by weight, or about 0.45% to 2% by weight.

[0020] In certain embodiments, the depot formulation is shear thinning when subjected to shear stress and may be injectable through a small needle (e.g., an 18-30 gauge needle, a 20-30 gauge needle, a 25-30 gauge needle, or a 27-30 gauge needle). In certain embodiments, the silica hydrogel composite is free flowing when subjected to shear stress and may be injected through a syringe with an attached needle. In certain embodiments, the needle is 25-30 gauge. In some embodiments, the needle is up to 25 gauge. In some embodiments, the needle is up to 30 gauge. In some embodiments, the needle is up to 35 gauge. In some embodiments, the needle is up to 40 gauge. In some embodiments, the needle is up to 50 gauge.

[0021] After injection (ie, at the end of application of shear stress), the silica hydrogel composite is non-flowable and structurally stable at 37°C.

[0022] In some embodiments, the depot formulations disclosed herein can include a range of pH values, hi some embodiments, the depot formulations have a pH of about 5.5 to about 7.5, or about 5.5 to about 7.0, or about 5.5 to about 6.5.

[0023] In some embodiments, the microparticle of the depot formulation can comprise a plurality of oligopeptide or peptidomimetic compounds, hi some embodiments, the oligopeptide or peptidomimetic compound of the microparticle is targeted to mitochondria.

[0024] In some embodiments, the microparticle comprises an oligopeptide or a pharmaceutically acceptable salt thereof.In some embodiments, the oligopeptide is selected from the group consisting of elamipretide, compound II, compound III, and compound IV, and pharmaceutically acceptable salts thereof.The structures of elamipretide, compound II, compound III, and compound IV (in the form of the free base equivalent) are shown below: [ka]

[0025] In certain embodiments, the oligopeptide is elamipretide or a pharma- ceutically acceptable salt thereof.

[0026] In certain embodiments, the oligopeptide is compound II or a pharma- ceutically acceptable salt thereof.

[0027] In certain embodiments, the oligopeptide is compound III or a pharma- ceutically acceptable salt thereof.

[0028] In certain embodiments, the oligopeptide is compound IV or a pharma- ceutically acceptable salt thereof.

[0029] In some embodiments, the microparticles of the depot formulation can include a peptidomimetic compound or a pharma- ceutically acceptable salt thereof. Compound I is an example of one such peptidomimetic compound (or a pharma- ceutically acceptable salt thereof) that can be used in the depot formulations disclosed herein: [ka]

[0030] The depot formulations disclosed herein may be formulated to be suitable for injection (or infusion), implantation, inhalation, or other means of parenteral administration. In some aspects, the depot formulations may be formulated to be suitable for subcutaneous, intramuscular, peritoneal, or ocular administration. In some embodiments, the depot formulations are formulated for intravitreal injection or local ocular delivery of the oligopeptide or peptidomimetic compound. In certain embodiments, the formulation is for intravitreal injection. In some embodiments, the depot formulations are formulated for intravenous or subcutaneous administration for systemic delivery of the oligopeptide or peptidomimetic compound.

[0031] In some embodiments, the depot formulation is for administration once a week to once a year, or once a month to once a year, or once a month to once every six months, or once a week to once a month, or once a month to once every three months, once every three months to once every six months, once every six months to once every nine months, or once every nine months to once a year. Alternatively, the depot formulation may be for administration once a month to once every six months, or once a month to once every five months, once every two months to once every five months, once every two months to once every four months, or once every two months to once every three months, once every three months to once every four months, or about once every three months.

[0032] Depot formulations that release drug over time are called "controlled release" depot formulations. Such depot formulations formulated for controlled release can help patient compliance, since some patients find it easier to inject the drug once over a long period of time than to take it daily. In further embodiments, controlled release depot formulations can be better tolerated by patients, for example, compared to daily injections (e.g., daily subcutaneous injections), since depot formulations are associated with less frequent injection site reactions.

[0033] In some embodiments, the formulation delivers a daily dose of 0.01 μg to 10 μg, 0.1 μg to 5 μg, 1 μg to 5 μg, or about 3 μg of the oligopeptide or peptidomimetic compound, where the recited amounts are calculated based on the mass of the free base equivalent of the oligopeptide or peptidomimetic compound (regardless of whether the free base or a salt form of the oligopeptide or peptidomimetic compound is used to prepare the formulation).

[0034] In some embodiments, the amount of the free base equivalent of the oligopeptide or peptidomimetic compound in the depot formulation is between 10 μg / 50 μL and 100 mg / 50 μL, between 10 μg / 50 μL and 1000 μg / 50 μL, between 10 μg / 50 μL and 500 μg / 50 μL, between 50 μg / 50 μL and 300 μg / 50 μL, or between 75 μg / 50 μL and 275 μg / 50 μL, wherein the recited amounts are calculated based on the mass of the free base equivalent of the oligopeptide or peptidomimetic compound. Alternatively, the amount of free base equivalent of the oligopeptide or peptidomimetic compound in the depot formulation is between 100 μg / 50 μL and 500 μg / 50 μL, between 150 μg / 50 μL and 450 μg / 50 μL, between 200 μg / 50 μL and 400 μg / 50 μL, between 250 μg / 50 μL and 350 μg / 50 μL, or about 270 μg / 50 μL.

[0035] In certain embodiments, the depot formulation is for intravitreal injection about every three months.

[0036] In certain embodiments, the depot formulation is sterilized, for example by gamma sterilization.

[0037] The pharmaceutical depot of the present application can include silica hydrogels having a range of solids contents. In some embodiments, the silica hydrogels have a solids content of 3% or less, 2% or less, or 1% or less by weight. In some embodiments, the silica particles comprise 0.1-70%, 0.3-50%, or 1-15% by weight of the oligopeptide or peptidomimetic compound.

[0038] In some embodiments of the depot formulation, the plurality of microparticles comprises silica particles having a range of diameters. In some embodiments, the silica particles have a diameter of 0.5 μm to 300 μm. In some embodiments, the silica particles have a diameter of 0.5 μm to 100 μm, 0.5 μm to 30 μm, or 0.5 μm to 20 μm.

[0039] The depot formulations of the present application may have a volume fraction of silica particles with a range of diameters. In some embodiments, the volume fraction of silica particles with a diameter of less than 1 μm is less than 3%, less than 2%, or less than 1%.

[0040] In some embodiments, the depot formulation may comprise 10% to 75%, 15% to 60%, or 25% to 55% composite solids by weight.

[0041] In some embodiments, the depot formulation comprises a range of complex modulus measured under small angle oscillatory shear in the linear viscoelastic region, hi some embodiments, the complex modulus is less than 2400 kPa, less than 1200 kPa, or less than 600 kPa.

[0042] In some embodiments, the formulation depot comprises a loss modulus (ie, viscous modulus / elastic modulus) value of less than 1, less than 0.8, or less than 0.6.

[0043] In some embodiments, the depot formulation is -1 Viscosity of about 10-50 Pas measured at a shear rate of 200-210 s -1 Viscosity of approximately 0.4 to 1.5 Pas measured at a shear rate of 600 to 610 s -1 It has a viscosity of about 0.1 to 0.4 Pas measured at a shear rate of 100 .mu.m.

[0044] In certain embodiments, the formulation is pre-packaged in a syringe and needle configuration for single-use intravitreal injection.

[0045] The present application also provides a pre-filled syringe comprising the depot formulation described herein. In some embodiments, the pre-filled syringe is sterilized by exposure to gamma radiation.

[0046] Microparticles can be prepared by many different methods. Microparticles (often, but not necessarily, spherical particles, microspheres) are small particles with diameters typically in the micrometer range (typically 1 μm to 1000 μm). There are several direct techniques that are usually used to prepare or manufacture microparticles with a controlled particle size distribution, such as spray drying (often followed by centrifugation of the microparticles in a cyclone), single emulsion, double emulsion, polymerization, coacervation phase separation and solvent extraction methods. By using these techniques, a small volume fraction of submicron particles (usually 0.5 to 1.0 μm) can be included in the resulting product, but their proportion is usually very low, often less than 1% by volume. Thus, their impact in, for example, controlled release microparticle formulations is small. It is also possible to prepare microparticles by casting or by crushing larger structures into microparticles, but in that case the size and morphology of the resulting particles may be diverse and additional preparation steps such as tumbling and particle sizing may be required.

[0047] In certain embodiments, a plurality of microparticles comprising silica and an oligopeptide or peptidomimetic compound, or a pharma- ceutically acceptable salt thereof, are formed using a continuous flow process.

[0048] In certain embodiments, the continuous flow process further comprises the use of a spray dryer to create a plurality of microparticles comprising silica and the oligopeptide or peptidomimetic compound or a pharma- ceutically acceptable salt thereof. In further embodiments, the continuous flow process further comprises the use of an in-line mixer.

[0049] The silica microparticles of the silica-silica composite play a role in the controlled release of the drug, while the hydrogel structure ensures both the stability and injectability of the resulting composite. Although stable at rest, e.g., when stored in a prefilled syringe, the hydrogel composite structure is so loose that it is shear thinning when the hydrogel composite is injected from a ready-to-use syringe through a fine needle (e.g., 20-30 gauge, such as 20 gauge, 24 gauge, 25 gauge, 28 gauge or 30 gauge). This combination of properties provides a minimally invasive, long-term treatment of, e.g., ocular diseases.

[0050] The present application further provides a method for treating, inhibiting, ameliorating, or delaying the onset of other diseases, disorders, or conditions, such as Fuchs endothelial corneal dystrophy (FECD), (wet or dry) age-related macular degeneration (AMD), glaucoma, optic neuropathy, retinopathy, diabetic macular edema, diabetic retinopathy, retinal telangiectasia, or retinitis pigmentosa. The method comprises administering to a subject in need thereof an effective amount of a depot formulation as described herein. In some embodiments, the AMD is AMD with neovascularization (wet AMD) or AMD with retinal geographic atrophy (dry AMD).

[0051] The present application further provides a method of treating, inhibiting, ameliorating or delaying the onset of a disease, comprising administering to a subject in need thereof an effective amount of a depot formulation described herein. The disease to be treated can be Leber's hereditary optic neuropathy.

[0052] In some embodiments, the disease may be associated with mitochondrial dysfunction.

[0053] The present application further provides a method of making the pharmaceutical depot described herein, comprising combining silica particles comprising an oligopeptide or peptidomimetic compound or a pharma- ceutically acceptable salt thereof, optionally as a suspension, with a silica hydrogel, the silica particles having a maximum diameter of 1000 μm or less and a molecular weight of the oligopeptide or peptidomimetic compound free base equivalent of less than 1,200 amu. In some embodiments, the silica hydrogel has a solids content of 50% or less by weight; the silica particles comprise a polymerized alkoxysilane; the pharmaceutical depot comprises up to 85% by weight of said silica particles; and the hydrogel composite is non-flowable and structurally stable during static storage and shear thinning when subjected to shear stress by injection.

[0054] In some embodiments, the method further comprises combining the silica sol with a solution comprising an oligopeptide or peptidomimetic compound or a pharmaceutical salt thereof, thereby producing a mixture comprising the silica sol and an oligopeptide or peptidomimetic compound or a pharma- ceutically acceptable salt thereof. The resulting mixture can be spray dried to form silica particles, which are finally combined with the silica hydrogel (see, e.g., FIG. 5).

