CN101137370B - Liquid formulations for the treatment of a disease or condition - Google Patents

Abstract

Diseases and disorders associated with tissues in the body, including but not limited to intraocular tissues, can be effectively treated, prevented, inhibited, delayed in their occurrence, or regressed by administering therapeutic agents to these tissues. Described herein are liquid formulations that deliver a variety of therapeutic agents (including but not limited to rapamycin) to a subject for an extended period of time; a liquid formulation that forms a non-dispersed mass when placed in an aqueous medium of a subject; a non-dispersed mass-forming liquid formulation that forms a gel or gel-like substance in an aqueous medium; a liquid formulation comprising a therapeutic agent and a plurality of polymers; and methods of using the liquid formulations to deliver therapeutic agents to a subject over an extended period of time. Liquid formulations may be placed (including but not limited to, by intraocular or periocular administration) in an aqueous medium of a subject, or in the vicinity of the site of a disease or disorder to be treated in a subject. May be used to administer rapamycin to treat or prevent angiogenesis, choroidal neovascularization, or age-related macular degeneration, or wet age-related macular degeneration in a subject. The liquid formulation may comprise rapamycin or other therapeutic agent.

Description

Liquid formulations for the treatment of a disease or condition

technical field

Described herein are liquid formulations for treating, preventing, inhibiting, delaying the onset, or causing resolution of a disease or condition by delivering a therapeutic agent to a subject, including but not limited to, a human subject, including but not limited to A liquid formulation comprising a therapeutic agent is delivered to the eye of a human subject, including but not limited to, to treat age-related macular degeneration (AMD). Non-limiting examples of such liquid formulations include solutions, suspensions, and gel-in-situ formulations. the

Cross-reference with related applications

This application is related to U.S. Provisional Patent Application Serial No. 60/664,040, filed March 21, 2005 entitled "Liquid Formulations For Treatment Of Diseases Or Conditions," and filed March 21, 2005, entitled "In Situ Gelling Formulations And Liquid U.S. Provisional Patent Application Serial No. 60/664,306 for "Formulations For Treatment of Diseases Or Conditions," and U.S. Provisional Patent Application Serial No. 60/651,790, filed February 9, 2005, entitled "Formulations For Ocular Treatment," relate to and claim their priority, each material is incorporated herein by reference in its entirety for all purposes. the

technical background

The retina of the eye contains the light-sensing cones and rods. In the center of the retina is the macula, which is about 1/3 to 1/2 cm in diameter. The macula provides detailed vision (especially in the center (fovea)) because of the higher density of cones. Blood vessels, ganglion cells, inner nuclear layer and cells, and plexiform layer all move to one side (rather than resting on the cones), allowing light to take a more direct path to the cones. the

Subretina is the choroid, consisting of a set of blood vessels embedded in fibrous tissue and a dark pigmented epithelium covering the choroid layer. Choroidal blood vessels provide nourishment to the retina, particularly its visual cells. the

There are a variety of retinal diseases for which there is currently no treatment or for which current therapies are suboptimal. Retinal diseases such as uveitis (inflammation of the pigmented layers: sclera, ciliary body, and choroid), central retinal vein occlusive disease (CRVO), branch retinal vein occlusion (BRVO), macular degeneration, macular edema, proliferative diabetic retinopathy Lesions and retinal detachments are generally all retinal diseases that are difficult to treat with conventional therapies. the

Age-related macular degeneration (AMD) is the leading cause of severe vision loss in individuals older than 60 years in the United States. AMD occurs in an atrophic or less commonly exudative form. The atrophic form of AMD is also known as "dry AMD" and the exudative form of AMD is also known as "wet AMD". the

In exudative AMD, blood vessels grow from the choriocapillary layer through defects in Bruch's membrane (and in some cases the underlying retinal pigment epithelium). Serous or hemorrhagic exudate tissue escaping from these vessels causes fibrovascular scarring of the macula with collateral degeneration of the neuroretina, detachment and tearing of the retinal pigment epithelium, vitreous hemorrhage, and permanent loss of central vision. This process accounts for more than 80% of cases of significant vision loss in subjects with AMD. Current or upcoming treatments include laser photocoagulation, photodynamic therapy, treatment with VEGF antibody fragments, treatment with pegylated aptamers, and treatment with certain small molecule agents. the

Several studies have recently described the use of laser photocoagulation in the treatment of primary or recurrent neovascular injury associated with AMD (Macular Photocoagulation Study Groups (1991) in Arch. Ophthal. 109: 1220; Arch. Ophthal 109: 1232 ; Arch. Ophthal. 109:1242). Unfortunately, laser-treated AMD patients with subfoveal lesions experienced very rapid visual acuity loss (average 3 lines) at 3-month follow-up. In addition, two years after treatment, the treated eyes showed only a small improvement in vision compared to their untreated eyes (average 20/320 and 20/400, respectively). Another disadvantage of this method is the immediate deterioration of vision following surgery. the

Photodynamic therapy (PDT) is a type of light therapy -- including all treatments that use light to produce a beneficial response in a subject. At its best, PDT destroys unwanted tissue and spares normal tissue. Compounds known as photosensitizers are typically administered to the subject. Usually the photosensitizer itself has little or no effect on the subject. When light (usually from a laser) is directed at the tissue containing the photosensitizer, the photosensitizer is activated and begins to destroy the target tissue. Since the light directed at the subject is confined to a specific target area, PDT can be used to selectively target abnormal tissue, thereby sparing surrounding healthy tissue. PDT is currently used to treat retinal diseases such as AMD. PDT is currently the main method for treating subfoveal choroidal neovascularization in AMD patients (Photodynamic Therapy for Subfoveal Choroidal Neovascularization in Age Related Macular Degeneration with Verteporfin (TAP Research Group) Arch Ophthalmol.1999 117:1329-1345). the

Choroidal neovascularization (CNV) proves to be treatment-resistant in most cases. Conventional laser therapy can excise the CNV and help preserve vision without involving the center of the retina, but this is limited to about 10% of cases. Unfortunately, even with successful conventional laser photocoagulation, neovascularization recurs in 50%-70% of eyes (50% at 3 years and >60% at 5 years). (Macular Photocoagulation Study Group, Arch. Ophthalmol. 204:694-701 (1986)). Additionally, many subjects with CNV are not good candidates for laser therapy because the CNV is too large for laser therapy, or the location cannot be determined so the doctor cannot precisely target the laser. Although photodynamic therapy is used in up to 50% of new cases of subfoveal CNV, it has only a small benefit on the natural history and often delays the progression of vision loss rather than improving vision already reduced after subfoveal damage. PDT is neither preventative nor definitive. Each subject typically requires several PDT treatments, and in addition, certain subtypes of CNV do not progress as well as others. the

Accordingly, there remains a long felt need for methods, compositions and formulations useful for optimally preventing or significantly inhibiting choroidal neovascularization and for preventing and treating wet AMD. the

In addition to AMD, choroidal neovascularization has been associated with retinal diseases such as presumed ocular histoplasma syndrome, myopic degeneration, angioid striae, idiopathic central serous chorioretinopathy, retinal and/or choroidal inflammatory disease Diseases and eye injuries. Angiogenic impairment associated with neovascularization occurs in a variety of diseases, including diabetic retinopathy, venous occlusions, sickle cell retinopathy, retinopathy of prematurity, retinal detachment, ocular ischemia, and trauma. the

Uveitis is another retinal disorder that has proven refractory to existing therapies. Uveitis is a general term used to designate inflammation of any component of the uvea. The uvea of the eye consists of the sclera, ciliary body, and choroid. Inflammation of the overlying retina (called retinitis) or of the optic nerve (called optic neuritis) may occur with or without uveitis. the

Uveitis is most commonly anatomically classified as anterior, intermediate, posterior, or diffuse. Posterior uveitis means any of the various forms of retinitis, choroiditis, or optic neuritis. Divergent uveitis is inflammation that involves all parts of the eye, including the front, middle, and back structures. the

Symptoms and signs of uveitis can be mild and vary widely depending on the location and severity of the inflammation. In the case of posterior uveitis, the most common symptoms include the presence of suspended matter and decreased vision. Cells in the vitreous humor, white or yellow-white lesions in the retina and/or underlying choroid, exudative retinal detachment, retinal vasculitis, and optic nerve edema may also be present in subjects with posterior uveitis. the

Ophthalmic complications of uveitis can produce significant and irreversible vision loss, especially when unrecognized or inappropriately treated. The most common complications of posterior uveitis include retinal detachment, neovascularization of the retina, optic nerve, or sclera, and cystoid macular edema. the

Macular edema (ME) occurs if swelling, leaks and hard exudates are noted in the macula (the central 5% of the retina most critical for vision) in background diabetic retinopathy (BDR). Background diabetic retinopathy (BDR) usually consists of small retinal aneurysms caused by changes in retinal microcirculation. These small aneurysms are often the earliest visible change in retinopathy seen with ophthalmoscopy and appear as scattered red spots in the retina in which tiny weakened blood vessels have bulged. Visual findings during background diabetic retinopathy progress to cotton wool spots, intraretinal hemorrhages, fluid seepage from retinal capillaries, and retinal exudates. Increased vascular permeability also involves increased levels of local growth factors, such as vascular endothelial growth factor. The macula is rich in cones, the nerve endings upon which color perception and daytime vision depend. When increased retinal capillary permeability affects the macula, central or only the sides of the central visual field appear blurred, as if looking through cellophane. Vision loss can develop over a period of months and can be very disturbing due to the inability to focus clearly. ME is a common cause of several visual impairments. the

There have been attempts to use drugs to treat CNV and its associated diseases and conditions, as well as other conditions such as macular edema and chronic inflammation. For example, the use of rapamycin to inhibit CNV and wet AMD is described in US Application No. 10/665,203, which is incorporated herein by reference in its entirety. The use of rapamycin for the treatment of inflammatory diseases of the eye is described in US Patent No. 5,387,589, entitled Method of Treating Ocular Inflammation, inventor Prassad Kulkarni, assigned to the University of Louisville Research Foundation, the contents of which are incorporated herein by reference in their entirety. the

Particularly for chronic diseases, including those described herein, there is a great need for methods for delivering therapeutic agents to the eye (e.g., to the posterior segment for the treatment of conditions such as AMD, macular edema, proliferative retinopathy, and chronic inflammation). CNV in similar diseases) long-acting method. Formulations that deliver the therapeutic agent for extended periods of time are more comfortable and convenient for the subject, as the frequency of ocular injections of the therapeutic agent is reduced. the

Delivery of a therapeutic agent directly to the eye rather than systemic administration can be advantageous because the concentration of the therapeutic agent at the site of action is increased relative to the concentration of the therapeutic agent in the subject's circulatory system. Additionally, systemic delivery of therapeutic agents to treat posterior segment disease can have undesired side effects. Thus, local drug delivery can increase efficiency while reducing side effects and systemic toxicity. the

Summary of the invention

The methods, compositions and liquid formulations described herein allow for the delivery of a therapeutic agent to a subject (including but not limited to a human subject) or to the subject's eye. Described herein are methods, compositions, and liquid formulations for the prolonged delivery of various therapeutic agents useful for treating, preventing, inhibiting, delaying the onset of, or causing regression of various conditions or diseases that Diseases include, but are not limited to eye diseases or disorders. Liquid formulations include, but are not limited to solutions, suspensions, and gel-in-situ formulations. the

Described herein are methods, compositions, and liquid formulations for administering rapamycin to a human subject in an amount effective to treat, prevent, inhibit, delay the onset, or cause regression of wet AMD. the

As described in further detail in the "Detailed Description of the Invention" section, the methods, compositions and liquid formulations can also be used for delivery to a subject (including but not limited to a human subject) or to the eye of a human subject. A therapeutically effective amount of rapamycin is used to treat, prevent, inhibit, delay its onset, or cause its regression. In some variations, wet AMD is treated using the methods, compositions and liquid formulations. In some variations, the methods, compositions and liquid formulations are used for the prevention of wet AMD. In some variations, the methods and formulations described herein are used to prevent conversion of dry AMD to wet AMD. The methods, compositions and liquid formulations can also be used to deliver a therapeutically effective amount of rapamycin to a subject (including but not limited to a human subject) or to the eye of a subject for the treatment, prevention, inhibition of CNV , delay its onset or cause its resolution. In some variations, CNV is treated using the methods, compositions and fluid formulations. The methods, compositions and liquid formulations can also be used to deliver a therapeutically effective amount of rapamycin to a subject (including but not limited to a human subject) or to the subject's eye for treatment, prevention, inhibition of ocular Angiogenesis, delaying its onset, or causing its regression. In some variations, the methods, compositions and liquid formulations are used for the treatment of angiogenesis. Other diseases and conditions for which rapamycin can be used to treat, prevent, inhibit, delay their onset, or cause their regression are described in the "Diseases and Conditions" section of the "Detailed Description of the Invention". the

As described in further detail in the "Detailed Description of the Invention" section, the methods, compositions and liquid formulations can also be used for delivering a treatment to a subject (including but not limited to a human subject) or to the eye of a subject An effective amount of a therapeutic agent other than rapamycin is used to treat, prevent, inhibit, delay its onset, or cause its regression. In some variations, the methods, compositions and liquid formulations are used for the treatment of wet AMD. Therapeutic agents that can be used are described in detail in the "Therapeutic Agents" section. Such therapeutic agents include, but are not limited to, compounds that bind immunophilin. Compounds that bind immunophilin that may be used include, but are not limited to, the limus family of compounds further described in the "Therapeutic Agents" section herein, including rapamycin, SDZ-RAD, tacrolimus, everolimus ), pimecrolimus, CCI-779, AP23841, ABT-578 and derivatives, analogs, prodrugs, salts and esters thereof. The methods, compositions and liquid formulations can also be used to deliver a therapeutically effective amount of a therapeutic agent to a subject (including but not limited to a human subject) or to the subject's eye for treating, preventing, inhibiting CNV , delay its onset or cause its resolution. In some variations, the methods, compositions and liquid formulations are used for the treatment of CNV. The methods, compositions and liquid formulations can also be used to deliver a therapeutically effective amount of a therapeutic agent to a subject (including but not limited to a human subject) or to the subject's eye for treating, preventing, inhibiting ocular Angiogenesis, delaying its onset, or causing its regression. In some variations, the methods, compositions and liquid formulations are used for the treatment of angiogenesis. Other diseases and conditions that can be treated, prevented, inhibited, delayed onset, or caused to regress using therapeutic agents other than rapamycin are described in the "Diseases and Conditions" section of the "Detailed Description of the Invention". the

A liquid formulation described herein comprises a solution containing a therapeutic agent dissolved in a solvent. Generally any solvent having the desired effect in which the therapeutic agent is dissolved and which can be administered to a subject (including but not limited to a human subject) or to the eye of a subject can be used. Generally any concentration of therapeutic agent that has the desired effect can be used. In some variations, the formulation is an unsaturated, saturated or supersaturated solution. The solvent may be a pure solvent or may be a mixture of liquid solvent components. In some variations, the resulting solution is an in situ gelling formulation. The types of solvents and solutions that can be used are well known to those skilled in the art of such drug delivery techniques. the

The liquid formulations described herein can form non-divergent masses when placed in a rabbit eye, including but not limited to the vitreous humor of a rabbit eye. In some variations, the non-dispersed mass comprises a gel. In some variations, a liquid formulation comprises a therapeutic agent and multiple polymers. In some variations, one of the polymers is polyacrylate or polymethacrylate. In some variations, one of the polymers is polyvinylpyrrolidone. the

In some variations, the non-divergent mass comprises a depot formulation. In some variations, the non-divergent mass consists of the depot formulation. the

For liquid formulations that form a non-divergent mass, the non-divergent mass may generally be of any geometry or shape. A liquid formulation that forms a non-divergent mass may, for example, appear as a dense spherical mass when placed in the vitreous. In some variations, the liquid formulations described herein, when placed in the vitreous, form an opalescent or white semi-continuous or semi-solid non-diffusing mass relative to the medium in which they are placed. the

Liquid formulations can generally be administered in any volume that has the desired effect. In one method, a volume of the liquid formulation is administered to the vitreous, the liquid formulation being less than half the volume of the vitreous. the

Routes of administration for administration of liquid formulations that may be used include, but are not limited to (1) placement of liquid formulations by placement (including injection) into media (including, but not limited to, aqueous media in the body), including, but not limited to, intraocular or periocular injections ; or (2) oral liquid preparation. Liquid formulations may be administered systemically, including but not limited to the following routes of administration: rectal, vaginal, infusion, intramuscular, intraperitoneal, intraarterial, intrathecal, intrabronchial, intracisternal, epidermal, subcutaneous, intradermal, transdermal, Intravenous, intracervical, intraperitoneal, intracranial, intrapulmonary, intrathoracic, intratracheal, nasal, buccal, sublingual, oral, parenteral or spray or aerosolized using an aerosol propellant. In some variations, the liquid formulation is administered subconjunctivally. In some variations, the liquid formulation is administered intravitreally. the

The liquid formulations described herein can be delivered to any medium, including but not limited to an aqueous medium of a subject, including but not limited to a human subject. the

One liquid formulation described herein comprises a liquid formulation of rapamycin or other therapeutic agent. Liquid formulations may include solutions, suspensions, gel-in-situ formulations or emulsions. The droplets in the emulsion can generally be of any size, including but not limited to as large as about 5,000 nm. the

In some formulations described herein, a liquid formulation may comprise a therapeutic agent (including but not limited to rapamycin) and one or more solubilizers or solvents. In some variations, the solubilizer or solvent is glycerol, DMSO, DMA, N-methylpyrrolidone, ethanol, benzyl alcohol, isopropanol, polyethylene glycols of various molecular weights (including but not limited to PEG 300 and PEG 400) , or propylene glycol, or a mixture of one or more of these substances. the

In some formulations described herein, the liquid formulation includes hyaluronic acid. the

The liquid formulations described herein can deliver therapeutic agents over extended periods of time. Non-limiting examples of such extended release delivery systems are those that deliver a therapeutic agent to a subject (including, but not limited to, a human subject) or to a subject in an amount sufficient to maintain an effective amount over an extended period of time. An ocular system, said effective amount being capable of treating, preventing, inhibiting, delaying the onset of, or causing regression of a disease or condition in a subject. In some variations, liquid formulations are used to treat a disease or condition in a subject, including but not limited to a human subject. In some variations, the liquid formulation delivers the therapeutic agent for at least about one month, about two months, about three months, about six months, about nine months, or about twelve months. the

The liquid formulations described herein can deliver rapamycin or other therapeutic agents for extended periods of time. A non-limiting example of such an extended release delivery system is the delivery of rapamycin to a subject (including but not limited to a human subject) or to a subject for an extended period of time in an amount sufficient to maintain an effective amount. The eyes of the subject, the effective amount can treat, prevent, inhibit, delay the occurrence of wet age-related macular degeneration or cause its regression in the subject. In some variations, the liquid formulation is used to treat wet age-related macular degeneration for an extended period of time. In some variations, wet age-related macular degeneration is prevented for an extended period of time using the liquid formulation. In some variations, the conversion of dry AMD to wet AMD is prevented for an extended period of time using a liquid formulation. In one non-limiting example, the liquid formulation delivers rapamycin to a subject in an amount sufficient to treat, prevent, inhibit, delay the onset of, or cause regression of wet age-related macular degeneration (including but not limited to At least about three months, about six months, about nine months, or about twelve months in the vitreous, sclera, retina, choroid, macula, or other tissue of a human subject). In some variations, the level of rapamycin is sufficient to treat AMD. In some variations, the level of rapamycin is sufficient to prevent the development of wet AMD. the

Other extended time releases are described in the "Detailed Description of the Invention". the

Overview of drawings

Figures 1A-1C illustrate the formation of a non-dispersed mass following intravitreal injection of a liquid formulation into the eye, as is believed to occur in some variations. the

Figure 2 depicts the vitreous body (ng/ml), retina choroid (ng/ml) and Levels of rapamycin in sclera (ng/ml). the

Figure 3 depicts the vitreous body (ng/ml), retinal choroid (ng/ml) and sclera (ng/ml) of rabbit eyes on days 14, 35, 62 and 85 after subconjunctival injection of 5% rapamycin PEG 400 and ethanol solution ) levels of rapamycin. the

Figure 4 depicts the vitreous body (ng/ml), retina choroid (ng/ml) and sclera (ng/ml) of rabbit eyes after intravitreal injection of 5% rapamycin PEG 400 and ethanol solution at 14, 35, 62 and 90 days ) levels of rapamycin. Rapamycin levels (ng/ml) present in the vitreous 2 days after injection are also shown. the

Figure 5 depicts images of rabbit eyes 8 days after intravitreal injection of 10 μl (Figure 4A), 20 μl (Figure 4B) and 40 μl (Figure 4C) of 6% rapamycin in PEG400 suspension. the

Figure 6 depicts the vitreous body (ng/ml), retinal choroid tissue (ng/mg ) and levels of rapamycin in the sclera (ng/ml). the

Figure 7 depicts the vitreous (ng/ml), retinal choroidal tissue (ng/mg) and sclera (ng/mg) of rabbit eyes on days 14, 42, 63 and 91 after subconjunctival injection of 3% rapamycin in PEG 400 suspension levels of rapamycin. the

Figure 8 depicts the vitreous (ng/ml), retinal choroidal tissue (ng/mg) and sclera (ng/mg) of rabbit eyes after intravitreal injection of 3% rapamycin in PEG 400 suspension on days 14, 42, 63 and 91 Rapamycin levels in the vitreous as well as intravitreal levels 63 and 91 days after injection. the

Figure 9 depicts the vitreous (ng/ml), retinal choroidal tissue (ng/mg) and sclera (ng/mg) of rabbit eyes after subconjunctival injection of 2% rapamycin ethanol and PEG 400 solution on day 14, 42, 63 and 91 levels of rapamycin. the

Figure 10 depicts the levels of rapamycin in retinal choroid tissue (ng/mg) and sclera (ng/mg) in rabbit eyes on day 14, 42, 63 and 91 after intravitreal injection of 2% rapamycin ethanol and PEG 400 solution . the

Figure 11 depicts the levels (ng/ml) of rapamycin in the vitreous of rabbit eyes on days 63 and 91 after intravitreal injection of 2% rapamycin ethanol and PEG 400 solution. the

Figure 12 depicts the concentration of rapamycin in the vitreous (ng/ml) of rabbit eyes at 5, 30, 60, 90 and 120 days after subconjunctival injection of 20 μl, 40 μl and 60 μl doses of 2% rapamycin in ethanol and PEG 400 solution. level. the

Figure 13 depicts rapamycin in the retina and choroid (ng/mg) of rabbit eyes on days 5, 30, 60, 90 and 120 after subconjunctival injection of 20 μl, 40 μl and 60 μl doses of 2% rapamycin in ethanol and PEG 400 s level. the

Figure 14 depicts 5, 30, 60, 90 and 120 days after intravitreal injection of 20 μl and 40 μl doses of 2% rapamycin in ethanol and PEG 400 and 100 μl dose of 0.4% rapamycin in ethanol and PEG 400 Levels of rapamycin in the vitreous (ng/ml) of rabbit eyes. the

Figure 15 depicts 5, 30, 60, 90 and 120 days after intravitreal injection of 20 μl and 40 μl doses of 2% rapamycin in ethanol and PEG 400 and 100 μl dose of 0.4% rapamycin in ethanol and PEG 400 Levels of rapamycin in retinal choroidal tissues (ng/mg) of rabbit eyes. the

Figure 16 depicts the vitreous humor (ng/ml) of rabbit eyes 5 and 14 days after subconjunctival injection of a single 10 μl dose, a single 60 μl dose, two 30 μl doses and three 30 μl doses of 2% rapamycin in ethanol and PEG 400 levels of rapamycin. the

Figure 17 depicts subconjunctival injection of a single 10 μl dose, a single 60 μl dose, two 30 μl doses and three 30 μl doses of 2% rapamycin in ethanol and PEG 400 solution for 5 and 14 days in rabbit eyes with retinal choroid tissue (ng/ mg) levels of rapamycin. the

Figure 18 depicts Rapamycin in the vitreous (ng/ml) of rabbit eyes 5, 14 and 30 days after subconjunctival injection of a single 10 μl dose, a single 30 μl dose and three 30 μl doses of 3% rapamycin in PEG 400 suspension prime level. the

Figure 19 depicts subconjunctival injection of a single 10 μl dose, a single 30 μl dose, and three 30 μl doses of 3% rapamycin in PEG 400 suspension 5, 14, and 30 days after the retinal choroidal tissue (ng/mg) of rabbit eyes. Pamycin levels. the

Figure 20 depicts intravitreal injection of 10 μl of 0.2% rapamycin in ethanol and PEG 400, 10 μl of 0.6% rapamycin in ethanol and PEG 400, and 10 μl of 2% rapamycin in ethanol and PEG 400 Levels of rapamycin in retinal choroid tissue (ng/mg) of rabbit eyes on day 5, 30 and 90. the

Figure 21 depicts intravitreal injection of 10 μl of 0.2% rapamycin in ethanol and PEG 400, 10 μl of 0.6% rapamycin in ethanol and PEG 400 and 10 μl of 2% rapamycin in ethanol and PEG 400 Levels of rapamycin in the vitreous (ng/ml) of rabbit eyes on day 5, 30 and 90. the

Figure 22 depicts subconjunctival injection of 40 μl of 2% rapamycin in ethanol and PEG 400 solution 1, 4, 7, 11, 14, 21, 28, 35, 54 and 56 days after rabbit eye aqueous humor (ng/ml), Rapamycin levels in cornea (ng/mg) and retinal choroidal tissue (ng/mg). the

Detailed description of the invention

Described herein are compositions, liquid formulations, and methods related to the delivery of therapeutic agents to a subject (including but not limited to a human subject) or to the eye of a subject. These compositions, liquid formulations, and methods are useful for treating, preventing, inhibiting, delaying the onset, or causing regression of ocular diseases and disorders, including, but not limited to, posterior segment diseases or disorders, including, but not limited to, choroidal neovascularization macular degeneration, age-related macular degeneration (including wet AMD and dry AMD), retinal angiogenesis, uveitis, and other retinal proliferative disorders. In some variations, the compositions, liquid formulations, and methods are used to treat the ocular diseases or conditions described above. the

Described herein are (1) therapeutic agents that can be delivered to a subject (including, but not limited to, a human subject) or to the eye of a subject using the compositions, liquid formulations, and methods described herein; (2) therapeutic agents that can be delivered by Diseases and conditions that treat, prevent, inhibit, delay onset, or cause regression; (3) liquid formulations that can be used to deliver therapeutic agents; (4) routes of administration that deliver liquid formulations; (5) prolonged delivery of therapeutic agents, so Such therapeutic agents include, but are not limited to, rapamycin; and (6) describe the treatment of CNV and wet AMD by administering the composition and liquid formulation to a subject (including, but not limited to, Human subjects) or ocular delivery of rapamycin to subjects. the

As used herein, the term "about" refers to the level of accuracy obtained when using the methods described herein (eg, in the Examples). However, a defined amount of a formulation component by "about" means 90-110% of the stated amount. the

therapeutic agent

In general, any compound and composition known or still to be discovered that is useful for treating, preventing, inhibiting, delaying the onset of, or causing regression of, the diseases and conditions described herein may be used in the compositions, liquid formulations described herein and methods of treatment. the

Therapeutic agents that may be used include compounds that act by binding members of the immunophilin family of cellular proteins. Such compounds are known as "immunophilin-binding compounds". Compounds that bind immunophilin include, but are not limited to, the "limus" family of compounds. Examples of limus compounds that can be used include, but are not limited to, cyclophilins and FK506 binding proteins (FKBPs), including sirolimus (rapamycin) and its water-soluble analog SDZ-RAD (Novartis), TAFA -93 (Isotechnika), tacrolimus, everolimus, RAD-001 (Novartis), pimecrolimus, temsirolimus, CCI-779 (Wyeth), AP23841 (Ariad), AP23573 (Ariad), and ABT-578 (Abbott Laboratories). Limus compound analogs and derivatives that may be used include, but are not limited to, those described in US Patent Nos. 5,527,907; 6,376,517 and 6,329,386 and US Patent Application No. 09/950,307, each of which is incorporated herein by reference in its entirety. Therapeutic agents also include analogs, prodrugs, salts and esters of the limus compounds. the

The terms rapamycin, rapa and sirolimus are used interchangeably herein. the

Other useful rapamycin derivatives include, but are not limited to, 7-epi-rapamycin, 7-thiomethyl-rapamycin, 7-epi-trimethoxyphenyl-rapamycin, 7 -Epi-thiomethyl-rapamycin, 7-demethoxy-rapamycin, 32-demethoxy-rapamycin, 2-demethyl-rapamycin, rapamycin Mono- and diester derivatives of ketones, rapamycin 27-oxime; 42-oxo analogs of rapamycin; bicyclic rapamycin; rapamycin dimer; rapamycin A Silane ethers; aryl sulfonates and sulfamates of rapamycin, mono- and diesters at positions 31 and 42, 30-desmethoxyrapamycin, and Vezina et al., "Rapamycin (AY-22 , 989), A New Antifungal Antibiotic.I.Taxonomy Of The Producing Streptomycete And Isolation Of The Active Principle "J.Antibiot.(Tokyo) 28:721-726(1975); Sehgal et al., "Rapamycin(AY-22,989) , A New Antifungal Antibiotic.II.Fermentation, Isolation AndCharacterization "J.Antibiotic.(Tokyo) 28:727-732(1975); Sehgal et al., Demethoxyrapamycin (AY-24,668), A New Antifungal Antibiotic"J.Antibiotic. (Tokyo) 36:351-354 (1983); and Paiva et al., "Incorporation Of Acetate, Propionate, And Methionine Into Rapamycin By Streptomyceteshygroscopicus" J Nat Prod 54:167-177 (1991), WO 92/05179, EP 467606, Caufield et al., "Hydrogenated Rapamycin Derivatives" U.S. Patent No. 5,023,262; Kao et al., "Bicyclic Rapamycins" U.S. Patent No. 5,120,725; Kao et al., "Rapamycin Dimers" U.S. Patent No. 5,120,727; Failli et al., "SilylEthers Of Rapamycin "U.S. Patent No. 5,120,842; Failli et al., "Rapamycin42- Sulfonates And 42-(N-carboalkoxy) Sulfamates Useful As Immunosuppressive Agents" U.S. Patent No. 5,177,203; Nicolaou et al., "Total Synthesis Of Rapamycin" J.Am.Chem.Soc.115:4419-4420 (1993); Romo et al., "Total Synthesis Of Rapamycin" Of(-)Rapamycin Using An Evans-Tishchenko Fragment Coupling” J.Am.Chem.Soc.115:7906-7907 (1993); and Hayward et al., “Total Synthesis Of Rapamycin Via A NovelTitanium-Mediated Aldol Macrocyclization Reaction” J.Am. . Chem. Soc., 115: 9345-9346 (1993), each of which is incorporated herein by reference in its entirety. the

Compounds of the Limus family are useful in compositions, liquid formulations and methods for treating, preventing, inhibiting, delaying the onset or regression of angiogenesis-mediated ocular diseases and conditions, including choroidal neovascularization. The limus family of compounds are useful for preventing, treating, inhibiting, delaying the onset or regressing of AMD, including wet AMD. Rapamycin and rapamycin derivatives and analogs are useful for preventing, treating, inhibiting, delaying the onset or regression of angiogenesis-mediated ocular diseases and conditions, including choroidal neovascularization. Rapamycin can be used to prevent, treat, inhibit, delay the occurrence of AMD (including wet AMD) or make it regress. In some variations, members of the limus family of compounds or rapamycin are used to treat wet AMD or angiogenesis-mediated ocular diseases and conditions, including choroidal neovascularization. the

Other therapeutic agents that may be used include those disclosed in the following patents and publications, each of which is incorporated herein by reference in its entirety: PCT Publication WO 2004/027027, published April 1, 2004, entitled Method of inhibiting choroidal neovascularization, assigned to Trustees of the University of Pennsylvania; U.S. Patent No. 5,387,589, issued February 7, 1995, and entitled Method of Treating Ocular Inflammation, inventor Prassad Kulkarni, awarded to University of Louisville Research Foundation; U.S. Patent No. 6,376,517, 2003 Published on April 23, 2004, entitled Pipecolic acid derivatives for vision and memory disorders, transferred to GPI NIL Holdings, Inc; PCT Publication WO 2004/028477, published on April 8, 2004, entitled Method subretinal administration of therapeutics including steroids: method for localizing pharmacodynamic action at the choroid and retina; and related methods for treatment and or prevention of retinal diseases, assigned to Innorx, Inc; U.S. Patent No. 6,416,777, filed July 9, 2002, and entitled Ophthalmic drug Uni delivery al device, assigned to Alcont L. ; U.S. Patent No. 6,713,081, issued March 30, 2004, entitled Ocular therapeutic agent delivery device and methods forming and using such devices, assigned to the Department of Health and Human Services; U.S. Patent No. 5,100,899, issued March 31, 1992, Entitled Methods of inhibiting transplant rejection in mammals using rapamycin and derivatives and prodrugs thereof. the

Other therapeutic agents that may be used include pyrrolidines, dithiocarbamates (NFκB inhibitors); squalamine; TPN 470 analogs and fumagillin; PKC (protein kinase C) inhibitors; Tie-1 and Tie-2 kinase inhibitors; VEGF receptor kinase inhibitors; proteosome inhibitors such as Velcade ^TM (bortezomib) for injection; ranibuzumab (Lucentis ^TM ) and other antibodies against the same target; pegaptanib (Macugen ^TM ); Protein receptor antagonists, such as cyclic peptide antagonists of vitronectin receptor-type integrins; alpha-v/beta-3 integrin antagonists; alpha-v/beta-1 integrin antagonists; thiazolidines Diketones, such as rosiglitazone or troglitazone; interferons, including gamma-interferon or interferons targeted to CNVs through the use of dextran and metal coordination; pigment epithelium-derived factor (PEDF); endostatin; angiostatin; tumistatin; canstatin; anecortave acetate; acetonide; triamcinolone; tetrathiomolybdate; RNA silencing of angiogenic factors or RNA interference (RNAi), including ribozymes targeting VEGF expression; Accutane ^TM (13-cis retinoic acid); ACE inhibitors, including but not limited to quinopril, captopril, and perindozril; mTOR inhibition (target of rapamycin in mammals); 3-aminothalidomide; pentoxifylline; 2-methoxyestradiol; colchicum AMG-1470; cyclooxygenase inhibitors, such as nepafenac, rofecoxib, diclofenac, rofecoxib, NS398, celecoxib, Vioxx and (E)-2-alkyl-2(4-methylsulfonylphenyl)-1-styrene; t-RNA synthase modulator; metalloproteinase 13 inhibitor; acetylcholinesterase inhibitor Potassium channel blocker; endorepellin; purine analogue of 6-thioguanine; cycloperoxide ANO-2; (recombinant) arginine deiminase (deiminase); epigallocatechin- 3-gallate (epigallocatechin-3-gallate); cerivastatin (cerivastatin); suramin analogue; VEGF trap molecule; apoptosis inhibitor; Visudyne ^{T M} , snET2 and other photosensitizers, which can be used in photodynamic therapy (PDT); hepatocyte growth factor inhibitors (growth factor antibodies or their receptors, small molecule inhibitors of c-met tyrosine kinase, cutoffs of HGF The short form is like HK4).