[0055] In certain such embodiments, the method further comprises combining tetraethyl orthosilicate (TEOS), water and hydrochloric acid, and optionally an organic co-solvent (e.g., ethanol), thereby creating a silica sol (see, e.g., Figures 5 and 6).

[0056] In further such embodiments, the method includes combining the oligopeptide or peptidomimetic compound, or a pharmaceutical salt thereof, with water and optionally an organic co-solvent (e.g., ethanol), thereby creating a solution comprising the oligopeptide or peptidomimetic compound, or a pharmaceutical salt thereof (e.g., an API solution; see, e.g., FIG. 7).

[0057] In certain embodiments, the step of combining the silica sol with the solution containing the oligopeptide or peptidomimetic compound or a pharmaceutical salt thereof is carried out in a continuous flow process, for example in a continuous flow reactor. Such a process may include the use of an in-line mixer and / or a pump (see, for example, Figures 5-7).

[0058] Continuous flow process is repeatable and allows for scale-up.Thus, in some embodiments of the method for making depot formulation, the formulation is made at 10x scale, 50x scale, 150x scale, 250x scale, 500x scale, 750x scale, 1000x scale, 1500x scale, 2000x scale or more.Compared to the manufacturing method using single batch reactor, continuous flow process can provide more consistent particle size (and size distribution) and API content.

[0059] In certain embodiments, the formulation is packaged in a syringe, optionally equipped with a needle.

[0060] The method of making the depot formulation may further comprise sterilizing the depot formulation, for example by radiation sterilization. Radiation sterilization may comprise gamma radiation sterilization.

[0061] In certain embodiments, the oligopeptide is elamipretide or a pharma- ceutically acceptable salt thereof.

[0062] In certain embodiments, the oligopeptide is compound II or a pharma- ceutically acceptable salt thereof.

[0063] In certain embodiments, the oligopeptide is compound III or a pharma- ceutically acceptable salt thereof.

[0064] In certain embodiments, the oligopeptide is compound IV or a pharma- ceutically acceptable salt thereof.

[0065] In certain embodiments, the peptidomimetic is Compound I or a pharma- ceutically acceptable salt thereof.

[0066] definition As used herein, "ameliorating" or "ameliorating" a disease, disorder or condition (e.g., a tauopathy) refers to the result of making the occurrence of the disease, disorder or condition (or a sign, symptom or condition thereof) better or more tolerable in a statistical sample or particular subject, in a sample or subject administered a therapeutic agent, as compared to a control sample or subject.

[0067] The term biodegradation refers to erosion, i.e. the gradual degradation of the matrix material (e.g. silica) within the body, preferably by dissolution in body fluids.

[0068] As used herein, the phrase "delay" or "delay the onset" means to postpone or prevent the onset of a disease, disorder or condition, or to cause one or more signs, symptoms or symptoms of a disease, disorder or condition to occur more slowly than normal in a statistical sample, in a sample or subject administered a therapeutic agent, as compared to a control sample or subject.

[0069] A depot formulation as referred to in this application is defined as the administration of a controlled release drug (active agent) formulation that allows for sustained release and slow absorption by the subject so that the active agent can act and be released in the subject's body over an extended period of time (i.e., days to months). Depot formulations are administered parenterally, either by implantation or injection into the subcutaneous, intramuscular, peritoneal or ocular.

[0070] A free base equivalent should be understood to refer to a compound (e.g., an oligopeptide or peptidomimetic) in its simplest, lowest molecular weight form, e.g., without a salt, or other atoms, molecules, or agents that may typically be associated or complexed with the compound. For example, elamipretide and compounds I-IV typically contain three basic nitrogen atoms that form complexes with negative ions when the nitrogens are protonated (and thus become positively charged). Free base equivalents of elamipretide and compounds I-IV are exemplified above.

[0071] A gel should be understood in the context of this application as a homogeneous mixture of at least one solid and one liquid phase, i.e. a colloidal dispersion, in which the solid phase (e.g. silica itself and / or partially or completely hydrolyzed) is the continuous phase and the liquid (e.g. water, ethanol and residues of the silica precursor) is homogeneously dispersed in the structure. Gels are viscoelastic, with elastic properties dominating, as shown by rheological measurements under small angle oscillatory shear. When the loss factor (or loss tangent) tan δ = (G" / G') is less than 1, the elastic properties dominate and the structure is non-flowable. The combined effect of the elastic modulus G' and the viscous modulus G" is expressed as a complex modulus (or complex shear modulus) of the form, G * It can also be expressed as =G'+G".

[0072] The gel point should be understood to mean the point at which the fluid sol turns into a gel, which is viscoelastic, and the elastic properties become dominant, which is indicated by rheological measurements under small angle oscillatory shear, where the elastic modulus G' is greater than the viscous modulus and the loss factor is less than 1. Viscoelastic properties are usually measured by oscillatory shear with small shear stresses (small deformation angles) using a rheometer (a measuring device for determining the correlation between deformation, shear stress and time). The total resistance in small oscillatory shear is determined by the complex modulus (G * The complex modulus (or complex shear modulus) contains two components (G * = G' + iG"): 1) The elastic modulus, also called the storage modulus G', describes that the material has some elastic properties characteristic of solid materials, i.e., gel systems gain energy from oscillatory motions, as long as the motions do not destroy the gel structure. This energy is stored in the sample and is described by the elastic modulus; 2) The viscous modulus G", also called the loss modulus, describes the flow properties, i.e., in a system, for example a silica sol, that gives rise to motions between the components of the sol, which accounts for the part of the energy lost as viscous dissipation in oscillatory shear. G * If = G', the material is called elastic, and G *= G”, the material is called viscous. At the gel point, the elastic modulus G' becomes greater than the viscous modulus G”. If G'>G”, the viscoelastic material is called semi-solid and correspondingly if G”>G', the viscoelastic material is called semi-liquid. The magnitude of the elastic and viscous moduli depends on the shear stress, which in turn depends on the applied strain (small angle deformation) and frequency (of oscillatory shear). Measurements are made by ensuring an appropriate signal for the particular measurement system, i.e. a strain sweep is usually made at a constant frequency to find the appropriate signal for the rheometer system and the linear viscoelastic region, and then the actual measurements are made at constant strain with varying frequency. The varying frequency gives varying elastic and viscous moduli and the measurement indicates whether the solid or liquid phase is dominant. It is also typical that the modulus increases rapidly after the gel point if the ambient conditions are not significantly changed; for example, for gels formed from acidic sols near room temperature, e.g., R15 sol (R = molar ratio of water to alkoxide) at pH = 2 that gels, it is typical for G' to increase 100-700 times within minutes of the gel point. For larger R values, such as R150 and R400, the modulus G' remains at a low level even after the gel point, and the increase in G' is not as fast, which allows to have a gel structure that remains injectable with a fine needle. In the form of gels after a defined gel point, the solid state predominates, but the system still contains a variable amount of liquid, and the material is typically soft and viscoelastic before drying and becomes hard and brittle when sufficiently dried. In the form of sols, the liquid state predominates, but the system still contains a variable amount of solid phase, and the system is still fluid. Prior to the gel point, a sharp increase in dynamic viscosity and modulus is typically observed, which continues to increase after the gel point as structure develops. In the context of this application, the gel point of the composite is reached before an injectable gel is obtained.

[0073] A hydrogel should be understood to be a gel in which the liquid phase is water or an aqueous system containing more than 50 weight percent (wt%) water. Typically, the liquid phase of a hydrogel contains more than 65 wt%, more typically more than 90 wt%, and most typically more than 95 wt% water. The liquid phase can further contain other liquids, typically organic solvents (e.g., ethanol). Typically, the concentration of such solvents, e.g., ethanol, is less than 10 wt%, more typically less than 3 wt%, and most typically less than 1 wt%. In the context of this application, the composites disclosed herein are considered hydrogels since they meet the basic criteria of a hydrogel. Thus, when referring to a hydrogel composite disclosed herein, this reference is equivalent to a reference to a composite.

[0074] As used herein, "inhibit" or "inhibiting" refers to an objectively measurable amount or degree of reduction in a sign, symptom or condition (e.g., risk factor) associated with a tauopathy-related disease, disorder or condition compared to a control. In one embodiment, inhibition or inhibiting refers to at least a statistically significant amount of reduction compared to a control (or control subject). In one embodiment, inhibition or inhibiting refers to at least a 5 percent reduction compared to a control (or control subject). In various individual embodiments, inhibition or inhibiting refers to at least a 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 33, 40, 50, 60, 67, 70, 75, 80, 90, 95, or 99 percent reduction compared to a control (or control subject).

[0075] Injectable, in the context of this application, means that it can be administered through a surgical administration device, such as a needle, a catheter, or a combination thereof.Shear thinning, in the context of this application, is a rheological property of a composition.Whenever the shear stress or shear rate of such a composition changes, the composition gradually moves toward its new equilibrium state, and at lower shear rates, the shear thinning composition becomes more viscous, and at higher shear rates, it becomes less viscous.Thus, shear thinning refers to the effect that the viscosity of a fluid, i.e., a measure of the resistance of a fluid to flow, decreases with increasing shear rate.

[0076] An injectable gel or hydrogel in the context of this application is a rheological property of the composition. When stored in a syringe and / or in aluminum foil prior to injection, e.g. at a temperature below 37°C, e.g. at room temperature (20-25°C) or at refrigerator temperature (3-6°C), the composition is a gel, i.e. the elastic modulus (measured under small angle oscillatory shear) G' is greater than the viscous modulus G" and the loss factor tan δ = (G" / G') is less than 1. Hydrogel composite structures are gel-like structures, the composite structure remains stable and non-flowable upon static storage, but the gel structure is very loose and shear thinning when shear stress is applied, e.g. in the form of injection through a syringe, e.g. with a 25G needle (0.50 mm x 25 mm).

[0077] Non-flowable and structurally stable at rest storage refers to a stable composite hydrogel structure composed of silica particles in a silica hydrogel. Stability is indicated by rheological measurements under small-angle oscillatory shear with an elastic modulus G' greater than the viscous modulus and a loss factor less than 1. If the elastic modulus is greater than the viscous modulus and the loss factor is less than 1, the structure is non-flowable. The non-flowable structure ensures the stability of the composite hydrogel structure by preventing phase separation of the silica particles. In other words, the silica particles are embedded in the silica hydrogel, and they do not settle or separate to the bottom of a container, e.g., a syringe, when the hydrogel composite is stored at temperatures typically below 25°C. Although the composite hydrogel structure is non-flowable at rest, e.g., when stored in a prefilled, ready-to-use syringe, the structure is very loose and shear thinning when shear stress is applied to the hydrogel composite by injection, and thus injectable through a thin needle.