Other therapeutic agents that may be used include anti-inflammatory agents, including but not limited to non-steroidal anti-inflammatory agents and steroidal anti-inflammatory agents. In some variations, active agents that may be used in liquid formulations are ace-inhibitors, endogenous cytokines, agents that affect basement membrane, agents that affect endothelial cell growth, adrenergic agonists or blockers, Cholinergic agonists or blockers, aldose reductase inhibitors, analgesics, anesthetics, antiallergics, antibacterials, antihypertensives, pressors, antiprotozoals, antihypertensives Viral, antifungal, antiinfective, antineoplastic, antimetabolite and antiangiogenic agents. the

Steroid treatments that may be used include, but are not limited to, 21-pregnenolone acetate, alclometasone, algestone, amcinonide, beclomethasone, betamethasone ( betamethasone), budesonide, chloroprednisone, clobetasol, clobetasone, clocortolone, cloprednol, Corticosterone, cortisone, cortivazol, deflazacort, desonide, desoximetasone, dexamethasone ), diflorasone, diflucortolone, difluprednate, enoxolone, fluazacort, flucloronide, Flumethasone, flunisolide, fluocinolone acetonide, fluocinonide, fluocortin butyl, fluocortolone, fluorometholone (fluorometholone), fluperolone acetate, fluprednidene acetate, fluprednisolone, flurandrenolide, fluticasone propionate, flu formocortal, halcinonide, clobetasol propionate, halometasone, halopredone acetate, hydrocortamate, hydrocortisone hydrocortisone, loteprednol ethyl carbonate, mazipredone, medrysone, meprednisone, methylprednisolone ednisolone), mometasone furoate, paramethasone, prednicarbate, prednisolone, prednisolone-25-diethylamino-acetate ( prednisolone 25-diethylamino-acetate), prednisolone sodium phosphate, prednisone, prednisolone valerate, prednylidene, limar rimexolone, tixocortol, triamcinolone, triamcinolone acetonide, triamcinolone benetonide, triamcinolone hexacetonide, and any derivative. the

In some variations, cortisone, dexamethasone, fluocinolone, hydrocortisone, methylprednisolone, prednisolone, prednisone, and triamcinolone and their derivatives may be used. Liquid formulations may contain a combination of two or more steroid therapeutics. the

In one non-limiting example, the steroid therapeutic agent may comprise from about 0.05% to about 50% by weight of the liquid formulation. In another non-limiting example, the steroid constitutes the liquid formulation by weight from about 0.05% to about 10%, between about 10% to 20%, between about 30% to about 40%, or from about 40% to about Between 50%. the

Other non-limiting examples of therapeutic agents that may be used include, but are not limited to, anesthetics, analgesics, cell transport/migration inhibitors such as colchicine, vincristine vincristine, cytochalasin B, and related compounds; carbonic acid Anhydrase inhibitors such as acetazolamide, methazolamide, dichlorphenamide, diamox and neuroprotective agents such as nimodipine and related compounds; antibiotics such as tetracyclines, Aureomycin, bacitracin, neomycin, polymyxin, gramicidin, cephalexin, oxytetracycline, chloramphenicol, rifampicin, ciprofloxacin, aminoglycoside, gentamicin, erythromycin Penicillins and penicillins, quinolones, ceftazidime, vancomycine imipeneme; antifungals such as amphotericin B, cloconazole, ketoconazole, and miconazole; antibacterials such as sulfonamides sulfonamides, sulfadiazine, sulfacetamide, sulfamethizole, and sulfamethizole sulfisoxazole, nitrofurazone, and sodium propionate; antiviral agents such as idoxuridine, trifluorothymidine, trifluorouridine, acyclovir, gancixi Ganciclovir, cidofovir, interferon, DDI, AZT, foscamet, vidarabine, irbavirin, protease inhibitors and anti-CMV agents; Allergic agents such as sodium cromoglycate, antazoline, methapyriline, chlorpheniramine, cetirizine, pyrilamine, and pheniramine (prophenpyridamine); synthetic glucocorticoids and mineralocorticoids and more prevalent hormone forms derived from cholesterol metabolism (DHEA, progesterone, estrogen); nonsteroidal anti-inflammatory drugs such as salicylates, Indomethacin, ibuprofen, diclofenac, flurbiprofen, piroxicam and COX2 inhibitors; antineoplastic agents such as carmustine, cisplatin ( cisplatin), fluorouracil; doxorubicin, asparaginase, azacitidine, azathioprine, bleomycin, busulfan, carboplatin, carlimus Chlorambucil, cyclophosphamide, cyclosporine, cytarabine, dacarbazine, dactinomycin, daunorubicin ), doxorubicin, estramustine, etoposide, etretinate, filgrastin, floxuridine, fluoride Fludarabine, fluorouracil, florxymesterone, flutamide, goserelin, hydroxyurea, ifosfamide, leuprolide, Imidazole (lev amisole), limustine, nitrogen mustard, melphalan, mercaptopurine, methotrexate, mitomycin, mitotane, Pentostatin, pipobroman, plicamycin, procarbazine, sargramostin, streptozocin, tamox Tamoxifen, taxol, teniposide, thioguanine, uracil mustard, vinblastine, vincristine, and vindesine ); immune drugs such as vaccines and immunostimulants; insulin, calcitonin, parathyroid hormone and peptide and vasopressin hypothalamic releasing factor; beta-adrenergic blocking agents such as timolol (timolol), levo Levobunolol and betaxolol; cytokines, interleukins and growth factors, epidermal growth factor, fibroblast growth factor, platelet-derived growth factor, transforming growth factor beta, ciliary neurotrophic growth factor , glial cell-derived neurotrophic factor, NGF, EPO, PLGF, brain nerve growth factor (BNGF), vascular endothelial growth factor (VEGF) and monoclonal antibodies or fragments thereof against such growth factors; anti-inflammatory agents such as hydrocortisone Dexamethasone, dexamethasone, fluocinolone, prednisone, prednisolone, methylprednisolone, fluorometholone, betamethasone, and triamcinolone; decongestants such as phenylephrine, namethazole Naphazoline and tetrahydrazoline; miotics and anticholinesterases such as pilocarpine, carbachol, di-isopropylfluorophosphates, diethoxyphosphorylthiocholine iodide Phospholine iodine and demecarium bromide; mydriatics such as atropine sulfate, cyclopentolate, homatropine, scopolamine, tropicamide (tropicamide), eucatropine; sympathomimetic agents such as epinephrine and vasoconstrictors and vasodilators, anticoagulants (anticlotting agents) such as heparin, antifibrin Alcohol, fibrinolysin, anticlotting activase, antidiabetic agents including acetohexamide, chlorpropamide, glipizide, gliben Urea (glyburide), tolazamide, tolbutamide, insulin and aldose reductase inhibitors, hormones, peptides, nucleic acids, sugars, lipids, glycolipids, glycoproteins and other macromolecules , including endocrine hormones such as pituitary hormone, insulin, insulin-related growth factor, thyroxine, growth hormone; heat shock proteins; immune response modifiers such as muramyl dipeptide, cyclosporin, interferon (including α-, beta- and gamma-interferons), interleukin 2, cytokines, FK506 (an epoxy-pyrido-oxazepine-tetraketone, also known as tacrolimus), tumor Necrosis factor, pentostatin, thymopentin, transforming factor β2, erythropoetin; anti-neoplastic proteins (such as anti-VEGF, interferon), antibodies (monoclonal, polyclonal, humanized, etc. ) or antibody fragments, oligomeric aptamers, aptamers and gene fragments (oligonucleotides, plasmids, ribozymes, small interfering RNA (SiRNA), nucleic acid fragments, peptides), immunomodulators such as endoxan, sand Lilidomide, tamoxifen; antithrombotics and vasodilators such as rtPA, urokinase, plasmin; nitric oxide donors, nucleic acids, dexamethasone, cyclosporine A, azathioprine, buquine Brequinar, gusperimus, 6-mercaptopurine, mizoribine, rapamycin, tacrolimus (FK-506), folate analogs (e.g. edatrexate, methotrexate, piritrexim, pteropterin, Tomudex , trimetrexate), purine analogs (e.g. cladribine, fludarabine, 6-mercaptopurine, thiamiprine, thiaguanine), pyrimidine analogs (e.g. Ancitabine, azacitidine, 6-azuridine, carmofur, cytarabine, doxifluridine, emitefur, behenaridine (enocitabine), fluorouracil deoxynucleoside, fluorouracil, gemcitabine, tegafur), fluocinolone, triaminolone, anecorta, fluorometholone, medrysone, and prednisolone. In some variations, the immunosuppressant is dexamethasone. In some variations, the immunosuppressant is cyclosporin A.

In some variations, formulations comprise a combination of one or more therapeutic agents. the

Other non-limiting examples of therapeutic agents that may be used in the formulations described herein include antibacterial antibiotics, aminoglycosides (such as amikacin, apramycin, arbekacin) , bambermycin (bambermycins), butirosin (butirosin), dibekacin (dibekacin), dihydrostreptomycin (dihydrostreptomycin), astemicin (fortimicin(s)), gentamicin, Isepamicin, kanamycin, micronomicin, neomycin, neomycin undecylenate, netilmicin, paromomycin ( paromomycin), ribostamycin, sisomicin, spectinomycin, streptomycin, tobramycin, trospectomycin), amphenicols (such as azidamfenicol, chloramphenicol, florfenicol, thiamphenicol), ansamycins (such as rifamide, rifampicin, rifamycin sodium ( rifamycin sv), rifapentine, rifaximin), beta-lactams (e.g. carbacephems (e.g. loracarbef), carbapenems (e.g. biapenem, imipenem, meropenem, panipenem), cephalosporins (eg, cefaclor, cefadroxil) , cefamandole, cefatrizine, cefazedone, cefazolin, cefcapene pivoxil, cefclidin, cefdinir , cefditoren, cefepime, cefetamet, cefixime, cefinenoxime, cefodizime, cefonicid, cefoperazone ), ceforanide, cefotaxime, cefotiam otiam), cefozopran, cefpimizole, cefpiramide, cefpirome, cefpodoxime proxetil, cefprozil, cefroxadine , cefsulodin, ceftazidime, cefteram, ceftezole, ceftibuten, ceftizoxime, ceftriaxone, cefuroxime Cefuroxime, cefuzonam, cephacetrile sodium, cephalexin, cephaloglycin, cephaloridine, cephalosporin, cephalothin ( cephalothin, cephapirinsodium, cephradine, pivcefalexin), cephamycins (eg, cefbuperazone, cefinetazole, cefminox, cefotetan, cephalexin cetin), monoamides (eg aztreonam, carumonam, tigemonam), oxacephems (flomoxef, Latamoxef (moxalactam)), penicillins (such as nitrogen amdinocillin, nitrogen amdinocillin pivoxil, amoxicillin, ampicillin, apalcillin, aspoxicillin, azidocillin, azlocillin, bacampicillin , benzylpenicillinic acid, benzylpenicillin sodium, carbenicillin, carindacillin, clometocillin, cloxacillin, cyclocillin ( cyclacillin, dicloxacillin, epicillin, fenbenicillin, floxacillin, hetacillin, lenampicillin, metampicillin ), methicillin sodium, mezlocillin, nafcillin sodium, oxacillin, penamecillin, penethamate hydroxide , penicillin g benethamine, benzathine penicillin G (penicillin g benzathine), penicillin g diphenylmethylamine, penicillin g calcium, penicillin g penicillin g, penicillin g potassium (penicillin g potassium), procaine g procaine), penicillin N (penicillin n), penicillin O (penicillin o), penicillin V (penicillin v), benzathine penicillin V (penicillin v benzathine), penicillin v hydrabamine, penicillin (penimepicycline), phenethicillin potassium, piperacillin, pivampicillin, propicillin, quinacillin, sulbenicillin, Sultacillin (sultamicillin), phthalimillin ( talampicillin), temocillin, ticarcillin), ritipenem, lincosamides (eg, clindamycin, lincomycin), macrocyclic Lactones (eg, azithromycin, carbomycin, clarithromycin, dirithromycin, erythromycin, erythromycin acistrate , erythromycin estolate, erythromycin glucoheptonate, erythromycin lactobionate, erythromycin propionate, erythromycin stearate stearate), josamycin, leucomycins, midecamycin, miokamycin, oleandomycin, primycin, rotamycin Rokitamycin, rosaramicin, roxithromycin, spiramycin, troleandomycin), polypeptides (such as amphomycin, bacitracin ), capreomycin, colistin, enduracidin, enviomycin, fusafungine, gramicidin S, gramicidin, rice Cardamycin, polymyxin, pristinamycin, ristocetin, teicoplanin, thiostrepton, tuberactinomycin, short Bacitracin, gramicillin, vancomycin, puromycin, virginiamycin, zinc bacitracin), tetracyclines (eg, apicycline, chlortetracycline ), clomocycline, demeclocycline, doxycycline, guanmethycycline ( Guamecycline), lymecycline, meclocycline, methacycline, minocycline, oxytetracycline, penicillin, pipacycline, rolitetracycline, sancycline, tetracycline) and others (eg, cycloserine, mupirocin, tuberin); synthetic antibacterials, 2, 4-Diaminopyrimidines (e.g. brodimoprim, tetroxoprim, trimethoprim), nitrofurans (e.g. furaltadone, fluazolium chloride ( furazolium chloride), nifuradene, nifuratel, nifurfoline, nifurpirinol, nifurprazine, nifurateinol ( nifurtoinol), nitrofurantoin), quinolones and analogues (e.g. cinoxacin, ciprofloxacin, clinafloxacin, difloxacin, enoxacin (enoxacin), fleroxacin, flumequine, grepafloxacin, lomefloxacin, miloxacin, nadifloxacin, nalidixic acid acid), norfloxacin, ofloxacin, oxolinic acid, pazufloxacin, pefloxacin, pipemidic acid, piromidic acid ( piromidic acid), rosoxacin, rufloxacin, sparfloxacin, temafloxacin, tosufloxacin, trovafloxacin )), sulfonamides (such as acetylsulfamethoxypyrazine, benzylsulfamide, chloramine-b, chloramine-t, dichloram inet), N2-formylsulfisomidine (n2-formylsulfisomidine), n4-β-d-glucosylsulfonamide, mafenide, 4′-(methylsulfamoyl)sulfanilanilide, norpro Noprylsulfamide, Phthalylsulfacetamide, Phthalylsulfathiazole, Salazosulfadimidine, Succinylsulfathiazole, Sulfabenzamide, Sulfacetamide, Sulfonamide sulfachlorpyridazine, sulfachrysoidine, sulfacytine, sulfadiazine, sulfapentene, sulfadimethoxine, sulfadoxine, Sulfaethidole, sulfaguanidine, sulfaguanol, sulfalene, sulfaloxic acid, sulfamerazine, sulfameter ), sulfamethazine, sulfamethizole, sulfamethomidine, sulfamethazine sulfamethoxazole, sulfamethoxypyridazine, sulfametrole, sulfamidochrysoidine, sulfamethoxypyridazine Sulfamoxole, sulfanilamide, 4-sulfanilamidosalicylic acid, n4-sulfanilylsulfanilamide, sulfanilylurea, n-sulfonyl-3,4-propaneglutamine, sulfonamide sulfanitran, sulfaperine, sulfaphenazole, sulfaproxyline, sulfapyrazine, sulfapyridine, sulfasomizole , sulfasymazine, sulfathiazole, sulfathiourea, sulfatolamide, sulfisomidine, sulfiso sulfisoxazole), sulfones (such as acedapsone, acediasulfone, acetosulfone sodium, dapsone, diathymosulfone ), glucosulfone sodium, solasulfone, succisulfone, sulfanilic acid, sulfanyl benzylamine, sulfanilamide Sodium sulfoxone (sulfoxone sodium, thiazolsulfone) and others (such as clofoctol, hexedine, methenamine, methenamine citrate methenamineanhydromethylene-citrate, methenamine hippurate, methenamine mandelate, methenamine sulfosalicylate, nitroxoline ), taurolidine, xibomol), antifungal antibiotics, polyenes (such as amphotericin B, candicidin, dermostatin , filipin, fungichromin, hachimycin, hamycin, lucensomycin, mepartricin, natal Natamycin, nystatin, pecilocin (perimycin), azaserine, griseofulvin, oligomycins, undecylenic acid Neomycin undecylenate, pyrrolnitrin, siccanin, tubercidin, viridin, synthetic antifungals, allylamines (eg, butycin butenafine, naftifine, terbinafine), imidazoles (such as bifonazole, butoco nazole, chlordantoin, chlormidazole, cloconazole, clotrimazole, econazole, enilconazole, fentecan Fenticonazole, flutrimazole, isoconazole, ketoconazole, lanoconazole, miconazole, omoconazole, Oxiconazole nitrate, sertaconazole, sulconazole, tioconazole), thiocarbamates (such as tolciclate, trol tolindate, tolnaftate), triazoles (e.g. fluconazole, itraconazole, saperconazole, terconazole), acridine acrisorcin, amorolfine, biphenamine, bromosalicylchloranilide, buclosamide, calcium propionate, chlorphenesin (chlorphenesin), ciclopirox, cloxyquin, coparaffinate, diamthazoledihydrochloride, exalarnide, flucytosine, amclofenthiazole ( halethazole, hexetidine, loflucarban, nifuratel, potassium iodide, propionic acid, pyrithione, salicylanilide, sodium propionate, Sulbentine, tenonitrozole, triacetin, ujothion, undecylenic acid, zinc propionate, antineoplastic agents, antibiotics and the like (eg aclacinomycins, actinomycin f1, anthramycin, Azaserine, bleomycins, cactinomycin, carubicin, carzinophilin, chromomycins, actinomycetes D (dactinomycin), daunorubicin (daunorubicin), 6-diazo-5-oxo-L-norleucine, doxorubicin, epirubicin (epirubicin), idarubicin (idarubicin) ), menogaril, mitomycins, mycophenolic acid, nogalamycin, olivomycins, peplomycin, pirarubicin, plicamycin, porfiromycin, puromycin, streptonigrin, streptozocin, tuberculosis tubercidin, zinostatin, zorubicin), antimetabolites (such as folate analogs (such as denopterin, edatrexate, methotrexate) methotrexate, piritrexim, pteropterin, Tomudex , trimetrexate), purine analogs (such as cladribine, fludarabine, 6-mercaptopurine, thiamiprine, thioguanine), pyrimidine analogs ( For example, ancitabine, azacitidine, 6-azuridine, carmofur, cytarabine, doxifluridine, emitefur, enoxitabine ( enocitabine), floxuridine, fluorouracil, gemcitabine, tagafur, anti-inflammatory agents, steroidal anti-inflammatory agents, acetoxypregnenolone, alclometasone, alclometasone Progesterone (algestone), amcinonide, beclomethasone, betamethasone, budesonide, cprednisone, clobetasol, clobetasone, clocotorone, cprednisol, corticosterone, Cortisone, Cortisone, Deflazacort, Desonide, Dexamethasone, Dexamethasone, Diflurasone, Diflucorone, Difluprednate, Glycyrrhetinic Acid, Fluoride Fluazacort, Fluazacort, Fluordiclosone, Diflumethasone, Flunisolide, Fluocinonide, Fluocinonide Acetate, Fluoxetyl, Fluocortolone, Fluorometholone, Meflolon Acetate, Fluoromethyl Acetate Trichosolone, fluprednisolone, fludroxycortisone, fludexone propionate, fluformylone, halcinonide, halbetasol propionate, halometasone, haloprednisolone acetate, hydrocortisone, hydrocortisone Cortisone, loteprednol ethyl carbonate, malpredone, medrison, methylprednisone, methylprednisolone, mometasone furoate, paramethasone, prednicate, prednisolone, Prednisolone 25-diethylamino-acetate, prednisolone sodium phosphate, prednisone, prednisolone valerate, prednidine, rimexolone, tecortisone ( tixocortol), triamcinolone, triamcinolone acetonide, benzolone acetonide, and triamcinolone hexacetonide (triamcinolone hexacetonide), nonsteroidal anti-inflammatory agents, aminoaryl carboxylic acid derivatives (e.g. (etofenamate), flufenamic acid, isonixin, meclofenamic acid, mefenamic acid, niflumic acid, talniflumate (talniflumate, terofenamate, tolfenamic acid), aryl acetic acid derivatives (eg, aceclofenac, acemetacin, aclofenac (al clofenac), amfenac, amtolmetin guacil, bromfenac, bufexamac, cinmetacin, clopirac, bis Diclofenac sodium, etodolac, felbinac, fenclozic acid, fentiazac, glucametacin, ibufenac (ibufenac), indomethacin, isofezolac, isoxepac, lonazolac, metiazinic acid, mofezolac ), oxametacine, pirazolac, proglumetacin, sulindac, tiaramide, tolmetin, tropesin, zomepirac) , aryl butyric acid derivatives (such as bumadizon, butibufen, fenbufen, xenbucin), aryl carboxylic acids (such as cyclochloride clidanac, ketorolac, tinoridine), arylpropionic acid derivatives (e.g. alminoprofen, benzene Benoxaprofen, bermoprofen, bucloxic acid, carprofen, fenoprofen, flunoprofen, flurbiprofen ( flurbiprofen, ibuprofen, ibuproxam, indoprofen, ketoprofen, loxoprofen, naproxen, oxaprozin (oxaprozin), piketoprolen, pirprofen, pranoprofen, protizinic acid, suprofen, tiaprofenic acid, ximoprofen ), zaltoprofen (zaltoprofen), pyrazoles (such as difenamizole, epirizole), pyrazolones (such as azapropazone, benziprone ( benzpiperylon), feprazone, mofebutazone, morazone, oxyphenbutazone, phenylbutazone, pipebuzone, iprofen Propyphenazone, ramifenazone, suxibuzone, thiazolinobutazone), salicylic acid derivatives (eg, acetaminosalol, aspirin, benorylate, brominated water bromosaligenin, calcium acetylsalicylate, diflunisal, etersalate, fendosal, gentisic acid, hydroxyethyl salicylate (glycol salicylate), imidazole salicylate, lysine acetylsalicylate, mesalamine, morpholine salicylate, 1-naphthyl salicylate ( 1-naphthyl salicylate), olsalazine, paxamide (parsalmide), phenyl acetylsalicylate, phenyl salicylate, salacetamide, salicylamide o-acetic acid, salicylamide salicylsulfuric acid, salsalate, sulfasalazine), thiazinecarboxamides (such as ampiroxicam, dr Droxicam, isoxicam, lomoxicam, piroxicam, tenoxicam), ε-etheraminocaproic acid, s-adenosylmethionine, 3-amino -4-Hydroxybutyric acid, amixetrine, bendazac, benzydamine, a-bisabolol, bucolome, bixenpyramide ( difenpiramide), ditazol, emorfazone, fepradinol, guaiazulene, nabumetone, nimesulide , oxaceprol, paranyline, perisoxal, proquazone, superoxide dismutase, tenidap, and zileuton ( zileuton).

The therapeutic agents may also be used in combination with other therapeutic agents and therapies, including but not limited to those for the treatment, prevention, inhibition, delay or regression of angiogenesis or neovascularization, particularly CNV. In some variations, angiogenesis or neovascularization (particularly CNV) is treated or regressed with an additional therapeutic agent or therapy. Non-limiting examples of such additional therapeutic agents and therapies include pyrrolidine, dithiocarbamate (NFκB inhibitor); squalamine; TPN 470 analogs and fumagillin; PKC (protein Kinase C) inhibitors; Tie-1 and Tie-2 kinase inhibitors; VEGF receptor kinase inhibitors; proteosome inhibitors such as Velcade ^™ (bortezomib for injection; ranibuzumab (Lucentis ^™ ) and other antibodies against the same target; pegaptanib (Macugen ^™ ); vitronectin receptor antagonists, such as cyclic peptide antagonists of vitronectin receptor-type integrins; alpha-v/beta-3 integrin antagonists; alpha-v/beta-1 Integrin antagonists; thiazolidinediones such as rosiglitazone or troglitazone; interferons, including gamma-interferon or interferons targeting CNVs by using dextran and metal coordination; pigment epithelium-derived Angiogenesis factor (PEDF); endostatin; angiostatin; tumistatin; canstatin; anecortal acetate; acetonide; triamcinolone; tetrathiomolybdate; (RNAi), including ribozymes targeting VEGF expression; Accutane ^TM (13-cis retinoic acid); ACE inhibitors, including but not limited to quinidine (quinopril), captopril, and perindozril; mTOR inhibitors ( Mammalian target of rapamycin); 3-aminothalidomide; pentoxifylline; 2-methoxyestradiol; colchicine; AMG- 1470; Cyclooxygenase inhibitors such as nepafenac, rofecoxib, diclofenac, rofecoxib, NS398, celecoxib, vioxx ) and (E)-2-alkyl-2(4-methylsulfonylphenyl)-1-styrene; t-RNA synthase modulator; metalloproteinase 13 inhibitor; acetylcholinesterase inhibitor; potassium channel Blocker; endorepellin; purine analogue of 6-thioguanine; cycloperoxide ANO-2; (recombinant) arginine deiminase; epigallocatechin-3-gallate (epigallocatechin -3-gallate); cerivastatin; suramin analogs; VEGF trap molecules; hepatocyte growth factor inhibitors (growth factor antibodies or their receptors, c-met tyrosine kinase Small molecule inhibitors, truncated forms of HGF such as HK4); apoptosis apoptosis inhibitors; Visudyne ^™ , snET2 and other photosensitizers for photodynamic therapy (PDT); and laser photocoagulation.

Diseases and conditions that can be treated, prevented, inhibited, delayed or caused to regress

Described herein are diseases and conditions that can be treated, prevented, inhibited, delayed onset, or caused to regress using the therapeutic agents and formulations, fluid formulations, and methods described herein. In some variations, the diseases and conditions are treated using the therapeutic agents and formulations, fluid formulations and methods described herein. Unless otherwise stated herein, all therapeutic methods are considered to be performed on subjects including, but not limited to, human subjects. the

In general, any ocular disease or condition susceptible to treatment, prevention, inhibition, delay of onset, or cause of regression using the therapeutic agents and formulations, fluid formulations, and methods described herein can be treated, prevented, inhibited, delayed onset of, or caused to resolve. Examples of ocular diseases or disorders include, but are not limited to, diseases or disorders associated with neovascularization, including retinal and/or choroidal neovascularization. the

Diseases or conditions associated with retinal and/or choroidal neovascularization that can be treated, prevented, inhibited, delayed onset, or caused to regress using the formulations, liquid formulations, and methods described herein include, but are not limited to, diabetic retinopathy, macular degeneration , wet and dry AMD, retinopathy of prematurity (retrolentic fibroplasia), infections causing retinitis or choroiditis, presumed ocular histoplasmosis, myopic degeneration, angioid streaks, and eye injury. Other non-limiting examples of ocular diseases and conditions that may be treated, prevented, inhibited, delayed onset, or caused to resolve using the formulations, liquid formulations, and methods described herein include, but are not limited to, pseudoxanthoma elasticum, venous occlusion (vein occlusion), arterial occlusion, carotid obstructive disease, sickle cell anemia, Eales disease, myopia, chronic retinal detachment ( chronic retinal detachment), hyperviscosity syndrome, toxoplasmosis, trauma, polypoid (polypoidal choroidal vasculopathy), post-laser complications, Complications of idiopathic central serous chorioretinopathy, complications of choroidal inflammatory conditions, rubeosis, diseases associated with redness (corner neovascularization) , neovascular glaucoma, uveitis and chronic uveitis, macular edema, proliferative retinopathy, and diseases or conditions caused by abnormal proliferation of fibrovascular or fibrous tissue, including all forms of hyperplastic vitreous Retinopathy (including postoperative proliferative vitreoretinopathy), whether or not associated with diabetes. the

In some variations, the formulations and pharmaceutical formulations described herein are used to prevent or delay the onset of an ocular disease or disorder in which a subject (including but not limited to a human subject) is at increased risk of developing the ocular disease or disorder. A subject at increased risk of developing a disease or condition is one who has one or more indications that the disease or condition is likely to occur in that particular subject. In some variations, the subject at increased risk of developing wet AMD is a subject with dry AMD in at least one eye. In some variations, the subject at increased risk of developing wet AMD in the fellow eye is a subject with wet AMD in the other eye. In some variations, the formulations and pharmaceutical formulations described herein are used to prevent or delay the occurrence of CNV in a subject at increased risk of developing CNV, including but not limited to preventing or delaying the occurrence of CNV in a subject (including but not limited to one Occurrence of CNV in the fellow eye of a human subject with AMD. In some variations, the formulations and pharmaceutical formulations described herein are used to prevent or delay the occurrence of CNV in the fellow eye in patients with wet AMD in one eye. In some variations, formulations and pharmaceutical formulations comprise limus compounds, including but not limited to rapamycin. In some variations, formulations and pharmaceutical formulations are administered periocular (including but not limited to subconjunctival) to human subjects with 20/40 or better vision. In some variations, formulations and pharmaceutical formulations are administered periocularly (including but not limited to subconjunctivally) to the eye of a human subject, wherein the eye to which the formulation is administered has visual acuity of 20/40 or better. the

In some variations, the formulations and pharmaceutical formulations described herein are used to treat, prevent or delay the onset of AMD. In some variations, the formulations and pharmaceutical formulations described herein are used to treat, prevent or delay the onset of dry AMD. In some variations, a subject having non-central geographic atrophy (including but not limited to a human subject) is administered a formulation or pharmaceutical formulation described herein to treat, prevent, or delay the onset of central geographic atrophy occur. In some variations, formulations and pharmaceutical formulations contain limus compounds, including but not limited to rapamycin. In some variations, formulations and pharmaceutical formulations are administered periocular (including but not limited to subconjunctival) to human subjects with 20/40 or better vision. In some variations, the formulations and pharmaceutical formulations described herein are administered and the subject (including but not limited to human subjects) is also treated with another therapy for the treatment of a disease or condition. In some variations, the formulations and pharmaceutical formulations described herein are used to treat, prevent or delay the onset of wet or dry AMD, and before, during or after treatment with the formulations and pharmaceutical formulations described herein, the subject ( Including but not limited to human subjects) are also treated with laser therapy such as photodynamic laser therapy. the

In some variations, the formulations and pharmaceutical formulations described herein are used to treat one or more of uveitis, allergic conjunctivitis, macular edema, glaucoma, or dry eye. the

In some variations, the formulation or pharmaceutical formulation comprises a limus compound, such as rapamycin, and is administered to treat, prevent, or delay the onset of dry eye. In some variations, the formulation or pharmaceutical formulation comprises a limus compound, such as rapamycin, and is administered to treat, prevent, or delay the onset of allergic conjunctivitis. the

In some variations, the formulations and pharmaceutical formulations described herein are used to treat glaucoma. In some variations, the formulations and pharmaceutical formulations described herein for treating glaucoma comprise a limus compound, such as rapamycin, and are used as a surgical adjuvant to prevent, reduce or delay surgical complications. In some variations, the formulations and pharmaceutical formulations described herein for treating glaucoma contain limus compounds, such as rapamycin, and are used to enhance or prolong surgical graft success. In some variations, the formulations and pharmaceutical formulations described herein for treating glaucoma contain a limus compound, such as rapamycin, and are used to facilitate or prolong the success of argon laser trabeculectomy or other glaucoma-related surgery. In some variations, the formulations and pharmaceutical formulations described herein are neuroprotective and are used to treat glaucoma. the

In some variations, the formulations and pharmaceutical formulations described herein are used to treat retinitis pigmentosa. In some variations, the formulations and pharmaceutical formulations described herein for treating glaucoma comprise a limus compound, such as rapamycin, and are used to treat, prevent, or delay the onset of retinitis pigmentosa. In some variations, the formulations and pharmaceutical formulations described herein are neuroprotective and are used to treat retinitis pigmentosa. the

In some variations, the formulations and pharmaceutical formulations described herein are used to treat one or more of central retinal vein occlusion (CRVO), branch retinal vein occlusion (BRVO), retinal vascular diseases and disorders, macular edema, diabetic macular edema , scleral neovascularization, diabetic retinopathy, or corneal transplant rejection. In some variations, formulations and pharmaceutical formulations contain a limus compound, such as rapamycin, and are administered to treat, prevent, or delay the onset of one or more such diseases or conditions. In some variations, the formulations and pharmaceutical formulations described herein are administered subconjunctivally to an eye with 20/40 or better vision. the

When used to treat, prevent, inhibit, delay onset or cause resolution of uveitis, the formulations and liquid formulations described herein can be administered by various routes known in the art, including but not limited to ophthalmic or oral administration . Other routes of administration are known and routine in the art. In some variations, the formulations described herein comprise rapamycin and are used to treat uveitis. the

One disease whose onset can be treated, inhibited, delayed or regressed using the formulations, fluid formulations and methods described herein is wet AMD. In some variations, wet AMD is treated using the formulations, liquid formulations, and methods described herein. Wet AMD is characterized by the growth of blood vessels from their usual location in the choroid into undesired locations under the retina. These new blood vessels leak and bleed, causing vision loss and possible blindness. the

The formulations, liquid formulations and methods described herein can also be used to prevent or delay the hygrotropism of dry AMD in which the retinal pigment epithelium or RPE degenerates and causes photoreceptor cell death and the formation of yellowish deposits called drusen under the retina Conversion of AMD. the

"Macular degeneration" is characterized by the overproduction of fibrous deposits and atrophy of the retinal pigment epithelium in the macula and retina. As used herein, an eye "affected" by macular degeneration is understood as the eye exhibiting at least one detectable physical characteristic associated with macular degeneration. Administration of rapamycin has been shown to limit and regress angiogenesis such as choroidal neovascularization in age-related macular degeneration (AMD), which occurs without treatment. As used herein, the term "angiogenesis" refers to the production of new blood vessels ("neovascularization") in a tissue or organ. An "angiogenesis-mediated disease or disorder" of the eye or retina is a disease or disorder in which new blood vessels form in the eye or retina in a disease-causing manner, causing reduced or lost vision or other problems, such as associated with AMD choroidal neovascularization. the

The formulations and liquid formulations described herein (including but not limited to formulations and liquid formulations containing rapamycin) can also be used to treat, prevent, inhibit, delay the onset of, or cause regression of various immune-related diseases and disorders, said Diseases and disorders include, but are not limited to, organ transplant rejection in the host, graft versus host disease, autoimmune diseases, inflammatory diseases, hyperproliferative vascular diseases, solid tumors, and fungal infections. In some variations, the formulations and liquid formulations described herein (including but not limited to rapamycin-containing formulations and liquid formulations) are used to treat a variety of immune-related diseases and disorders, including but not limited to host Organ transplant rejection, graft-versus-host disease, autoimmune disease, inflammatory disease, hyperproliferative vascular disease, solid tumor and fungal infection. The formulations and liquid formulations described herein, including but not limited to rapamycin-containing formulations and liquid formulations, are useful as immunosuppressants. The formulations and liquid formulations described herein, including but not limited to rapamycin-containing formulations and liquid formulations, are useful for treating, preventing, inhibiting, or delaying the occurrence or resolution of rejection of a transplanted organ or tissue that Organs or tissues include, but are not limited to, transplanted hearts, livers, kidneys, spleens, lungs, small intestines, pancreas, and bone marrow. In some variations, the formulations and liquid formulations described herein are used to treat the occurrence of rejection of a transplanted organ or tissue including, but not limited to, a transplanted heart, liver, kidney, spleen, lung, small intestine, pancreas and bone marrow. When used for the treatment, prevention, suppression, delay or regression of immune-related diseases (including but not limited to transplant rejection), the formulations and liquid formulations described herein can be administered by various routes known in the art, including but not only Limited to oral administration. the

Systemic administration can be accomplished through oral liquid formulations. Other routes of systemic administration are routine in the art. Some examples of these are listed in the Detailed Description of the Invention section. the

As used herein, "inhibition" of a disease or condition by administration of a therapeutic agent refers to the reduction in at least one detectable physiological characteristic or symptom of the disease or condition following administration of the therapeutic agent compared to the progression of the disease or condition in the absence of administration of the therapeutic agent. Development is delayed or stopped. the

As used herein, "prevention" of a disease or condition by administration of a therapeutic agent refers to the absence of development of detectable physical characteristics or symptoms of the disease or condition following administration of the therapeutic agent. the

As used herein, "delaying" the "onset" of a disease or condition by administering a therapeutic agent means that following administration of the therapeutic agent, at least one detectable physiological symptom of the disease or condition is reduced compared to the progression of the disease or condition without the administration of the therapeutic agent. Features or symptoms develop later. the

As used herein, "treating" a disease or condition by administering a therapeutic agent refers to the progression of at least one detectable physiological characteristic or symptom of the disease or condition following administration of the therapeutic agent as compared to the progression of the disease or condition without the administration of the therapeutic agent be delayed, stopped or reversed. the

As used herein, "causing" a disease or disorder "regression" by administration of a therapeutic agent means that the progression of at least one detectable physiological characteristic or symptom of the disease or disorder is reversed to some extent following administration of the therapeutic agent. the

Subjects with a predisposition or in need of prophylaxis, including but not limited to human subjects, can be identified by a skilled physician using methods and criteria established in the art according to the teachings herein. Individuals in need of inhibition or treatment in light of the teachings herein can also readily be diagnosed by a skilled physician based on established criteria in the art for identifying angiogenesis and/or neovascularization. the

A "subject" as used herein is generally any animal that would benefit from administration of a therapeutic agent described herein. In some variations, a therapeutic agent is administered to a mammalian subject. In some variations, the therapeutic agent is administered to a human subject. In some variations, a therapeutic agent may be administered to a veterinary animal subject. In some variations, a therapeutic agent may be administered to a model animal subject. the

Other diseases and conditions that may be treated, prevented, inhibited, delayed, or caused to regress using the methods described herein include those disclosed in the following patents and publications, the contents of each of which are incorporated herein by reference in their entirety: PCT Publication WO 2004/027027, published April 1, 2004, entitled Method of inhibiting choroidal neovascularization, assigned to Trustees of the University of Pennsylvania; U.S. Patent No. 5,387,589, published February 7, 1995, entitled Method of Treating Ocular Inflammation, inventor Prassad Kulkarni, assigned to University of Louisville Research Foundation; U.S. Patent No. 6,376,517, published April 23, 2003, entitled Pipecolic acidderivatives for vision and memory disorders, assigned to GPI NIL Holdings, Inc; PCT Publication WO 2004 /028477, published April 8, 2004, entitled Method subretinal administration of therapeutics including steroids: method for localizing pharmacodynamic action at the choroid and retina; and related methods for treatment and or prevention of retinal diseases, assigned to U.S. Patent No.; Innorx, Inc. 6,416,777, issued July 9, 2002, entitled Ophthalmic drug delivery device, assigned to Alcon Universal Ltd; U.S. Patent No. 6,713,081, issued March 30, 2004, entitled Ocular therapeutic agent delivery device and methods for making and using such devices, Assigned to the Department of Health and Human Services; and U.S. Patent No. 5,536,729, filed July 16, 1996, and entitled Rapamycin Formulati ons for Oral Administration, assigned to AmericanHome Products Corp., and U.S. Patent Application Nos. 60/503,840 and 10/945,682. the

liquid preparation

Liquid formulations described herein contain a therapeutic agent and may generally be any liquid formulation including, but not limited to, solutions, suspensions, and emulsions. In some variations, the liquid formulation forms a non-dispersed mass relative to the surrounding medium when placed in the vitreous of a rabbit eye. the

When administering a volume of liquid formulation, it is understood that there are certain imprecisions in the accuracy of the various devices available for administering liquid formulations. When a volume is specified, it is understood to be the volume of interest. However, certain devices such as insulin syringes are inaccurate by greater than 10%, and sometimes inaccurate by greater than 20% or more. Hamilton HPLC type syringes are generally considered accurate to within 10% and are recommended for volumes to be injected of less than 10 μl. the

In some variations, the volume of a liquid formulation described herein administered to the vitreous of a rabbit eye or a subject's eye, including but not limited to a human subject's eye, is less than about 500 μl, less than about 400 μl, less than about 300 μl, less than about 200 μl , less than about 100 μl, less than about 90 μl, less than about 80 μl, less than about 70 μl, less than about 60 μl, less than about 50 μl, less than about 40 μl, less than about 30 μl, less than about 20 μl, less than about 10 μl, less than about 5 μl, less than about 3 μl, or less than About 1 μl. In some variations, the volume of a liquid formulation described herein administered to the vitreous of a rabbit eye or a subject's eye, including but not limited to a human subject's eye, is less than about 20 μl. In some variations, the volume of a liquid formulation described herein administered to the vitreous of a rabbit eye or a subject's eye, including but not limited to a human subject's eye, is less than about 10 μl. In some variations, the volume of the liquid formulation described herein administered to the vitreous of a rabbit eye or a subject's eye, including but not limited to a human subject's eye, is between about 0.1 μl and about 200 μl, between about 50 μl and about 200 μl, between about 50 μl and about 200 μl, at Between about 50 μl and about 150 μl, between about 0.1 μl and about 100 μl, between about 0.1 μl and about 50 μl, between about 1 μl and about 40 μl, between about 1 μl and about 30 μl, between about 1 μl and about 20 μl, between about 1 μl and about 10 μl or between about 1 μl and about 10 μl Between about 1 μl and about 5 μl. In some variations, the volume of a liquid formulation described herein administered into the vitreous of a rabbit eye or a subject's eye, including but not limited to a human subject's eye, is between about 1 μl and about 10 μl. In some variations, the volume of a liquid formulation described herein administered to the intravitreous of a rabbit eye or a subject's eye, including but not limited to a human subject's eye, is between about 1 μl and about 5 μl. In some variations, the volume of a liquid formulation described herein administered to the intravitreous of a rabbit eye or a subject's eye, including but not limited to a human subject's eye, is between about 1 μl and about 5 μl. In some variations, the volume of the liquid formulation described herein administered to the intravitreal of a rabbit eye or eye of a subject is between about 0.1 μl and about 200 μl. the

In some variations, the total volume of the liquid formulations described herein administered subconjunctivally to the eye of a rabbit or a subject (including but not limited to an eye of a human subject) is less than about 1000 μl, less than about 900 μl, less than about 800 μl, less than About 700 μl, less than about 600 μl, less than about 500 μl, less than about 400 μl, less than about 300 μl, less than about 200 μl, less than about 100 μl, less than about 90 μl, less than about 80 μl, less than about 70 μl, less than about 60 μl, less than about 50 μl, less than about 40 μl , less than about 30 μl, less than about 20 μl, less than about 10 μl, less than about 5 μl, less than about 3 μl, or less than about 1 μl. In some variations, the volume of a liquid formulation described herein that is subconjunctivally administered to a rabbit eye or a subject's eye, including but not limited to a human subject's eye, is less than about 20 μl. In some variations, the volume of a liquid formulation described herein that is subconjunctivally administered to a rabbit eye or a subject's eye, including but not limited to a human subject's eye, is less than about 10 μl. In some variations, the volume of the liquid formulation described herein is between about 0.1 μl to about 200 μl, between about 50 μl and about 200 μl, subconjunctivally administered to the eye of a rabbit or subject (including but not limited to the eye of a human subject) , between about 200 μl and about 300 μl, between about 300 μl and about 400 μl, between about 400 μl and about 500 μl, between about 600 μl and about 700 μl, between about 700 μl and about 800 μl, between about 800 μl and about 900 μl, between about 900 μl and about 1000 μl, Between about 50 μl and about 150 μl, between about 0.1 μl and about 100 μl, between about 0.1 μl and about 50 μl, between about 1 μl and about 40 μl, between about 1 μl and about 30 μl, between about 1 μl and about 20 μl, between about 1 μl and about 10 μl or Between about 1 μl and about 5 μl. In some variations, the volume of the liquid formulation described herein is between about 1 μl and about 10 μl administered subconjunctivally to the eye of a rabbit or subject, including but not limited to the eye of a human subject. In some variations, the volume of the liquid formulation described herein is between about 1 μl and about 5 μl administered subconjunctivally to the eye of a rabbit or subject, including but not limited to the eye of a human subject. In some variations, the volume of the liquid formulation described herein is between about 1 μl and about 5 μl administered subconjunctivally to the eye of a rabbit or subject, including but not limited to the eye of a human subject. In some variations, the volume of the liquid formulation described herein is between about 0.1 μl and about 200 μl administered subconjunctivally to the eye of a rabbit or subject, including but not limited to the eye of a human subject. the

In some variations, the liquid formulations described herein are administered at multiple subconjunctival locations within a period of time, including but not limited to, within one hour of each other. Without being bound by theory, it is believed that such multiple administrations (eg, multiple injections) allow a larger total dose to be administered subconjunctivally compared to a single dose, since the ability of local ocular tissue to absorb larger volumes may be limited. the

One liquid formulation described herein is a gel-in-situ formulation. In situ gelling formulations as described herein comprise a therapeutic agent and a variety of polymers that give a formulation that forms a gel or gel-like substance when placed in an aqueous medium, including but not limited to that of the eye. the

In some variations of the liquid formulations described herein, the therapeutic agent is a solution or suspension of rapamycin in the liquid medium. Liquid media include, but are not limited to, solvents including, but not limited to, those of the "Therapeutic Agent Solubilization" section. the

The liquid formulations described herein may contain a solubilizing agent component. In some variations, the solubilizer component is a surfactant. Note that there is overlap between ingredients that can be solvents and solubilizers, so the same ingredient can act as either solvent or solubilizer in some systems. Liquid formulations contain a therapeutic agent and an ingredient that may be considered a solvent or solubilizer or a surfactant if the ingredient acts as a solvent is considered a solvent; if the ingredient does not act as a solvent then the ingredient may be considered a Solubilizer or Surfactant. the

Liquid formulations may also optionally contain stabilizers, excipients, gelling agents, adjuvants, antioxidants, and/or other ingredients described herein. the

In some variations, all ingredients of the liquid formulation, except the therapeutic agent, are liquid at room temperature. the

In some variations, the liquid formulation includes a release modifier. In some variations, the release modifier is a film-forming polymer component. The film-forming polymer composition may comprise one or more film-forming polymers. Any film-forming polymer can be used in the excipient component. In some variations, the film-forming polymer component contains a water-insoluble film-forming polymer. In some variations, the release modifier composition comprises an acrylate polymer, including but not limited to polymethacrylates, including but not limited to Eudragit RL. the

Compositions and liquid formulations for the delivery of therapeutic agents described in the "Therapeutic Agents" section are described herein. Delivery of therapeutic agents using the compositions and liquid formulations described herein can be used to treat, prevent, inhibit, delay the onset of, or cause regression of the diseases and conditions described in the "Diseases and Conditions" section. Compositions and liquid formulations described herein may contain any of the therapeutic agents described in the "Therapeutic Agents" section, including but not limited to rapamycin. The compositions and liquid formulations described herein may contain one or more than one therapeutic agent. Compositions and liquid formulations other than those expressly described herein may be used. the

When the therapeutic agent is rapamycin, the compositions and liquid formulations are useful for maintaining within the vitreous an amount of rapamycin effective to treat wet AMD. In one non-limiting example, it is believed that a liquid formulation for delivering rapamycin that maintains about 10 pg/ml to about 2 μg/ml of rapamycin in the vitreous for a period of time is believed to be useful in the treatment of wet AMD concentration. When rapamycin is contained in a liquid formulation that forms a non-dispersed mass, the stated concentration of rapamycin represents an amount effective to treat an ocular disease or condition, rather than being present only in the non-dispersed mass form . In another non-limiting example, it is believed that a delivery system for the delivery of rapamycin capable of maintaining about 0.01 pg/mg to about 10 ng/mg in retinal choroidal tissue over a period of time is useful for the treatment of wet AMD concentration of rapamycin. Other therapeutically effective amounts of therapeutic agents are also possible and can be readily determined by one of skill in the art from the teachings herein. the

When the therapeutic agent is rapamycin, the compositions and liquid formulations described herein are useful for delivering a dose of rapamycin to a subject (including but not limited to a human subject) or to the eye of the subject. In one non-limiting example, a liquid formulation containing a dose of about 20 μg to about 4 mg is believed to be useful in the treatment of wet AMD. the

In some variations, the therapeutic agent in the liquid formulation comprises between about 0.01% and about 30%; between about 0.05% and about 15%; between about 0.1% and about 10%; between about 1% and about 5% by weight of the total composition or between about 5% and about 15%; between about 8% and about 10%; between about 0.01% and about 1%; between about 0.05% and about 5%; between about 0.1% and about 0.2%; about Between 0.2% and about 0.3%; between about 0.3% and about 0.4%; between about 0.4% and about 0.5%; between about 0.5% and about 0.6%; between about 0.6% and about 0.7%; between about 0.7% and about 1% Between about 1% and about 5%; between about 5% and about 10%; between about 15% and about 30%, between about 20% and about 30%; or between about 25% and about 30%. the

One skilled in the art can determine what amount or concentration of a given therapeutic agent is equivalent to a certain amount or concentration of rapamycin based on the teachings herein, for example, by administering the therapeutic agent in different amounts or concentrations to a disease model system (e.g., in vivo or in vitro model system) and compare the results in the model system relative to the results of various amounts or concentrations of rapamycin. One skilled in the art can also determine, based on the teachings herein, what amount or concentration of a given therapeutic agent is equivalent to a certain amount or concentration of rapamycin by reviewing experiments performed in the scientific literature comparing rapamycin with other therapeutic agents. It is understood that even the same therapeutic agent may have different equivalent levels of rapamycin when, for example, different diseases or conditions are being assessed, or when different types of formulations are used. A non-limiting example of the scientific literature comparing rapamycin and other therapeutic agents for ocular diseases is Ohia et al., Effects of steroids andimmunosuppressive drugs on endotoxin-uveitis in rabbits, J.Ocul.Pharmacol.8(4):295-307 (1992); Kulkarni, Steroidal and nonsteroidal drugs in endotoxin-induced uveitis, J.Ocul.Pharmacol.10(1):329-34(1994); Hafizi et al., Differential effects of rapamycin, cyclosporine A, and FK506 on humany coronary artery Muscle cell proliferation and signaling, Vascul Pharmacol. 41(4-5):167-76 (2004); and US 2005/0187241. the

For example, in a wet AMD model, if a therapeutic agent is found to be 10-fold less potent or effective than rapamycin in treating wet AMD, then a therapeutic agent concentration of 10 ng/ml should be equivalent to a rapamycin concentration of 1 ng/ml. Or if the therapeutic agent is found to be 10-fold less potent or effective than rapamycin in treating wet AMD, then a 10-fold amount of therapeutic agent should be administered relative to the amount of rapamycin. the

The solvent component, for example, may constitute between about 0.01% and about 99.9%; between about 0.1% and about 99%; between about 25% and about 55%; between about 30% and about 50%; or about 35% of the total weight of the composition Between about 0.1% and about 10%; Between about 10% and about 20%; Between about 20% and about 30%; Between about 30% and about 40%; Between about 40% and about 45% between about 40% and about 45%; between about 45% and about 50%; between about 50% and about 60%; between about 50% and about 70%; between about 70% and about 80%; between about 80% and about Between 90%; or between about 90% and about 100%. the

The solubilizer component, for example, may constitute between about 0.01% and about 30% of the total weight of the composition; between about 0.1% and about 20%; between about 2.5% and about 15%; between about 10% and about 15%; or about 5% Between about 8% and about 12%; between about 10% and about 20%; between about 20% and about 30%. the

In some variations, the liquid formulations described herein have a viscosity between 40% and 120% centipoise. In some variations, the liquid formulations described herein have a viscosity of between 60% and 80% centipoise. the

In some variations, the liquid formulations described herein contain a therapeutic agent and a solvent component. The solvent component may contain a single solvent or a combination of solvents. The therapeutic agent composition may contain a single therapeutic agent or a combination of therapeutic agents. In some variations, the solvent is glycerol, dimethylsulfoxide, N-methylpyrrolidone, dimethylacetamide (DMA), dimethylformamide, glycerol formal, ethoxydiglycol, triethylene glycol Glycol Dimethyl Ether, Triacetin, Glyceryl Diacetate, Corn Oil, Acetyl Triethyl Citrate (ATC), Ethyl Lactate, Pegylated Caprylic Glycerides, Gamma Butyrolactone, Dimethyl Isosorbide, benzyl alcohol, ethanol, isopropanol, polyethylene glycols of various molecular weights (including but not limited to PEG 300 and PEG 400) or propylene glycol, or a mixture of one or more of these. the

In some variations, the liquid formulations described herein are solutions and comprise a therapeutic agent and a solvent component. In some variations, the solvent component includes ethanol. In some variations, the solvent component comprises ethanol and polyethylene glycol, including but not limited to liquid polyethylene glycol (including but not limited to one or more of PEG 300 or PEG 400). the

In some variations, the liquid formulations described herein contain no more than about 250 μl of polyethylene glycol. In some variations, the liquid formulations described herein contain no more than about 250 μl, no more than about 200 μl, no more than about 150 μl, no more than about 125 μl, no more than about 100 μl, no more than about 75 μl, no more than about 50 μl, not more than about 25 μl, not more than about 20 μl, not more than about 15 μl, not more than about 10 μl, not more than about 7.5 μl, not more than about 5 μl, not more than about 2.5 μl, not more than About 1.0 μl or no more than about 0.5 μl of polyethylene glycol. Formulations containing polyethylene glycol may contain, for example, PEG 300 or PEG 400. the

In some variations, the liquid formulations described herein are suspensions and contain a therapeutic agent and a diluent component. In some variations, the diluent component contains one or more components listed herein as solvents or solubilizers, wherein the resulting mixture is a suspension. the

In some variations, a liquid formulation is part solution and part suspension. the

In some variations, the liquid formulation is an in situ gelling formulation and comprises a therapeutic agent and a polymeric component, where the polymeric component can contain multiple polymers. In some variations, the liquid formulations contain polymethacrylate polymers. In some variations, the liquid formulation contains polyvinylpyrrolidone polymer. the

Some variations of liquid formulations contain between about 0.01% and about 20% by total weight of therapeutic agent (such as but not limited to rapamycin), between about 5% and about 15% by total weight of solvent, by about 5% by total weight % and about 15% of solubilizers (including but not limited to surfactants), and water as the main retention component. In some variations, the formulation further comprises between about 0 and about 40% by total weight of a stabilizer, excipient, adjuvant, or antioxidant. the

In some variations, the liquid formulation contains up to about 5% by total weight of a therapeutic agent (including but not limited to rapamycin); and up to about 99.9% by total weight of a solvent component. In some variations, the liquid formulation contains up to about 5% by weight of the total therapeutic agent (including but not limited to rapamycin); and up to about 99.9% of the diluent component. the

In some variations, the liquid formulation contains up to about 5% by total weight of a therapeutic agent (including but not limited to rapamycin); up to about 10% by total weight of a solvent component; and up to about 85% by total weight of an additive soluble ingredients. In some variations, the solubilizing component is an aqueous solution of surfactant. the

The various polymer components may constitute, for example, between about 0.01% and about 30% of the total weight of the composition; between about 0.1% and about 20%; between about 2.5% and about 15%; between about 10% and about 15%; about Between 3% and about 5%; between about 5% and about 10%; between about 8% and about 12%; between about 10% and about 20%; or between about 20% and about 30%. the

Some variations of liquid formulations contain between about 0.01% and about 20% by total weight of a therapeutic agent (including but not limited to rapamycin), between about 60% and about 98% by total weight of a solvent component, and various polymeric substances in a combined percentage of between about 0.1% and about 15% by weight of the total. In some variations, the formulation further comprises between about 0 and about 40% by total weight of a stabilizer, excipient, adjuvant, or antioxidant. the

In some variations, the liquid formulation may contain about 4% by total weight of therapeutic agent (including but not limited to rapamycin); about 91% by total weight of solvent; and about 5% by total weight of polymeric component. the

Some examples and variations of the liquid formulations described herein were prepared and listed in Table 1. The listed formulations are referred to according to their type as one or more solutions ("S"), suspensions ("SP"), emulsions ("E") or in situ gels ("ISG"). Some suspensions list median particle sizes. As described herein, some liquid formulations form non-dispersed masses upon injection, for example, into an aqueous environment such as the vitreous of the eye. For these formulations injected into the vitreous of rabbit eyes, the right-hand column of Table 1 indicates whether non-dispersed masses (NDM) formed after the specified volume was injected into the vitreous of rabbit eyes. the

The following references show one or more formulations, including but not limited to rapamycin formulations, which are incorporated herein in their entirety and describe various dosages of rapamycin and other therapeutic agents for the treatment of various diseases or Uses for Conditions: US 60/651,790, filed 2/9/2005, entitled FORMULATIONS FOR OCULAR TREATMENT, attorney docket number 57796-30002.00; US 60/664,040, filed 2/9/2005, attorney docket number 57796 -30004.00, entitled LIQUID FORMULATIONS FOR TREATMENTOF DISEASES OR CONDITIONS; US 60/664,119, filed 3/21/2005, Attorney Docket No. 57796-30005.00, entitled DRUG DELIVERY SYSTEMS FOR TREATMENT OF 30 DISEASES OR CONDITIONS; 6,6,6,6 Filed on /21/2005, attorney docket number 57796-30006.00, titled IN SITU GELLINGFORMULATIONS AND LIQUID FORMULATIONS FOR TREATMENTOF DISEASES OR CONDITIONS; US_/_, filed 2/9/2006, titled FORMULATIONS FOR OCULAR TREATMENT, attorney docket number 6-579 20002.00; _/_, filed 2/9/2006, Attorney Docket No. 57796-20004.00, entitled LIQUID FORMULATIONS FOR TREATMENT OF DISEASES ORCONDITIONS; US 2005/0187241 and US 2005/0064010. the

Liquid formulations that form non-dispersed masses

One class of liquid formulations described herein forms non-dispersible masses when placed in an aqueous medium. A "non-dispersed mass" is used herein to mean, when a liquid formulation is placed in an environment, it forms or assumes a shape relative to the environment in which it is placed. In general, a non-dispersed mass of a liquid formulation is anything other than a uniform distribution of the liquid formulation in the surrounding medium. Non-dispersed clumps can be indicated, for example, by observing the liquid formulation being administered and characterizing its appearance relative to the surrounding environment. the

In some variations, the aqueous medium is water. In some variations, the aqueous medium is deionized water, distilled water, sterile water, or tap water (including but not limited to tap water available under the purview of MacuSight in Union City, California). the

In some variations, the aqueous medium is the aqueous medium of the subject. In some variations, the aqueous medium is the aqueous medium of the subject's eye, including but not limited to the vitreous of the subject's eye. In some variations, the subject is a human subject. In some variations, the subject is a rabbit. the

In some variations, the liquid formulation is exposed to a temperature or range of temperatures (including but not limited to about room temperature, about ambient temperature, about 30°C, about 37°C, or about the temperature of the subject's aqueous medium) form non-dispersed clumps. the

In some variations, the liquid formulation forms non-dispersible masses when exposed to a pH or range of pH including, but not limited to, a pH between about 6 and about 8. the

In some variations, the non-dispersed mass includes a gel or gel-like substance. the

In some variations, the non-dispersed mass includes a polymer matrix. In some variations, the non-dispersed mass includes a polymer matrix in which the therapeutic agent is dispersed. the

Liquid formulations described herein may generally be of any geometry or shape following administration to a subject or eye of a subject, including but not limited to a human subject. In some variations, the non-dispersed clumps are between about 0.1 mm and about 5 mm. In some variations, the non-dispersed clumps are between about 1 mm and about 3 mm. Liquid formulations that form non-dispersed masses may appear, for example, as dense spherical masses when applied to the vitreous. In some cases, liquid formulations may appear as non-dispersed masses relative to the surrounding medium, wherein the non-dispersed masses are less well defined and more amorphous in geometry than spherical. the

A liquid formulation described herein that forms a non-dispersed mass may form a non-dispersed mass immediately upon placement in the medium, or a non-dispersed mass may form over a period of time after the liquid formulation has been placed. In some variations, the non-dispersed clumps form in about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, or about 7 days. In some variations, non-dispersed clumps are formed in about 1 week, about 2 weeks, or about 3 weeks. the

In some variations, a liquid formulation described herein that forms a non-dispersed mass appears as a milky or whitish semi-continuous or semi-solid non-dispersed mass relative to the medium in which it is placed. the

A liquid formulation described herein forms a non-dispersed mass that forms a solid when the formulation is injected into any or all of the water (vitreous humor of the rabbit eye or between the sclera and conjunctiva of the rabbit eye) repository. A liquid formulation described herein forms a non-dispersed mass that forms a semi-dispersed mass when the formulation is injected into any or all of the water (vitreous humor of the rabbit eye or between the sclera and conjunctiva of the rabbit eye). solid. A liquid formulation described herein forms non-dispersed masses that form aggregates when the formulation is injected into any or all of the water (vitreous humor of the rabbit eye or between the sclera and conjunctiva of the rabbit eye) Substrate. A liquid formulation described herein forms a non-dispersed mass with coagulation when the formulation is injected into any or all of the water (vitreous humor of the rabbit eye or between the sclera and conjunctiva of the rabbit eye). In the form of a glue, hydrogel or gel-like substance. the

In some variations described herein, when the surrounding medium is aqueous, the liquid formulation forms a non-dispersed mass relative to the surrounding medium. An "aqueous medium" or "aqueous environment" is a medium or environment that contains at least about 50% water. Examples of aqueous media include, but are not limited to, water, vitreous, extracellular fluid, conjunctiva, sclera, between sclera and conjunctiva, aqueous humor, gastric juice, and any tissue or body fluid that contains at least about 50% water. Aqueous media include, but are not limited to, gel structures, including but not limited to those of the conjunctiva and sclera. the

In some variations, the liquid formulations described herein form non-dispersed masses when a test volume of the liquid formulation is placed in the vitreous of a rabbit's eye. In some variations, a test volume is administered to a rabbit eye, and the test volume is equal to the volume of the liquid formulation administered to a subject, including but not limited to the eye of a human subject. the

In some variations, the test volume administered to the rabbit's eye is equal to the volume administered to the subject's eye multiplied by a scaling factor, and the scaling factor is equal to the average volume of the rabbit's eye divided by the average volume of the subject's eye. "Average volume" of the eye as used herein generally refers to the average eye volume of similarly aged members of the species under consideration, relative to the average eye volume of any particular individual. the

In some variations, the test volume administered to the rabbit eye is between about 10 μl and about 50 μl. In some variations, the test volume administered to the rabbit eye is between about 1 μl and about 30 μl. In some variations, the test volume administered to the rabbit eye is between about 50 μl and about 100 μl. In some variations, the test volume administered to the rabbit eye is between about 25 μl and about 75 μl. In some variations, the test volume administered to a rabbit eye is about 30 μl. the

In some variations, the liquid formulation that forms a non-dispersible mass when placed in a medium comprises one or more therapeutic agents at a concentration between about 0.01% and about 10% by weight of the total, and between about 10% and about 10% by weight of the total. 99% between solvents. In some variations, liquid formulations also contain solubilizing agents including, but not limited to, surfactants. In some variations, the liquid formulation also contains between about 0 and about 40% by total weight of stabilizers, excipients, adjuvants, or antioxidants, among others. In some variations, the therapeutic agent comprises about 5% by weight of the total, and the solvent component comprises about 95% by weight of the total. the

Whether a liquid formulation exhibits a non-dispersed mass relative to the surrounding medium when present in a subject (including but not limited to a human subject) or in the subject's eye can be determined by mixing the therapeutic agent with a solvent, administering to The vitreous of the eye of a subject, including but not limited to a human subject, and compare the liquid formulation to the surrounding medium. the

A liquid formulation useful for treating, preventing, inhibiting, delaying the onset of, or causing regression of diseases and conditions in subjects, including but not limited to human subjects, forms a non-dispersed mass when placed in the vitreous of a rabbit eye liquid formulations. When used to treat, prevent, inhibit, delay the onset of, or cause regression of a disease or condition in a subject, the liquid formulation is administered to a subject. Liquid formulations may or may not form non-dispersed masses in a subject. A liquid formulation described herein forms non-dispersed clumps when administered to a subject and forms non-dispersed clumps when administered to a rabbit eye. the

Without being bound by theory, it is believed that the low solubility of rapamycin in the vitreous contributes to the formation of non-dispersed masses in some of the rapamycin-containing liquid formulations described herein. The vitreous is a clear gel consisting essentially entirely of water (up to 99%). Without being bound by theory, it is believed that when rapamycin in the injected formulation comes into contact with the vitreous, precipitation of rapamycin occurs. the

Without being bound by theory, it is believed that factors affecting the formation and shape of non-dispersed masses include the concentration of rapamycin in the formulation, the viscosity of the formulation, the ethanol content of the formulation, and the injection volume. It is believed that maintaining a higher concentration of rapamycin after injecting the formulation contributes to the formation of non-dispersed clumps, as opposed to the lower local concentration of rapamycin following injecting the formulation. Formation of non-dispersed masses is less favored as the volume of a given dose increases. The formation of non-dispersed clumps can be more favored when the concentration of rapamycin is increased and/or the viscosity is increased. The ethanol content affects the solubility of rapamycin in the formulation and the viscosity of the formulation. the

In a comparison, 100 μl of a solution of 0.4% rapamycin, 4.0% ethanol and 95.6% PEG 400 (400 μg dose) did not form non-dispersed clumps after injection into rabbit eyes. In contrast, 20 μl of a solution of 2.00% rapamycin, 4.0% ethanol, and 94% PEG 400 (also a 400 μg dose) formed dense spherical non-dispersed masses after injection into rabbit eyes. the

Without being bound by theory, in the latter example it is assumed that the formation of non-dispersed agglomerates occurs as described in Figures 1A-1C and below. When injected, the liquid formulation forms a spheroid 100 in the vitreous 110 due to its viscosity. Ethanol then diffuses from the pellet, forming a localized precipitate 120 of rapamycin in the pellet. Finally, polyethylene glycol also diffuses from the pellets, resulting in solid, dense, non-dispersed masses 130 of rapamycin. the

In some variations, the non-dispersed agglomerates described herein consist of at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80% by volume. %, at least about 90%, or at least about 95% of the composition of the therapeutic agent when injected into the vitreous of a rabbit eye. the

In some variations, the rapamycin-containing non-dispersed mass is delivered at a substantially constant rate over an extended period of time. Without being bound by any theory, it is believed that delivery of rapamycin from non-dispersed masses in the vitreous depends on the dissolution of the rapamycin in the vitreous, which in turn depends on the clearance of the drug from the vitreous to other tissues. Without being bound by any theory, it is believed that this release process maintains a steady state concentration of rapamycin in the vitreous. the

In some variations, the formation of a non-dispersed mass reduces the toxicity of the injected liquid formulation compared to an equivalent dose that does not form a non-dispersed mass. In variations where the liquid formulation injected into the vitreous does not form non-dispersed clumps, the drug (eg rapamycin) appears dispersed within the vitreous. In some variations, this can interfere with vision. the

In some variations, liquid formulations that are suspensions form a non-dispersed mass upon injection into the vitreous. Formation of non-dispersed masses from injected suspensions may be more advantageous as the particle size of the suspension is increased. the

In some variations, liquid formulations are believed to form visually visible non-dispersed masses when injected into the eye of a subject, including but not limited to a human subject. the

In some variations, liquid formulations are believed to form non-dispersed masses when injected subconjunctivally. In some variations, subconjunctival administration of the liquid formulation is believed to form a depot in the scleral tissue. That is, it is believed that the therapeutic agent is absorbed into the sclera near the injection site and creates a local concentration of drug in the sclera. the

in situ gelling preparation

Described herein are liquid formulations that form non-dispersed masses that form a gel or gel-like mass when placed in an aqueous medium. In some variations, the non-dispersed mass comprises a gel; in some variations the gel is a hydrogel. the