[0078] As used herein, the term "pharmaceutical acceptable carrier" refers to a diluent, adjuvant, excipient, or vehicle with which a compound / medicine / composition (including a medicament) is administered or formulated for administration. Non-limiting examples of such pharmaceutical acceptable carriers include liquids such as water, saline, oil, and solids such as gum arabic, gelatin, starch paste, talc, keratin, colloidal silica, silica particles (nanoparticles or microparticles) urea. In addition, auxiliary agents, stabilizers, thickeners, lubricants, flavoring agents, and coloring agents may be used. Other examples of suitable pharmaceutical carriers are described in Remington's Pharmaceutical Sciences by EW Martin, which is incorporated herein by reference in its entirety.

[0079] As used herein, the term "pharmaceutical acceptable salts" refers to salts of therapeutically active compounds that can be prepared using relatively non-toxic acids or bases, depending on the specific substituents found on the compounds described herein. When a compound contains a relatively acidic functional group, a base addition salt can be obtained by contacting a neutral form of such a compound with a sufficient amount of the desired base, neat or in a suitable inert solvent. Examples of pharmaceutical acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino or magnesium salts, or similar salts. When a compound contains a relatively basic functional group, an acid addition salt can be obtained by contacting a neutral form of such a compound with a sufficient amount of the desired acid, neat or in a suitable inert solvent. Salts derived from pharmaceutical acceptable inorganic bases include ammonium, calcium, copper, ferric, ferrous, lithium, magnesium, manganous, manganous, potassium, sodium, and zinc salts, and the like. Salts derived from pharma- ceutically acceptable organic bases include, for example, arginine, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-methylmorpholine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethylamine (NEt 3 ), trimethylamine, tripropylamine, tromethamine, and the like, including substituted amines, cyclic amines, naturally occurring amines, and the like, for example, salts of protonated forms of organic bases (e.g., [HNEt 3 ] +). Salts derived from pharma- ceutically acceptable inorganic acids include salts of boric acid, carbonic acid, hydrohalic acids (hydrobromic acid, hydrochloric acid, hydrofluoric acid, or hydroiodic acid), nitric acid, phosphoric acid, sulfamic acid, and sulfuric acid. Salts derived from pharma-ceutically acceptable organic acids include salts of aliphatic hydroxyl acids (e.g., citric acid, gluconic acid, glycolic acid, lactic acid, lactobionic acid, malic acid, and tartaric acid), aliphatic monocarboxylic acids (e.g., acetic acid, butyric acid, formic acid, propionic acid, and trifluoroacetic acid), amino acids (e.g., aspartic acid and glutamic acid), aromatic carboxylic acids (e.g., benzoic acid, p-chlorobenzoic acid, diphenylacetic acid, gentisic acid, hippuric acid, and triphenylacetic ... and aromatic carboxylic acids (e.g., benzoic acid, p-chlorobenzoic acid, diphenylacetic acid, gentisic acid, hippuric acid, and triphenylacetic acid). Examples of the salts of aromatic hydroxyl acids (e.g., o-hydroxybenzoic acid, p-hydroxybenzoic acid, l-hydroxynaphthalene-2-carboxylic acid, and 3-hydroxynaphthalene-2-carboxylic acid), ascorbic acid, dicarboxylic acids (e.g., fumaric acid, maleic acid, oxalic acid, and succinic acid), glucuronic acid, mandelic acid, mucic acid, nicotinic acid, orotic acid, pamoic acid, pantothenic acid, sulfonic acids (e.g., benzenesulfonic acid, camphorsulfonic acid, edisylic acid, ethanesulfonic acid, isothionic acid, methanesulfonic acid, naphthalenesulfonic acid, naphthalene-1,5-disulfonic acid, naphthalene-2,6-disulfonic acid, p-toluenesulfonic acid (PTSA)), xinafoic acid, and the like. In some embodiments, the pharma- ceutically acceptable counterion is selected from the group consisting of acetate, benzoate, besylate, bromide, camphorsulfonate, chloride, chlorotheophyllinate, citrate, ethanedisulfonate, fumarate, gluceptate, gluconate, glucuronic acid, hippurate, iodide, isethionate, lactate, lactobionic acid, lauryl sulfate, malate, maleate, mesylate, methylsulfate, naphthoate, sapsylate, nitrate, octadecanoate, oleate, oxalate, pamoate, phosphate, polygalacturonic acid, succinate, sulfate, sulfosalicylate, tartaric acid, tosylate, and trifluoroacetate.In some embodiments, the salt is a tartrate, fumarate, citrate, benzoate, succinate, suberate, lactate, oxalate, phthalate, methanesulfonate, benzenesulfonate, maleate, trifluoroacetate, hydrochloride, or tosylate. Also included are salts of amino acids such as arginate, and salts of organic acids such as glucuronic acid or galacturonic acid (see, for example, Berge et al, Journal of Pharmaceutical Science 66: 1-19 (1977)). Certain compounds of the present application may contain both basic and acidic functionalities, which allows the compounds to be converted into either base or acid addition salts or to exist in zwitterionic form. These salts can be prepared by methods known to those skilled in the art. Other pharma-ceutically acceptable carriers known to those skilled in the art are also suitable for the present technology.

[0080] The term silica refers to amorphous SiO that can be prepared by the sol-gel method. 2Sol-gel derived silica refers to silica prepared by the sol-gel method, where the silica is prepared from liquid-phase precursors such as alkoxides, alkylalkoxides, aminoalkoxides, or inorganic silicate solutions, which form a sol by hydrolysis and condensation reactions that convert to a gel or form a stable sol. The liquid in the stable silica sol can be evaporated, resulting in the formation of a powder, typically consisting of colloidal silica particles. The resulting gel / particles can be optionally aged, dried, and heat treated, preferably at a temperature below 700°C, if heat treated. Sol-gel derived silica prepared below 700°C is usually amorphous. If the gel contains biologically active agents, such as drugs and active pharmaceutical ingredients (e.g., oligopeptides or peptidomimetics as described herein), the heat treatment is typically skipped. The formed gel is then typically only aged (typically below 40°C) and dried (typically below 40°C). The sol can be gelled in a mold for shaping. Sol-gel derived silica can also be processed and prepared into different morphologies by simultaneous gelation, aging, drying and shaping, e.g., by spray drying into microparticles, by dip / drain / spin coating into films, by extrusion into monolithic structures, or by spinning into fibers.

[0081] The term silica sol refers to a suspension, i.e., a mixture of a liquid (continuous phase) and a solid phase (dispersed phase), where the solid phase consists of silica particles and / or aggregated silica particles, the particle size of which is typically less than 1 μm, i.e., the silica particles and / or particle aggregates are colloidal. Silica sols are usually prepared from alkoxides or inorganic silicates, which, via hydrolysis, form either partially hydrolyzed silica species or fully hydrolyzed silicic acid. The liquid phase is typically composed of water, as well as hydrolyzates and condensation products such as ethanol. Subsequent condensation reactions of the SiOH-containing species result in the formation of larger silica species with increasing amounts of siloxane bonds. These species form nano-sized colloidal particles and / or particle aggregates. Depending on the conditions, the silica sol either remains as a stable colloidal suspension or becomes a gel.

[0082] A sol should be understood as a homogeneous mixture of at least one liquid phase and one solid phase, i.e. a colloidal dispersion, in which the liquid phase (e.g. water, ethanol, and residues of silica precursor) is the continuous phase and the solid phase (e.g. colloidal particles of silica and / or partially or completely hydrolyzed silica and / or aggregates of said particles) is homogeneously dispersed in said liquid phase, the sol being characterized by having clear flow properties and being dominated by the liquid phase. A suspension can also be specifically called a sol when the solid particles are colloidal and have a diameter smaller than 1 μm. However, in the context of this application, the term sol refers to a colloidal dispersion in which the solid particles are <50 nm, and the term suspension refers to a dispersion in which the solid particles are >50 nm.

[0083] As used herein, the terms "subject," "individual," or "patient" may be an individual organism, vertebrate, mammal, or human. Subjects include, but are not limited to, rats, mice, chickens, cows, monkeys, rabbits, primates, etc., prior to testing in human subjects. The subject may be a human.

[0084] As used herein, the term "treat" or "treatment" refers to therapeutic treatment, where the purpose is to alleviate, alleviate, or slow down (reduce) an existing disease, disorder, or condition, or its associated signs, symptoms, or conditions. By way of example and not limitation, a subject is successfully "treated" for a disease if, after administration of an effective amount of a compound / composition / pharmaceutical product or a pharmaceutically acceptable salt thereof, the subject exhibits an observable and / or measurable reduction or absence of one or more signs, symptoms, or conditions associated with the disease, disorder, or condition.

[0085] Pharmaceutical Compositions In certain embodiments, the present application is directed to a pharmaceutical composition. In certain embodiments, the pharmaceutical composition comprises a depot formulation disclosed herein. In some embodiments, the depot formulation further comprises an additional therapeutic compound (i.e., a drug) and a pharma- ceutically acceptable carrier. The at least one additional therapeutic agent may refer to an agent useful for treating a disease such as Leber's hereditary optic neuropathy. The pharmaceutical composition may be a medicine.

[0086] Pharmaceutical compositions can be prepared by combining a compound, eg, an oligopeptide or peptidomimetic compound, with a pharma- ceutically acceptable carrier and optionally one or more additional therapeutic agents.

[0087] "Effective amount" refers to any amount sufficient to achieve a desired biological effect (e.g., an amount that treats, inhibits, ameliorates, or delays the onset of a disease, disorder, or condition, or physiological signs, symptoms, or conditions of a disease or disorder). By selecting from among various active compounds, in combination with the teachings provided herein, and weighing factors such as potency, relative bioavailability, subject body weight, severity of adverse side effects, and mode of administration, one can design an effective prophylactic (i.e., preventative) or therapeutic treatment regimen that is effective in treating a particular subject's condition, disorder, or disease without causing substantial undesirable toxicity. The effective amount for any particular indication may vary depending on factors such as the disease, disorder, or condition being treated, the particular compound of the present application being administered, the size of the subject, or the severity of the disease, disorder, or condition. Effective amounts can be determined by methods familiar to physicians and clinicians during preclinical and clinical trials. Those of ordinary skill in the art can empirically determine the effective amounts of particular compounds of the present application and / or other therapeutic agents without necessitating undue experimentation. A maximum dose, i.e., the maximum safe dose according to some medical judgment, may be used. Appropriate systemic levels may be determined, for example, by measuring peak or sustained plasma levels in a subject of the drug. A locally delivered dose may be determined, for example, by levels in a particular compartment of the body, such as the vitreous humor of the eye. "Dose" and "dosage" are used interchangeably herein. A dose may be administered by oneself, by another, or by a device (e.g., a pump or syringe). A compound / composition / medicine may be administered in combination with one or more additional therapeutic compounds / agents (e.g., so-called "co-administration" in which the additional therapeutic agents may be administered simultaneously, sequentially, or by separate administration).

[0088] Compounds for use in treatment or prevention can be tested in suitable animal model systems.Similarly, for in vivo testing, any animal model system known in the art can be used before administration to human subjects.Suitable animal model systems include, but are not limited to, rats, mice, chickens, cows, monkeys, rabbits, etc., before testing in human subjects.

[0089] The therapeutic compound and optionally other therapeutic agents can be administered in free base equivalent form or in the form of pharmaceutically acceptable salt.When used in medicine, salt should be pharmaceutically acceptable, but pharmaceutically unacceptable salts can be conveniently used to prepare pharmaceutically acceptable salts.Pharmaceutically acceptable salt forms can include forms in which the ratio of salt-containing molecules is not 1:1.For example, a salt can include more than one inorganic or organic acid molecule per molecule of base, such as two hydrochloric acid molecules per molecule of compound.As another example, a salt can include less than one inorganic or organic acid molecule per molecule of base, such as two compound molecules per molecule of tartaric acid.

[0090] The pharmaceutical compositions of the present application contain an effective amount of a therapeutic compound described herein (e.g., elamipretide, Compound I, Compound II, Compound III or Compound IV), which can be optionally distributed in a pharma- ceutically acceptable carrier. The components of the pharmaceutical compositions can also be commingled with the compounds of the present application, and with each other, in a manner such that there is no interaction which would substantially impair the desired pharmaceutical efficiency.

[0091] The dosage, toxicity and therapeutic efficacy of any therapeutic compound, composition (e.g., formulation or pharmaceutical), other therapeutic agent, or mixture thereof can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, for example, to determine the LD50 (the dose lethal to 50% of the population) and the ED50 (the dose therapeutically effective in 50% of the population). The dose ratio between the toxic effect and the therapeutic effect is the therapeutic index, which can be expressed as the ratio LD50 / ED50. Compounds that exhibit high therapeutic indices are advantageous.

[0092] Data obtained from cell culture assays and animal studies can be used in formulating a range of dosages for use in humans. The dosage of such compounds can be within a range of circulating concentrations that include the ED50 with little or no toxicity. Dosages can vary within this range depending on the dosage form used and the route of administration utilized. Doses can be formulated in animal models to achieve a circulating plasma concentration range that includes the IC50 (i.e., the concentration of the test compound that achieves half (50%) inhibition of symptoms). Such information can be used to accurately determine useful doses in humans. Levels of active pharmaceutical ingredients / therapeutic compounds (e.g., oligopeptides or peptidomimetics) in the plasma of animals or humans can be measured, for example, by high performance liquid chromatography.

[0093] Exemplary treatment regimes may involve administration once a week to once a month, or once a month to once a year, or once a month to once every six months, or once a month to once every three months, once every three months to once every six months, once every six months to once every nine months, or once every nine months to once a year. In some embodiments, the treatment regimen is an intravitreal injection of the depot formulation about once every three months, once every six months, once every nine months, or about once a year. In therapeutic applications, relatively high dosages at relatively short intervals are sometimes required until the progression of the disease, disorder, or condition is alleviated or terminated, or until the subject shows partial or complete improvement of the symptoms of the disease, disorder, or condition. The subject may then be administered a prophylactic regimen.

[0094] For use in treatment, an effective amount of the formulations disclosed herein (e.g., depot formulations) can be administered to a subject by any mode that delivers the compound to the desired surface. Administration of pharmaceutical compositions can be achieved by any means known to those skilled in the art. Routes of administration include, but are not limited to, oral, topical, intranasal, systemic, subcutaneous, intraperitoneal, intradermal, intraocular, ocular, transmucosal, intravitreal, or intramuscular administration. Administration includes self-administration, administration by another person, and administration by a device.

[0095] For ocular or intraocular indications, any suitable mode of delivering therapeutic compounds or pharmaceutical compositions to the eye or the area near the eye can be used.For ophthalmic formulations, see generally Mitra (ed.), Ophthalmic Drug Delivery Systems, Marcel Dekker, Inc., New York, NY (1993), and Havener, WH, Ocular Pharmacology, CV Mosby Co., St. Louis (1983).Non-limiting examples of pharmaceutical compositions suitable for administration in or near the eye include, but are not limited to, ocular inserts, minitablets, depot formulations, and topical formulations such as eye drops, ointments, and in situ gels.In one embodiment, contact lenses are coated with pharmaceutical compositions comprising the therapeutic compounds disclosed herein.

[0096] For local ocular administration, the therapeutic compound or pharmaceutical composition may be formulated as a solution, gel, ointment, cream, suspension, etc., as is known in the art. Ophthalmic gels may include silica and may be sol-gels or hydrogels. Ointments are semi-solid dosage forms for external use, such as topical application to the eye or skin. In some embodiments, the ointment comprises a solid or semi-solid hydrocarbon base with a melting or softening point close to the core temperature of humans. In some embodiments, the ointment applied to the eye breaks down into small droplets that remain in the conjunctival sac for extended periods of time, thus enhancing bioavailability.

[0097] Ophthalmic inserts are solid or semi-solid dosage forms that do not suffer from the drawbacks of conventional ophthalmic drug forms. They are less susceptible to defense mechanisms such as outflow via the nasolacrimal duct, exhibit the ability to remain in the conjunctival sac for extended periods, and are more stable than conventional dosage forms. They also offer advantages such as accurate dosing of one or more therapeutic compounds, sustained release of one or more therapeutic compounds at a constant rate, and limited systemic absorption of one or more therapeutic compounds. In some embodiments, the ophthalmic insert comprises one or more therapeutic compounds and one or more polymeric materials disclosed herein. The polymeric materials can include, but are not limited to, methylcellulose and its derivatives (e.g., hydroxypropylmethylcellulose (HPMC)), ethylcellulose, polyvinylpyrrolidone (PVPK-90), polyvinyl alcohol, chitosan, carboxymethylchitosan, gelatin, and various mixtures of the aforementioned polymers. The ophthalmic insert can include silica, including in the form of a sol-gel or hydrogel.

[0098] Ophthalmic or intraocular preparations and medicaments may contain non-toxic auxiliary substances such as antimicrobial ingredients that are harmless upon use, such as thimerosal, benzalkonium chloride, methyl and propylparabens, benzyldodecinium bromide, benzyl alcohol, or phenylethanol; buffer ingredients such as sodium chloride, sodium borate, sodium acetate, sodium citrate, or gluconate buffer; and other conventional ingredients, such as, for example, sorbitan monolaurate, triethanolamine, polyoxyethylene sorbitan monopalmitate, ethylenediaminetetraacetic acid, and the like.

[0099] In some embodiments, the viscosity of the ophthalmic formulation containing one or more therapeutic compounds is increased to improve the contact with the cornea and the bioavailability in the eye.The viscosity can be increased by adding a high molecular weight hydrophilic polymer that does not diffuse through biological membranes and forms a three-dimensional network in water.Non-limiting examples of such polymers include polyvinyl alcohol, poloxamer, hyaluronic acid, carbomer, and polysaccharides, cellulose derivatives, gellan gum, and xanthan gum.

[0100] In some embodiments, the ophthalmic formulation can be injected into the eye, for example as a sol-gel. In some embodiments, the ophthalmic formulation is a depot formulation, such as a controlled release formulation. Such controlled release formulations can include particles, such as microparticles or nanoparticles.

[0101] The therapeutic compound or other therapeutic agent or mixture thereof can be formulated in a carrier system. The carrier can be a colloidal system. The colloidal system can be a sol-gel or a hydrogel. In one embodiment, the therapeutic compound or other therapeutic agent or mixture thereof can be encapsulated in a sol-gel or hydrogel while maintaining the integrity of the therapeutic compound or other therapeutic agent or mixture thereof.

[0102] The carrier or colloidal system may be a sol-gel that includes silica nanoparticles or microparticles that contain or encapsulate the therapeutic agent.

[0103] In some embodiments, the therapeutic compound or other therapeutic agents, or mixtures thereof, are prepared with carriers that will protect the therapeutic compound, other therapeutic agents, or mixtures thereof against rapid elimination from the body, such as a controlled release formulation, including implants and microencapsulated delivery systems.

[0104] The therapeutic compound may be included in a controlled release system. The term "controlled release" is intended to refer to any drug-containing formulation in which the manner and profile of drug release from the formulation is controlled. It refers to immediate release formulations and non-immediate release formulations, including but not limited to sustained release formulations and delayed release formulations. The term "sustained release" (also called "extended release") is used in its conventional sense to refer to a drug formulation that provides a gradual release of drug over an extended period of time, preferably, but not necessarily, providing a substantially constant blood level of drug over an extended period of time. The term "delayed release" is used in its conventional sense to refer to a drug formulation in which there is a time delay between administration of the formulation and the release of the drug. "Delayed release" may or may not include a gradual release of drug over an extended period of time, and therefore may or may not be a "sustained release".

[0105] The use of long-term sustained release implants or depot formulations may be particularly suitable for the treatment of chronic conditions. The terms "implant" and "depot formulation" are intended to include a single composition (such as a mesh), or a composition comprising multiple components (e.g., a fiber mesh constructed from several individual pieces of mesh material), or multiple individual compositions (e.g., microparticles or nanoparticles), where the multiple compositions remain localized and provide a long-term sustained release resulting from the aggregation of the multiple compositions. As used herein, "long-term" release means that the implant or depot formulation is constructed and arranged to deliver a therapeutic or prophylactic level of the active ingredient for at least 2 days. In some embodiments, the implant or depot formulation is constructed and arranged to deliver a therapeutic or prophylactic level of the active ingredient for at least 7 days. In some embodiments, the implant or depot formulation is constructed and arranged to deliver a therapeutic or prophylactic level of the active ingredient for at least 14 days. In some embodiments, the implant or depot formulation is constructed and arranged to deliver a therapeutic or prophylactic level of the active ingredient for at least 30 days. In some embodiments, the implant or depot formulation is constructed and arranged to deliver a therapeutic or prophylactic level of the active ingredient for at least 60 days. In some embodiments, the implant or depot formulation is constructed and arranged to deliver therapeutic or prophylactic levels of the active ingredient for at least 90 days. In some embodiments, the implant or depot formulation is constructed and arranged to deliver therapeutic or prophylactic levels of the active ingredient for at least 180 days. In some embodiments, the implant or depot formulation is constructed and arranged to deliver therapeutic or prophylactic levels of the active ingredient for at least one year. In some embodiments, the implant or depot formulation is constructed and arranged to deliver therapeutic or prophylactic levels of the active ingredient for 15-30 days. In some embodiments, the implant or depot formulation is constructed and arranged to deliver therapeutic or prophylactic levels of the active ingredient for 30-60 days.In some embodiments, the implant or depot formulation is constructed and arranged to deliver therapeutic or prophylactic levels of the active ingredient for 60 to 90 days. In some embodiments, the implant or depot formulation is constructed and arranged to deliver therapeutic or prophylactic levels of the active ingredient for 90 to 120 days. In some embodiments, the implant or depot formulation is constructed and arranged to deliver therapeutic or prophylactic levels of the active ingredient for 120 to 180 days. In some embodiments, long-term sustained release implants or depot formulations are well known to those skilled in the art and include some of the release systems described above. In some embodiments, such implants or depot formulations may be administered surgically. In some embodiments, such implants or depot formulations may be administered topically or by injection.

[0106] Sterilization Procedure In certain embodiments, the depot formulation is a sterile formulation.