As used herein, "gelling in situ formulation" means that a liquid formulation forms a gel-like non-dispersible formulation when placed in an aqueous medium (including, but not limited to, water, the vitreous humor of the rabbit eye, and the aqueous medium between the sclera and conjunctiva of the rabbit eye). Liquid formulations of lumps. In some variations, the in situ gelling formulation forms a gel-like non-dispersed mass when placed in tap water. the

In some variations, the gel-in-situ formulation is a suspension prior to placing in the aqueous medium and forms a gel when placed in the aqueous medium. In some variations, the gel-in-situ formulation is in solution prior to placement in the aqueous medium and forms a gel in situ when placed in the aqueous medium. In some variations, the gel-in-situ formulation is an emulsion before being placed in an aqueous medium and forms a gel when placed in an aqueous medium. In some variations, the in situ gelling formulation forms a gel-like non-dispersed mass upon placement in an aqueous medium (including, but not limited to, any or all of water, the vitreous, or between the sclera and conjunctiva of the eye). In some variations, the in situ gel is formed from a polymer matrix. In some variations the therapeutic agent is dispersed in the polymer matrix. the

Described herein are in situ gelling formulations useful for treating, preventing, inhibiting, delaying the onset of, or causing regression of diseases and conditions in a subject, including but not limited to a human subject. When used to treat, prevent, inhibit, delay the onset of, or cause regression of a disease or condition in a subject, the in situ gelling formulation is administered to a subject. A liquid formulation described herein comprises an in situ gelling formulation that forms a non-dispersed mass when applied to a subject and forms a non-dispersed mass when applied to a rabbit eye. the

In some variations, the gel-in-situ formulation comprises one or more polymers. Various types of polymers are described herein, including polymers that are solvents, polymers that are solubilizers, polymers that are release modifiers, polymers that are stabilizers, and the like. In some variations, any combination of polymers is used wherein, when placed in an aqueous medium (including, but not limited to, water, the vitreous, or any or all between the sclera and conjunctiva), the polymers form a non-dispersed Any or all of agglomerates, gels, hydrogels or polymer matrices. the

In some variations, the in situ gelling formulation delivers extended release of the therapeutic agent to the subject when administered to the subject. the

In some variations, the liquid formulation comprises a therapeutic agent and multiple polymers, wherein one of the polymers is polymethacrylate. Polymethacrylate is known by many names and is available in various formulations including but not limited to polymethacrylate, methacrylic acid-ethyl acrylate copolymer (1:1), 30% dispersion Methacrylic acid-ethyl acrylate copolymer (1:1), methacrylic acid-methyl methacrylate copolymer (1:2), methacrylic acid-methyl methacrylate copolymer (1:1), acidum methacrylicum et ethylis acrylas polymerisatum 1∶1、acidummethacrylicum et ethylis acrylas polymerisatum 1∶1 dispersio 30 percentum、acidum methacrylicum et methylis methacrylas polymerisatum1∶1、acidum methacrylicum et methylis methacrylas polymerisatum 1∶2、USPNF：异丁烯酸铵共聚物、甲Acrylic acid copolymer, methacrylic acid copolymer dispersion. the

In some variations, one of the polymers is polyvinylpyrrolidone. Polyvinylpyrrolidone is known by various names and is available in various formulations, including but not limited to povidone, povidonum, kollidon; plasdone; poly[1-(2-oxo-1-pyrrolidinyl)ethylene]; polyvidone; PVP; 1-vinyl-2-pyrrolidinyl polymer and 1-vinyl-2-pyrrolidinone homopolymer. the

A liquid formulation described herein comprises a therapeutic agent and a solvent component. The solvent component may comprise a single solvent or a combination of solvents. the

In some variations, the solvent is glycerol, dimethylsulfoxide, N-methylpyrrolidone, ethanol, isopropanol, polyethylene glycols of various molecular weights (including but not limited to PEG 300 and PEG 400), or propylene glycol, or A mixture of one or more of them. the

In some variations, the solvent is polyethylene glycol. Polyethylene glycol is known by various names and is available in a variety of formulations, including but not limited to polyethylene glycol, macrogol 400, macrogol 1500, macrogol 4000, macrogol 6000, macrogol 20000, macrogola, breox PEG; carbowax; carbowax sentry; Hodag PEG; Lipo; Lipoxol; Lutrol E; PEG; 1,2-ethanediyl). the

Compositions and liquid formulations for delivery of therapeutic agents

The compositions and liquid formulations described herein can be used to deliver a therapeutic agent in an amount effective to treat, prevent, inhibit, delay the onset of, or cause regression of the diseases and conditions described in the "Diseases and Conditions" section. In some variations the compositions and liquid formulations described herein deliver one or more therapeutic agents over an extended period of time. the

An "effective amount" of a therapeutic agent for administration described herein (which is also referred to herein as a "therapeutically effective amount") refers to an amount that, when administered to a subject (including but not limited to a human subject), seeks to provide the therapeutic Effect of the amount of therapeutic agent. Achieving different therapeutic effects may require different effective amounts of the therapeutic agent. For example, a therapeutically effective amount of a therapeutic agent used to prevent a disease or condition may be different from a therapeutically effective amount used to treat, inhibit, delay onset or cause regression of a disease or condition. Additionally, a therapeutically effective amount may depend on the subject's age, weight and other medical conditions known to those skilled in the art of managing the disease or condition. Accordingly, a therapeutically effective amount can vary in each subject to which the therapeutic agent is administered. the

An effective amount of a therapeutic agent for treating, preventing, inhibiting, delaying the onset of, or causing regression of a particular disease or condition also refers herein to an amount of a therapeutic agent effective to treat, prevent, inhibit, delay the onset of, or cause its regression . the

In order to determine whether the level of therapeutic agent is a "therapeutically effective amount" to treat, prevent, inhibit, delay the onset of, or cause regression of the diseases and conditions described in the "Diseases and Conditions" section, the liquid formulation may be administered to an animal model of the disease or condition of interest , and observe the effect. In addition, dosages within the range of human clinical trials can be performed to determine a therapeutically effective amount of a therapeutic agent. the

In general, a therapeutic agent can be formulated in any composition or liquid formulation capable of delivering a therapeutically effective amount of the therapeutic agent to a subject or subject's eye for a desired delivery time. Compositions include liquid formulations. the

Solubilization of therapeutic agents

One composition or liquid formulation that can be used is one in which the therapeutic agent is dissolved in a solvent component. In general, any solvent having the desired effect in which the therapeutic agent is dissolved can be used. In some variations the solvent is aqueous. In some variations the solvent is non-aqueous. An "aqueous solvent" is a solvent that contains at least about 50% water. the

In general, any concentration of dissolved therapeutic agent that has the desired effect can be used. The solvent component may be a single solvent or a mixture of solvents. The types of solvents and solutions are well known to those skilled in the art of drug delivery. See e.g. Remington: The Science and Practice of Pharmacy, 20th ed., Lippincott Williams &Wilkins; 20th ed. (Dec. 15, 2000); Ansel's Pharmaceutical Dosage Forms and Drug Delivery Systems, 8th ed., Lippincott Williams & Wilkins (2004 August); HandbookOf Pharmaceutical Excipients 2003, American Pharmaceutical Association, Washington, DC, USA and Pharmaceutical Press, London, UK; and Strickley, Solubilizing Excipients in Oral and Injectable Formulations, Pharmaceutical Research.4, Vol. 220, No. February. the

As previously mentioned, some solvents can also be used as solubilizers. the

Solvents that can be used include, but are not limited to, DMSO, ethanol, methanol, isopropanol; castor oil, propylene glycol, glycerin, polysorbate 80, benzyl alcohol, dimethylacetamide (DMA), dimethylformamide (DMF) , triacetin, glyceryl diacetate, corn oil, acetyl triethyl citrate (ATC), ethyl lactate, glycerin formal, ethoxydiglycol (Transcutol, Gattefosse), triethylene glycol dimethyl Ether (Triglyme), dimethylisosorbide (DMI), γ-butyrolactone, N-methyl-2-pyrrolidone (NMP), polyethylene glycols of various molecular weights (including but not limited to PEG 300 and PEG 400) and pegylated caprylic glycerides (Labrasol, Gattefosse), a combination of any one or more of the foregoing, or an analog or derivative of any one or more of the foregoing. the

In some variations, the solvent is polyethylene glycol. Polyethylene glycol is known by various names and is available in a variety of formulations, including but not limited to polyethylene glycol, macrogol 400, macrogol 1500, macrogol 4000, macrogol 6000, macrogol 20000, macrogola, breox PEG; carbowax; carbowax sentry; Hodag PEG; Lipo; Lipoxol; Lutrol E; PEG; 1,2-ethanediyl). the

In some variations, the polyethylene glycol is a liquid PEG and is one or more of PEG 300 or PEG 400. the

Other solvents include an amount of _C6 - _C24 fatty acid sufficient to solubilize the therapeutic agent.

Phospholipid solvents can also be used, such as lecithin, phosphatidylcholine, or a mixture of various diglycerides (diglycerides of stearic, palmitic, and oleic acids) linked to phosphorylcholine esters; hydrogenated soybean phospholipids Acylcholine (HSPC), Distearoylphosphatidylglycerol (DSPG), L-α-Dimyristoylphosphatidylcholine (DMPC), L-α-Dimyristoylphosphatidylglycerol (DMPG). the

Other examples of solvents include ingredients such as alcohols, propylene glycol, polyethylene glycols of different molecular weights, propylene glycol esters, propylene glycol esterified with fatty acids such as oleic acid, stearic acid, palmitic acid, capric acid, linoleic acid, etc.; Medium chain mono-, di- or triglycerides, long chain fatty acids, naturally occurring oils and mixtures thereof. The oil component of the solvent system includes commercially available oils and naturally occurring oils. The oil may also be a vegetable oil or a mineral oil. The oils can be characterized as non-surface active oils, which generally do not have a hydrophilic-lipophilic balance. Commercially available materials comprising heavy chain triglycerides include, but are not limited to, Captex 100, Captex 300, Captex 355, Miglyol 810, Miglyol 812, Miglyol 818, Miglyol 829, and Dynacerin 660. Commercially available propylene glycol ester compositions include Captex 200 and Miglyol 840, among others. The commercially available product, Capmul MCM, contains one of many possible medium-chain mixtures containing mono- and diglycerides. the

Other solvents include naturally occurring oils such as peppermint and seed oils. Exemplary natural oils include oleic acid, castor oil, safflower seed oil, soybean oil, olive oil, sunflower oil, sesame oil, and peanut oil. Soy fatty acid can also be used. Examples of fully saturated non-aqueous solvents include, but are not limited to, esters of medium to long chain fatty acids (eg, fatty acid triglycerides having chain lengths of about _C6 to about _C24 ). Hydrogenated soybean oil and other vegetable oils can also be used. Mixtures of fatty acids can be isolated and refined from natural oils such as coconut oil, palm kernel oil, carnauba oil, and the like. In some embodiments, medium chain (about _C8 to about _C12 ) triglycerides such as caprylic/capric triglycerides from coconut oil or palm seed oil may be used. Medium chain mono- or diglycerides may also be used. Other fully saturated non-aqueous solvents include, but are not limited to, saturated coconut oil (which typically includes a mixture of lauric, myristic, palmitic, capric and caproic acids), including that sold under the trademark Miglyol ^™ from Huls and under the tradename Those of 810, 812, 829 and 840. The NeoBee ^(TM) product sold by Drew Chemicals is also mentioned. Non-aqueous solvents include isopropyl myristate. Examples of synthetic oils include triglycerides and propylene glycol esters of saturated or unsaturated fatty acids having from 6 to 24 carbon atoms, such as caproic acid, caprylic acid (caprylic acid), nonanoic acid (nonanoic acid), capric acid (capric acid), undecanoic acid, dodecanoic acid, tridecanoic acid, myristic acid (myristic acid), pentadecanoic acid, hexadecanoic acid (palmitic acid), heptadecanoic acid, tendecanoic acid Octanoic acid (stearic acid), nonadecanoic acid, heptadecanoic acid, eicosanoic acid, eicosanoic acid, behenic acid, and lignoceric acid, etc. Examples of unsaturated carboxylic acids include oleic acid, linoleic acid, linolenic acid, and the like. Non-aqueous solvents may include mono-, diglycerides and triglycerides or mixed glycerides and/or propylene glycol mono- or di-esters of fatty acids in which at least one molecule of glycerol is esterified with fatty acids having different carbon atom lengths. A non-limiting example of a "non-oil" used as a solvent is polyethylene glycol.

Exemplary vegetable oils include cottonseed oil, corn oil, sesame oil, soybean oil, olive oil, fractionated coconut oil, peanut oil, sunflower oil, safflower oil, almond oil, avocado oil, palm oil, palm kernel oil, carnauba oil , beech nut oil, linseed oil, canola oil, etc. Mono-, diglycerides, and triglycerides of vegetable (including, but not limited to, corn) oils can also be used. the

Crosslinked or uncrosslinked polyvinylpyrrolidone (PVP) can also be used as solvent. Other solvents include but not limited to C ₆ -C ₂₄ fatty acids, oleic acid, Imwitor 742, Capmul, F68, F68 (Lutrol), PLURONICS (including but not limited to PLURONICS F108, F127 and F68, Poloxamers, Jeffamines), Tetronics, F127; cyclodextrins such as α-cyclodextrin, β-cyclodextrin, hydroxypropyl-β-cyclodextrin, sulfobutyl ether-β-cyclodextrin (Captisol); CMC, polysorbitan 20, Cavitron, polyethylene glycols of various molecular weights including but not limited to PEG300 and PEG400.

Beeswax and d-alpha-tocopherol (vitamin E) can also be used as solvents. the

Solvents for liquid formulations can be determined by a variety of methods known in the art, including but not limited to (1) theoretically estimating their solubility parameter values using standard equations in the art and selecting a solvent that matches the therapeutic agent. the solvent; and (2) experimentally determine the saturation solubility of the therapeutic agent in the solvent, and select a solvent that exhibits the desired solubility. the

Solubilization of rapamycin

When the therapeutic agent is rapamycin, solvents that can be used to prepare rapamycin solutions or suspensions include but are not limited to any solvents described herein, including but not limited to DMSO, glycerol, ethanol, methanol, isopropanol ; Castor Oil, Propylene Glycol, Polyethylene Propylene, Glycerin, Polysorbate 80, Benzyl Alcohol, Dimethylacetamide (DMA), Dimethylformamide (DMF), Glycerin Formal, Ethoxydiglycol (Transcutol , Gattefosse), triethylene glycol dimethyl ether (Triglyme), dimethyl isosorbide (DMI), γ-butyrolactone, N-methyl-2-pyrrolidone (NMP), polyethylene glycol with various molecular weights Any one or more of alcohol (including but not limited to PEG 300 and PEG 400) and pegylated caprylic glycerides (Labrasol, Gattefosse). the

Other solvents include but not limited to _C6 - _C24 fatty acids, oleic acid, Imwitor 742, Capmul, F68, F68 (Lutrol), PLURONICS (including but not limited to PLURONICS F108, F127 and F68, Poloxamers, Jeffamines), Tetronics , F127, beta-cyclodextrin, CMC, polysorbitan 20, Cavitron, softigen 767, captisol and sesame oil.

Other methods that can be used to solubilize rapamycin are described in Solubilization of Rapamycin, P. Simamora et al. Int'l J. Pharma 213 (2001) 25-29, the entire contents of which are incorporated herein by reference. the

As non-limiting examples, rapamycin can be dissolved in 5% DMSO or methanol in equilibrated saline solution. The rapamycin solution can be an unsaturated, saturated or supersaturated rapamycin solution. The rapamycin solution can be contacted with solid rapamycin. In one non-limiting example, rapamycin can be dissolved at a concentration of up to about 400 mg/ml. Rapamycin can also be dissolved, for example, in propylene glycol esterified with fatty acids, such as oleic acid, stearic acid, palmitic acid, capric acid, linoleic acid, and the like. the

Many other solvents are possible. Those of ordinary skill in the art will find it routine to identify solvents for rapamycin in light of the teachings herein. the

Solubilizers

Generally, any solubilizing agent or combination of solubilizing agents can be used in the liquid formulations described herein. the

In some variations, the solubilizing agent is a surfactant or a combination of surfactants. Many surfactants are possible. Combinations of surfactants, including combinations of types of surfactants, may also be used. For example, nonionic, anionic (eg soap, sulfonate), cationic (eg CTAB), zwitterionic, polymeric or amphoteric surfactants may be used. the

Useful surfactants can be determined by mixing the therapeutic agent of interest with a putative solvent and a putative surfactant, and observing the properties of the formulation after exposure to the medium. the

Examples of surfactants include, but are not limited to, fatty acid esters or amides or ether analogs, or hydrophilic derivatives thereof; monoesters or diesters, or hydrophilic derivatives thereof; or mixtures thereof; monoglycerides or glycerol Diesters, or hydrophilic derivatives thereof; or mixtures thereof; mixtures with enriched monoglycerides and/or diglycerides, or hydrophilic derivatives thereof; surfactants partially derivatized with hydrophilic moieties; Monoesters or diesters or polyesters of other alcohols, polyols, sugars or oligosaccharides or polysaccharides, their oxyalkylene oligomers or polymers or block polymers, or their hydrophilic derivatives, or their amide analogues; Fatty acid derivatives of amines, polyamines, polyimines, aminoalcohols, aminosugars, hydroxyalkylamines, hydroxypolyamines, peptides, polypeptides or ether analogs thereof. the

Hydrophile-lipophile balance ("HLB") represents the relative simultaneous attraction of a surfactant to water and oil (or to the two phases of the emulsion system). the

Surfactants are characterized according to the balance between the hydrophilic and lipophilic parts of their molecules. The hydrophilic-lipophilic balance (HLB) number indicates the polarity of a molecule in an arbitrary range of 1-40, with most commonly used emulsifiers having values between 1-20. HLB increases with increasing hydrophilicity. the

Surfactants that may be used include, but are not limited to, surfactants having an HLB greater than 10, 11, 12, 13, or 14. Examples of surfactants include polyoxyethylene products of hydrogenated vegetable oils, polyethoxylated castor oil or polyethoxylated hydrogenated castor oil, polyoxyethylene-sorbitan fatty acid esters, polyoxyethylene castor oil Derivatives, etc., such as Nikkol HCO-50, NikkolHCO-35, Nikkol HCO-40, Nikkol HCO-60 (from Nikko Chemicals Co.Ltd.); Cremophor (from BASF) such as Cremophor RH40, Cremophor RH60, Cremophor EL, TWEENs ( from ICI Chemicals), such as Tween 20, Tween 21, Tween 40, Tween 60, Tween 80, Tween 81, Cremophor RH 410, Cremophor RH455, etc. the

The surfactant component may be selected from compounds having at least one ether formed from at least about 1 to 100 ethylene oxide units and at least one fatty alcohol chain having at least about 12 to about 22 carbon atoms; Compounds having esters of from about 1 to 100 ethylene oxide units and at least one fatty acid chain having at least about 12 to 22 carbon atoms; compounds having at least one ether formed of from at least about 1 to 100 ethylene oxide units, compounds of esters or amides and at least one vitamin or vitamin derivative; and combinations thereof consisting of not more than two surfactants. the

Other examples of surfactants include Lumulse GRH-40, TGPS, Polysorbate-80 (Tween-80), Polysorbate-20 (Tween-20), Polyoxyethylene (20) sorbitan mono Oleate, glyceryl glycol ester, polyethylene glycol ester, polyglycolyzed glycerides, etc. or mixtures thereof; polyethylene sorbitan fatty acid ester, polyoxyethylene glyceride such as Tagat TO, Tagat L, Tagat I, tagat I2 and Tagat 0 (commercially available from Goldschmidt Chemical Co., Essen, Germany); ethylene glycol esters such as ethylene glycol stearate and ethylene glycol distearate; propylene glycol esters , such as propylene glycol myristate; fatty acid glycerides, such as glyceryl stearate and glyceryl monostearate; sorbitan esters, such as Spans and Tweens; polyglycerides, such as polyglyceryl 4 - Oleates; fatty alcohol ethoxylates such as Brij type emulsifiers; ethoxylated propoxylated block copolymers such as poloxamers; polyethylene glycol esters of fatty acids such as PEG 300 Glyceryl Linoleate or Labrafil 2125 CS, PEG 300 Glyceryl Oleate or Labrafil M 1944 CS, PEG 400 Caprylic/Capric Glycerides or Labrasol and PEG 300 Caprylic/Capric Glycerides or Softigen 767; cremophors such as Cremophor E , Polyoxyl 35 Castor Oil or Cremophor EL, Cremophor EL-P, Cremophor RH 4OP, Polyoxyl 40 Hydrogenated Castor Oil, Cremophor RH40; Polyoxyl 60 Hydrogenated Castor Oil or Cremophor RH 60, Monocaprylic/Capric Glycerides , such as Campmul CM 10; polyoxyethylated fatty acids (PEG-stearate, PED-laurate, Brij ), polyoxyethylated fatty acid glycerides, polyoxyethylated glycerin fatty acid esters, such as Solutol HS-15; PEG-ether (Mirj ), sorbitan derivatives (Tween), sorbitan monooleate or Span 20, aroma compounds (Tritons ), PEG-Glycerides (PECEOL ^TM ), PEG-PPG (Polypropylene Glycol) Copolymers (PLURONICS, including but not limited to PLURONICS F108, F127 and F68, Poloxamers, Jeffamines), Tetronics, Polyglycerol, PEG-Tocopherol Phenol, PEG-LICOL 6-oleate; propylene glycol derivatives, alkyl and acyl derivatives of sugars and polysaccharides (octyl sucrose, sucrose stearate, lauroyl dextran, etc.) and/or mixtures thereof; based on Surfactants of oleic or lauric esters of polyols copolymerized with ethylene oxide; Labrasol Gelucire 44/14; polyoxyethylene stearate; saturated polyethylene glycolated glycerides; or poloxamers; All are commercially available. The polyoxyethylene sorbitan fatty acid ester may include polysorbates such as polysorbate 20, polysorbate 40, polysorbate 60, and polysorbate 80. The polyoxyethylene stearate may include polyoxyethylene 6 stearate, polyoxyethylene 8 stearate, polyoxyethylene 12 stearate, and polyoxyethylene 20 stearate. Saturated PEGylated glycerides are eg GELUCIRE 44/14 or GELUCIRE ^™ 50/13 (Gattefosse, Westwood, NJ, USA). Poloxamer as used herein includes Poloxamer 124 and Poloxamer 188.

Surfactants include d-alpha-tocopheryl polyethylene glycol 1000 succinate (TPGS), polyoxyethylene 8 stearate (PEG 400 monostearate), polyoxyethylene 40 stearate ( PEG 1750 monostearate) and peppermint oil. the

In some variations, a surfactant with an HLB below 10 is used. Such surfactants may optionally be used in combination with other surfactants as co-surfactants. Some surfactants, mixtures and other equivalent compositions having an HLB less than or equal to 10 are propylene glycol, glyceryl fatty acids, glyceryl fatty acid esters, polyethylene glycol esters, glyceryl glycol esters, polyethylene glycolated glycerol esters and polyoxyethylene sterol ethers. Propylene glycol esters or partial esters form a component of commercial products such as Lauroglycol FCC (which contains propylene glycol laurate). The commercially available excipient Maisine 35-1 contains long chain fatty acids such as glyceryl linoleate. Products containing polyoxyethylene stearate, such as Acconon E, may also be used. Labrafil M 1944 CS is an example of a surfactant where the composition contains a mixture of glyceryl glycol esters and polyethylene glycol esters. the

Solubilizer of rapamycin

A number of solubilizing agents can be used with rapamycin, including but not limited to those described above in the "Solubilizing Agents" section. the

In some variations, the solubilizing agent is a surfactant. Non-limiting examples of surfactants that can be used with rapamycin include, but are not limited to, surfactants with an HLB of greater than 10, 11, 12, 13, or 14. A non-limiting example is Cremophor EL. In some variations, the surfactant can be a polymeric surfactant, including but not limited to PLURONICS F108, F127 and F68 and Tetronics. Some of the solvents used herein may also function as surfactants. Those of ordinary skill in the art will find it routine to identify which solubilizers and surfactants are useful for rapamycin in light of the teachings herein. the

viscosity modifier

The liquid formulations described herein may be administered with or also contain a viscosity modifier. the

One exemplary viscosity modifier that can be used is hyaluronic acid. Hyaluronic acid is a glycosaminoglycan. It consists of repeating sequences of glucuronic acid and glucosamine. Hyaluronic acid is present in many tissues and organs of the body and contributes to the viscosity and consistency of such tissues and organs. Hyaluronic acid is present in the eye (including the vitreous of the eye) and together with collagen contributes to its viscosity. The liquid formulations described herein may also comprise or be administered with hyaluronic acid. the

Other non-limiting examples of viscosity modifiers include polyalkylene oxides, glycerin, carboxymethylcellulose, sodium alginate, chitosan, dextran, dextran sulfate, and collagen. These viscosity modifiers can be chemically modified. the

Other viscosity modifiers that may be used include, but are not limited to, carrageenan, cellulose gel, colloidal silica, gelatin, propylene carbonate, carbonic acid, alginic acid, agar, carboxyvinyl polymer, or carbomer and polyacrylamide, gum arabic, ester gum, guar gum, acacia gum, ghatti, caraya gum, tragacanth gum, terra, pectin, tamarind, larch arabinogalactan, alginate, sophora Soy, xanthan gum, starch, magnesium aluminum silicate, tragacanth gum, polyvinyl alcohol, gellan gum, hydrocolloid blend, and povidone. Other viscosity modifiers known in the art may also be used, including but not limited to sodium carboxymethylcellulose, sodium alginate, carrageenan, galactomannan, hydroxypropylmethylcellulose, hydroxypropylcellulose Polyethylene glycol, polyvinylpyrrolidone, sodium carboxymethyl chitosan, sodium carboxymethyl dextran, sodium carboxymethyl starch, xanthan gum and zein. the

Other components of liquid preparations

The formulations described herein may also contain various other ingredients, such as stabilizers. Stabilizers that can be used in the formulations described herein include, but are not limited to, those that will (1) increase the compatibility of excipients with encapsulating materials such as gelatin, (2) increase the compatibility of therapeutic agents such as rapamycin and/or rapamycin. Stability of derivatives (eg, prevention of crystal growth of therapeutic agents such as rapamycin), and/or (3) agents that increase formulation stability. Note that there is overlap between ingredients that are stabilizers and ingredients that are solvents, solubilizers, or surfactants, and that the same ingredient can serve more than one role. the

Stabilizers may be selected from fatty acids, fatty alcohols, alcohols, long-chain fatty acid esters, long-chain ethers, hydrophilic derivatives of fatty acids, polyvinylpyrrolidone, polyvinyl ether, polyvinyl alcohol, hydrocarbons, hydrophobic polymers, hygroscopic polymers and combinations thereof. Amide analogs of the aforementioned stabilizers may also be used. Stabilizers chosen can alter the hydrophobicity of the formulation (e.g. oleic acid, waxes) or facilitate mixing of ingredients in the formulation (e.g. ethanol), control the moisture level of the formulation (e.g. PVP), control phase mobility (with high Substances with a melting point at room temperature, such as long-chain fatty acids, alcohols, esters, ethers, amides, etc., or mixtures thereof; waxes) and/or to promote compatibility of the formulation with the encapsulating material (such as oleic acid or waxes). Some of these stabilizers are available as solvents/co-solvents (e.g. ethanol). Stabilizers may be present in sufficient amounts to inhibit crystallization of the therapeutic agent (eg, rapamycin). the

Examples of stabilizers include, but are not limited to, saturated, monoene, polyene, branched, ring-containing, alkyne, dicarboxylic and functional group-containing fatty acids such as oleic acid, caprylic acid, capric acid, caproic acid, lauryl Acids, myristic acid, palmitic acid, stearic acid, behenic acid, linoleic acid, linolenic acid, eicosapentaenoic acid (EPA), DHA; fatty alcohols such as stearyl alcohol, cetyl alcohol, ceteryl alcohol; Other alcohols such as ethanol, isopropanol, butanol; long-chain fatty acid esters, ethers or amides such as glyceryl stearate, cetyl stearate, oleyl ether, stearyl ether, cetyl ether , oleylamide, stearamide; hydrophilic derivatives of fatty acids, such as polyglyceryl fatty acids, polyethylene glycol fatty acid esters; polyvinylpyrrolidone, polyvinyl alcohol (PVA), wax, docosahexaenoic acid and dehydroabietic acid, etc. the

The formulations may also contain gelling agents which modify the texture of the final formulation by forming a gel. the

A therapeutic agent as described herein (such as rapamycin) may be subjected to conventional pharmaceutical manipulations (e.g., sterilization), and compositions containing the therapeutic agent may also contain conventional adjuvants such as preservatives, stabilizers, wetting agents , emulsifier, buffer, etc. Therapeutic agents can also be formulated with clinically used pharmaceutically acceptable excipients for the manufacture of pharmaceutical compositions. Formulations for ophthalmic administration may be present as solutions, suspensions, particles of solid material, discrete pieces of solid material, incorporated into polymer matrices, liquid formulations, or any other form for ophthalmic administration. Therapeutic agents are useful in the manufacture of medicaments for treating, preventing, inhibiting, delaying the onset of, or causing regression of any of the conditions described herein. In some variations, a therapeutic agent may be used in the manufacture of a medicament to treat any of the conditions described herein. the

Compositions containing a therapeutic agent such as rapamycin may contain one or more adjuvants suitable for the indicated route of administration. Adjuvants with which the therapeutic agent may be mixed include, but are not limited to, lactose, dextrose, starch powder, cellulose alkanoate, stearic acid, talc, magnesium stearate, magnesium oxide, sodium and calcium salts of phosphoric and sulfuric acids , gum arabic, gelatin, sodium alginate, polyvinylpyrrolidine and/or polyvinyl alcohol. When a solubilized formulation is desired, the therapeutic agent can be in a solvent including, but not limited to, polyethylene glycol of various molecular weights, propylene glycol, carboxymethylcellulose colloidal solution, methanol, ethanol, DMSO, corn oil, Peanut oil, cottonseed oil, sesame oil, tragacanth and/or various buffers. Other adjuvants and modes of administration are well known in the art of pharmacy and may be used in the practice of the methods, compositions and liquid formulations described herein. The carrier or diluent may include a time delay material such as glyceryl monostearate or glyceryl distearate, alone or with a wax, or other materials well known in the art. Formulations for the uses described herein may also include gel formulations, erodible and non-erodible polymers, microspheres and liposomes. the

Other adjuvants and excipients that may be used include, but are not limited to, _C8 - _C10 fatty acid esters such as softigen 767, polysorbate 80, PLURONICS, Tetronics, Miglyol and Transcutol.