[0107] In certain embodiments, the method of making a depot formulation further comprises sterilizing the depot formulation. The sterilization step can be performed according to any of the procedures described herein, including, but not limited to, radiation sterilization (e.g., alpha (α), beta (β), gamma (γ), neutron, electron beam, or X-ray sterilization), chemical sterilization (e.g., using ethylene oxide or hydrogen peroxide), or physical sterilization (e.g., using heat, filtration, or retort canning, etc.). In certain embodiments, the method of making a depot formulation further comprises sterilizing the formulation with gamma irradiation.

[0108] In certain embodiments, the method of making the depot formulation comprises a terminal sterilization step, where the depot formulation is optionally sealed in a container (such as a syringe) and then subjected to a sterilization step. Preferably, the sterilization is by irradiation, such as gamma irradiation.

[0109] Various procedures are known in the art for sterilizing chemical compositions such as the depot formulations of the present invention.Sterilization can be achieved by chemical, physical, or irradiation techniques.Examples of chemical methods include exposure to ethylene oxide or hydrogen peroxide vapor.

[0110] Examples of physical methods include sterilization by heating (dry or wet), retort canning, and filtration. For effective sterilization, the British Pharmacopoeia recommends heating at least 160°C for at least 2 hours, at least 170°C for at least 1 hour, and at least 180°C for at least 30 minutes. For examples of heat sterilization, see U.S. Patent No. 6,136,326, which is incorporated herein by reference. Compositions can be sterilized using passing a chemical composition through a membrane. For example, the composition is filtered through a small pore filter, such as a 0.22 micron filter, that contains a material that is inert to the composition being filtered. In certain instances, filtration is performed in a clean room of class 100,000 or higher.

[0111] Irradiation methods include alpha, beta, gamma irradiation, neutron irradiation, electron beam (or electron beam) irradiation, microwave irradiation, and irradiation using visible light. One preferred method is gamma ray (γ-ray) irradiation.

[0112] There are several sources for electron beam irradiation. The two main groups of electron beam accelerators are: (1) dynamitrons, which use an insulated core transformer, and (2) radio frequency (RF) linear accelerators (linacs). Dynamitrons are particle accelerators designed to energize electrons (4.5 MeV). High energy electrons are generated and accelerated by electrostatic fields of accelerator electrodes located within a length of a glass insulated beam tube (accelerator tube). These electrons, traveling through an evacuated beam tube and an extension of the beam transport (drift pipe), are subjected to a magnet deflection system to generate a "scanned" beam before exiting the vacuum vessel through a beam window. Dose can be regulated by control of percent scan, beam current, and conveyor speed. In a specific example, the electron beam radiation used is at least about 2 microcuries (μCi) / cm. 2 , at least about 5 μCi / cm 2 , at least about 8 μCi / cm 2 , or at least about 10 μCi / cm 2 In certain instances, the electron beam radiation used may be maintained at an initial fluence of about 2 to about 25 μCi / cm 2 In certain instances, the electron beam dose is about 5 to 50 kilograys (kGray), or about 15 to about 20 kGray, with the particular dose being selected relative to the density of the material being exposed to the electron beam radiation as well as the amount of bioburden estimated to be therein. Such factors are within the skill of one of ordinary skill in the art.

[0113] Sterilization procedures using visible light are described in U.S. Patent No. 6,579,916, which is incorporated herein by reference. Visible light for sterilization can be generated using any conventional generator of sufficient power and wavelength width to effect sterilization. Generators are commercially available under the trade name PureBright® In-line Sterilization System (PurePulse Technologies, Inc. 4241 Ponderosa Ave, San Diego, Calif. 92123, USA). The PureBright® In-line Sterilization System uses visible light to sterilize transparent liquids at an intensity approximately 90,000 times greater than surface sunlight. If the amount of UV light transmission is a concern, conventional UV absorbers can be used to filter the UV light.

[0114] One method of sterilizing the formulations provided herein is by radiation sterilization. Such radiation includes alpha radiation, beta radiation, gamma radiation, neutron radiation, electron beam, and X-rays. In certain preferred embodiments, the radiation is gamma radiation (γ-ray) sterilization. Exemplary radiation doses include: about 0.5-10 MRad (5-100 kGy), about 1.5-5.0 MRad (10-50 kGy), or about 2-4 MRad (20-40 kGy) of radiation. For example, gamma radiation sterilization can be performed at one of the following doses (kGY): 10, 15, 20, 25, 30, 35, 40, 45, or 50. Generally, the dose of electron beam or gamma radiation ranges from about 10-50 kGy or 20-40 kGy.

[0115] The dose of radiation may be determined based on the bioburden level (initial contamination) of the composition prior to sterilization. Exposure time depends on the nature of the composition, the intensity of the beam, and the conveyor speed, among other variables, and is typically less than a minute; generally, it ranges from 1 / 10 of a second to several seconds. A dosimeter may be used to determine the optimal exposure time for the particular sample or composition being irradiated.

[0116] The temperature of the sterilization procedure may range from about 2° C. to 50° C., or from about 15° C. to 40° C., or from about 20° C. to 30° C. Typically, irradiation is carried out under ambient conditions.

[0117] In certain embodiments, the depot formulation is sterilized and -3 , or at least 10 -4 , or even at least 10 -5 Sterility assurance levels (SAL) of at least 10. Measurement standards for sterility assurance levels are described, for example, in ISO / CD 14937, the entire disclosure of which is incorporated herein by reference. In some examples, the sterile compositions provided herein provide a sterility assurance level (SAL) of at least 10. -6 (greater than 1 in 1 million chance of nonsterile units).

[0118] The depot formulation may be contained in any type of at least partially e-beam or gamma-ray transparent container, including but not limited to glass, plastic, foil and film-forming packages.The container may be sealed or may have an opening.Examples of glass and possibly plastic containers include, but are not limited to, ampoules, vials, syringes (single, dual or multiple), pipettes, applicators, tubes, etc.

[0119] For example, the container may be a syringe, or a syringe that optionally includes a vented syringe cap. Examples of vented syringe caps include Vented FLL Cap 3 micron filter (Qosina, P / N 12089), Female Luer Cap Vented Hex Luer Fit (Qosina, P / N 6570), and Luer Tip Syringe Cap (Value Plastics, Fort Collins, Colo., P / N VPM0480201N).

[0120] Containers such as but not limited to those described above may be further packaged and sealed in a pouch, for example, a sealed foil pouch.Packaging materials for use in the pharmaceutical and medical industries are well known.As applicable to the compositions of the types provided herein, the containers described in "Pharmaceutical Packaging Technology", Dean, DA, et al., Ed., Taylor & Francis, Inc., 2000 are contemplated for use in the present method and for the compositions and kits provided herein.

[0121] Penetration of e-beam or gamma radiation is generally a function of the packaging material selected. In certain embodiments, if the penetration from the side of a stationary e-beam or gamma radiation is determined to be insufficient, the container can be inverted or rotated to achieve proper penetration. Alternatively, the e-beam or gamma radiation source can be moved around the stationary package or container. Dose maps can be performed to determine dose distribution and dose penetration in the product load. Dose maps can be used to identify minimum and maximum dose zones for a particular product, package or container. In certain embodiments, after the container containing the depot formulation is sterilized, for example with e-beam or gamma radiation, the container can be further sterilized. For example, the container is placed in a kit with other components that may require sterilization. In this process, the entire kit can be sterilized. For example, the entire kit can be further sterilized by chemical (e.g., with ethylene oxide or hydrogen peroxide vapor), physical (e.g., dry heat), or other techniques such as microwave irradiation, e-beam irradiation, or gamma irradiation.

[0122] Having now generally described the invention, it will be more readily understood by reference to the following examples, which are included merely for the purpose of illustrating certain aspects and embodiments of the invention and are not intended to limit the invention. EXAMPLES

[0123] Methods for preparing the formulations in Examples 1 and 2 below were carried out essentially as described in WO2014 / 207304 and WO2017 / 068245 (each of which is incorporated by reference herein for all purposes), but modified as specifically described in each example below.

[0124] Example 1: Preparation of Elamipretide-containing Silica-API-Silica Hydrogel Composite (Depot) Formulation Formulation Specifications: Total drug dose: 270 μg API in 50 μL injection Daily drug dose: 3μg In vivo release profile: 3-month sustained release Administration route: Intravitreal injection Injection volume: 50μL or less Needle size: 30 gauge Injection frequency: Repeat once every 3 months

[0125] The information in Table 1 provides details of the formulations prepared and their analysis. 1) Microparticle formulations used to produce injectable hydrogel composites 2) Mass (g) of microparticles per mL of hydrogel precursor (R400) 3) Weight percent of particulates in the depot (500 mg of particulates per gram of depot) 4) The amount of API measured in the final depot formulation, the density of the depot was estimated to be 1.2 g / mL 5) pH of the final depot formulation 6) Manual injectability testing was performed with a 100 μL injection (after priming).

[0126] The in vitro dissolution data under sink conditions for the composites listed in Table 1 are shown in Figures 1 and 2.

[0127] discussion: FIG. 1 shows that formulation #05D releases the API (i.e., elamipretide) over a period of approximately 8-9 days, which should equate to approximately 9 months in vivo.

[0128] FIG. 2 shows that formulation #08D releases the API (i.e., elamipretide) over a period of about 4-5 days, which should equate to about 5 months in vivo.

[0129] The data in this Example 1 and in Figures 1 and 2 demonstrate that by adjusting the conditions under which the particles are formed (eg, formation pH), the release rate of the API can be tuned to a desired release criteria.

[0130] Example 2: Preparation of Silica-API-Silica Hydrogel Composite (Depot) Containing Compound I 1) Microparticle formulations used to produce injectable hydrogel composites 2) Mass (g) of microparticles per mL of hydrogel precursor (R400) 3) Weight percent of particulates in the depot (500 mg of particulates per gram of depot) 4) The amount of API measured in the final depot formulation, the density of the depot was estimated to be 1.2 g / mL 5) pH of the final depot formulation 6) A manual injectability test was performed with a 100 μL injection (after priming).

[0131] The in vitro dissolution data under sink conditions for the composites listed in Table 2 are shown in Figures 3 and 4.

[0132] Summary of Results The hydrogel component reduces the release rate of both silica and API in formulation #05D. The hydrogel component had no significant effect on the release rate of formulation #08D compared to #05D. All depot formulations were injected through either a 25-gauge or 27-gauge needle. The hydrogel composite was easy to fabricate. The particles were quickly wetted and the suspension was easy to work with. Data for Compound I formulations #05D and #08D, in particular, were similar to those observed for elamipretide: - Figure 3 shows that formulation #05D releases the API (i.e., Compound I) over a period of about 9-10 days, which would equate to about 10 months in vivo. - Figure 4 shows that formulation #08D releases the API (i.e., Compound I) over a period of about 4-5 days, which would equate to about 5 months in vivo. The data in this Example 2 demonstrates that by adjusting the conditions under which the particles are formed (e.g., formation pH), the release rate of the API can be tuned to the desired release criteria.

[0133] [Table 1]

[0134] [Table 2]

[0135] Example 3: Release of Compound I and Elamipretide in Ammonium Bifluoride: 0.1M NH 4 HF 2 Add to silica microparticles to a particle concentration of 1 mg / mL (i.e., 10 mg of particles for 10 mL of solution). · Silica-placebo microparticles spiked with API at concentrations of 16μg / mL or 40μg / mL. Dissolve the silica particles at room temperature. Excess silica-placebo microparticles were mixed with inert, non-reactive NH 4 HF 2 Add to (16mg). Blank sample: the same formulation without the API. · Centrifuge 1.5mL of sample solution at 14000rpm for 10 minutes. The supernatant is analyzed by a stability-indicating RP-HPLC method. · Control / Standard → 15μg / mL or 40μg / mL API in water.

[0136] [Table 3]

[0137] [Table 4]

[0138] Conclusion: The relative amounts of impurities in the fresh API samples and in the samples with API released from the microparticle formulation were at similar levels. Both elamipretide and Compound I were not significantly modified by the formulation process and no adverse effects were observed in the formulations making them unsuitable for the development of these formulations for controlled release of the tested API.

[0139] Example 5: Preparation of sol for microparticles and preparation of microparticles An overview of the manufacturing process for the silica-API microparticle-silica hydrogel depot is shown in Figure 5.

[0140] Silica sol for producing spray-dried microparticles can be made by hydrolysis of tetraethyl orthosilicate (TEOS) in water using hydrochloric acid (HCl) as a catalyst at pH 2. The molar ratio of water:TEOS determines the first R value (first R) of the microparticle formulation. The active pharmaceutical ingredient (API) is dissolved in a diluent (ethanol or water, or a mixture thereof), and then this solution is combined with the sol. As a result, the TEOS in the formed solution is further diluted, which is shown as a second R value (second R), calculated as if the diluent were water. Prior to spray drying, the pH of the sol can be adjusted and then the sol can be spray-dried to form microparticles. These microparticles mainly determine the properties of the formulation. The specific experimental procedures are described below.

[0141] Silica sols for making microparticles were generated by hydrolysis of tetraethyl orthosilicate (TEOS) in water as follows: First, TEOS was mixed with water; the initial molar ratio of water to TEOS (= the first R in the standard nomenclature used to describe formulation recipes, e.g., R10-100) was varied from 3 to 10. Since TEOS is immiscible in water, it was a two-phase system before hydrolysis. The water-TEOS mixture was then adjusted to pH 2 with 0.1 M HCl, resulting in a homogeneous aqueous solution. The amount of water in the HCl is taken into account when calculating the R value. After the exothermic hydrolysis reaction, the silica sol was cooled in an ice bath. A scheme showing the molecular polymerization is shown in Figure 6.

[0142] The cold silica sol was then added to the API-water solution, denoted as the dilution factor R, i.e., the molar ratio of water to TEOS after dilution (= the second R in the standard nomenclature used to describe formulation recipes, e.g., R10-100). The second R value was 100 for all microparticles produced.

[0143] The effect of API load-% was also investigated, where API load-% was varied from 2% to 5% relative to the silica content in the sol. Finally, the sol was adjusted to a final pH with 0.1 M NaOH solution, and the pH was varied from 4.0 to 5.9 before spray drying. In general, the pH of the sol before spray drying affects the condensation degree of silica and thus the dissolution rate of the microparticles.

[0144] Finally, the silica-API microparticles were prepared by spray drying the sol containing the API with filtered ambient air. The spray drying parameters are given below: [Table 5]

[0145] Example 6: Preparation of Silica-API Microparticle Silica Hydrogel Depot Formulation To prepare an injectable depot, a silica sol with high water content (R400 sol, water content 98-99%) is also made by hydrolyzing TEOS in water using HCl as a catalyst at pH 2. Spray-dried silica-API microparticles with the first and second R values ​​are mixed with the R400 sol. This silica microparticle-silica hydrogel suspension is used to fill a syringe. The syringe can be kept homogenous by rotating in a rotary mixer until a gel is formed. The specific experimental procedures are described below.

[0146] Silica sol with high water content (R400, water content 98-99%) was prepared by hydrolysis of TEOS and deionized water using 0.1 M HCl as a catalyst. After hydrolysis, the pH of the sol (i.e., the sol for hydrogel) was adjusted to about pH 6 with 0.1 M NaOH solution.

[0147] Spray dried silica-API microparticles from Example 5 were mixed manually with R400 sol at a 1:1 ratio (w:v). The suspension of silica sol and microparticles were mixed by pipetting the suspension up and down, with batch sizes of approximately 1-3 ml. The suspension was transferred to a plastic syringe (1 ml disposable BD syringe) by drawing the suspension directly into the syringe (no needle) and removing potential air bubbles by tapping the syringe. The suspension was then allowed to gel at ambient temperature for 1-2 days, keeping the silica-API microparticle-silica hydrogel suspension homogenous by gently mixing the capped syringe in a tube rotator to prevent particle settling from occurring.

[0148] Example 7: In vitro dissolution test The properties of the formulation are mainly determined by the particulates. The first R value determines the dissolution rate of the silica. In general, the smaller the first R, the faster the silica matrix dissolves in water. The second R value, on the other hand, affects the encapsulation efficiency. The reported pH is the pH of the sol before spray drying. The pH affects the degree of condensation of the silica and therefore the dissolution rate, which depends on the formulation and API. The API loading % indicates the amount of API in the formulation relative to the silica content. The API loading rate affects the structure and dissolution rate of the silica. The diluent is a solvent / diluent used to dissolve the API before adding it in the sol. The diluent can improve the solubility of the API and prevent the precipitation of the API in spray drying, for example. The inlet temperature (IT) in spray drying plays a role in particle formation by controlling the evaporation rate of the solvent.

[0149] In vitro dissolution of silica and release of API were measured in 50 mM TRIS containing 0.01% (v / v) TWEEN® 80 at pH 7.4 (internally referred to as TW) at 37°C. The analyzed microparticle or hydrogel samples were approximately 10-30 mg in size and were dissolution tested in a shaking water bath (60 strokes / min) for up to 168 hours (7 days). Silica and API concentrations were measured in the dissolution medium under sink conditions (SiO 2 Free dissolution of the matrix was maintained, i.e. silica was kept below 20% of saturation concentration). The dissolution medium was replaced with fresh medium at each time point to keep the silica concentration below 30 ppm, i.e. to ensure sink conditions were maintained. At the 1 hour sampling time point and every 24 hours thereafter, samples were centrifuged before replacing the dissolution medium. Three replicate samples were taken at each time point and the average values ​​are presented in the results.

[0150] The total silica content of the samples was measured separately by dissolving the samples (10-25 mg) in 0.5 M NaOH solution, and elution was continued until the cumulative amount of eluted silica was equal to the total silica content. The total content of Compound I was measured by dissolving the samples (10 mg microparticles or 20 mg depot formulation) in 0.1 M ammonium bifluoride, while elamipretide (10 mg microparticles or 20 mg depot formulation) was dissolved in 150 ml of 50 mM glycine buffer (pH 9.4) supplemented with 0.01% (v / v) TWEEN® 80 (internally referred to as GW). In addition, to assess how much of the samples remained after the 168 hour time point, total dissolution was performed in 0.5 M NaOH solution on the sink condition elution samples after their final sampling time point.

[0151] The encapsulation efficiency of the microparticle formulation was defined as the mass ratio of API to silica calculated after considering the non-encapsulated API and was measured in ethanol. The mass of the analyzed sample was about 20-30 mg, and ethanol solution was added so that the final mass concentration of microparticles in ethanol was 1 mg / ml. The ethanol dissolution test was performed at room temperature for up to 24 hours. The rationale here is that the silica matrix of the silica-API microparticles is insoluble in ethanol. Therefore, only the non-encapsulated API dissolves under these conditions.

[0152] It should be noted that dissolution testing under sink conditions is an accelerated dissolution test method, and the dissolution profiles of the API and silica do not correspond to their in vivo dissolution characteristics without appropriate in vitro-in vivo correlation (IVIVC) factors. For intravitreal injection, it has been previously shown that an IVIVC factor of 30 can be used to estimate the in vivo dissolution rate of a silica matrix from in vitro dissolution data under sink conditions. This means that the dissolution rate of the silica matrix is ​​30 times slower in vivo compared to sink test conditions.

[0153] Example 8: Formulations containing Compound I I. Silica-API Microparticle Formulations Containing Compound I The compositions of the silica-API microparticle formulations produced in this study are shown in the table below, and the cumulative in vitro Compound I release and dissolution of the silica matrix under sink conditions for the microparticle formulations are shown in Figures 8-10. The figures show the effect of the studied process parameters on the release profile of the formulations.

[0154] [Table 6]

[0155] II. In vitro dissolution of silica-API microparticle formulations containing Compound I under sink conditions In vitro dissolution tests were performed up to 168 hours (7 days) under sink conditions. All formulations tested (#01-#08) had low burst release (less than 5% of the total content), indicating good encapsulation of the API. This was also confirmed by ethanol dissolution tests, which showed that the API content was below the limit of quantification after 24 hours of dissolution.

[0156] A summary of the in vitro cumulative silica dissolution and API release from silica-API microparticle formulations #01-#03 is shown in Figure 8. The API loading in these microparticle formulations was 2% and the pH was kept constant at 5.9. The first R value of the microparticle formulations had a significant effect on the silica dissolution and API release profiles. A decrease in the first R value resulted in a faster dissolution rate of the silica matrix because more hydroxyl groups remained in the silica matrix. Within 7 days, approximately 5% of the silica matrix was dissolved from the slowest formulation (#01) and approximately 84% was dissolved from the fastest formulation (#03). The API release profile correlated with the silica dissolution profile, with 12-87% of the total API content being released within 7 days.

[0157] A summary of the in vitro cumulative silica dissolution and cumulative API release from silica-API microparticle formulations #03-#06 at various pH values ​​is shown in Figure 9. The API loading in these microparticle formulations was 2%, the first R value was kept constant at 3, and the second R value was kept constant at 100.

[0158] In this study, significant differences in the release rate of API and dissolution of silica matrix are found between formulations #03 and #04, but not between #05 and #06 (Figure 9). Within 7 days, approximately 47% of the silica matrix was dissolved from the slowest formulation (#04) and 100% was dissolved from the fastest formulations (#05 and #06). The API release profile correlated with the silica dissolution profile, with 48-100% of the total API content released within 7 days. The in vitro dissolution profile of formulation #03 under sink conditions is unusual as it should be the slowest formulation due to the highest pH (more condensed silica matrix). API release correlates with silica dissolution for all studied microparticle formulations.

[0159] Increasing API loading % from 2% to 5% was investigated in R3-100 based formulations (Figure 10). Formulations #05 and #08 are faster formulations and identical to each other, while formulations #03 and #07 are slower formulations. Dissolution results show that increasing API loading from 2% to 5% does not significantly affect the dissolution rate. In contrast, pH has a more significant effect on the dissolution profile.

[0160] III. Particle size distribution The particle size distributions of the developed silica-Compound I microparticle formulations are summarized below. [Table 7]

[0161] The D values ​​are the 10%, 50%, and 90% intercepts of the cumulative mass of the analyzed sample. For example, the D value is the diameter at which 10% of the mass of the sample is composed of particles with a diameter equal to or less than this value. Here, the particle size of all the microparticle formulations is obviously small, since the D value is less than 10 μm (except #06). Silica-API microparticle formulations #05 and #08 showed the most desirable characteristics in terms of silica dissolution, API release, and particle size distribution, and therefore they were selected as the lead formulations to be used in the depot formulation development.