Additives and diluents commonly used in the pharmaceutical field may be optionally added to pharmaceutical compositions and liquid preparations. These additives and diluents include thickening agents, granulating agents, dispersing agents, flavoring agents, sweeteners, colorants and stabilizers (including pH stabilizers), other excipients, antioxidants (e.g., tocopherols, BHA, BHT, TBHQ, tocopheryl acetate, ascorbyl palmitate, propyl ascorbyl gallate, etc.), preservatives (such as parabens), etc. Exemplary preservatives include, but are not limited to, benzyl alcohol, ethanol, benzalkonium chloride, phenol, chlorobutanol, and the like. Some useful antioxidants provide oxygen or peroxide inhibitors to the formulation and include, but are not limited to, butylated hydroxytoluene, butylated hydroxyanisole, propyl gallate, ascorbyl palmitate, alpha-tocopherol, and the like. Thickeners such as lecithin, hydroxypropylcellulose, aluminum stearate, etc. can improve the texture of the preparation. the

In some variations, the therapeutic agent is rapamycin formulated as rapamycin in solid or liquid form. In some variations, rapamycin is formulated as an oral dose. the

Additionally, viscous polymers may be added to the suspension to aid in positioning and ease of placement and handling. In some uses of the liquid formulation, a pocket may be surgically formed in the sclera to receive infusion of the liquid formulation. The hydrogel structure of the sclera acts as a rate-controlling membrane. The therapeutic agent substance particles used to form the suspension can be produced by known methods including, but not limited to, for example, by ball milling using ceramic beads. For example, a Cole Parmer ball mill such as the Labmill 8000 can be used with 0.8mm YTZ ceramic beads from Tosoh or Norstone Inc. the

The formulations may conveniently be presented in unit dosage form and may be prepared by conventional techniques of pharmacy. Such techniques include the step of bringing into association the therapeutic agent and a pharmaceutical carrier or excipient. The formulations can be prepared by uniformly and intimately bringing into association the active ingredient with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product. the

In some variations, the formulations described herein are presented in one or more unit dosage forms, wherein the unit dosage forms contain an amount of a liquid formulation described herein effective to treat or prevent the disease or condition for which it is administered. In some variations, the formulations described herein are provided in one or more unit dosage forms comprising an amount of a liquid rapamycin formulation described herein effective to treat or prevent the disease or condition. the

In some embodiments, a unit dosage form is formulated at its concentration to be administered. In some variations, unit dosage forms are diluted prior to administration to a subject. In some variations, liquid formulations described herein are diluted in an aqueous medium prior to administration to a subject. In some variations the aqueous medium is isotonic. In some variations, liquid formulations described herein are diluted in a non-aqueous medium prior to administration to a subject. the

Another aspect herein provides kits comprising one or more unit dosage forms described herein. In some embodiments, the kit comprises one or more packages and instructions for treating one or more diseases or conditions. In some embodiments, the kit comprises a diluent that does not physically contact the formulation or pharmaceutical formulation. In some embodiments, the kit comprises any one or more unit dosage forms described herein in one or more sealed tubes. In some embodiments, the kit comprises any one or more sterile unit dosage forms. the

In some variations, the unit dosage form is a container, including but not limited to a sterile sealed container. In some variations, the container is a vial, ampoule, or small volume applicator, including but not limited to a syringe. In some variations, the small volume applicator is prefilled with rapamycin for the treatment of an ophthalmic disease or condition, including but not limited to the limus compound for the treatment of age-related macular degeneration. Described herein are prefilled small volume applicators prefilled with formulations containing therapeutic agents, including but not limited to rapamycin. In some variations, the small volume applicator is prefilled with a solution containing a therapeutic agent (including but not limited to rapamycin) and polyethylene glycol, and optionally also contains one or more additional ingredients, including but not only limited to ethanol. In some variations, prefilled small volume applicators are prefilled with a solution comprising about 2% rapamycin, about 94% PEG-400, about 4% ethanol. the

Kits comprising one or more containers are described herein. In some variations, a kit comprising one or more small volume applicators is prefilled with a therapeutic agent-containing formulation described herein, including but not limited to formulations comprising rapamycin, formulations comprising rapamycin and polyethylene glycol Formulations of diol and optionally one or more additional ingredients including, but not limited to, ethanol, and formulations comprising about 2% rapamycin, about 94% PEG-400, about 4% ethanol Preparations in liquid form. In some variations, the kit contains one or more containers (including, but not limited to, pre-filled small volume applicators) and instructions for their use. In another variation, the kit comprises one or more small volume applicators prefilled with rapamycin and instructions for their use in the treatment of an ocular disease or condition. In some variations, containers described herein are in secondary packaging. the

route of administration

The compositions, methods and liquid formulations described herein deliver one or more therapeutic agents to a subject (including but not limited to a human subject). the

In some variations, the compositions, methods and liquid formulations described herein deliver one or more therapeutic agents to an aqueous medium in a human subject. the

In some variations, the compositions, methods, and liquid formulations described herein deliver one or more therapeutic agents to an aqueous solution in or near the area of a disease or condition to be treated, prevented, inhibited, delayed, or resolved. medium. the

In some variations, the compositions, methods, and liquid formulations described herein deliver one or more therapeutic agents to the subject's eye, including macular and retinal choroidal tissue, in an amount and for a duration of It is effective in treating, preventing, inhibiting, delaying the onset of or regressing the diseases and disorders described in the "Diseases and disorders" section. the

As used herein, "retinal choroid" and "retinal choroidal tissue" are synonymous and refer to the combined retinal and choroidal tissue of the eye. the

As a non-limiting example, the compositions, liquid formulations, and methods described herein may be administered directly or via the periocular route to Tissue: Vitreous, aqueous humor, sclera, conjunctiva, between sclera and conjunctiva, retinal choroidal tissue, macula, or other areas in or near the subject's eye. Effective amounts and durations can vary for each treatment, prevention, inhibition of the onset or regression of CNV and wet AMD, and can vary for each different delivery site. the

Intravitreal administration is more invasive than some other types of ocular procedures. Because of the possible risk of adverse effects, intravitreal administration is not optimal for treating relatively healthy eyes. In contrast, periocular administration (eg, subconjunctival administration) is much less invasive than intravitreal administration. When the therapeutic agent is delivered by the periocular route, patients with healthier eyes can be treated than those who can be treated using intravitreal administration. In some variations, subconjunctival injection is used to prevent or delay onset of the disease or condition of the eye, wherein the subject's eye has visual acuity of 20/40 or better. the

"Subconjunctival" placement or injection as used herein refers to placement or injection between the sclera and conjunctiva. Subconjunctival administration is sometimes referred to herein as "sub-conj". the

Routes of administration that can be used to administer a liquid formulation include, but are not limited to, placing the liquid formulation in an aqueous medium in a subject, such as by injection, including, but not limited to, placing the liquid formulation in a subject, including but not limited to, by injection. not limited to the eyes of human subjects). Liquid formulations may be administered systemically, including but not limited to the following routes of delivery: rectal, vaginal, infusion, intramuscular, intraperitoneal, intraarterial, intrathecal, intrabronchial, intracisternal, epidermal, subcutaneous, intradermal, transdermal, intravenous , intracervical, intraperitoneal, intracranial, intraocular, intrapulmonary, intrathoracic, intratracheal, nasal, buccal, sublingual, oral, parenteral or sprayed or atomized using an aerosol propellant. the

Compositions and liquid formulations containing therapeutic agents can be administered directly to the eye using a variety of procedures including, but not limited to, procedures in which (1) the therapeutic agent is administered by injection using a syringe and hypodermic needle, (2) using a special The device is designed to inject the therapeutic agent, and (3) prior to the injection of the therapeutic agent, a pocket is surgically formed in the sclera to serve as a container for the therapeutic agent or composition of therapeutic agents. For example, in one administration procedure, a surgeon forms a pocket within the sclera of the eye and then injects a solution or liquid formulation containing the therapeutic agent into the pocket. the

Other administration procedures include, but are not limited to, procedures in which (1) the formulation of the therapeutic agent is injected through a specially designed curved cannula, placing the therapeutic agent directly against the portion of the eye, (2) the therapeutic agent is placed in compressed form directly Partially placed on the eye, (3) the therapeutic agent is inserted into the sclera via a specially designed syringe or inserter, (4) a liquid formulation containing the therapeutic agent is incorporated into the polymer, (5) the surgeon makes a small conjunctival incision, Sutures and any therapeutic agent delivery structures are threaded through the incision, securing the structure adjacent to the sclera, (6) needles are used for direct injection into the vitreous of the eye or into any other site described. the

The liquid formulations described herein can be used directly (eg, by injection), as elixirs for topical administration (including but not limited to, by eye drops), or contained in soft or hard gelatin or starch capsules. Capsules may be tied to prevent leakage. the

delivered by injection

One method that can be used to deliver the compositions and liquid formulations described herein is by injection. In this method, the compositions and liquid formulations may be injected into a subject (including but not limited to a human subject), or into a location in or near the eye of a subject, for delivery to the subject or subject the eye of the reader. Injections include, but are not limited to, intraocular and periocular injections. Non-limiting examples of locations in or near the eyes of a subject are as follows. the

Injection of the therapeutic agent into the vitreous can provide high local concentrations of the therapeutic agent within the vitreous and retina. In addition, the elimination half-life of drugs in the vitreous has been found to increase with molecular weight. the

Intracameral injections or injections into the anterior chamber may also be used. In one example, up to 100 μl may be injected intracamerally. the

The periocular route of delivery can deliver therapeutic agents to the retina without some of the risks of intravitreal delivery. Periocular routes include, but are not limited to, subconjunctival, subtenon, retrobulbar, periocular, and postjuxtascleral delivery. A "periocular" route of delivery refers to placement near or around the eye. For an exemplary description of the periocular route of retinal drug delivery see Pericular routes for retinal drug delivery, Raghava et al. (2004), Expert Opin. Drug Deliv. 1(1):99-114, which is incorporated herein by reference in its entirety. the

In some variations, the liquid formulations described herein are administered intraocularly. Intraocular administration includes placement or injection into the eye (including but not limited to the vitreous). the

Subconjunctival injections may be performed by injecting the therapeutic agent under the conjunctiva, or between the sclera and the conjunctiva. In one example, up to about 500 μl may be injected subconjunctivally. As a non-limiting example, a needle of about 25 to 30 gauge and about 30 mm long may be used. Local pressure at the subconjunctival site of therapeutic agent administration can enhance delivery of therapeutic agent to the posterior segment by reducing local choroidal blood flow. the

Subtenon injections can be made by injecting the therapeutic agent into the tenon's capsule around the upper part of the eye, and into the "belly" of the superior rectus muscle. In one example, up to about 4 ml may be injected subtenon. As a non-limiting example, a blunt cannula approximately 2.5 cm long can be used. the

Retrobulbar injection refers to the injection into the conical compartment of the four rectus muscles behind the eyeball and their intermuscular septum. In one example, up to about 5 ml may be injected retrobulbarally. As a non-limiting example, blunt needles of about 25 or about 27 gauge may be used. the

Periocular injections can be at locations outside the region of the four rectus muscles and their intermuscular membranes (i.e., outside the cone of the muscle). Periocular injections may be made, for example, in volumes up to about 10 ml. As a non-limiting example, a blunt cannula about 1.25 inches long and about 25 gauge can be used. the

Juxtascleral post-delivery refers to placing the therapeutic near or on the macula, in direct contact with the outer surface of the sclera, and without puncturing the eyeball. In one example, up to about 500 ml may be post juxtascleral injected. As a non-limiting example, a blunt curved cannula (specifically a 56° design) is used to place the therapeutic agent in the scleral incision. the

In some variations, the liquid formulations described herein are injected intraocularly. Intraocular injection includes injection into the eye. the

Sites where the compositions and liquid formulations may be administered include, but are not limited to, the vitreous, aqueous humor, sclera, conjunctiva, between the sclera and the conjunctiva, the retinal choroidal tissue, the macula, or other areas in or near the subject's eye. Methods that can be used to deliver compositions and liquid formulations include, but are not limited to, injection. the

In one method that can be used, the therapeutic agent is dissolved in a solvent or mixture of solvents and injected into the subject's vitreous, aqueous humor, sclera, conjunctiva, between the sclera and conjunctiva, retinal choroidal tissue, macula or Other areas in or near the eye, or other media in or near the subject. In one such method that can be used, the therapeutic agent is rapamycin in a liquid formulation. the

When the therapeutic agent is rapamycin, the compositions and liquid formulations are useful for delivering or maintaining an amount of rapamycin in ocular tissue, including but not limited to retina, choroid, or vitreous, in an amount effective Treat AMD. In one non-limiting example, it is believed that a liquid formulation of rapamycin in an amount capable of delivering about 0.1 pg/ml to about 2 μg/ml of rapamycin in the vitreous is useful for the treatment of wet AMD. concentration. In some non-limiting examples, liquid formulations that deliver a concentration of about 0.1 pg/mg to about 1 μg/mg rapamycin in retinal choroidal tissue are believed to be useful in the treatment of wet AMD. Other effective concentrations can be readily determined by one of skill in the art based on the teachings described herein. the

Methods of preparing liquid formulations

A non-limiting method that can be used to prepare the liquid formulations described herein, including but not limited to liquid formulations containing rapamycin, is by mixing a solvent and therapeutic agent at room temperature or slightly elevated temperature (optionally using Sonicator) together until a solution or suspension is obtained, then cool the formulation. Other ingredients, including but not limited to those described above, can then be mixed with the formulation. Other preparative methods that may be used are described herein, including in the Examples, and those skilled in the art will be able to select other preparative methods based on the teachings herein. the

Extended Delivery of Therapeutics

Described herein are compositions and liquid formulations that exhibit in vivo delivery or clearance profiles having one or more of the following characteristics. The delivery or clearance profile is the clearance profile of the therapeutic agent following subconjunctival injection or injection of the composition or liquid formulation into the vitreous of a rabbit eye. In some variations, the delivery or clearance profile is the clearance profile of rapamycin in vivo following subconjunctival injection or injection of the composition or liquid formulation into the vitreous of a rabbit eye. The volume of the rabbit vitreous is about 30-40% of the volume of the human vitreous. The amount of therapeutic agent is measured using techniques as described in Example 2, without limitation the formulation and therapeutic agent described in Example 2. the

In some variations, therapeutic agents having in vivo delivery or clearance profiles described herein include, but are not limited to, those described in the "Therapeutic Agents" section. In some variations the therapeutic agent is rapamycin. In some variations, the liquid formulations described herein are used to deliver the therapeutic agent at an equivalent concentration to rapamycin. The liquid formulations described herein may contain any therapeutic agent equivalent to rapamycin (including but not limited to the concentrations described herein included in the Examples), including but not limited to those described in the "Therapeutic Agents" section. the

"In vivo average percent" levels refer to the average concentration of therapeutic agent obtained from multiple rabbit eyes for a given time point and are divided by the average concentration of therapeutic agent at one time point by the average concentration of therapeutic agent at another time point. In some variations of the average percentage level in vivo, the therapeutic agent is rapamycin. the

The average concentration of therapeutic agent in rabbit eye tissue at a given time after administration of the formulation containing the therapeutic agent can be measured according to the following method. When injecting volumes less than 10 μl, use a Hamilton syringe. the

Liquid preparations are stored at a temperature of 2-8°C until use. the

The experimental animals were New Zealand White rabbits with specific pathogen free (SPF). A mixed population of about 50% males, about 50% females was used. Rabbits are at least 12 weeks old, usually at least 14 weeks old at the time of dosing. Each rabbit weighs at least 2.2 kg, usually at least 2.5 kg at the time of dosing. Animals were quarantined for at least one week prior to the study and checked for general health parameters. No unhealthy animals of any kind were used in the research. For a given time point, at least 6 eyes were measured and averaged. the

House and sanitize according to standard procedures used in industry. Animals were provided with approximately 150 g of Teklad Certified Hi-Fiber Rabbit Diet per day, and tap water was provided ad libitum. The water was known to be free of contamination and no additional analysis was performed beyond that provided by the local water community. Monitor environmental conditions. the

Each animal underwent a pre-treatment ophthalmic examination (slit-lamp and ophthalmoscopy) by a board-certified veterinary ophthalmologist. Eye examination findings were scored according to the McDonald and Shadduck scoring system described in Dermatoxicology, F.N. Marzulli and H.I. Maibach, 1977 "Eye Irritation," T.O. McDonald and J.A. Shadduck (pp. 579-582). Record observations using standardized data collection forms. Acceptance criteria for the study were as follows: conjunctival hyperemia and swelling scores ≤ 1; all other observed variables scored 0. the

Gentamicin eye drops were administered to both eyes of each animal twice a day on the day before the administration, on the day of the administration (Day 1) and on the day after the administration (Day 2). Dosing was performed in two phases, the first phase involving one group of animals and the second phase involving the other animals. Animals were randomized into masked treatment groups according to a modified Latin formula prior to each dosing period. Animals were fasted for at least 8 hours prior to injection. Record the time of initiation of fasting and the time of injection. the

Animals were weighed and anesthetized with an intravenous injection of a ketamine/xylazine mixture (87 mg/mL ketamine, 13 mg/mL xylazine) at a volume of 0.1-0.2 mL/kg. Both eyes of each animal were prepared for injection as follows: approximately 5 minutes prior to injection, the eyes were moistened with ophthalmic betadine. After five minutes the povidone-iodine was washed out of the eye with sterile saline. 0.5% (1-2 drops) of Proparacaine hydrochloride was delivered to each eye. For eyes to be injected intravitreally, 1% Tropicamide (1 drop) was delivered to each eye. the

On day 1, the eyes of each animal were injected with the test article or the control article. Animals in selected groups were dosed a second time on day 90±1. Subconjunctival or intravitreal administration. Actual treatments, injection sites, and dose volumes were masked and revealed at the end of the study. the

Subconjunctival injections were given using an insulin syringe and a 30 gauge x 1/2 inch needle. Use forceps to elevate the bulbar conjunctiva in the dorsotemporal quadrant. The test article is injected into the subconjunctival space. the

Intravitreal injections were performed using an insulin syringe and a 30 gauge x 1/2 inch needle. For each injection, the needle was introduced through the ventral-nasal quadrant of the eye, 2-3 mm posterior to the limbus, with the bevel of the needle pointing in a downward and posterior direction, avoiding the lens. Inject the test article into the vitreous near the retina with a single bolus injection. the

Animals were observed twice daily for mortality/morbidity. Animals determined to be moribund were euthanized with a commercially available euthanasia solution intravenously. Both eyes were removed and stored frozen at -70°C for possible future evaluation. If the animal was found dead prior to post mortem cataract, both eyes were enucleated and stored frozen at -70°C for possible future evaluation. Animals found dead after postmortem rigidity were not subjected to necropsy. the

Animals were weighed randomly on day 1 before dosing and before euthanasia. the

Ophthalmic observations (slit-lamp and ophthalmoscope indirect examination) were performed on all animals on days 5±1, 30±1, 60±1, 90±1 (in some variants at later dates). Observations were performed by a board-certified veterinary ophthalmologist. For animals to be dosed on Day 90 ± 1, ophthalmic observations were performed prior to dosing. Ophthalmic examination findings were scored according to the McDonald and Shadduck scoring system described in Dermatoxicology, F.N.Marzulli and H.I. Maibach, 1977, "Eye Irritation," T.O. McDonald and J.A. Shadduck (pp. 579-582), and observations were recorded using a standardized data collection form result. the

Whole blood samples (1-3 mL per sample) were collected from each animal prior to necropsy in EDTA-containing vacuumtainer tubes. Each tube was at least 2/3 full and mixed thoroughly for at least 30 seconds. Tubes were stored frozen until shipped on dry ice. the

Euthanize the animal with a commercially available euthanasia solution intravenously. Euthanasia was performed according to standard procedures used in industry. the

For treatment groups with intravitreally or subconjunctivally administered placebo, all eyes from each of these groups were placed in Davidsons solution for approximately 24 hours. After 24 hours the eyes were transferred to 70% ethanol; these eyes underwent masked pathological evaluation by a board-certified veterinary pathologist. The time the eye was placed in the Davidsons and the time it was removed was recorded. the

For groups treated with intravitreal or subconjunctival administration of the test article, some eyes from each group were frozen at -70°C and subjected to pharmacokinetic analysis. The remaining eyes from each group were placed in Davidsons solution for approximately 24 hours. After 24 hours, the eyes were transferred to 70% ethanol; these eyes underwent masked histopathological evaluation by a board-certified veterinary pathologist. The time the eye was placed in the Davidsons and the time it was removed was recorded. the

Frozen samples for pharmacokinetic analysis were dissected with disposable tools. Use one set of tools per eye, then discard. The samples were thawed at room temperature for 1 to 2 minutes to ensure that the frost around the tissue was removed. Dissect the sclera into 4 quadrants and remove the vitreous. If non-dispersed masses (NDM) were clearly visible within the vitreous, the vitreous was separated into two sections. Sections with NDM are about two-thirds of the way down the vitreous. The slice without NDM is the part of the vitreous that is furthest away from the NDM. Separate the aqueous humor, lens, iris and cornea. Retinochoroidal tissue was removed using forceps and collected for analysis. The conjunctiva is separated from the sclera. the

Multiple tissue types were collected into pre-weighed separate vials, and the tubes were capped and weighed. Tissue vials were stored at -80°C until analysis. the

Determination of sirolimus in retinachoroid, sclera, vitreous humor, and anticoagulated whole blood by high pressure liquid chromatography/tandem mass spectrometry (HPLC/MS/MS) using 32-O-desmethoxyrapamycin as an internal standard content. When NDM was observed in the vitreous, vitreous sections with and without NDM were analyzed separately. the

The average therapeutic agent concentration over a period of time is the average concentration at each time point for a representative time point referring to the time period. For example, if the time period is 30 days, the average concentration can be measured at 5-day intervals: for the average concentration on day 5, the mean of the multiple concentration measurements on day 5 should be calculated; for the average concentration on day 10, the Average of multiple concentration measurements over 10 days, etc. the

In some variations, the liquid formulations described herein may have an in vivo delivery profile to the vitreous that is characterized below for the in vivo delivery of a therapeutic agent following injection of the liquid formulation between the sclera and conjunctiva of a rabbit eye. A non-limiting variation of the in vivo delivery profile to the vitreous is shown in FIG. 2 . the

At day 40 post-injection, the mean percent vitreous level in vivo may be between about 70% and about 100%, and more often between about 80% and about 90%, relative to the level at day 20 post-injection. At day 40 post-injection, the mean percent vitreous level in vivo may be greater than about 70%, and more often greater than about 80%, relative to the level at day 20 post-injection. the

At day 67 post-injection, the mean percent vitreous level in vivo may be between about 75% and about 115%, and more often between about 85% and about 105%, relative to the level at day 20 post-injection. At day 67 post-injection, the mean percent vitreous level in vivo may be greater than about 75%, and more often greater than about 85%, relative to the level at day 20 post-injection. the

At day 90 post-injection, the mean percent vitreous level in vivo may be between about 20% and about 50%, and more often between about 30% and about 40%, relative to the level at day 20 post-injection. At day 90 post-injection, the mean percent vitreous level in vivo may be greater than about 20%, and more often greater than about 30%, relative to the level at day 20 post-injection. the

In some variations, the mean percent vitreous level in vivo relative to the level expressed at day 20 post-injection is characterized by: it is less than about 100% at day 40 post-injection; it is less than about 115% at day 67 post-injection; and After day 90 it was less than about 50%. the

In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit's eye, resulting in at least about 30 days, at least about 60 days, or at least about 90 days after application of the liquid formulation to the rabbit's eye. A mean concentration of therapeutic agent of at least about 0.01 ng/mL. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit's eye, resulting in a vitreous in the rabbit's eye within at least about 30 days, at least about 60 days, or at least about 90 days after the liquid formulation is administered to the rabbit's eye. A mean concentration of therapeutic agent of at least about 0.1 ng/mL. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit's eye, resulting in a vitreous in the rabbit's eye within at least about 30 days, at least about 60 days, or at least about 90 days after the liquid formulation is administered to the rabbit's eye. A mean concentration of therapeutic agent of at least about 1 ng/mL. the

In some variations, the liquid formulations described herein may have an in vivo delivery profile to the retina choroid characterized by delivery of a therapeutic agent delivered in vivo following injection of the liquid formulation between the sclera and conjunctiva of the rabbit eye Spectrum. the

At day 40 post-injection, the mean percent retinochoroidal level in vivo may be between about 350% and about 410%, and more often between about 360% and about 400%, relative to the level expressed at day 20 post-injection. At day 40 post-injection, the mean percent retinochoroidal level in vivo may be greater than about 350%, and more often greater than about 360%, relative to the level expressed at day 20 post-injection. the

At day 67 post-injection, the mean percent retinochoroidal level in vivo may be between about 125% and about 165%, and more often between about 135% and about 155%, relative to the level expressed at day 20 post-injection. At day 67 post-injection, the mean percent retinochoroidal level in vivo may be greater than about 125%, and more often greater than about 135%, relative to the level expressed at day 20 post-injection. the

At day 90 post-injection, the mean percent retinochoroidal level in vivo may be between about 10% and about 50%, and more often between about 20% and about 40%, relative to the level expressed at day 20 post-injection. At day 90 post-injection, the mean percent retinochoroidal level in vivo may be greater than about 10%, and more often greater than about 20%, relative to the level expressed at day 20 post-injection. the

In some variations, the in vivo mean percent retinal choroidal level relative to the level expressed at day 20 post-injection is characterized by: at day 40 post-injection, it is less than about 410%; at day 67 post-injection, it is less than about 165%; and It was less than about 50% by day 90 post-injection. the

In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit eye, resulting in the retinal choroid of the rabbit eye within at least about 30 days, at least about 60 days, or at least about 90 days after application of the liquid formulation to the rabbit eye. A mean concentration of therapeutic agent in the tissue of at least about 0.001 ng/mL. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit eye, resulting in the vitreous retina of the rabbit eye within at least about 30 days, at least about 60 days, or at least about 90 days after application of the liquid formulation to the rabbit eye. A mean concentration of therapeutic agent in the choroidal tissue of at least about 0.01 ng/mL. the

In some variations, the level of therapeutic agent present in the retinal choroidal tissue first increases, then peaks and decreases. The peak can occur, for example, at about day 40 after injection. the

In some variations, the liquid formulations described herein can have an in vivo scleral clearance profile characterized by the clearance profile of the in vivo clearance of the therapeutic agent following injection of the liquid formulation between the sclera and conjunctiva of a rabbit's eye. When injected between the sclera and the conjunctiva, the level of the sclera is considered to include the injected liquid formulation. the

At day 40 post-injection, the mean percent sclera level in vivo may be between about 150% and about 230%, and more often between about 170% and about 210%, relative to the level expressed at day 20 post-injection. At day 40 post-injection, the mean percent sclera level in vivo may be greater than about 150%, and more often greater than about 170%, relative to the level expressed at day 20 post-injection. the

At day 67 post-injection, the mean percent sclera level in vivo may be between about 30% and about 70%, and more often between about 40% and about 60%, relative to the level expressed at day 20 post-injection. At day 67 post-injection, the mean percent sclera level in vivo may be greater than about 30%, and more often greater than about 40%, relative to the level expressed at day 20 post-injection. the

At day 90 post-injection, the mean percent sclera level in vivo may be between about 110% and about 160%, and more often between about 125% and about 145%, relative to the level expressed at day 20 post-injection. At 90 days post-injection, the mean percent sclera level in vivo may be greater than about 110%, and more often greater than about 125%, relative to the level expressed at day 20 post-injection. the

In some variations, the in vivo mean percent sclera level relative to the level expressed at day 20 post-injection is characterized by: at day 40 post-injection, it is less than about 230%; at day 67 post-injection, it is less than about 70%; and After day 90 it was less than about 160%. the

In some variations, the level of therapeutic agent present in the sclera first increases, then peaks and decreases. The peak can occur, for example, at about day 40 after injection. the

In some variations, the liquid formulations described herein may have an in vivo delivery profile to the vitreous, wherein the delivery profile is that of delivering a therapeutic agent in vivo following injection of the liquid formulation between the sclera and conjunctiva of a rabbit eye . the

At day 14 post-injection, the mean percent vitreous level in vivo can be between about 1350% and about 1650%, and more often between about 1450% and about 1550%, relative to the level expressed at day 2 post-injection. At day 14 post-injection, the mean percent vitreous level in vivo may be greater than about 1350%, and more often greater than about 1450%, relative to the level expressed at day 2 post-injection. the

At day 35 post-injection, the mean percent vitreous level in vivo can be between about 200% and about 300%, and more often between about 225% and about 275%, relative to the level expressed at day 2 post-injection. At day 35 post-injection, the mean percent vitreous level in vivo may be greater than about 200%, and more often greater than about 225%, relative to the level expressed at day 2 post-injection. the

At day 62 post-injection, the mean percent vitreous level in vivo may be between about 100% and about 160%, and more often between about 115% and about 145%, relative to the level expressed at day 2 post-injection. At day 62 post-injection, the mean percent vitreous level in vivo may be greater than about 100%, and more often greater than about 115%, relative to the level expressed at day 2 post-injection. the

At day 85 post-injection, the mean percent vitreous level in vivo may be between about 5% and about 30%, and more often between about 10% and about 25%, relative to the level expressed at day 2 post-injection. At day 85 post-injection, the mean percent vitreous level in vivo may be greater than about 5%, and more often greater than about 10%, relative to the level expressed at day 2 post-injection. the

In some variations, the mean percent vitreous level in vivo relative to the level expressed on day 2 post-injection is characterized by: it is less than about 1600% on day 14 post-injection; it is less than about 300% on day 35 post-injection; It was less than about 160% on day 62 post-injection and less than about 30% on day 85 post-injection. the

In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit's eye, resulting in the intravitreal of the rabbit's eye within at least about 30 days, at least about 60 days, or at least about 85 days after the liquid formulation is administered to the rabbit's eye. A mean concentration of therapeutic agent of at least about 0.01 ng/mL. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit's eye, resulting in the intravitreal of the rabbit's eye within at least about 30 days, at least about 60 days, or at least about 85 days after the liquid formulation is administered to the rabbit's eye. A mean concentration of therapeutic agent of at least about 0.1 ng/mL. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit's eye, resulting in at least about 1 ng/mL in the vitreous of the rabbit's eye for at least about 30 days, at least about 60 days after the liquid formulation is administered to the rabbit's eye. The average concentration of the therapeutic agent. the

In some variations, the level of therapeutic agent present in the vitreous first increases, then peaks and decreases. The peak can occur, for example, at about day 14 after injection. the

In some variations, the liquid formulations described herein may have an in vivo delivery profile to the retina choroid characterized by delivery of a therapeutic agent delivered in vivo following injection of the liquid formulation between the sclera and conjunctiva of the rabbit eye Spectrum. the

At day 35 post-injection, the mean percent retinochoroidal level in vivo may be between about 320% and about 400%, and more often between about 340% and about 380%, relative to the level expressed at day 14 post-injection. At day 35 post-injection, the mean percent retinochoroidal level in vivo may be greater than about 320%, and more often greater than about 340%, relative to the level expressed at day 14 post-injection. the

At day 62 post-injection, the mean percent retinochoroidal level in vivo may be between about 3% and about 25%, and more often between about 6% and about 20%, relative to the level expressed at day 14 post-injection. At day 62 post-injection, the mean percent retinochoroidal level in vivo may be greater than about 3%, and more often greater than about 6%, relative to the level expressed at day 14 post-injection. the

At day 85 post-injection, the mean percent retinochoroidal level in vivo may be between about 0.1% and about 6%, and more often between about 0.5% and about 4%, relative to the level expressed at day 14 post-injection. At day 85 post-injection, the mean percent retinochoroidal level in vivo may be greater than about 0.1%, and more often greater than about 0.5%, relative to the level expressed at day 14 post-injection. the

In some variations, the in vivo mean percent retinal choroidal level relative to the level expressed at day 14 post-injection is characterized by: it is less than about 400% at day 35 post-injection; it is less than about 25% at day 62 post-injection; and It was less than about 6% at day 85 post-injection. the

In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit eye, resulting in the retinal choroid of the rabbit eye within at least about 30 days, at least about 60 days, or at least about 85 days after application of the liquid formulation to the rabbit eye. A mean concentration of therapeutic agent in the tissue of at least about 0.001 ng/mL. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit eye, resulting in the retinal choroid of the rabbit eye within at least about 30 days, at least about 60 days, or at least about 85 days after application of the liquid formulation to the rabbit eye. A mean concentration of therapeutic agent in the tissue of at least about 0.01 ng/mL. the

In some variations, the liquid formulations described herein can have an in vivo scleral clearance profile characterized by the clearance profile of the in vivo clearance of the therapeutic agent following injection of the liquid formulation between the sclera and conjunctiva of a rabbit's eye. When injected between the sclera and the conjunctiva, the level of the sclera is considered to include the injected liquid formulation. the

At day 35 post-injection, the mean percent sclera level in vivo may be between about 0.1% and about 0.7%, and more often between about 0.2% and about 0.6%, relative to the level expressed at day 14 post-injection. At day 35 post-injection, the mean percent sclera level in vivo may be greater than about 0.1%, and more often greater than about 0.2%, relative to the level expressed at day 14 post-injection. the

At day 62 post-injection, the mean percent sclera level in vivo may be between about 0.05% and about 0.35%, and more often between about 0.07% and about 0.3%, relative to the level expressed at day 14 post-injection. At day 62 post-injection, the mean percent sclera level in vivo may be greater than about 0.05%, and more often greater than about 0.07%, relative to the level expressed at day 14 post-injection. the

At day 85 post-injection, the mean percent sclera level in vivo may be between about 0.1% and about 0.9%, and more often between about 0.3% and about 0.7%, relative to the level expressed at day 14 post-injection. At day 85 post-injection, the mean percent sclera level in vivo may be greater than about 0.1%, and more often greater than about 0.3%, relative to the level expressed at day 14 post-injection. the

In some variations, the in vivo mean percent sclera level relative to the level expressed at day 14 post-injection is characterized by: at day 35 post-injection, it is less than about 0.7%; at day 62 post-injection, it is less than about 0.35%; and After 85 days it was less than about 0.9%. the

In some variations, the liquid formulations described herein can have an in vivo vitreous clearance profile characterized by the clearance profile of in vivo clearance of a therapeutic agent following injection of the liquid formulation into the vitreous of a rabbit's eye. When injected into the vitreous, the measured vitreous levels are considered to include the injected formulation. the

At day 35 post-injection, the mean percent vitreous level in vivo can be between about 1% and about 40%, and more often between about 1% and about 10%, relative to the level expressed at day 14 post-injection. At day 35 post-injection, the mean percent vitreous level in vivo may be greater than about 1% relative to the level expressed at day 14 post-injection. the

At day 62 post-injection, the mean percent vitreous level in vivo may be between about 1% and about 40%, and more often between about 5% and about 25%, relative to the level expressed at day 14 post-injection. At day 62 post-injection, the mean percent vitreous level in vivo may be greater than about 1% relative to the level expressed at day 14 post-injection, and more often greater than about 5% relative to the level expressed at day 14 post-injection. the

At day 90 post-injection, the mean percent vitreous level in vivo may be between about 1% and about 40%, and more often between about 10% and about 30%, relative to the level expressed at day 14 post-injection. At day 90 post-injection, the mean percent vitreous level in vivo may be greater than about 1% relative to the level expressed at day 14 post-injection, and more often greater than about 10% relative to the level expressed at day 14 post-injection. the

In some variations, the level of therapeutic agent present in the vitreous first increases, then peaks and decreases. The peak can occur, for example, at about day 14 after injection. the

In some variations, the liquid formulations described herein may have an in vivo delivery profile to the retina and choroid, wherein the delivery profile is that of delivering a therapeutic agent in vivo following injection of the liquid formulation into the vitreous of a rabbit eye. the

At day 35 post-injection, the mean percent retinochoroidal level in vivo may be between about 3400% and about 5100%, and more often between about 3750% and about 4750%, relative to the level expressed at day 14 post-injection. At day 35 post-injection, the mean percent retinochoroidal level in vivo may be greater than about 3400%, and more often greater than about 3750%, relative to the level expressed at day 14 post-injection. the

At 62 days post-injection, the mean percent retinochoroidal level in vivo may be between about 0.1% and about 5%, and more often between about 1% and about 3%, relative to the level expressed at day 14 post-injection. At day 62 post-injection, the mean percent retinochoroidal level in vivo may be greater than about 0.1%, and more often greater than about 1%, relative to the level expressed at day 14 post-injection. the

At 90 days post-injection, the mean percent retinochoroidal level in vivo may be between about 10% and about 50%, and more often between about 20% and about 40%, relative to the level expressed at day 14 post-injection. At day 90 post-injection, the mean percent retinochoroidal level in vivo may be greater than about 10%, and more often greater than about 20%, relative to the level expressed at day 14 post-injection. the

In some variations, the in vivo mean percent retinal choroidal level relative to the level expressed at day 14 post-injection is characterized by: at day 35 post-injection, it is less than about 5100%; at day 62 post-injection, it is less than about 5%; and It was less than about 50% by day 90 post-injection. the

In some variations, the liquid formulations described herein may have an in vivo delivery profile to the sclera characterized by delivery of a therapeutic agent in vivo following injection of the liquid formulation into the vitreous of a rabbit eye. the

At day 35 post injection, the mean percent sclera level in vivo may be between about 1700% and about 2600%, and more often between about 1900% and about 2400%, relative to the level expressed at day 14 post injection. At day 35 post-injection, the mean percent sclera level in vivo may be greater than about 1700%, and more often greater than about 1900%, relative to the level expressed at day 14 post-injection. the

At day 62 post-injection, the mean percent sclera level in vivo may be between about 120% and about 180%, and more often between about 140% and about 160%, relative to the level expressed at day 14 post-injection. At day 62 post-injection, the mean percent sclera level in vivo may be greater than about 120%, and more often greater than about 140%, relative to the level expressed at day 14 post-injection. the

At day 90 post-injection, the mean percent sclera level in vivo may be between about 95% and about 155%, and more often between about 115% and about 135%, relative to the level expressed at day 14 post-injection. At 90 days post-injection, the mean percent sclera level in vivo may be greater than about 95%, and more often greater than about 115%, relative to the level expressed at day 14 post-injection. the

In some variations, the in vivo mean percent sclera level relative to the level expressed at day 14 post-injection is characterized by: at day 35 post-injection, it is less than about 2600%; at day 62 post-injection, it is less than about 180%; and After 90 days it was less than about 155%. the

In some variations, the therapeutic agent is delivered when the liquid formulation is injected into the vitreous of the rabbit eye, resulting in at least about 0.001 ng in the sclera of the rabbit eye within at least about 30 days, at least about 60 days, or at least about 90 days after the liquid formulation is administered to the rabbit eye. /mL mean concentration of therapeutic agent. In some variations, the therapeutic agent is delivered when the liquid formulation is injected into the vitreous of the rabbit eye, resulting in at least about 0.01 ng in the sclera of the rabbit eye within at least about 30 days, at least about 60 days, or at least about 90 days after the liquid formulation is administered to the rabbit eye. /mL mean concentration of therapeutic agent. In some variations, the therapeutic agent is delivered when the liquid formulation is injected into the vitreous of the rabbit eye, resulting in at least about 0.1 ng in the sclera of the rabbit eye within at least about 30 days, at least about 60 days, or at least about 90 days after the liquid formulation is administered to the rabbit eye. /mL mean concentration of therapeutic agent. the

In some variations, the level of therapeutic agent present in the vitreous first increases, then peaks and decreases. The peak can occur, for example, at about day 35 after injection. the

In some variations, the in situ gelling formulations described herein may have an in vivo delivery profile to the vitreous that delivers the therapeutic agent in vivo following injection of the liquid formulation between the sclera and conjunctiva of the rabbit eye delivery spectrum. the

At day 32 post-injection, the mean percent vitreous level in vivo may be between about 25% and about 85%, and more often between about 45% and about 65%, relative to the level expressed at day 7 post-injection. At day 40 post-injection, the mean percent vitreous level in vivo may be greater than about 25%, and more often greater than about 45%, relative to the level expressed at day 7 post-injection. the

At day 45 post-injection, the mean percent vitreous level in vivo may be between about 2% and about 50%, and more often between about 8% and about 20%, relative to the level expressed at day 7 post-injection. At day 67 post-injection, the mean percent vitreous level in vivo may be greater than about 2%, and more often greater than about 5%, relative to the level expressed at day 7 post-injection. the

At day 90 post-injection, the mean percent vitreous level in vivo may be between about 40% and about 100%, and more often between about 60% and about 80%, relative to the level expressed at day 7 post-injection. At day 90 post-injection, the mean percent vitreous level in vivo may be greater than about 40%, and more often greater than about 60%, relative to the level expressed at day 7 post-injection. the

In some variations, the mean percent vitreous level in vivo relative to the level expressed on day 7 post-injection is characterized by: it is less than about 80% at day 32 post-injection; it is less than about 30% at day 45 post-injection; and After day 90 it was less than about 100%. the

In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit's eye, resulting in a vitreous in the rabbit's eye within at least about 30 days, at least about 60 days, or at least about 90 days after the liquid formulation is administered to the rabbit's eye. A mean concentration of therapeutic agent of at least about 0.1 pg/mL. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit's eye, resulting in a vitreous in the rabbit's eye within at least about 30 days, at least about 60 days, or at least about 90 days after the liquid formulation is administered to the rabbit's eye. A mean concentration of therapeutic agent of at least about 0.01 ng/mL. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit's eye, resulting in a vitreous in the rabbit's eye within at least about 30 days, at least about 60 days, or at least about 90 days after the liquid formulation is administered to the rabbit's eye. A mean concentration of therapeutic agent of at least about 0.1 ng/mL. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit's eye, resulting in a vitreous in the rabbit's eye within at least about 30 days, at least about 60 days, or at least about 90 days after the liquid formulation is administered to the rabbit's eye. A mean concentration of therapeutic agent of at least about 1 ng/mL. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit's eye, resulting in a vitreous in the rabbit's eye within at least about 30 days, at least about 60 days, or at least about 90 days after the liquid formulation is administered to the rabbit's eye. A mean concentration of therapeutic agent of at least about 10 ng/mL. the

In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit eye, resulting in the vitreous humor of the rabbit eye within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after the liquid formulation is administered to the rabbit eye. Mean concentration of therapeutic agent within at least 0.001 ng/mL. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and conjunctiva of the rabbit's eye, resulting in at least about 30 days, at least about 60 days, at least about 90 days, or at least about 120 days after application of the liquid formulation to the rabbit's eye. The average concentration of therapeutic agent in the vitreous of rabbit eyes is at least 0.01 ng/mL within 1 day. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit eye, resulting in the vitreous humor of the rabbit eye within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after the liquid formulation is administered to the rabbit eye. Mean concentration of therapeutic agent within at least 0.1 ng/mL. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit eye, resulting in the vitreous humor of the rabbit eye within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after the liquid formulation is administered to the rabbit eye. Mean concentration of therapeutic agent within at least 0.5 ng/mL. the

In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit eye, resulting in the vitreous humor of the rabbit eye within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after the liquid formulation is administered to the rabbit eye. The average concentration of therapeutic agent within 0.001 ng/mL and 10.0 ng/mL. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit eye, resulting in the vitreous humor of the rabbit eye within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after the liquid formulation is administered to the rabbit eye. The average concentration of therapeutic agent within 0.01 ng/mL and 10.0 ng/mL. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit eye, resulting in the vitreous humor of the rabbit eye within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after the liquid formulation is administered to the rabbit eye. Mean concentrations of therapeutic agent within 0.1 ng/mL and 10 ng/mL. the

In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit eye, resulting in the vitreous humor of the rabbit eye within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after the liquid formulation is administered to the rabbit eye. Mean concentrations of therapeutic agent within 0.5 ng/mL and 10.0 ng/mL. the

In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit eye, resulting in the vitreous humor of the rabbit eye within 30 days to at least 60 days, at least 90 days, or at least 120 days after administration of the liquid formulation to the rabbit eye. The ratio of the maximum average concentration of the therapeutic agent in the rabbit eye to the minimum average concentration of the therapeutic agent in the vitreous of the rabbit eye is less than 100. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit eye, resulting in a vitreous humor of the rabbit eye within 30 days to at least 60 days, at least 90 days, or at least 120 days after application of the liquid formulation to the rabbit eye. The ratio of the maximum average concentration of the therapeutic agent in the rabbit eye to the minimum average concentration of the therapeutic agent in the vitreous of the rabbit eye is less than 50. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit eye, resulting in the vitreous humor of the rabbit eye within 30 days to at least 60 days, at least 90 days, or at least 120 days after administration of the liquid formulation to the rabbit eye. The ratio of the maximum mean concentration of the therapeutic agent in the rabbit eye to the minimum mean concentration of the therapeutic agent in the vitreous of the rabbit eye is less than 10. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit eye, resulting in the vitreous humor of the rabbit eye within 30 days to at least 60 days, at least 90 days, or at least 120 days after administration of the liquid formulation to the rabbit eye. The ratio of the maximum average concentration of the therapeutic agent in the rabbit eye to the minimum average concentration of the therapeutic agent in the vitreous of the rabbit eye is less than 5. the

As used herein, "approximately constant" means that the average level does not change by more than one order of magnitude over an extended period of time, i.e. the difference between the maximum and minimum values of the average concentration measured at different times in the relevant time period is less than 10 times. the

In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit's eye, resulting in a reduction in the vitreous of the rabbit's eye within 30 days to at least 60 days, at least 90 days, or at least 120 days after application of the solution to the rabbit's eye. The average concentration of therapeutic agent was roughly constant at values greater than 0.001 ng/mL. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit eye, resulting in the vitreous humor of the rabbit eye within 30 days to at least 60 days, at least 90 days, or at least 120 days after administration of the liquid formulation to the rabbit eye. The mean concentration of the internal therapeutic agent was roughly constant at values greater than 0.01 ng/mL. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit's eye, resulting in a reduction in the vitreous of the rabbit's eye within 30 days to at least 60 days, at least 90 days, or at least 120 days after application of the solution to the rabbit's eye. The average concentration of therapeutic agent was roughly constant at values greater than 0.1 ng/mL. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit eye, resulting in the vitreous humor of the rabbit eye within 30 days to at least 60 days, at least 90 days, or at least 120 days after administration of the liquid formulation to the rabbit eye. The mean concentration of internal therapeutic agent was approximately constant at a value of 1.0 ng/mL. the

In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit's eye, resulting in a rabbit eye within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after application of the liquid formulation to the rabbit eye. Mean concentration of therapeutic agent in retinal choroidal tissue of at least 0.001 ng/mg. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit's eye, resulting in a rabbit eye within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after application of the liquid formulation to the rabbit eye. Mean concentration of therapeutic agent in retinal choroidal tissue of at least 0.005 ng/mg. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit's eye, resulting in a rabbit eye within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after application of the liquid formulation to the rabbit eye. Mean concentration of therapeutic agent in retinal choroidal tissue of at least 0.01 ng/mg. the

In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit's eye, resulting in a rabbit eye within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after application of the liquid formulation to the rabbit eye. Mean concentrations of therapeutic agent between 0.001 ng/mg and 1.0 ng/mg in retinal choroidal tissue. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit's eye, resulting in a rabbit eye within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after application of the liquid formulation to the rabbit eye. Mean concentrations of therapeutic agent between 0.001 ng/mg and 0.50 ng/mg in retinal choroidal tissue. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit's eye, resulting in a rabbit eye within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after application of the liquid formulation to the rabbit eye. Mean concentrations of therapeutic agent between 0.001 ng/mg and 0.15 ng/mg in retinal choroidal tissue. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit's eye, resulting in a rabbit eye within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after application of the liquid formulation to the rabbit eye. Mean concentrations of therapeutic agent between 0.001 ng/mg and 0.1 ng/mg in retinal choroidal tissue. the

In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit's eye, resulting in a rabbit eye within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after application of the liquid formulation to the rabbit eye. Mean concentrations of therapeutic agent between 0.005 ng/mg and 1.0 ng/mg in retinal choroidal tissue. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit's eye, resulting in a rabbit eye within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after application of the liquid formulation to the rabbit eye. Mean concentrations of therapeutic agent between 0.005 ng/mg and 0.50 ng/mg in retinal choroidal tissue. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit's eye, resulting in a rabbit eye within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after application of the liquid formulation to the rabbit eye. Mean concentrations of therapeutic agent between 0.005 ng/mg and 0.15 ng/mg in retinal choroidal tissue. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit's eye, resulting in a rabbit eye within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after application of the liquid formulation to the rabbit eye. Mean concentrations of therapeutic agent between 0.005 ng/mg and 0.1 ng/mg in retinal choroidal tissue. the

In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit's eye, resulting in a rabbit eye within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after application of the liquid formulation to the rabbit eye. Mean concentrations of therapeutic agent between 0.01 ng/mg and 1.0 ng/mg in retinal choroidal tissue. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit's eye, resulting in a rabbit eye within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after application of the liquid formulation to the rabbit eye. Mean concentrations of therapeutic agent between 0.01 ng/mg and 0.50 ng/mg in retinal choroidal tissue. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit's eye, resulting in a rabbit eye within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after application of the liquid formulation to the rabbit eye. Mean concentrations of therapeutic agent between 0.01 ng/mg and 0.15 ng/mg in retinal choroidal tissue. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit's eye, resulting in a rabbit eye within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after application of the liquid formulation to the rabbit eye. Mean concentrations of therapeutic agent between 0.01 ng/mg and 0.1 ng/mg in retinal choroidal tissue. the

In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit eye, resulting in retinal damage in the rabbit eye within 30 days to at least 60 days, at least 90 days, or at least 120 days after application of the liquid formulation to the rabbit eye. The ratio of the maximum average concentration of the therapeutic agent in the choroid tissue to the minimum average concentration of the therapeutic agent in the retina choroid tissue of the rabbit eye is less than 100. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit eye, resulting in retinal damage in the rabbit eye within 30 days to at least 60 days, at least 90 days, or at least 120 days after application of the liquid formulation to the rabbit eye. The ratio of the maximum average concentration of the therapeutic agent in the choroid tissue to the minimum average concentration of the therapeutic agent in the retina choroid tissue of the rabbit eye is less than 50. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit eye, resulting in retinal damage in the rabbit eye within 30 days to at least 60 days, at least 90 days, or at least 120 days after application of the liquid formulation to the rabbit eye. The ratio of the maximum average concentration of the therapeutic agent in the choroid tissue to the minimum average concentration of the therapeutic agent in the retina choroid tissue of the rabbit eye is less than 10. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit eye, resulting in retinal damage in the rabbit eye within 30 days to at least 60 days, at least 90 days, or at least 120 days after application of the liquid formulation to the rabbit eye. The ratio of the maximum average concentration of the therapeutic agent in the choroid tissue to the minimum average concentration of the therapeutic agent in the retina choroid tissue of the rabbit eye is less than 5. the

In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit eye, resulting in retinal damage in the rabbit eye within 30 days to at least 60 days, at least 90 days, or at least 120 days after application of the liquid formulation to the rabbit eye. The average concentration of therapeutic agent in the choroidal tissue was approximately constant at values greater than 0.001 ng/mg. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit eye, resulting in retinal damage in the rabbit eye within 30 days to at least 60 days, at least 90 days, or at least 120 days after application of the liquid formulation to the rabbit eye. The average concentration of therapeutic agent in the choroidal tissue was approximately constant at values greater than 0.005 ng/mg. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit eye, resulting in retinal damage in the rabbit eye within 30 days to at least 60 days, at least 90 days, or at least 120 days after application of the liquid formulation to the rabbit eye. The average concentration of therapeutic agent in the choroidal tissue was approximately constant at values greater than 0.01 ng/mg. the

In some variations, the therapeutic agent is delivered when the liquid formulation is injected into the vitreous of the rabbit eye such that within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after administration of the liquid formulation to the rabbit eye, at least The mean concentration of therapeutic agent was 100 ng/mL. In some variations, the therapeutic agent is delivered when the liquid formulation is injected into the vitreous of the rabbit eye such that within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after administration of the liquid formulation to the rabbit eye, at least The mean concentration of therapeutic agent was 1000 ng/mL. In some variations, the therapeutic agent is delivered when the liquid formulation is injected into the vitreous of the rabbit eye such that within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after administration of the liquid formulation to the rabbit eye, at least The mean concentration of therapeutic agent was 10,000 ng/mL. the

In some variations, the therapeutic agent is delivered when the liquid formulation is injected into the vitreous of the rabbit's eye such that 100 ng/day of the liquid formulation is administered to the rabbit's eye within 30 days to at least 60 days, at least 90 days, or at least 120 days. Average concentration of therapeutic agent between mL and 100,000 ng/mL. In some variations, the therapeutic agent is delivered when the liquid formulation is injected into the vitreous of the rabbit's eye, resulting in a 100 ng/kg intravitreal of the rabbit's eye within 30 days to at least 60 days, at least 90 days, or at least 120 days after the liquid formulation is administered to the rabbit eye. Average concentration of therapeutic agent between mL and 50,000 ng/mL. the

In some variations, the therapeutic agent is delivered when the liquid formulation is injected into the vitreous of the rabbit's eye, resulting in 1000 ng/kg in the vitreous of the rabbit's eye within 30 days to at least 60 days, at least 90 days, or at least 120 days after the liquid formulation is administered to the rabbit's eye. Average concentration of therapeutic agent between mL and 100,000 ng/mL. In some variations, the therapeutic agent is delivered when the liquid formulation is injected into the vitreous of the rabbit's eye, resulting in 1000 ng/kg in the vitreous of the rabbit's eye within 30 days to at least 60 days, at least 90 days, or at least 120 days after the liquid formulation is administered to the rabbit's eye. Average concentration of therapeutic agent between mL and 50,000 ng/mL. the

In some variations, the therapeutic agent is delivered when the liquid formulation is injected into the vitreous of the rabbit's eye, such that the therapeutic agent is released within the vitreous of the rabbit's eye within 30 days to at least 60 days, at least 90 days, or at least 120 days after administration of the liquid formulation to the rabbit's eye. The ratio of the maximum mean concentration to the minimum mean concentration of therapeutic agent in the rabbit eye vitreous was less than 100. In some variations, the therapeutic agent is delivered when the liquid formulation is injected into the vitreous of the rabbit's eye, such that the therapeutic agent is released within the vitreous of the rabbit's eye within 30 days to at least 60 days, at least 90 days, or at least 120 days after administration of the liquid formulation to the rabbit's eye. The ratio of the maximum mean concentration to the minimum mean concentration of therapeutic agent in the rabbit eye vitreous was less than 50. In some variations, the therapeutic agent is delivered when the liquid formulation is injected into the vitreous of the rabbit's eye, such that the therapeutic agent is released within the vitreous of the rabbit's eye within 30 days to at least 60 days, at least 90 days, or at least 120 days after administration of the liquid formulation to the rabbit's eye. The ratio of the maximum mean concentration to the minimum mean concentration of therapeutic agent in the rabbit eye vitreous was less than 10. the

In some variations, the therapeutic agent is delivered when the liquid formulation is injected into the vitreous of the rabbit's eye, such that the therapeutic agent is released within the vitreous of the rabbit's eye within 30 days to at least 60 days, at least 90 days, or at least 120 days after administration of the liquid formulation to the rabbit's eye. The average concentration of is roughly constant at values greater than 100 ng/mL. In some variations, the therapeutic agent is delivered when the liquid formulation is injected into the vitreous of the rabbit's eye, such that the therapeutic agent is released within the vitreous of the rabbit's eye within 30 days to at least 60 days, at least 90 days, or at least 120 days after administration of the liquid formulation to the rabbit's eye. The average concentration of is roughly constant at values greater than 1000 ng/mL. In some variations, the therapeutic agent is delivered when the liquid formulation is injected into the vitreous of the rabbit's eye, such that the therapeutic agent is released within the vitreous of the rabbit's eye within 30 days to at least 60 days, at least 90 days, or at least 120 days after administration of the liquid formulation to the rabbit's eye. The average concentration of the drug is roughly constant at values greater than 10,000 ng/mL. the

In some variations, the therapeutic agent is delivered when the liquid formulation is injected into the vitreous of the rabbit eye, resulting in retinal choroidal tissue in the rabbit eye within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after administration of the liquid formulation to the rabbit eye. Mean concentration of therapeutic agent within at least 0.001 ng/mg. In some variations, the therapeutic agent is delivered when the liquid formulation is injected into the vitreous of the rabbit eye, resulting in retinal choroidal tissue in the rabbit eye within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after administration of the liquid formulation to the rabbit eye. Mean concentration of therapeutic agent within at least 0.01 ng/mg. In some variations, the therapeutic agent is delivered when the liquid formulation is injected into the vitreous of the rabbit eye, resulting in retinal choroidal tissue in the rabbit eye within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after administration of the liquid formulation to the rabbit eye. Mean concentration of therapeutic agent within at least 0.05 ng/mg. In some variations, the therapeutic agent is delivered when the liquid formulation is injected into the vitreous of the rabbit eye, resulting in retinal choroidal tissue in the rabbit eye within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after administration of the liquid formulation to the rabbit eye. Mean concentration of therapeutic agent within at least 0.10 ng/mg. the

In some variations, the therapeutic agent is delivered when the liquid formulation is injected into the vitreous of the rabbit eye, resulting in retinal choroidal tissue in the rabbit eye within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after administration of the liquid formulation to the rabbit eye. Mean concentrations of therapeutic agent within 0.001 ng/mg and 10.00 ng/mg. In some variations, the therapeutic agent is delivered when the liquid formulation is injected into the vitreous of the rabbit eye, resulting in retinal choroidal tissue in the rabbit eye within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after administration of the liquid formulation to the rabbit eye. Mean concentrations of therapeutic agent within 0.001 ng/mg and 5.00 ng/mg. In some variations, the therapeutic agent is delivered when the liquid formulation is injected into the vitreous of the rabbit eye, resulting in retinal choroidal tissue in the rabbit eye within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after administration of the liquid formulation to the rabbit eye. Mean concentrations of therapeutic agent within 0.001 ng/mg and 1.00 ng/mg. the

In some variations, the therapeutic agent is delivered when the liquid formulation is injected into the vitreous of the rabbit eye, resulting in retinal choroidal tissue in the rabbit eye within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after administration of the liquid formulation to the rabbit eye. Mean concentrations of therapeutic agent within 0.01 ng/mg and 10.00 ng/mg. In some variations, the therapeutic agent is delivered when the liquid formulation is injected into the vitreous of the rabbit eye, resulting in retinal choroidal tissue in the rabbit eye within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after administration of the liquid formulation to the rabbit eye. Average concentration of therapeutic agent between 0.01 ng/mg and 5.00 ng/mg. In some variations, the therapeutic agent is delivered when the liquid formulation is injected into the vitreous of the rabbit eye, resulting in retinal choroidal tissue in the rabbit eye within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after administration of the liquid formulation to the rabbit eye. Mean concentrations of therapeutic agent within 0.01 ng/mg and 1.00 ng/mg. the

In some variations, the therapeutic agent is delivered when the liquid formulation is injected into the vitreous of the rabbit eye, resulting in retinal choroidal tissue in the rabbit eye within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after administration of the liquid formulation to the rabbit eye. Mean concentrations of therapeutic agent within 0.05 ng/mg and 10.00 ng/mg. In some variations, the therapeutic agent is delivered when the liquid formulation is injected into the vitreous of the rabbit eye, resulting in retinal choroidal tissue in the rabbit eye within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after administration of the liquid formulation to the rabbit eye. Mean concentrations of therapeutic agent within 0.05 ng/mg and 5.00 ng/mg. In some variations, the therapeutic agent is delivered when the liquid formulation is injected into the vitreous of the rabbit eye, resulting in retinal choroidal tissue in the rabbit eye within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after administration of the liquid formulation to the rabbit eye. Mean concentrations of therapeutic agent within 0.05 ng/mg and 1.00 ng/mg. the

In some variations, the therapeutic agent is delivered when the liquid formulation is injected into the vitreous of the rabbit eye, resulting in retinal choroidal tissue in the rabbit eye within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after administration of the liquid formulation to the rabbit eye. Mean concentrations of therapeutic agent within 0.10 ng/mg and 10.00 ng/mg. In some variations, the therapeutic agent is delivered when the liquid formulation is injected into the vitreous of the rabbit eye, resulting in retinal choroidal tissue in the rabbit eye within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after administration of the liquid formulation to the rabbit eye. Mean concentrations of therapeutic agent within 0.10 ng/mg and 5.00 ng/mg. In some variations, the therapeutic agent is delivered when the liquid formulation is injected into the vitreous of the rabbit eye, resulting in retinal choroidal tissue in the rabbit eye within at least 30 days, at least 60 days, at least 90 days, or at least 120 days after administration of the liquid formulation to the rabbit eye. Mean concentrations of therapeutic agent within 0.10 ng/mg and 1.00 ng/mg. the

In some variations, the therapeutic agent is delivered when the liquid formulation is injected into the vitreous of the rabbit eye, resulting in a reduction in the retinal choroidal tissue of the rabbit eye within 30 days to at least 60 days, at least 90 days, or at least 120 days after administration of the liquid formulation to the rabbit eye. The ratio of the maximum average concentration of the therapeutic agent to the minimum average concentration of the therapeutic agent in the retinal choroid tissue of the rabbit eye is less than 100. In some variations, the therapeutic agent is delivered when the liquid formulation is injected into the vitreous of the rabbit eye, resulting in a reduction in the retinal choroidal tissue of the rabbit eye within 30 days to at least 60 days, at least 90 days, or at least 120 days after administration of the liquid formulation to the rabbit eye. The ratio of the maximum average concentration of the therapeutic agent to the minimum average concentration of the therapeutic agent in the retinal choroid tissue of the rabbit eye is less than 50. the

In some variations, the in situ gelling formulations described herein may have an in vivo delivery profile to retinal choroidal tissue characterized by delivery in vivo following injection of the liquid formulation between the sclera and conjunctiva of a rabbit eye. Delivery profiles of therapeutic agents. the

At day 32 post-injection, the percent in vivo vitreous level can be between about 20% and about 80%, and more often between about 40% and about 60%, relative to the level expressed at day 7 post-injection. At day 40 post-injection, the percent in vivo vitreous level may be greater than about 20%, and more often greater than about 40%, relative to the level expressed at day 7 post-injection. the

At day 45 post-injection, the percent in vivo vitreous level may be between about 15% and about 55%, and more often between about 25% and about 45%, relative to the level expressed at day 7 post-injection. At day 67 post-injection, the percent in vivo vitreous level may be greater than about 15%, and more often greater than about 25%, relative to the level expressed at day 7 post-injection. the

At day 90 post-injection, the percent in vivo vitreous level may be between about 60% and about 100%, and more often between about 70% and about 90%, relative to the level expressed at day 7 post-injection. At 90 days post-injection, the percent in vivo vitreous level may be greater than about 60%, and more often greater than about 70%, relative to the level expressed at day 7 post-injection. the

In some variations, the percent in vivo vitreous level relative to the level expressed on day 7 post-injection is characterized by: it is less than about 80% at day 32 post-injection; it is less than about 60% at day 45 post-injection; and On day 90 it was less than about 100%. the

In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit eye, resulting in the retinal choroid of the rabbit eye within at least about 30 days, at least about 60 days, or at least about 90 days after application of the liquid formulation to the rabbit eye. A mean concentration of therapeutic agent in the tissue of at least about 0.1 pg/mg. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit's eye, resulting in a vitreous in the rabbit's eye within at least about 30 days, at least about 60 days, or at least about 90 days after the liquid formulation is administered to the rabbit's eye. A mean concentration of therapeutic agent of at least about 0.01 ng/mg. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit's eye, resulting in the intravitreal of the rabbit's eye within at least about 30 days, at least about 60 days, or at least about 90 days after the liquid formulation is administered to the rabbit's eye. A mean concentration of therapeutic agent of at least about 0.1 ng/mg. In some variations, the therapeutic agent is delivered when the liquid formulation is injected between the sclera and the conjunctiva of the rabbit's eye, resulting in a vitreous in the rabbit's eye within at least about 30 days, at least about 60 days, or at least about 90 days after the liquid formulation is administered to the rabbit's eye. A mean concentration of therapeutic agent of at least about 1 ng/mg. the

In some variations, after the in situ gelling formulation is placed in or near the eye, the mean levels of the therapeutic agent in two or more of the retinal choroidal tissue, the sclera, and the vitreous are expressed in logarithmic base-decade ratios over an extended period of time. largely unchanged in the segment. In some variations, after the in situ gelling formulation is placed between the sclera and conjunctiva of the eye, the mean levels of the therapeutic agent in two or more retinal choroidal tissues, the sclera, and the vitreous are prolonged by a logarithmic base-decade ratio. roughly unchanged over the period of time. In some variations, after the in situ gelling formulation is placed between the sclera and conjunctiva of the eye, the mean base-decade ratio of the therapeutic agent levels in the vitreous and sclera is approximately constant over an extended period of time. the

In some variations, the ratio of the base ten logarithm of the mean level of the therapeutic agent in the vitreous and retinal choroidal tissue is approximately constant over an extended period of time. In other words, when considered on a logarithmic scale, as the level of therapeutic agent in the vitreous increases, the level of therapeutic agent in the retinal choroidal tissue increases to a similar extent, and vice versa. the

In some variations, the ratio of the base ten logarithm of the mean level of therapeutic agent in vitreous versus retinal choroidal tissue is approximately constant over an extended period of about 7 days, about 30 days, about 60 days, or about 90 days. In some variations, after placement of the in situ gelling formulation between the sclera and conjunctiva, the ratio of the mean level of therapeutic agent in the vitreous relative to the mean level of therapeutic agent in the retinal choroidal tissue was constant at about 37:1 on day 7, About 40:1 on day 32, about 10:1 on day 45 and about 34:1 on day 90. the

In some variations, the ratio of the average level of therapeutic agent in the vitreous to the average level of therapeutic agent in the retinal choroidal tissue is constant at about 40:1 at about 7 days, about 32 days, about 45 days, or about 90 days. the

In some variations, the average level of any or all of the therapeutic agent in the retinal choroidal tissue, sclera, and vitreous is substantially unchanged over an extended period of time after the in situ gelling formulation is placed in or near the eye. the

In some variations, the mean level of therapeutic agent in the vitreous is approximately constant at about 8.1 ng/ml following placement of the gel-in-situ formulation between the sclera and conjunctiva. In some variations, the mean level of the therapeutic agent in the retinochoroidal tissue is approximately constant at about 0.25 ng/mg following placement of the in situ gelling formulation between the sclera and conjunctiva. In some variations, the mean concentration of the therapeutic agent in the sclera is approximately constant at about 1930 ng/mg following placement of the gel-in-situ formulation between the sclera and the conjunctiva. the

In some variations, the in situ gelling formulation when injected between the sclera and conjunctiva of the rabbit eye maintains the intravitreal treatment of the rabbit eye for at least about 30 days, at least about 60 days, or at least about 90 days after the liquid formulation is administered to the rabbit eye The average level of the dose was roughly constant at about 0.1 pg/mL. In some variations, the in situ gelling formulation when injected between the sclera and conjunctiva of the rabbit eye maintains the intravitreal treatment of the rabbit eye for at least about 30 days, at least about 60 days, or at least about 90 days after the liquid formulation is administered to the rabbit eye The average level of the dose was roughly constant at about 0.001 ng/mL. In some variations, the in situ gelling formulation when injected between the sclera and conjunctiva of the rabbit eye maintains the intravitreal treatment of the rabbit eye for at least about 30 days, at least about 60 days, or at least about 90 days after the liquid formulation is administered to the rabbit eye The average level of the dose was roughly constant at about 0.01 ng/mL. In some variations, the in situ gelling formulation when injected between the sclera and conjunctiva of the rabbit eye maintains the intravitreal treatment of the rabbit eye for at least about 30 days, at least about 60 days, or at least about 90 days after the liquid formulation is administered to the rabbit eye The average level of the dose was roughly constant at about 0.1 ng/mL. In some variations, the in situ gelling formulation when injected between the sclera and conjunctiva of the rabbit eye maintains the intravitreal treatment of the rabbit eye for at least about 30 days, at least about 60 days, or at least about 90 days after the liquid formulation is administered to the rabbit eye The average level of the dose was roughly constant at about 1 ng/mL. In some variations, the in situ gelling formulation when injected between the sclera and conjunctiva of the rabbit eye maintains the intravitreal treatment of the rabbit eye for at least about 30 days, at least about 60 days, or at least about 90 days after the liquid formulation is administered to the rabbit eye The average level of the dose was roughly constant at about 10 ng/mL. In some variations, the in situ gelling formulation when injected between the sclera and conjunctiva of the rabbit eye maintains the intravitreal treatment of the rabbit eye for at least about 30 days, at least about 60 days, or at least about 90 days after the liquid formulation is administered to the rabbit eye The average level of the dose was roughly constant at about 100 ng/mL. the

In some variations, when injected between the sclera and the conjunctiva of the rabbit eye, the in situ gelling formulation remains within the retinal choroidal tissue for at least about 30 days, at least about 60 days, or at least about 90 days after the liquid formulation is administered to the rabbit eye The average level of therapeutic agent was roughly constant at about 0.1 pg/mg. In some variations, when injected between the sclera and conjunctiva of the rabbit eye, the in situ gelling formulation remains in the retinal choroidal tissue for at least about 30 days, at least about 60 days, or at least about 90 days after the liquid formulation is administered to the rabbit eye The average level of therapeutic agent was roughly constant at about 0.001 ng/mg. In some variations, when injected between the sclera and the conjunctiva of the rabbit eye, the in situ gelling formulation remains within the retinal choroidal tissue for at least about 30 days, at least about 60 days, or at least about 90 days after the liquid formulation is administered to the rabbit eye Mean levels of therapeutic agents were roughly constant at about 0.01 ng/mg. In some variations, when injected between the sclera and the conjunctiva of the rabbit eye, the in situ gelling formulation remains within the retinal choroidal tissue for at least about 30 days, at least about 60 days, or at least about 90 days after the liquid formulation is administered to the rabbit eye The average level of therapeutic agent was roughly constant at about 0.1 ng/mg. In some variations, when injected between the sclera and the conjunctiva of the rabbit eye, the in situ gelling formulation remains within the retinal choroidal tissue for at least about 30 days, at least about 60 days, or at least about 90 days after the liquid formulation is administered to the rabbit eye The average level of therapeutic agent was roughly constant at about 1 ng/mg. In some variations, when injected between the sclera and the conjunctiva of the rabbit eye, the in situ gelling formulation remains within the retinal choroidal tissue for at least about 30 days, at least about 60 days, or at least about 90 days after the liquid formulation is administered to the rabbit eye The average level of therapeutic agent was roughly constant at about 10 ng/mg. the

In some variations, when injected between the sclera and the conjunctiva of the rabbit's eye, the in situ gelling formulation maintains the intrascleral therapeutic agent for at least about 30 days, at least about 60 days, or at least about 90 days after administration of the liquid formulation to the rabbit's eye The average level of about 0.1pg/mg is roughly constant. In some variations, when injected between the sclera and the conjunctiva of the rabbit's eye, the in situ gelling formulation maintains the intrascleral therapeutic agent for at least about 30 days, at least about 60 days, or at least about 90 days after administration of the liquid formulation to the rabbit's eye The average level of is roughly constant at about 0.001 ng/mg. In some variations, when injected between the sclera and the conjunctiva of the rabbit's eye, the in situ gelling formulation maintains the intrascleral therapeutic agent for at least about 30 days, at least about 60 days, or at least about 90 days after administration of the liquid formulation to the rabbit's eye The average level of is roughly constant at about 0.01 ng/mg. In some variations, when injected between the sclera and the conjunctiva of the rabbit's eye, the in situ gelling formulation maintains the intrascleral therapeutic agent for at least about 30 days, at least about 60 days, or at least about 90 days after administration of the liquid formulation to the rabbit's eye The average level of is roughly constant at about 0.1 ng/mg. In some variations, when injected between the sclera and the conjunctiva of the rabbit's eye, the in situ gelling formulation maintains the intrascleral therapeutic agent for at least about 30 days, at least about 60 days, or at least about 90 days after administration of the liquid formulation to the rabbit's eye The average level of is roughly constant at about 1 ng/mg. In some variations, when injected between the sclera and conjunctiva of the rabbit's eye, the in situ gelling formulation maintains the intrascleral therapeutic agent for at least about 30 days, at least about 60 days, or at least about 90 days after the liquid formulation is administered to the rabbit's eye The average level of is roughly constant at about 10 ng/mg. In some variations, when injected between the sclera and the conjunctiva of the rabbit's eye, the in situ gelling formulation maintains the intrascleral therapeutic agent for at least about 30 days, at least about 60 days, or at least about 90 days after administration of the liquid formulation to the rabbit's eye The average level of is roughly constant at about 100 ng/mg. In some variations, when injected between the sclera and the conjunctiva of the rabbit's eye, the in situ gelling formulation maintains the intrascleral therapeutic agent for at least about 30 days, at least about 60 days, or at least about 90 days after administration of the liquid formulation to the rabbit's eye The average level of is roughly constant at about 1 μg/mg. In some variations, when injected between the sclera and the conjunctiva of the rabbit's eye, the in situ gelling formulation maintains the intrascleral therapeutic agent for at least about 30 days, at least about 60 days, or at least about 90 days after administration of the liquid formulation to the rabbit's eye The average level of is roughly constant at about 10 μg/mg. the

In order to treat, prevent, inhibit, delay the onset of, or cause regression of certain diseases or conditions, it may be desirable to maintain delivery of a therapeutically effective amount of a therapeutic agent over an extended period of time. The extended period of time may be at least about 1 week, at least about 2 weeks, at least about 3 weeks, at least about 1 month, at least about 3 weeks based on the disease or condition to treat, prevent, inhibit, delay its occurrence, or cause its regression. months, at least about 6 months, at least about 9 months, or at least about 1 year. In general however any extended delivery time may be possible. A therapeutically effective amount of a therapeutic agent can be delivered for an extended period of time via a liquid formulation or composition that maintains, for an extended period of time, a concentration of the therapeutic agent in the subject or in the subject's eye sufficient for prolonged periods of time. A therapeutically effective amount of the therapeutic agent is delivered within a time period. the

Delivery of a therapeutically effective amount of a therapeutic agent over an extended period of time can be accomplished by placing one composition or liquid formulation, or by applying two or more doses of the composition or liquid formulation. As a non-limiting example of such multiple applications, maintaining a therapeutic amount of rapamycin for three months to treat, prevent, inhibit, delay the onset of, or cause regression of wet AMD can be delivered by applying the therapeutic amount for 3 months one liquid formulation or composition, or by sequential application of multiple liquid formulations or compositions. The optimal dosing strategy will depend on the therapeutic amount of the therapeutic agent that needs to be delivered, and when it needs to be delivered. Those skilled in the art of prolonging the delivery of therapeutic agents will understand how to determine useful dosing strategies based on the teachings herein. the

When using certain therapeutic agents or for treating, preventing, inhibiting, delaying the onset or regressing of certain diseases, it may be desirable that the delivery of the therapeutic agent not begin immediately when the liquid formulation or composition is placed in the ocular area, but rather is to start delivery after a certain delay. For example, without limitation, such delayed release may be useful when the therapeutic agent inhibits or delays wound healing, and a delayed release is desired to allow any wound healing that occurs when the liquid formulation or composition is placed. Depending on the therapeutic agent being delivered and/or the disease and condition being treated, prevented, inhibited, delayed, or caused to regress, the delay period before the delivery of the therapeutic agent begins can be about 1 hour, about 6 hours, about 12 hours, about 18 hours, about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, about 10 days, about 11 days, about 12 days , about 13 days, about 14 days, about 21 days, about 28 days, about 35 days, or about 42 days. Other delay periods may also be possible. Delayed release formulations that can be used are known to those skilled in the art. the

Intravitreal and subconjunctival delivery of rapamycin for the treatment, prevention, suppression, delay or regression of AMD

In one method described herein, a liquid formulation comprising rapamycin is delivered subconjunctivally or into the vitreous of the eye to treat, prevent, inhibit, delay, or cause regression of angiogenesis in the eye, including but not limited to treatment such as in CNVs observed in AMD. In some variations, liquid formulations are used to treat angiogenesis in the eye, including but not limited to treating CNV as observed in AMD. Rapamycin has been shown to inhibit CNV in rabbit and mouse models as described in US Application No. 10/665,203, which is incorporated herein by reference in its entirety. Rapamycin has been observed to inhibit Matrigel ^™ and laser-induced CNV when administered systemically and subretinally. Additionally, periocular injection of rapamycin suppressed laser-induced CNV.