[0162] IV. Silica-API Microparticles-Silica Hydrogel Depot Formulation The composition of the silica-Compound I microparticle-silica hydrogel (HG) formulation (ie, silica-Compound I depot formulation) is shown below. [Table 8]

[0163] The cumulative dissolution and release rate of API from the depot formulations under sink conditions for silica matrix are shown in Figures 11 and 12. The release rates of the corresponding microparticle formulations are also shown in the figures, showing the effect of the HG component on the dissolution rate of the microparticle formulations. When the microparticles are incorporated into the silica-HG component, the cumulative amount of API released from (microparticle) formulation #05 in the first 24 hours decreases from about 44% to 5% and from 34% to 10% for (microparticle) formulation #08. The total release time increases, since both microparticle formulations released more than 96% of the API in 120 hours, whereas 78% (#05D) and 96% (#08D) of the API in the depot formulations were released in 168 hours. This phenomenon is expected since the silica hydrogel surrounding the silica microparticles acts as a barrier and locks-in-place the microparticles. Thus, the depot formulation dissolves the microparticles as a single solid rather than dispersing them in the dissolution buffer. This reduces the reactive surface area of ​​the formulation and also reduces the dissolution rate.

[0164] It is clear from Figures 11 and 12 that the HG component reduces the dissolution rate of the microparticle formulations at least to some extent. A short lag time is observed in the dissolution of silica and release of API for both depot formulations up to the 24 hour time point.

[0165] V. Rheological measurements and injectability Oscillatory measurements were performed to demonstrate the presence of a three-dimensional gel structure in the prefilled syringe depot formulation. For viscoelastic materials, if the loss factor (tan δ=G” / G') is greater than 1, the material behaves more like a liquid. On the other hand, if the loss factor is less than 1, the material behaves more like a semi-solid, e.g., a gel.

[0166] For depot formulations #05D and #08D, the loss factor is less than 1 in the entire frequency range shown in Figure 13. Thus, a three-dimensional gel structure is formed, preventing particle settling in the syringe at rest, i.e., during storage. Furthermore, the depot formulation samples induce appropriate storage modulus (G') values ​​that are frequency independent, indicating good mechanical strength of the depot formulations.

[0167] The depot formulation induces shear thinning properties as the viscosity decreases as a function of increasing shear rate (Figure 14). The viscosity is high at low shear rates since the material is a solid at rest. As the shear rate increases, the material begins to flow. This shear thinning property indicates that the material may be injected from a prefilled syringe through a thin needle. During the injection procedure, the material experiences shear rates ranging from 10 000 1 / s to 100 000 1 / s. In essence, the depot formulation is a stable solid-like material at rest that begins to flow as a liquid when subjected to stress.

[0168] Furthermore, to demonstrate that the shear thinning phenomenon shown in Figure 14 parallels good injectability of the depot formulation, a manual injectability test through a fine hypodermic needle was performed. The results of the injectability test are shown below. [Table 9]

[0169] Example 9: Formulations containing Elamipretide I. Silica-API Microparticle Formulations Containing Elamipretide The composition of the silica-elamipretide microparticles produced in this study is shown below, and the cumulative release at sink conditions of API from the microparticle formulation is shown in Figures 15-17. These figures show the effect of the studied process parameters on the release profile of the formulation.

[0170] [Table 10]

[0171] II. In Vitro Dissolution of Silica-API Microparticle Formulations Containing Elamipretide Under Sink Conditions In vitro dissolution tests were performed up to 168 hours (7 days) under sink conditions. All formulations tested (#01-#08) had low burst release (less than 5% of the total API content), indicating good encapsulation of the API. This was also confirmed by ethanol dissolution tests, which showed that the API content was below the detection limit after 24 hours of dissolution.

[0172] A summary of the in vitro cumulative silica dissolution and API release from silica-API microparticle formulations #01-#03 is shown in Figure 15. The API loading in these microparticle formulations was 2% and the pH was kept constant at 5.9. The first R value of the microparticle formulations influenced the silica dissolution and API release profiles. A decrease in the first R value resulted in a faster dissolution rate of the silica matrix because more hydroxyl groups remained in the silica matrix. Within 7 days, approximately 10% of the silica matrix was dissolved from the slowest formulations (#01 and #02) and approximately 61% was dissolved from the fastest formulation (#03). The API release profile correlated with the silica dissolution profile, with 15-75% of the total API content being released within 7 days.

[0173] A summary of the in vitro cumulative silica dissolution and cumulative API release from silica-API microparticle formulations #03-#06 at various pH values ​​is shown in Figure 16. In this study, significant differences in API release rate and silica matrix dissolution are observed between formulations #03 and #06. Within 7 days, approximately 61% of the silica matrix was dissolved from formulation #03, whereas approximately 100% was dissolved from formulation #06. API release correlates with silica dissolution in all studied formulations.

[0174] Increasing API loading % from 2% to 5% was investigated in R3-100 based formulations (Figure 17). Formulations #05 and #08 are faster formulations and their dissolution profiles appear similar, however the difference between formulations #03 and #07 is significant. The dissolution results show that increasing API loading from 2% to 5% has a significant effect on the dissolution rate when the pH of the sol prior to spray drying is higher (pH=5.9). In contrast, for the lower pH sols, API loading does not have a significant effect on the dissolution profile.

[0175] III. Particle size distribution The particle size distribution of the developed silica-elamipretide microparticle formulations is summarized below. [Table 11]

[0176] All microparticle formulations have small particle size, with D50 values ​​less than 10 μm (except #06). Silica-API microparticle formulations #05 and #08 showed the most desirable characteristics in terms of silica dissolution, API release and particle size distribution, and therefore they were selected as lead formulations to be used in the depot formulation development.

[0177] IV. Silica-API Microparticles-Silica Hydrogel Depot Formulation The composition of the silica-elamipretide microparticle-silica hydrogel (HG) formulation (ie, silica-elamipretide depot formulation) is shown below. [Table 12]

[0178] The cumulative dissolution and API release rates at sink conditions for silica matrices from the depot formulations are shown in Figures 18 and 19. The release rates of the corresponding microparticle formulations are also shown in the figures, showing the effect of the HG component on the dissolution rate of the microparticle formulations. When the microparticles are incorporated into the silica HG component, the cumulative amount of API released from microparticle formulation #05 within the first 24 hours decreases from about 47% to 14%, and from 34% to 14% for microparticle formulation #08. The total release time increases for formulation #05D, as 89% of the API in the depot formulation is released at 168 hours, whereas the microparticle formulation releases more than 96% of the API at 120 hours. This phenomenon is expected since the silica hydrogel surrounding the silica microparticles acts as a barrier and locks-in-place the microparticles. Thus, the depot formulation dissolves the microparticles as a single solid, rather than dispersing them in the dissolution buffer. This reduces the reactive surface area of ​​the formulation, which also reduces the dissolution rate.

[0179] It is clear from Figures 18 and 19 that the HG component reduces the dissolution rate of the microparticle formulations at least to some extent. A short lag time is observed in silica dissolution and API release for both depot formulations up to the 24 hour time point.

[0180] V. Rheological measurements and injectability Oscillatory measurements were performed to demonstrate the presence of a three-dimensional gel structure within the depot formulation loaded into the prefilled syringe. For viscoelastic materials, if the loss factor (tan δ=G” / G') is greater than 1, the material behaves more like a liquid. On the other hand, if the loss factor is less than 1, the material behaves more like a semi-solid, e.g., a gel.

[0181] For depot formulations #5D and #8D, the loss factor is less than 1 in the entire frequency range shown in Figure 20. Thus, a three-dimensional gel structure is formed, preventing particle settling in the syringe at rest, i.e., during storage. Furthermore, the depot formulation samples induce appropriate storage modulus (G') values ​​that are frequency independent, indicating good mechanical strength of the depot formulations.

[0182] The depot formulation induces shear thinning properties as the viscosity decreases as a function of increasing shear rate (Figure 21). The viscosity is high at low shear rates since the material is a solid at rest. As the shear rate increases, the material begins to flow. This shear thinning property indicates that the material may be injected from a prefilled syringe through a thin needle. During the injection procedure, the material experiences shear rates ranging from 10000 1 / s to 100000 1 / s. In essence, the depot formulation is a stable solid-like material at rest that begins to flow as a liquid when subjected to stress.

[0183] Furthermore, to demonstrate that the shear thinning phenomenon shown in Figure 21 parallels good injectability of the depot formulation, a manual injectability test through a fine hypodermic needle was performed. The results of the injectability test are shown below. [Table 13]

[0184] Example 10: Production using a continuous flow reactor system A continuous flow reactor system was utilized to scale up the production of elamipretide-loaded silica-API-silica hydrogel composites (depots) with API loadings of 2% and 5%. A schematic diagram is shown in Figure 7.

[0185] In the continuous flow process, the pH and concentration of the silica and API remained consistent throughout the manufacturing process.

[0186] [Table 14] The particle size distribution of the manufactured consistency batches of formulation #09 is similar and the consistency batches of formulation #10-5 have the same particle size distribution. · Continuous flow processes allow better control of reaction times leading to narrower particle size distributions. Particle size distribution was measured using a Sympatec HELOS laser diffraction instrument. Generally, the silica microparticles produced by our system have an average size of 5±2 μm (D50, i.e. 50% of the particles are below the stated size). - Figure 22 shows a graph of particle size distribution from a consistency batch. - Figure 23 shows the in vitro cumulative dissolution of elamipretide microparticle formulations #09 (2% API load) and #10-5 (5% API load) under sink conditions. The manufacturing process in the continuous flow reactor system is scalable and has been demonstrated with up to ten-fold (10x) scale-up, as shown in Figures 24 and 25. · The process was successfully modified from a batch to a continuous flow process using 2% and 5% API payload materials. · The pH and concentrations of silica and API remained consistent throughout the manufacturing process for the 2% and 5% API payload materials. All manufactured batches of microparticle formulations #09 and #10-5 have nearly identical PSD profiles (except for a slight variation in the size of the right shoulder for the first manufactured batch). The batch size was successfully increased by 10-fold (10x) (compared to the feasibility study batch size) and the dissolution profile of the manufactured formulation remained the same as the consistency batch of the 2% API formulation. For the 5% API formulation, the 10x batch had a slightly lower dissolution profile. Temperature control of the collection vessel prevented the changes in elution seen with the 5% payload material. The continuous flow process is a repeatable process for producing silica-API microparticles containing 2% and 5% API payload.

[0187] Example 11: Phase 1 Clinical Trial Data of Elamipretide in Moderate Age-Related Macular Degeneration and High-Risk Drusen A Phase 1 study was conducted to evaluate the safety, tolerability, and feasibility of subcutaneous administration of elamipretide in patients with moderate age-related macular degeneration (AMD) and high-risk drusen (HRD). The results of the study, entitled ReCLAIM, were published in Ophthalmology Science (MJ Allingham, PS Mettu, and SW Cousins; Ophthalmology Science, Vol. 2, Issue 1, March 2022, 100095; https: / / doi.org / 10.1016 / j.xops.2021.100095; incorporated herein by reference in its entirety).