Other therapeutic agents that can be delivered to the eye (particularly the vitreous of the eye) for the treatment, prevention, inhibition, delay or regression of ocular angiogenesis such as CNV are compounds of the limus family other than rapamycin, including but Not limited to everolimus and tacrolimus (FK-506). the

As described herein, the dosage of a therapeutic agent depends on the condition being treated (whether the condition is to be treated, prevented, inhibited, delayed or caused to regress), the specific therapeutic agent, and other clinical factors, such as the subject's Body weight and condition and route of administration of therapeutic agent. It is to be understood that the methods, liquid formulations and compositions described herein are applicable to human and veterinary use and possibly other animals. As described herein, tissue concentrations of therapeutic agents expressed in units of mass/volume generally refer to substantially aqueous tissue, such as the vitreous. Tissue concentrations of therapeutic agents expressed in mass/mass units generally refer to other tissues, such as sclera or retinal choroidal tissue. the

One concentration of rapamycin useful in the methods described herein is a concentration that provides tissue levels of rapamycin of about 0.01 pg/ml or pg/mg or more. Another concentration that can be used is one that provides about 0.1 pg/ml or ng/mg or more at the tissue level. Another concentration that can be used is one that provides about 1 pg/ml or ng/mg or more rapamycin at the tissue level. Another concentration that can be used is that which provides about 0.01 ng/ml or ng/mg or more at the tissue level. Another concentration that can be used is that which provides about 0.1 ng/ml or ng/mg or more at the tissue level. Another concentration that can be used is that which provides about 0.5 ng/ml or ng/mg or more at the tissue level. Another concentration that can be used is that which provides about 1 ng/ml or more at the tissue level. Another concentration that can be used is that which provides about 2 ng/ml or more at the tissue level. Another concentration that can be used is that which provides about 3 ng/ml or more at the tissue level. Another concentration that can be used is that which provides about 5 ng/ml or more at the tissue level. Another concentration that can be used is that which provides about 10 ng/ml or more at the tissue level. Another concentration that can be used is that which provides about 15 ng/ml or more at the tissue level. Another concentration that can be used is that which provides about 20 ng/ml or more at the tissue level. Another concentration that can be used is that which provides about 30 ng/ml or more at the tissue level. Another concentration that can be used is that which provides about 50 ng/ml or more at the tissue level. Those of skill in the art, given the teachings herein, will know how to achieve appropriate concentrations depending on the route and duration of administration used. the

Generally, the amount of rapamycin administered in a liquid formulation is an amount sufficient to treat, prevent, inhibit, delay the onset of, or cause regression of the ocular disease or condition for a desired amount of time. In some variations, the amount of rapamycin administered in the liquid formulation is an amount sufficient to treat the ocular disease or condition for the desired amount of time. the

In some variations, less than about 5 mg total rapamycin is administered subconjunctivally. In some variations, less than about 5.0 mg total rapamycin is administered subconjunctivally. In some variations, less than about 4.5 mg total rapamycin is administered subconjunctivally. In some variations, less than about 4.0 mg total rapamycin is administered subconjunctivally. In some variations, less than about 3.5 mg total rapamycin is administered subconjunctivally. In some variations, less than about 3.0 mg total rapamycin is administered subconjunctivally. In some variations, less than about 2.5 mg total rapamycin is administered subconjunctivally. In some variations, less than about 2 mg of the total amount of rapamycin is administered subconjunctivally. In some variations, less than about 1.2 mg total rapamycin is administered subconjunctivally. In some variations, less than about 1.0 mg total rapamycin is administered subconjunctivally. In some variations, less than about 0.8 mg total rapamycin is administered subconjunctivally. In some variations, less than about 0.6 mg total rapamycin is administered subconjunctivally. In some variations, less than about 0.4 mg total rapamycin is administered subconjunctivally. In some variations, a formulation volume comprising the amount of rapamycin described herein is administered. the

In some variations, a liquid formulation comprising between about 0.5% and about 6% by weight of the total weight of rapamycin is subconjunctivally administered to a human subject by administering between about 0.1 μl and about 200 μl of the liquid formulation described herein . In some variations, a liquid formulation comprising between about 0.5% and about 4% by weight of the total weight of a rapamycin is subconjunctivally administered to a human subject by administering between about 1 μl and about 50 μl of the liquid formulation described herein. In some variations, a liquid formulation comprising between about 1.5% and about 3.5% by weight of the total weight of a rapamycin is subconjunctivally administered to a human subject by administering between about 1 μl and about 15 μl of the liquid formulation described herein. In some variations, the human subject is subconjunctivally administered a liquid formulation comprising a rapamycin concentration of about 2% by weight of the total weight by administering between about 1 μl and about 15 μl of the liquid formulation described herein. the

In some variations, a liquid formulation comprising between about 0.2 μg and about 4 mg of rapamycin is subconjunctivally administered to a human subject by administering between about 0.1 μl and about 200 μl of the liquid formulation described herein. In some variations, a liquid formulation comprising between about 20 μg and about 2 mg of rapamycin is subconjunctivally administered to a human subject by administering between about 0.1 μl and about 100 μl of the liquid formulation described herein. In some variations, a liquid formulation comprising between about 20 μg and about 1 mg of rapamycin is subconjunctivally administered to a human subject by administering between about 1 μl and about 50 μl of the liquid formulation described herein. In some variations, a liquid formulation comprising between about 20 μg and about 500 μg of rapamycin is subconjunctivally administered to a human subject by administering between about 1 μl and about 25 μl of the liquid formulation described herein. In some variations, a liquid formulation comprising between about 20 μg and about 300 μg of rapamycin is subconjunctivally administered to a human subject by administering between about 1 μl and about 15 μl of the liquid formulation described herein. the

In some variations, a total amount of rapamycin of less than about 200 μg is administered intravitreally. In some variations, a total amount of rapamycin of less than about 200 μg is administered intravitreally. In some variations, a total amount of rapamycin of less than about 300 μg is administered intravitreally. In some variations, a total amount of rapamycin of less than about 400 μg is administered intravitreally. In some variations, less than about 500 μg of total rapamycin is administered intravitreally. In some variations, less than about 600 μg of total rapamycin is administered intravitreally. In some variations, a total amount of rapamycin of less than about 800 μg is administered intravitreally. In some variations, less than about 1 mg of the total amount of rapamycin is administered intravitreally. In some variations, less than about 2 mg of the total amount of rapamycin is administered intravitreally. In some variations, less than about 2.5 mg of the total amount of rapamycin is administered intravitreally. In some variations, less than about 3 mg of the total amount of rapamycin is administered intravitreally. In some variations, less than about 3.5 mg of the total amount of rapamycin is administered intravitreally. In some variations, less than about 4 mg of the total amount of rapamycin is administered intravitreally. In some variations, a formulation volume containing the amount of rapamycin described herein is administered. the

In some variations, a liquid formulation comprising between about 0.5% and about 6% by weight of the total weight of rapamycin is intravitreally administered to a human subject by administering between about 0.1 μl and about 200 μl of the liquid formulation described herein . In some variations, a liquid formulation comprising between about 0.5% and about 4% by weight of the total weight of rapamycin is intravitreally administered to a human subject by administering between about 1 μl and about 50 μl of the liquid formulation described herein. In some variations, a liquid formulation comprising between about 1.5% and about 3.5% by weight of the total weight of rapamycin is intravitreally administered to a human subject by administering between about 1 μl and about 15 μl of the liquid formulation described herein. In some variations, a liquid formulation comprising a concentration of about 2% by weight of the total weight of rapamycin is administered intravitreally to a human subject by administering between about 1 μl and about 15 μl of the liquid formulation described herein. the

In some variations, a liquid formulation comprising between about 0.2 μg and about 4 mg of rapamycin is administered intravitreally to a human subject by administering between about 0.1 μl and about 200 μl of the liquid formulation described herein. In some variations, a liquid formulation comprising between about 20 μg and about 2 mg of rapamycin is administered intravitreally to a human subject by administering between about 1 μl and about 100 μl of the liquid formulation described herein. In some variations, a liquid formulation comprising between about 20 μg and about 1 mg of rapamycin is administered intravitreally to a human subject by administering between about 1 μl and about 50 μl of the liquid formulation described herein. In some variations, a liquid formulation comprising between about 20 μg and about 500 μg of rapamycin is administered intravitreally to a human subject by administering between about 1 μl and about 25 μl of the liquid formulation described herein. In some variations, a liquid formulation comprising between about 20 μg and about 300 μg of rapamycin is administered intravitreally to a human subject by administering between about 1 μl and about 15 μl of the liquid formulation described herein. the

In some variations, a liquid formulation described herein comprising an amount of rapamycin between about 1 μg and about 5 mg for treating wet AMD is administered to a human subject. In some variations, a liquid formulation described herein comprising an amount of rapamycin between about 20 μg and about 4 mg for treating wet AMD is administered to a human subject. In some variations, a liquid formulation described herein comprising an amount of rapamycin between about 20 μg and about 1.2 mg for treating wet AMD is administered to a human subject. In some variations, a liquid formulation described herein comprising an amount of rapamycin between about 10 μg and about 0.5 mg for treating wet AMD is administered to a human subject. In some variations, a liquid formulation described herein comprising an amount of rapamycin between about 10 μg and about 90 μg for treating wet AMD is administered to a human subject. In some variations, a liquid formulation described herein comprising an amount of rapamycin between about 60 μg and about 120 μg for treating wet AMD is administered to a human subject. In some variations, a liquid formulation described herein comprising an amount of between about 100 μg and about 400 μg of rapamycin for treating wet AMD is administered to a human subject. In some variations, a liquid formulation described herein comprising an amount of rapamycin between about 400 μg and about 1 mg for treating wet AMD is administered to a human subject. In some variations, a liquid formulation described herein comprising an amount of rapamycin between about 1 mg and about 5 mg for treating wet AMD is administered to a human subject. In some variations, a liquid formulation described herein comprising an amount of rapamycin between about 3 mg and about 7 mg for treating wet AMD is administered to a human subject. In some variations, a liquid formulation described herein comprising an amount of rapamycin between about 5 mg and about 10 mg for treating wet AMD is administered to a human subject. the

In some variations, a human subject is administered a liquid formulation described herein comprising an amount of rapamycin between about 1 μg and about 5 mg for preventing wet AMD. In some variations, a human subject is administered a liquid formulation described herein comprising an amount of rapamycin between about 20 μg and about 4 mg for preventing wet AMD. In some variations, a human subject is administered a liquid formulation described herein comprising an amount of rapamycin between about 20 μg and about 1.2 mg for preventing wet AMD. In some variations, a human subject is administered a liquid formulation described herein containing rapamycin in an amount between about 10 μg and about 0.5 mg for preventing wet AMD. In some variations, a human subject is administered a liquid formulation described herein comprising an amount of rapamycin between about 10 μg and about 90 μg for the prevention of wet AMD. In some variations, a liquid formulation described herein comprising an amount of rapamycin between about 60 μg and about 120 μg is administered to a human subject for the prevention of wet AMD. In some variations, a human subject is administered a liquid formulation described herein comprising an amount of rapamycin between about 100 μg and about 400 μg for the prevention of wet AMD. In some variations, a human subject is administered a liquid formulation described herein comprising an amount of rapamycin between about 400 μg and about 1 mg for preventing wet AMD. In some variations, a human subject is administered a liquid formulation described herein comprising an amount of rapamycin between about 1 mg and about 5 mg for preventing wet AMD. In some variations, a human subject is administered a liquid formulation described herein comprising an amount of rapamycin between about 3 mg and about 7 mg for preventing wet AMD. In some variations, a human subject is administered a liquid formulation described herein comprising an amount of rapamycin between about 5 mg and about 10 mg for preventing wet AMD. the

In some variations, a human subject is administered a liquid formulation described herein comprising an amount of rapamycin between about 1 μg and about 5 mg for the treatment of dry AMD. In some variations, a liquid formulation described herein comprising an amount of rapamycin between about 20 μg and about 4 mg for treating dry AMD is administered to a human subject. In some variations, a human subject is administered a liquid formulation described herein comprising an amount of rapamycin between about 20 μg and about 1.2 mg for the treatment of dry AMD. In some variations, a human subject is administered a liquid formulation described herein comprising an amount of rapamycin between about 10 μg and about 0.5 mg for the treatment of dry AMD. In some variations, an amount of rapamycin comprising between about 10 μg and about 90 μg is administered to a human subject for the treatment of dry AMD. In some variations, an amount of rapamycin comprising between about 60 μg and about 120 μg is administered to a human subject for the treatment of dry AMD. In some variations, an amount of rapamycin comprising between about 100 μg and about 400 μg is administered to a human subject for the treatment of dry AMD. In some variations, an amount comprising between about 400 μg and about 1 mg of rapamycin is administered to a human subject for the treatment of dry AMD. In some variations, an amount comprising between about 1 mg and about 5 mg of rapamycin is administered to a human subject for the treatment of dry AMD. In some variations, an amount comprising between about 3 mg and about 7 mg of rapamycin is administered to a human subject for the treatment of dry AMD. In some variations, an amount comprising between about 5 mg and about 10 mg of rapamycin is administered to a human subject for the treatment of dry AMD. the

In some variations, a liquid formulation described herein comprising an amount of rapamycin between about 1 μg and about 5 mg is administered to a human subject for the treatment of angiogenesis, including but not limited to choroidal neovascularization. In some variations, rapamycin is administered to a human subject in an amount between about 20 μg and about 4 mg; between about 20 μg and about 1.2 mg; between about 10 μg and about 0.5 mg of rapamycin is administered to a human subject For the treatment of wet AMD, between about 10 μg and 90 μg, between about 60 μg and 120 μg is administered to a human subject; between about 100 μg and 400 μg, between about 400 μg and 1 mg is administered to a human subject; in some variations wherein, the human subject is administered rapamycin in an amount between about 1 mg and 5 mg; in some variations, the human subject is administered rapamycin in an amount between about 3 mg and 7 mg; in some variations, Rapamycin is administered to a human subject in an amount of between about 5 mg and 10 mg for the treatment of angiogenesis, including but not limited to choroidal neovascularization. the

In one approach, the liquid formulation described herein contains an amount of therapeutic agent equivalent to an amount of rapamycin. the

In one method, a human subject is administered an amount of a therapeutic agent described herein comprising an amount of a therapeutic agent equivalent to an amount of rapamycin between about 1 μg and about 5 mg for the treatment of wet AMD. In some variations, the human subject is administered an amount of therapeutic agent equivalent to between about 1 μg and about 5 mg of rapamycin; between about 20 μg and about 1.2 mg; between about 10 μg and about 0.5 mg to the human subject For the treatment of wet AMD, administering to a human subject between about 10 μg and 90 μg, between about 60 μg and 120 μg; administering to a human subject between about 100 μg and 400 μg, between about 400 μg and 1 mg; in some variations , administering to a human subject an amount of therapeutic agent equivalent to an amount of rapamycin between about 1 mg and 5 mg; in some variations, administering to a human subject an amount of rapamycin equivalent to between about 3 mg and 7 mg In some variations, the human subject is administered an amount of therapeutic agent equivalent to an amount of rapamycin between about 5 mg and 10 mg. the

In some variations, a liquid formulation described herein is administered to a human subject in an amount of a therapeutic agent equivalent to an amount of rapamycin between about 1 μg and about 5 mg for the treatment of dry AMD. In some variations, the human subject is administered an amount of therapeutic agent equivalent to between about 20 μg and about 4 mg of rapamycin; between about 20 μg and about 1.2 mg; between about 10 μg and about 0.5 mg to the human subject For the treatment of wet AMD, administering to a human subject between about 10 μg and 90 μg, between about 60 μg and 120 μg; administering to a human subject between about 100 μg and 400 μg, between about 400 μg and 1 mg; in some variations , administering to a human subject an amount of therapeutic agent equivalent to an amount of rapamycin between about 400 μg and 1 mg; in some variations, administering to a human subject an amount of rapamycin equivalent to between about 1 mg and 5 mg In some variations, the human subject is administered an amount of the therapeutic agent equivalent to an amount of rapamycin between about 3 mg and 7 mg; in some variations, the human subject is administered an amount equivalent to about 5 mg An amount of rapamycin equivalent therapeutic agent between 10 mg and 10 mg is used to treat dry AMD. the

In some variations, a human subject is administered an amount of a therapeutic agent described herein containing an amount of a therapeutic agent equivalent to an amount of rapamycin between about 1 μg and about 5 mg for preventing wet AMD. In some variations, the human subject is administered an amount of therapeutic agent equivalent to between about 20 μg and about 4 μg of rapamycin; between about 20 μg and about 1.2 mg; between about 10 μg and about 0.5 mg to the human subject For the prophylaxis of wet AMD, between about 10 μg and 90 μg, between about 60 μg and 120 μg is administered to a human subject; between about 100 μg and 400 μg, between about 400 μg and 1 mg is administered to a human subject; in some variations , administering to a human subject an amount of therapeutic agent equivalent to an amount of rapamycin between about 400 μg and 1 mg; in some variations, administering to a human subject an amount of rapamycin equivalent to between about 1 mg and 5 mg In some variations, the human subject is administered an amount of the therapeutic agent equivalent to an amount of rapamycin between about 3 mg and 7 mg; in some variations, the human subject is administered an amount equivalent to about 5 mg An amount of rapamycin equivalent therapeutic agent for prophylaxis of wet AMD. the

In some variations, every 3 or more months, every 6 or more months, every 9 or more months, or every 12 or more months or more, any one or more of the compounds described herein is administered intravitreally. A variety of preparations are used to treat one or more of choroidal neovascularization, wet AMD, and dry AMD, to prevent wet AMD or to prevent dry AMD from developing into wet AMD. In some variations, subconjunctival administration of any one or more of the compounds described herein is performed every 3 or more months, every 6 or more months, every 9 or more months, or every 12 or more months or more. A variety of preparations for treating one or more of choroidal neovascularization, wet AMD, dry AMD, or for preventing wet AMD. the

In some variations, every 3 or more months, every 6 or more months, every 9 or more months, or every 12 or more months or more, any one or more of the compounds described herein is administered intravitreally. A variety of rapamycin preparations are used to treat one or more of choroidal neovascularization, wet AMD, and dry AMD, to prevent wet AMD or to prevent dry AMD from developing into wet AMD. In some variations, subconjunctival administration of any one or more of the compounds described herein is performed every 3 or more months, every 6 or more months, every 9 or more months, or every 12 or more months or more. Various formulations of rapamycin for treating one or more of choroidal neovascularization, wet AMD, dry AMD, or for preventing wet AMD. In some variations, the effect of rapamycin persists beyond the period during which it is present in ocular tissue. the

Delivery of a therapeutic agent described herein can be, for example, at a dosage range between about 1 ng/day and about 100 μg/day, or at a dosage above or below this range, depending on the route and duration of administration. In some variations of the liquid formulations or compositions used in the methods described herein, the therapeutic agent may be delivered in a dosage range between about 0.1 μg/day and about 10 μg/day. In some variations of the liquid formulations or compositions used in the methods described herein, the therapeutic agent may be delivered in a dosage range between about 1 μg/day and about 5 μg/day. Dosages of the various therapeutic agents used to treat, prevent, inhibit, delay the onset or cause regression of the various diseases and conditions described herein can be optimized through the use of clinical trials. the

Liquid formulations including, but not limited to, solutions, suspensions, emulsions, and in situ gelling formulations and compositions described herein can be used to deliver a therapeutically effective amount of rapamycin to the eye over an extended period of time (as A non-limiting example, by ocular administration or periocular administration), for the treatment, prevention, inhibition, delay or regression of CNV, and thus can be used for the treatment, prevention, inhibition, delay of the occurrence of wet AMD or Regression or conversion of dry AMD to wet AMD. It is believed that by altering certain characteristics of the liquid formulations described herein, including, but not limited to, the composition of the liquid formulations, where the liquid formulations are delivered into the eye, including but not limited to subconjunctival or intravitreal placement, the liquid formulations can be used Delivering a therapeutically effective amount of rapamycin to the eye over various extended periods of time, including delivering a therapeutically effective amount for more than about 1 week, for more than about 2 weeks, for more than about 3 weeks, for more than about 1 months, more than about 3 months, more than about 6 months, more than about 9 months, more than about 1 year. the

Rapamycin treats, inhibits, or causes regression of wet AMD when a therapeutically effective amount of rapamycin is administered to a subject with wet AMD. Different therapeutically effective amounts may be required to treat, inhibit or cause regression. Subjects with wet AMD may have CNV lesions, and it is believed that administration of a therapeutically effective amount of rapamycin may have various effects including, but not limited to, causing regression of CNV lesions, stabilizing CNV lesions, and preventing the development of active CNV lesions. the

When a therapeutically effective amount of rapamycin is administered to a subject with dry AMD, it is believed that rapamycin prevents or delays progression from dry AMD to wet AMD. the

Example

Unless context dictates otherwise, error bars in graphs show one standard deviation. When ethanol was used, it was 200 proof ethanol from Gold Shield Distributors, Hayward, CA. When using rapamycin, it was from LC laboratories, Woburn, MA or Chunghwa Chemical Synthesis & Biotech Co., LTD (CCSB), Taipei Hsien, Taiwan, ROC. When PEG 400 was used, it was obtained from The Dow Chemical Company, New Milford, CT. Some figures use the expression "uL" or "ug" to refer to μL or μL, respectively. When administering volumes of 10 μL or less, use a Hamilton HPLC syringe. the

Example 1 - Preparation and Characterization of Rapamycin-Containing Solutions

1.256% rapamycin (percentage of total weight) was dissolved in 9.676% ethanol (percentage of total weight). A 15% aqueous solution of F127 (Lutrol) in sterile water was slowly added with constant stirring. The final concentration was about 78.57% sterile water (percentage of total weight) and about 10.50% F127 (Lutrol) (percentage of total weight). This solution is listed in Table 1 as Formulation #32. The solution was kept at 2°C until use. the

Example 2-subconjunctival injection of a solution containing rapamycin

50 µl of the solution described in Example 1 was injected between the sclera and conjunctiva of New Zealand white rabbits. the

Figure 2 depicts the logarithmic mean concentrations of rapamycin present in the vitreous (ng/ml), retinachoroid (ng/mg) and sclera (ng/mg) 20, 40, 67 and 90 days after injection. the

Analysis was performed by liquid chromatography mass spectrometry (LCMS) using an internal standard. the

At each time point, the mean concentration of rapamycin was calculated by summing the obtained rapamycin concentrations per eye for each rabbit and dividing the number of eyes analyzed by the total amount. In this experiment, each time point represents the average of two individual eyes of two rabbits (four eyes at the time point) or the average of both eyes of one rabbit (two eyes at the time point). the

Whole vitreous bodies were homogenized and analyzed. The mean concentration in the vitreous was calculated by dividing the measured mass of rapamycin by the analyzed vitreous volume. The samples did not include the site of application; thus, this measurement indicates the level of rapamycin delivered into the vitreous by the solution. the

Mean levels of intravitreal rapamycin were approximately 4.425, 3.800, 4.100 and 1.500 ng/ml at 20, 40, 67 and 90 days after subconjunctival injection, respectively. the

Whole retinal choroids were homogenized and analyzed. The average concentration of the retinochoroid was calculated by dividing the measured mass of rapamycin by the mass of the analyzed retinochoroid. The samples did not include the site of administration; thus, this measurement indicated the level of rapamycin delivered into the retinachoroid by the solution. the

Mean levels of rapamycin in the retina and choroid were approximately 0.055, 0.209, 0.080 and 0.017 ng/mg at 20, 40, 67 and 90 days after subconjunctival injection, respectively. the

The sclera was analyzed in the same manner as the retinachoroid. Sclera samples included the site of injection; thus, this measurement indicates the rate of clearance of rapamycin from the sclera. the

The mean levels of intrascleral rapamycin were approximately 0.141, 0.271, 0.067 and 0.192 ng/ml at 20, 40, 67 and 90 days after subconjunctival injection, respectively. the

Example 3 - Preparation and Characterization of Rapamycin-Containing Solutions

5.233% rapamycin (% of total formulation weight after addition of all ingredients) was dissolved in 0.4177g EtOH; the amount of EtOH was reduced to 0.1296g (6.344%, w/w) by forced evaporation (heating). Add PEG 400 with constant stirring. The final concentrations as total weight percentages are approximately: rapamycin 5.233%, ethanol 6.344% and PEG 400 88.424%. On contact with the vitreous, the formulation forms non-dispersed masses relative to the surrounding medium. This solution is listed in Table 1 as Formulation #34. the

Example 4-subconjunctival injection of a solution containing rapamycin

25 [mu]l of the solution described in Example 3 was injected between the sclera and conjunctiva of New Zealand white rabbits. the

Figure 3 depicts the logarithmic scale of rapamycin levels present in the vitreous (ng/ml), retinochoroid (ng/mg) and sclera (ng/mg) 14, 35, 62 and 85 days after injection. the

The vitreous was homogenized and analyzed as described in Example 2, except that a single eye from each of three rabbits was analyzed on day 2; both eyes from each of two rabbits were analyzed on day 14; Both eyes of one rabbit; both eyes of one rabbit were analyzed on day 62; and one eye from one rabbit and both eyes from the other rabbit were analyzed on day 85. the

The vitreous samples did not include the site of application; thus, this measurement indicated the level of rapamycin delivered into the vitreous by the solution. Mean levels of intravitreal rapamycin were approximately 3.57, 53.65, 9.00, 4.700 and 0.600 ng/ml at 2, 14, 35 and 85 days after subconjunctival injection, respectively. the

Retinochoroids were homogenized and analyzed as described in Example 2, samples were taken on the dates described above for the vitreous. Day 2 analysis was not performed. Retinochoroid samples did not include the site of administration; thus, this measurement indicated the level of rapamycin delivered into the retinochoroid by solution. Mean levels of rapamycin in the retina and choroid were approximately 0.4815, 1.725, 0.057 and 0.009 ng/mg at 14, 35, 62 and 85 days after subconjunctival injection, respectively. the

Sclera samples were analyzed as described in Example 2, sampled on the dates described for the retinachoroid. The sclera samples included the site of application; thus, this measurement indicates the rate of clearance of rapamycin from the sclera. The mean levels of intrascleral rapamycin were approximately 34.5815, 0.135, 0.042, and 0.163666667 ng/mg 14, 35, 62, and 85 days after subconjunctival injection, respectively. the

Example 5-intravitreal injection of a solution containing rapamycin

25 [mu]l of the solution described in Example 3 was injected into the vitreous of New Zealand White rabbit eyes. Figure 4 depicts the logarithmic scale levels of rapamycin present in the vitreous (ng/ml), retinachoroid (ng/mg) and sclera (ng/mg) 14, 35, 62 and 90 days after injection. The level of rapamycin present in the vitreous (ng/ml) 2 days after injection is also shown. the

The vitreous was homogenized and analyzed as described in Example 2, except that approximately 1 μl of each eye of three rabbits was analyzed on day 2; both eyes of each of two rabbits were analyzed on day 14; Both eyes of one rabbit were analyzed; both eyes of one rabbit were analyzed on day 62; and both eyes from each of the two rabbits were analyzed on day 90. the

Vitreous samples included the site of application except for Day 2 samples. Avoid the applied solution when possible. However, the accuracy of the measured rapamycin levels may be affected by sampling error due to inadvertent inclusion of the administered solution. the

The mean levels of intravitreal rapamycin at 2, 14, 35, 62 and 90 days after intravitreal injection were approximately 11.4, 136538, 2850.3, 21820.35 and 27142.75 ng/ml, respectively. the

Retinochoroids were homogenized and analyzed as described in Example 2, and samples were taken on the days described above for the vitreous. Day 2 analysis was not performed. Retinochoroid samples did not include the site of administration; thus, this measurement indicated the level of rapamycin delivered into the retinochoroid by solution. The mean levels of rapamycin in the retina and choroid were approximately 5.78975, 244.485, 0.105 and 1.782 ng/mg 14, 35, 62 and 90 days after intravitreal injection, respectively. the

Sclera samples were analyzed as described in Example 2, sampled on the dates described above for the retinachoroid. Sclera samples did not include the injection site; therefore, this measurement indicates the clearance of rapamycin from the sclera. The mean levels of rapamycin in the sclera were approximately 0.5695, 12.34, 0.8505 and 0.71175 ng/mg at 14, 35, 62 and 90 days after intravitreal injection, respectively. the

Example 6 - Preparation and Characterization of Rapamycin-Containing Suspensions

6% rapamycin (percentage of total weight) was dispersed in 94% PEG400 (percentage of total weight). This suspension is listed in Table 1 as Formulation #55. the

Example 7 - Intravitreal injection of a suspension containing rapamycin

The solution prepared in Example 6 was injected intravitreally into the eyes of New Zealand white rabbits. Figure 5 depicts images of rabbit eyes intravitreally injected with 10 μl (Figure 5A), 20 μl (Figure 5B) and 40 μl (Figure 5C) of 6% rapamycin suspended in PEG400. This resulted in injected doses of about 0.6, about 1.2 and about 2.4 mg. The image is focused on the applied suspension. These images show that the suspension forms non-dispersed masses relative to the surrounding vitreous medium. the

Example 8 - Preparation and Characterization of In Situ Gelling Formulations Containing Rapamycin

4.2% rapamycin (from LC laboratories in Woburn, MA and Chunghwa Chemical Synthesis & BioTech.Co, Ltd in Taiwan), 4.3% ethanol (from GoldShield Chemical in Hayward, CA), 2.2% PVP K90 (from BASF ), 87.1% PEG 400 (from DOW Chemical) and 2.2% Eudragit RL 100 (from RohmPharma Polymers), wherein all percentages are percentages of total weight. the

Dissolve Eudragit RL 100 in ethanol. This step may require sonication and heating. Ethanol - Eudragit added to PEG 400. PVP was slowly added to the Eudragit-ethanol-PEG solution to obtain a homogeneously mixed solution. This step may require intensive mixing. the

Rapamycin was added and dissolved in the Eudragit-ethanol-PEG-PVP mixture. Heat and sonication may be used. The formulations were mixed thoroughly (using a vortexer or mixer) to achieve homogeneity. This formulation is listed in Table 1 as #37. the

Liquid formulations formed non-dispersible clumps when placed in deionized or tap water. The non-dispersed mass appeared as a gel-like mass. the

Example 9 - Subconjunctival Injection of a Formulation Containing Rapamycin in Non-Dispersed Masses

50 µl of the solution described in Example 8 was injected between the sclera and conjunctiva of New Zealand white rabbits. the