[0188] The ReCLAIM trial demonstrated that elamipretide is safe and well tolerated in the treatment of AMD and HRD.

[0189] Example 12: Elamipretide treatment can slow or reverse age-related visual decline. Elamipretide has been studied for its ability to treat age-related visual impairment, as detailed in NM Alam, RM Douglas, and GT Prusky, Disease Models & Mechanisms (2021) 14, dmm048256. doi:10.1242 / dmm.048256, which is incorporated by reference in its entirety.

[0190] The authors determined that elamipretide is effective in treating, i.e., preventing and reversing, photopic age-related loss of spatial vision and improving the visual system's ability to process photopic temporal information. Indeed, the study revealed behavioral evidence that elamipretide treatment rather selectively ameliorates age-related loss of photopic function.

[0191] INCORPORATION BY REFERENCE All U.S. patents and U.S. and PCT published patent applications cited herein are hereby incorporated by reference.

[0192] Equivalent The above specification is believed to be sufficient to enable one skilled in the art to practice the invention. The present invention is not limited in scope by the examples provided, as the examples are intended as merely illustrative of one aspect of the invention, and other functionally equivalent embodiments are within the scope of the present invention. Various modifications of the present invention, in addition to those shown and described herein, will become apparent to those skilled in the art from the foregoing description and are included within the scope of the appended claims. The advantages and objectives of the present invention are not necessarily encompassed by each embodiment of the present invention.

Claims

1. A depot formulation comprising a biodegradable silica hydrogel composite, the hydrogel composite comprising: (i) silica hydrogel; and (ii) a plurality of microparticles comprising silica and an oligopeptide or peptidomimetic compound, or a pharma- ceutically acceptable salt thereof; Including, the free base equivalent of said oligopeptide or peptidomimetic compound has a molecular weight of less than 1,200 amu; The hydrogel composite is a depot formulation that is non-flowable and structurally stable during static storage and is shear thinning when subjected to shear stress.

2. said oligopeptide or peptidomimetic compound is positively charged in an aqueous buffer solution at neutral pH, and / or The pharmaceutical depot of claim 1, wherein the oligopeptide or peptidomimetic compound is water-soluble.

3. 2. The pharmaceutical depot of claim 1, wherein the microparticles are present in the pharmaceutical depot in an amount of about 30 to about 95% by weight.

4. 10. The pharmaceutical depot of claim 1, wherein the free base equivalent of said oligopeptide or peptidomimetic is present in the pharmaceutical depot in an amount of about 0.1% to about 10% by weight.

5. 10. The pharmaceutical depot of claim 1, wherein said silica hydrogel composite is free-flowing when subjected to shear stress and can be injected from a syringe equipped with a needle.

6. The depot preparation according to claim 5, wherein the needle has a gauge of 25 to 30.

7. The pharmaceutical depot according to claim 1, wherein the silica hydrogel composite is non-flowable and structurally stable at 37°C upon termination of application of shear stress.

8. The pharmaceutical depot of claim 1, wherein the pH of the formulation is from about 4.0 to about 7.

5.

9. The depot formulation of claim 1, wherein the microparticles comprise a plurality of oligopeptide or peptidomimetic compounds.

10. the microparticles comprise an oligopeptide or a pharma- ceutically acceptable salt thereof; the oligopeptide is selected from the group consisting of elamipretide, compound II, compound III, and compound IV, and pharma- ceutically acceptable salts thereof; Elamipretide has the following structure: 【Chemistry 1】 having Compound I has the following structure: 【Chemistry 2】 having Compound II has the following structure: 【Chemistry 3】 having Compound III has the following structure: 【Chemistry 4】 The pharmaceutical depot of claim 1, having the formula:

11. the microparticle comprises a peptidomimetic compound or a pharma- ceutically acceptable salt thereof; The peptidomimetic compound comprises 【Chemistry 5】 The pharmaceutical depot according to claim 1,

12. The pharmaceutical depot of claim 1, wherein the oligopeptide or peptidomimetic is targeted to mitochondria.

13. The pharmaceutical depot of claim 1, which is injectable or implantable.

14. The pharmaceutical depot according to claim 1, which is for parenteral administration.

15. 15. The pharmaceutical depot of claim 14, for subcutaneous, intramuscular, peritoneal or ocular administration.

16. The depot formulation according to claim 13, which is for intravitreal injection or topical ophthalmic administration.

17. 2. The pharmaceutical depot according to claim 1, which is for administration once a week to once a year, or once a month to once a year, or once a month to once every 6 months, or once a month to once every 3 months, or once every 3 months to once every 6 months, or once every 6 months to once every 9 months, or once every 9 months to once a year, or once a month to once every 6 months, or once a month to once every 5 months, or once every 2 months to once every 5 months, or once every 2 months to once every 4 months, or once every 2 months to once every 3 months, or once every 3 months to once every 4 months, or about once every 3 months.

18. 2. The pharmaceutical depot of claim 1, wherein the amount of the free base equivalent of the oligopeptide or peptidomimetic compound in the pharmaceutical depot is from 10 μg / 50 μL to 100 mg / 50 μL.

19. 10. The pharmaceutical depot of claim 1, which delivers a daily dose of between 0.01 μg and 10 μg of the free base equivalent of the oligopeptide or peptidomimetic compound.

20. The depot formulation according to claim 1, for intravitreal injection approximately once every three months.

21. 2. The pharmaceutical depot of claim 1, which is a sterile formulation.

22. 22. The pharmaceutical depot of claim 21, wherein the sterile formulation is a gamma-sterilized formulation.

23. 2. The pharmaceutical depot of claim 1, wherein the silica hydrogel has a solids content of 3% by weight or less.

24. The pharmaceutical depot according to claim 1, wherein the microparticles comprise 0.1 to 70% by weight of the oligopeptide or peptidomimetic compound.

25. The pharmaceutical depot according to claim 1, wherein the microparticles have a diameter of from 0.5 μm to 300 μm.

26. 2. The pharmaceutical depot of claim 1, wherein the volume fraction of microparticles having a diameter of less than 1 μm is less than 3%.

27. The pharmaceutical depot according to claim 1, wherein the solids content of the composite is from 10% to 75% by weight.

28. and / or a complex modulus measured under small angle oscillatory shear in the linear viscoelastic region of less than 2400 kPa; and / or a loss factor (i.e. viscous / elastic modulus) less than 1; and / or 2. The pharmaceutical depot according to claim 1, having a viscosity of 10-50 Pas when measured at a shear rate of 10-50 sec −1 , 0.4-1.5 Pas when measured at a shear rate of 200-210 sec −1 , and 0.1-0.4 Pas when measured at a shear rate of 600-610 sec −1 .

29. 10. The pharmaceutical depot of claim 1, prepackaged in a syringe and needle configuration for single-use intravitreal injection.

30. A composition for treating, inhibiting, ameliorating or delaying the onset of a disease, disorder or condition, said composition comprising a pharmaceutical depot according to any one of claims 1 to 29.

31. 31. The composition of claim 30, wherein the disease, disorder or condition is Leber's hereditary optic neuropathy.

32. 31. The composition of claim 30, wherein the disease, disorder or condition is Fuchs endothelial corneal dystrophy (FECD), age-related macular degeneration (AMD), glaucoma, optic neuropathy, retinopathy, diabetic macular edema, diabetic retinopathy, retinal telangiectasia, or retinitis pigmentosa.

33. 31. The composition of claim 30, wherein the disease, disorder or condition is AMD, including AMD with neovascularization (wet AMD) and AMD with geographic atrophy (dry AMD).

34. A pre-filled syringe comprising the pharmaceutical depot according to any one of claims 1 to 29, which may or may not be sterilized by exposure to gamma radiation.

35. A method for producing a pharmaceutical depot according to any one of claims 1 to 29, comprising the steps of: a) silica particles comprising an oligopeptide or peptidomimetic compound or a pharma- ceutically acceptable salt thereof, the silica particles having a maximum diameter of 1000 μm or less and a molecular weight of the free base equivalent of the oligopeptide or peptidomimetic compound of less than 1,200 amu; and b) Silica hydrogel [0033] i) the silica hydrogel has a solids content of 50% by weight or less; ii) the silica particles comprise a polymerized alkoxysilane; iii) said depot formulation comprises up to 85% by weight of said silica particles; iv) The method, wherein the hydrogel composite is non-flowable and structurally stable upon static storage and is shear thinning upon application of shear stress by injection.

36. 36. The method of claim 35, further comprising combining the silica sol with a solution comprising the oligopeptide or peptidomimetic compound or a pharma- ceutically acceptable salt thereof, thereby producing a mixture comprising the silica sol and the oligopeptide or peptidomimetic compound or a pharma- ceutically acceptable salt thereof.

37. 37. The method of claim 36, further comprising combining tetraethyl orthosilicate (TEOS), water and hydrochloric acid, or tetraethyl orthosilicate (TEOS), water, hydrochloric acid and an organic co-solvent, thereby producing a silica sol.

38. 37. The method of claim 36, further comprising combining the oligopeptide or peptidomimetic compound, or a pharma- ceutically acceptable salt thereof, with water or water and an organic co-solvent, thereby creating a solution comprising the oligopeptide or peptidomimetic compound, or a pharma- ceutically acceptable salt thereof.

39. 37. The method of claim 36, wherein combining the silica sol with the solution comprising the oligopeptide or peptidomimetic compound or a pharma- ceutically acceptable salt thereof is performed in a continuous flow process.

40. 37. The method of claim 36, wherein combining the silica sol with the solution comprising the oligopeptide or peptidomimetic compound or a pharma- ceutically acceptable salt thereof comprises the use of an in-line mixer and / or the use of a pump.

41. 37. The method of claim 36, further comprising spray drying a mixture of the silica sol and a solution comprising the oligopeptide or peptidomimetic compound or a pharma- ceutically acceptable salt thereof, thereby forming silica particles.

42. 36. The method of claim 35, wherein the formulation is packaged in a syringe with or without a needle attached.

43. 36. The method of claim 35, further comprising sterilizing the depot formulation.

44. 44. The method according to claim 43, wherein sterilizing the depot formulation is performed by radiation sterilization.

45. 45. The method of claim 44, wherein the radiation sterilization comprises gamma radiation sterilization.

46. ​​(i) The silica particles comprise an oligopeptide or a pharma- ceutically acceptable salt thereof; The oligopeptide is elamipretide having the structure: 【Chemistry 6】 or a pharma- ceutically acceptable salt thereof, or The oligopeptide is Compound II having the structure: 【Chemistry 7】 or a pharma- ceutically acceptable salt thereof, or The oligopeptide is compound III having the structure: 【Chemistry 8】 or a pharma- ceutically acceptable salt thereof, or The oligopeptide has the structure: 【Chemistry 9】 or a pharma- ceutically acceptable salt thereof; (ii) the silica particles comprise a peptidomimetic compound or a pharma- ceutically acceptable salt thereof; The peptidomimetic compound is Compound I having the structure: 【Chemistry 10】 or a pharma- ceutically acceptable salt thereof.

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