Figure 6 depicts the mean concentrations of rapamycin present in the vitreous (ng/ml), retinachoroid (ng/mg) and sclera (ng/mg) 7, 32, 45 and 90 days after injection of the in situ gelling formulation . the

Analysis by LCMS (liquid chromatography-mass spectrometry). the

When more than a single eye was analyzed, the mean concentration of rapamycin was calculated by adding the rapamycin concentrations obtained from each eye of each rabbit and dividing the total by the number of eyes analyzed. In this experiment, the vitreous day 7 and scleral day 7, 32, and 45 time points represent monocular relative to the mean. The remaining 7, 32 and 45 day time points represent the average of both eyes of one rabbit, while the 90 day time point represents the average of both eyes of each of two rabbits (total of four eyes). the

Whole vitreous bodies were homogenized and analyzed. The mean concentration in the vitreous was calculated by dividing the measured mass of rapamycin by the analyzed vitreous volume. The samples did not include the site of administration; therefore, this measurement indicates the level of rapamycin delivered into the vitreous by the in situ gelling formulation. the

Mean levels of rapamycin in the vitreous were about 13.9, about 7.4, about 1.35 and about 9.9 ng/ml at 7, 32, 45 and 90 days after subconjunctival injection, respectively. the

Whole retinal choroids were homogenized and analyzed. The average concentration of retinochoroid was calculated by dividing the measured mass of rapamycin by the mass of retinochoroid tissue analyzed. The samples did not include the site of administration; therefore, this measurement indicates the level of rapamycin delivered into the retinachoroidal tissue by the in situ gelling formulation. the

Mean levels of rapamycin in the retina-choroidal tissue were about 0.376, about 0.1875, about 0.136 and about 0.29 ng/mg at 7, 32, 45 and 90 days after subconjunctival injection, respectively. the

The sclera was analyzed in the same manner as the retinal choroidal tissue. Sclera samples may include injected liquid formulations; thus, this measurement indicates the rate of clearance of rapamycin from the sclera. the

The mean levels of intrascleral rapamycin were about 2033, about 1653, about 3626 and about 420.5 ng/mg at 7, 32, 45 and 90 days after subconjunctival injection, respectively. the

Example 10 - Preparation and Characterization of Rapamycin-Containing Suspensions

A rapamycin-containing suspension was prepared by dispersing 150.5 mg rapamycin (3.004% by weight) in 4860.3 mg PEG400 (96.996% by weight). This formulation is listed in Table 1 as #49. 150.5 mg rapamycin (3.004% by weight) and 4860.3 mg PEG 400 (96.996% by weight) were placed in an amber tube. 3mm diameter High Wear Resistant Zirconia Grinding Media (beads) were added up to three quarters of the total volume. The tubes were sealed and placed in a Cole-Parmer milling apparatus for 48 hours. The median particle size of rapamycin was 2.8386mm, and the mean value was 3.1275mm. The formulation was stored at 4°C until use. Volumes of 20 μl and 40 μl each formed non-dispersed clumps when placed in the vitreous of rabbit eyes. the

Example 11 - Subconjunctival injection of suspension containing rapamycin

40 µl of the suspension described in Example 10 was injected between the sclera and conjunctiva of New Zealand white rabbits. Figure 7 depicts the log scale levels of rapamycin in the vitreous (ng/ml), retinachoroid (ng/mg) and sclera (ng/mg) 14, 42, 63 and 91 days post-injection. the

Vitreous bodies were homogenized and analyzed as described in Example 2. Both eyes from each of two rabbits were analyzed at each time point except day 91, when both eyes from one rabbit were analyzed. Vitreous samples did not include the site of administration; thus, this measurement indicated the level of rapamycin delivered into the vitreous. Mean levels of intravitreal rapamycin were approximately 4.031, 23.11, 53.27 and 13.94 ng/ml 14, 42, 63 and 91 days after subconjunctival injection, respectively. the

Retinochoroids were homogenized and analyzed as described in Example 2, and samples were taken on the days described above for the vitreous. The retinochoroid does not include the site of administration; thus, this measurement indicates the level of rapamycin delivered into the retinochoroid. Mean levels of rapamycin in the retina and choroid were approximately 0.1577, 4.965, 0.385 and 0.05 ng/mg 14, 42, 63 and 91 days after subconjunctival injection, respectively. the

Sclera samples were homogenized and analyzed as described in Example 2, and samples were taken as described above for the vitreous. Sclera samples included injection sites. The mean levels of intrascleral rapamycin were approximately 1283, 476.3, 854.2, and 168.5 ng/mg 14, 42, 63, and 91 days after subconjunctival injection, respectively. the

Example 12 - Intravitreal injection of a suspension containing rapamycin

20 [mu]l of the suspension described in Example 10 was injected into the vitreous of New Zealand White rabbit eyes. The injected suspension forms a non-dispersed mass relative to the surrounding medium. Figure 8 depicts rapamycin on a logarithmic scale in the retina choroid (ng/mg) and sclera (ng/mg) at 14, 42, 63 and 91 days post-injection and in the vitreous (ng/ml) at 63 and 91 days post-injection prime level. the

Vitreous bodies were homogenized and analyzed as described in Example 2. Both eyes from each of the two rabbits were analyzed at each time point. A vitreous sample can comprise an application site. Mean levels of intravitreal rapamycin were approximately 381,600 and 150,400 ng/ml at 63 and 91 days after intravitreal injection, respectively. the

Retinochoroids were homogenized and analyzed as described in Example 2. Both eyes from each of the two rabbits were analyzed at each time point. The retinochoroid does not include the site of administration, so this measurement indicates the level of rapamycin delivered into the retinochoroid. The mean levels of rapamycin in the retina and choroid were approximately 2.588, 4.249, 21.42 and 0.922 ng/mg 14, 42, 63 and 91 days after intravitreal injection, respectively. the

Sclera samples were homogenized and analyzed as described in Example 2, and samples were taken as described above for the retinachoroid. Sclera samples did not include the injection site; thus, this measurement indicates the level of rapamycin delivered to the sclera. The mean levels of rapamycin in the sclera were approximately 0.7327, 6.053, 1.373 and 17.49 ng/mg 14, 42, 63 and 91 days after intravitreal injection, respectively. the

Example 13 - Preparation and Characterization of Rapamycin-Containing Solutions

A rapamycin-containing solution was formed by dissolving 116.6 mg of rapamycin in ethanol and storing the mixture at 4°C for 6 hours. This solution was then mixed with 4647.5 mg PEG 400 to obtain a solution having a final concentration by weight of 2.29% rapamycin, 6.05% ethanol and 91.66% PEG400. This solution is listed in Table 1 as Formulation #51. A volume of 30 μl formed non-dispersed clumps when placed in the vitreous of a rabbit eye. the

Example 14 - Subconjunctival injection of a solution containing rapamycin

40 µl of the solution described in Example 13 was injected between the sclera and conjunctiva of New Zealand White rabbit eyes. Figure 9 depicts rapamycin levels on a linear scale in the vitreous (ng/ml), retina choroid (ng/mg) and sclera (ng/mg) 14, 42, 63 and 91 days after injection. the

Vitreous bodies were homogenized and analyzed as described in Example 2. Both eyes from each of two rabbits were analyzed at each time point except day 91, when both eyes from one rabbit were analyzed. Vitreous samples did not include the site of administration; thus, this measurement indicated the level of rapamycin delivered into the vitreous. Mean levels of intravitreal rapamycin were approximately 1.804, 1.854, 1.785 and 1.255 ng/ml at 14, 42, 63 and 91 days after subconjunctival injection, respectively. the

Retinochoroids were homogenized and analyzed as described in Example 2, and samples were taken as described above for the vitreous. The retinochoroid does not include the site of administration; thus, this measurement indicates the level of rapamycin delivered into the retinochoroid. Mean levels of rapamycin in the retina and choroid were approximately 1.221, 4.697, 0.1075 and 0.02 ng/mg 14, 42, 63 and 91 days after subconjunctival injection, respectively. the

Sclera samples were homogenized and analyzed as described in Example 2, and samples were taken as described above for the vitreous. Sclera samples included injection sites. The mean levels of intrascleral rapamycin were approximately 1.987, 1.884, 0.56 and 10.84 ng/mg 14, 42, 63 and 91 days after subconjunctival injection, respectively. the

Example 15 - Intravitreal injection of a solution containing rapamycin

30 [mu]l of the solution described in Example 13 was injected into the vitreous of New Zealand White rabbit eyes. The injected solution forms a non-dispersed mass relative to the surrounding medium. Figure 10 depicts rapamycin levels on a linear scale in the retina choroid (ng/mg) and sclera (ng/mg) 14, 42, 63 and 91 days post-injection. the

Retinochoroids were homogenized and analyzed as described in Example 2. Both eyes from each of the two rabbits were analyzed at each time point. The retinochoroid does not include the site of administration, so this measurement indicates the level of rapamycin delivered into the retinochoroid. Mean levels of rapamycin in the retina and choroid were approximately 5.515, 5.388, 0.3833 and 11.52 ng/mg 14, 42, 63 and 91 days after intravitreal injection, respectively. the

Sclera samples were homogenized and analyzed as described in Example 2, and samples were taken as described above for the retinachoroid. The sclera samples did not include the injection site, so this measurement indicates the level of rapamycin delivered into the sclera. The mean levels of intrascleral rapamycin were approximately 1.077, 0.9239, 0.0975 and 2.0825 ng/mg at 14, 42, 63 and 91 days after intravitreal injection, respectively. the

Figure 11 depicts rapamycin levels in the vitreous (ng/ml) on a linear scale at 63 and 91 days post-injection. Vitreous bodies were homogenized and analyzed as described in Example 2. Both eyes from each of the two rabbits were analyzed at each time point. A vitreous sample can comprise an application site. Mean levels of intravitreal rapamycin were approximately 299,900 and 196,600 ng/ml at 63 and 91 days after intravitreal injection, respectively. the

Example 16 - Preparation and Characterization of Rapamycin-Containing Solutions

About 320 g of ethanol was flushed with _N2 for about 10 minutes, and then about 40 g of sirolimus was added to the ethanol. The mixture was sonicated for about 20 minutes, at the end of which all the sirolimus had gone into solution, forming a stock solution of sirolimus. The dilution solvent was prepared by sonicating about 1880 g of PEG 400 for about 60 minutes, then flushing the solvent with nitrogen for about 10 minutes.

The sirolimus stock solution and PEG 400 were then rotated at about room temperature for about 10 minutes in a rotary evaporator to mix the stock solution and dilution solvent. After mixing the solution was flushed with nitrogen for about 10 minutes and blanketed with nitrogen for about 5 minutes. After the solution was flushed and flooded with nitrogen, about 240 g of excess ethanol was evaporated from the solution by increasing the temperature of the solution, maintaining a temperature not exceeding 40° C. for an extended period of time, and continuing to swirl the solution for about 2.5 hours. the

The resulting solution contained about 40 g sirolimus (about 2% by weight), about 80 g ethanol (about 4% by weight), and about 1880 g PEG 400 (about 94% by weight). The solution was flushed with nitrogen for about 10 minutes and blanketed with nitrogen for about 5 minutes. The solution was then filtered through a 0.2 micron filter. HPLC vials were filled with 2 ml of each filtered solution, leaving approximately 400 [mu]l headspace in each vessel. The headspace was filled with nitrogen and capped. the

Example 17 - Preparation and Characterization of Rapamycin-Containing Solutions

The rapamycin, ethanol, and PEG 400 were placed in the container to give a final concentration of about 2.00% rapamycin, about 4.00% ethanol, and about 94.00% PEG 400 by weight. The mixture was capped and sonicated for 1-2 hours. Sonication generates heat, with temperatures as high as about 40 or 50°C. This solution is listed in Table 1 as Formulation #100. Volumes of 1 μl, 3 μl, 20 μl and 40 μl formed non-dispersed masses in the vitreous of rabbit eyes. the

Example 18 - Subconjunctival injection of a solution containing rapamycin

20 µl of the solution described in Example 17 was injected between the sclera and conjunctiva of New Zealand white rabbit eyes. Figure 12 depicts the levels of rapamycin present in the vitreous on a logarithmic scale at 5, 30, 60, 90 and 120 days post-injection. Figure 13 depicts rapamycin levels on a logarithmic scale in the retina and choroid at the same time points. For comparison, Figures 12 and 13 also depict the results of similar studies performed with 40 μl and 60 μl injections, described in Examples 19 and 20 below. the

In Figures 12-15 discussed in this and following examples, some outliers are omitted. Individual data points from the same study at the same time point were compared to each other. Data points above or below an order of magnitude were considered outliers when their arithmetic mean was below their standard deviation. the

Vitreous bodies were homogenized and analyzed as described in Example 2. Two to five rabbit eyes were analyzed per time point. Vitreous samples did not include the site of administration; thus, this measurement indicated the level of rapamycin delivered into the vitreous. Mean levels of rapamycin in the vitreous were approximately 1.81, 0.45, 0.39, 1.85 and 1.49 ng/ml at 5, 30, 60, 90 and 120 days after subconjunctival injection, respectively. the

Retinochoroids were homogenized and analyzed as described in Example 2, and samples were taken as described above for the vitreous. The retinochoroid does not include the site of administration; thus, this measurement indicates the level of rapamycin delivered into the retinochoroid. Mean levels of rapamycin in the retina and choroid were approximately 0.14, 0.03, 0.02, 0.02 and 0.01 ng/mg at 5, 30, 60, 90 and 120 days after subconjunctival injection, respectively. the

Example 19 - Subconjunctival injection of a solution containing rapamycin

40 µl of the solution described in Example 17 was injected between the sclera and conjunctiva of New Zealand white rabbit eyes. Figure 12 depicts the levels of rapamycin present in the vitreous on a logarithmic scale at 5, 30, 60, 90 and 120 days post-injection. Figure 13 depicts rapamycin levels on a logarithmic scale in the retina and choroid at the same time points. the

Vitreous bodies were homogenized and analyzed as described in Example 2. Two to five rabbit eyes were analyzed per time point. Vitreous samples did not include the site of administration; thus, this measurement indicated the level of rapamycin delivered into the vitreous. Mean levels of intravitreal rapamycin were approximately 2.39, 0.65, 0.54, 2.07 and 1.92 ng/ml at 5, 30, 60, 90 and 120 days after subconjunctival injection, respectively. the

Retinochoroids were homogenized and analyzed as described in Example 2, and samples were taken as described above for the vitreous. The retinochoroid does not include the site of administration; thus, this measurement indicates the level of rapamycin delivered into the retinochoroid. Mean levels of rapamycin in the retina and choroid were approximately 0.47, 0.04, 0.01, 0.05 and 0.0 ng/mg at 5, 30, 60, 90 and 120 days after subconjunctival injection, respectively. the

Example 20 - Subconjunctival injection of a solution containing rapamycin

60 µl of the solution described in Example 17 was injected between the sclera and conjunctiva of New Zealand white rabbit eyes. Figure 12 depicts the levels of rapamycin present in the vitreous on a logarithmic scale at 5, 30, 60, 90 and 120 days post-injection. Figure 13 depicts rapamycin levels on a logarithmic scale in the retina and choroid at the same time points. the

Vitreous bodies were homogenized and analyzed as described in Example 2. Two to five rabbit eyes were analyzed per time point. Vitreous samples did not include the site of administration; thus, this measurement indicated the level of rapamycin delivered into the vitreous. Mean levels of rapamycin in the vitreous were approximately 8.65, 0.29, 0.18, 2.00, 1.41 ng/ml at 5, 30, 60, 90 and 120 days after subconjunctival injection, respectively. the

Retinochoroids were homogenized and analyzed as described in Example 2, and samples were taken as described above for the vitreous. The retinochoroid does not include the site of administration; thus, this measurement indicates the level of rapamycin delivered into the retinochoroid. Mean levels of rapamycin in the retina and choroid were approximately 0.63, 0.02, 0.02, 0.06 and 0.01 ng/mg at 5, 30, 60, 90 and 120 days after subconjunctival injection, respectively. the

Example 21 - Intravitreal injection of a solution containing rapamycin

20 [mu]l of the solution described in Example 17 was injected into the vitreous of New Zealand White rabbit eyes. The injected solution forms a non-dispersed mass relative to the surrounding medium. Figure 14 depicts rapamycin levels in the vitreous on a logarithmic scale at 5, 30, 60, 90 and 120 days post-injection. Figure 15 depicts rapamycin levels on a logarithmic scale in the retina and choroid at the same time points. For comparison, Figures 14 and 15 also depict the results of other studies described in Example 22 and Example 24 below. the

Vitreous bodies were homogenized and analyzed as described in Example 2. Two to five rabbit eyes were analyzed per time point. A vitreous sample can include an application site. Mean levels of intravitreal rapamycin were approximately 162,100; 18,780; 57,830; 94,040 and 13,150 ng/ml at 5, 30, 60, 90 and 120 days after intravitreal injection, respectively. the

Retinochoroids were homogenized and analyzed as described in Example 2, and samples were taken as described above for the vitreous. Retinochoroid samples did not include the site of administration; thus, this measurement indicated the level of rapamycin delivered into the retinochoroid. Mean levels of rapamycin in the retina and choroid were approximately 2.84, 2.26, 0.17, 0.22 and 0.05 ng/mg at 5, 30, 60, 90 and 120 days after intravitreal injection, respectively. the

Example 22 - Intravitreal injection of a solution containing rapamycin

40 µl of the solution described in Example 17 was injected into the vitreous of New Zealand White rabbit eyes. The injected solution forms a non-dispersed mass relative to the surrounding medium. Figure 14 depicts rapamycin levels in the vitreous on a logarithmic scale at 5, 30, 60, 90 and 120 days post-injection. Figure 15 depicts rapamycin levels on a logarithmic scale in the retina and choroid at the same time points. the

Vitreous bodies were homogenized and analyzed as described in Example 2. Two to five rabbit eyes were analyzed per time point. A vitreous sample can include an application site. Mean levels of intravitreal rapamycin were approximately 415,600; 4,830; 74,510; 301,300 and 7,854 ng/ml at 5, 30, 60, 90 and 120 days after intravitreal injection, respectively. the

Retinochoroids were homogenized and analyzed as described in Example 2, and samples were taken as described above for the vitreous. Retinochoroid samples did not include the site of administration; thus, this measurement indicated the level of rapamycin delivered into the retinochoroid. Mean levels of rapamycin in the retina and choroid were approximately 5.36, 0.23, 1.27, 1.08 and 0.08 ng/mg at 5, 30, 60, 90 and 120 days after intravitreal injection, respectively. the

Example 23 - Preparation and characterization of solutions containing rapamycin. the

The rapamycin, ethanol, and PEG400 were placed in the container to give a final concentration of about 0.4% rapamycin, about 4.0% ethanol, and about 95.6% PEG400 by weight. The mixture was sonicated for 1-2 hours. Sonication causes the temperature to rise up to about 40 to 50°C. This solution is listed in Table 1 as Formulation #99. the

Example 24 - Intravitreal injection of a solution containing rapamycin

100 [mu]l of the solution described in Example 23 was injected into the vitreous of New Zealand White rabbit eyes. The injected solution does not form non-dispersed masses relative to the surrounding medium. Figure 14 depicts rapamycin levels in the vitreous on a logarithmic scale at 5, 30, 60 and 90 days post-injection. Figure 15 depicts rapamycin levels on a logarithmic scale in the retina and choroid at the same time points. the

Vitreous bodies were homogenized and analyzed as described in Example 2. Two to five rabbit eyes were analyzed per time point. A vitreous sample can include an application site. Mean levels of intravitreal rapamycin were approximately 151,000; 14,890; 4,743 and 1620 ng/ml at 5, 30, 60 and 90 days after intravitreal injection, respectively. the

Retinochoroids were homogenized and analyzed as described in Example 2, and samples were taken as described above for the vitreous. Retinochoroid samples did not include the site of administration; thus, this measurement indicated the level of rapamycin delivered into the retinochoroid. Mean levels of rapamycin in the retina and choroid were approximately 1.21, 1.84, 0.04, and 0.71 ng/mg at 5, 30, 60, and 90 days after intravitreal injection, respectively. the

Example 25 Preparation and Characterization of Solutions Containing Rapamycin. the

A rapamycin-containing solution was formed by dissolving 102.4 mg rapamycin in ethanol, adding 4719.3 mg PEG400 and vortexing. The resulting solution had a final concentration by weight of 2.036% rapamycin, 4.154% ethanol, and 93.81% PEG400. This solution is listed in Table 1 as Formulation #139. the

Example 26 Subconjunctival injection of a solution containing rapamycin

10 [mu]l of the solution described in Example 25 was injected as a single dose between the sclera and conjunctiva of New Zealand white rabbit eyes. Figure 16 depicts the levels of rapamycin present in the vitreous on a logarithmic scale at 5 and 14 days post-injection. Figure 17 depicts rapamycin levels on a logarithmic scale in the retina and choroid at the same time points. For comparison, Figures 16 and 17 also depict the results of other studies described in Examples 27-29 below. the

Vitreous bodies were homogenized and analyzed as described in Example 2. Four rabbit eyes were analyzed per time point. Vitreous samples did not include the site of administration, therefore, this measurement indicates the level of rapamycin delivered into the vitreous. Mean levels of rapamycin in the vitreous were approximately 2.45 and 20.13 ng/ml at 5 and 14 days after subconjunctival injection, respectively. the

Retinochoroids were homogenized and analyzed as described in Example 2, and samples were taken as described above for the vitreous. The retinochoroid does not include the site of administration, therefore, this measurement indicates the level of rapamycin delivered into the retinochoroid. Mean levels of rapamycin in the retina and choroid were approximately 0.13 and 0.19 ng/mg 5 and 14 days after subconjunctival injection, respectively. the

Example 27 - Subconjunctival injection of a solution containing rapamycin

60 [mu]l of the solution described in Example 25 was injected as a single dose between the sclera and conjunctiva of New Zealand white rabbit eyes. Figure 16 depicts the levels of rapamycin present in the vitreous on a logarithmic scale at 5 and 14 days post-injection. Figure 17 depicts rapamycin levels on a logarithmic scale in the retina and choroid at the same time points. the

Vitreous bodies were homogenized and analyzed as described in Example 2. Four rabbit eyes were analyzed per time point. Vitreous samples did not include the site of administration, therefore, this measurement indicates the level of rapamycin delivered into the vitreous. Mean levels of rapamycin in the vitreous were approximately 17.98 and 87.03 ng/ml at 5 and 14 days after subconjunctival injection, respectively. the

Retinochoroids were homogenized and analyzed as described in Example 2, and samples were taken as described above for the vitreous. The retinochoroid does not include the site of administration, therefore, this measurement indicates the level of rapamycin delivered into the retinochoroid. Mean levels of rapamycin in the retina and choroid were approximately 0.27 and 0.21 ng/mg 5 and 14 days after subconjunctival injection, respectively. the

Example 28 - Subconjunctival injection of a solution containing rapamycin

60 μl of the solution described in Example 25 was injected as two 30 μl doses into two sites between the sclera and the conjunctiva of New Zealand white rabbit eyes. Figure 16 depicts the levels of rapamycin present in the vitreous on a logarithmic scale at 5 and 14 days post-injection. Figure 17 depicts rapamycin levels on a logarithmic scale in the retina and choroid at the same time points. the

Vitreous bodies were homogenized and analyzed as described in Example 2. Four rabbit eyes were analyzed per time point. Vitreous samples did not include the site of administration, therefore, this measurement indicates the level of rapamycin delivered into the vitreous. The mean levels of rapamycin in the vitreous were approximately 502.2 and 31.80 ng/ml at 5 and 14 days after subconjunctival injection, respectively. the

Retinochoroids were homogenized and analyzed as described in Example 2, and samples were taken as described above for the vitreous. The retinochoroid does not include the site of administration, therefore, this measurement indicates the level of rapamycin delivered into the retinochoroid. Mean levels of rapamycin in the retina and choroid were approximately 0.80 and 0.15 ng/mg at 5 and 14 days after subconjunctival injection, respectively. the

Example 29 - Subconjunctival injection of a solution containing rapamycin

90 [mu]l of the solution described in Example 25 was injected as three 30 [mu]l doses at three sites between the sclera and the conjunctiva of New Zealand white rabbit eyes. Figure 16 depicts the levels of rapamycin present in the vitreous on a logarithmic scale at 5 and 14 days post-injection. Figure 17 depicts rapamycin levels on a logarithmic scale in the retina and choroid at the same time points. the

Vitreous bodies were homogenized and analyzed as described in Example 2. Four rabbit eyes were analyzed per time point. Vitreous samples did not include the site of administration, therefore, this measurement indicates the level of rapamycin delivered into the vitreous. Mean levels of rapamycin in the vitreous were approximately 39.05 and 13.63 ng/ml at 5 and 14 days after subconjunctival injection, respectively. the

Retinochoroids were homogenized and analyzed as described in Example 2, and samples were taken as described above for the vitreous. The retinochoroid does not include the site of administration, therefore, this measurement indicates the level of rapamycin delivered into the retinochoroid. Mean levels of rapamycin in the retina and choroid were approximately 0.83 and 0.10 ng/mg at 5 and 14 days after subconjunctival injection, respectively. the

Example 30 - Preparation and Characterization of Rapamycin-Containing Suspensions

A rapamycin-containing suspension was prepared by dissolving 1201.6 mg rapamycin (3.000% by weight) in 6518.8 mg PEG400 (97.000% by weight) and vortexing. The size of the resulting particles was not quantified, but was large, estimated to be around 10 μm. This suspension is listed in Table 1 as Formulation #147. the

Example 31 - Subconjunctival injection of a suspension containing rapamycin

10 [mu]l of the suspension described in Example 30 was injected as a single dose between the sclera and conjunctiva of New Zealand white rabbit eyes. Figure 18 depicts the levels of rapamycin present in the vitreous on a logarithmic scale at 5, 14 and 30 days post-injection. Figure 19 depicts rapamycin levels on a logarithmic scale in the retina and choroid at the same time points. For comparison, Figures 18 and 19 also depict the results of other studies described in Example 32 and Example 33 below. the

Vitreous bodies were homogenized and analyzed as described in Example 2. Eyes of four rabbits were analyzed per time point. Vitreous samples did not include the site of administration, therefore, this measurement indicates the level of rapamycin delivered into the vitreous. Mean levels of intravitreal rapamycin were approximately 2.68, 0.90 and 5.43 ng/ml at 5, 14 and 30 days after subconjunctival injection, respectively. the

Retinochoroids were homogenized and analyzed as described in Example 2, and samples were taken as described above for the vitreous. Retinochoroid samples did not include the site of administration, therefore, this measurement indicates the level of rapamycin delivered into the retinochoroid. Mean levels of rapamycin in the retina and choroid were approximately 0.20, 0.06 and 1.23 ng/mg at 5, 14 and 30 days after subconjunctival injection, respectively. the

Example 32 - Subconjunctival injection of a suspension containing rapamycin

30 [mu]l of the solution described in Example 30 was injected as a single dose between the sclera and conjunctiva of New Zealand white rabbit eyes. Figure 18 depicts the levels of rapamycin present in the vitreous on a logarithmic scale at 5, 14 and 30 days post-injection. Figure 19 depicts rapamycin levels on a logarithmic scale in the retina and choroid at the same time points. the

Vitreous bodies were homogenized and analyzed as described in Example 2. Eyes of four rabbits were analyzed per time point. Vitreous samples did not include the site of administration, therefore, this measurement indicates the level of rapamycin delivered into the vitreous. Mean levels of intravitreal rapamycin were approximately 84.55, 11.23 and 66.35 ng/ml at 5, 14 and 30 days after subconjunctival injection, respectively. the

Retinochoroids were homogenized and analyzed as described in Example 2, and samples were taken as described above for the vitreous. Retinochoroid samples did not include the site of administration, therefore, this measurement indicates the level of rapamycin delivered into the retinochoroid. Mean levels of rapamycin in the retina and choroid were approximately 1.09, 0.19 and 1.02 ng/mg at 5, 14 and 30 days after subconjunctival injection, respectively. the

Example 33 - Subconjunctival injection of a suspension containing rapamycin

90 μl of the solution described in Example 30 was injected as three 30 μl doses into three sites between the sclera and conjunctiva of New Zealand white rabbit eyes. Figure 18 depicts the levels of rapamycin present in the vitreous on a logarithmic scale at 5, 14 and 30 days post-injection. Figure 19 depicts rapamycin levels on a logarithmic scale in the retina and choroid at the same time points. the

Vitreous bodies were homogenized and analyzed as described in Example 2. Eyes of four rabbits were analyzed per time point. Vitreous samples did not include the site of administration, therefore, this measurement indicates the level of rapamycin delivered into the vitreous. Mean levels of intravitreal rapamycin were approximately 29.95, 15.30 and 49.20 ng/ml at 5, 14 and 30 days after subconjunctival injection, respectively. the

Retinochoroids were homogenized and analyzed as described in Example 2, and samples were taken as described above for the vitreous. Retinochoroid samples did not include the site of administration, therefore, this measurement indicates the level of rapamycin delivered into the retinochoroid. Mean levels of rapamycin in the retina and choroid were approximately 0.55, 1.31 and 5.74 ng/mg at 5, 14 and 30 days after subconjunctival injection, respectively. the

Example 34 Preparation and Characterization of Rapamycin-Containing Solutions.

10.3 mg rapamycin was taken in ethanol, 4995.8 mg PEG400 was added, and the mixture was vortexed to obtain a solution with a final concentration by weight of 0.205% rapamycin, 0.544% ethanol and 99.251% PEG 400. This solution is listed in Table 1 as Formulation #140. A volume of 10 μl of this solution formed non-dispersed masses when placed in the vitreous of a rabbit eye. the

Example 35 - Intravitreal injection of a solution containing rapamycin

10 µl of the solution described in Example 34 was injected into the vitreous of New Zealand White rabbit eyes. The injected solution forms a non-dispersed mass relative to the surrounding medium. Figure 20 depicts rapamycin levels on a logarithmic scale in the retina choroid at 5 and 30 days post-injection. Figure 21 depicts rapamycin levels in the vitreous on a logarithmic scale at the same time points. For comparison, Figures 20 and 21 also depict the results of other studies described in Example 37 and Example 39 below. the

Vitreous bodies were homogenized and analyzed as described in Example 2. Five rabbit eyes were analyzed per time point. A vitreous sample can include an application site. The mean levels of intravitreal rapamycin were approximately 12.02 and 0.92 ng/ml at 5 and 30 days after intravitreal injection, respectively. the

Retinochoroids were homogenized and analyzed as described in Example 2, and samples were taken as described above for the vitreous. The retinochoroid does not include the site of administration, therefore, this measurement indicates the level of rapamycin delivered into the retinochoroid. The mean levels of rapamycin in the retina and choroid were approximately 0.08 and 0.02 ng/mg at 5 and 30 days after intravitreal injection, respectively. the

Example 36 Preparation and Characterization of Rapamycin-Containing Solutions

31.5 mg rapamycin was placed in ethanol, 4918.9 mg PEG400 was added, and the solution was vortexed. The final concentration by weight was 0.628% rapamycin, 1.337% ethanol and 98.035% PEG400. The formulation was stored at 4°C until use. This solution is listed in Table 1 as Formulation #142. A volume of 10 μl of this solution formed non-dispersed masses when placed in the vitreous of a rabbit eye. the

Example 37 - Intravitreal injection of a solution containing rapamycin

10 µl of the solution described in Example 36 was injected into the vitreous of New Zealand White rabbit eyes. The injected solution forms a non-dispersed mass relative to the surrounding medium. Figure 20 depicts rapamycin levels on a logarithmic scale in the retina choroid at 5 and 30 days post-injection. Figure 21 depicts rapamycin levels in the vitreous on a logarithmic scale at the same time points. the

Vitreous bodies were homogenized and analyzed as described in Example 2. Five rabbit eyes were analyzed per time point. A vitreous sample can include an application site. The mean levels of intravitreal rapamycin were about 87.46 and 44.34 ng/ml at 5 and 30 days after intravitreal injection, respectively. the

Retinochoroids were homogenized and analyzed as described in Example 2, and samples were taken as described above for the vitreous. The retinochoroid does not include the site of administration, therefore, this measurement indicates the level of rapamycin delivered into the retinochoroid. The mean levels of rapamycin in the retina and choroid were approximately 1.40 and 0.01 ng/mg at 5 and 30 days after intravitreal injection, respectively. the

Example 38 Preparation and Characterization of Rapamycin-Containing Solutions

103.5 mg rapamycin was taken in ethanol, 4720.8 mg PEG 400 was added, and the mixture was vortexed to obtain a solution with a final concentration by weight of 2.057% rapamycin, 4.116% ethanol and 93.827% PEG 400. This solution is listed in Table 1 as Formulation #144. A volume of 10 [mu]l of this solution formed non-dispersed masses in the vitreous of rabbit eyes. the

Example 39 - Intravitreal injection of a solution containing rapamycin

10 µl of the solution described in Example 38 was injected into the vitreous of New Zealand White rabbit eyes. The injected solution forms a non-dispersed mass relative to the surrounding medium. Figure 20 depicts rapamycin levels on a logarithmic scale in the retina and choroid at 5, 30 and 90 days post-injection. Figure 21 depicts rapamycin levels in the vitreous on a logarithmic scale at the same time points. the

Vitreous bodies were homogenized and analyzed as described in Example 2. Four rabbit eyes were analyzed per time point. A vitreous sample can include an application site. Mean levels of intravitreal rapamycin were approximately 120,500; 55,160 and 0.55 ng/ml at 5, 30 and 90 days after intravitreal injection, respectively. the

The retina and choroid were homogenized and analyzed as described in Example 2, and samples were taken as described above for the vitreous, except that the five rabbit eyes were analyzed at the 5-day and 30-day time points. Retinochoroid samples did not include the site of administration, therefore, this measurement indicates the level of rapamycin delivered into the retinochoroid. Mean levels of rapamycin in the retina and choroid were approximately 4.75, 0.17 and 0.01 ng/mg at 5, 30 and 90 days after intravitreal injection, respectively. the

Example 40 - Subconjunctival injection of a solution containing rapamycin

40 µl of the solution described in Example 17 was injected between the sclera and conjunctiva of New Zealand white rabbit eyes. Figure 22 depicts rapamycin levels (ng/ml) on a logarithmic scale in aqueous humor at 1, 4, 7, 11, 14, 21, 28, 35, 54, and 56 days post-injection, and 4, 14, 54, and 56 days post-injection. Rapamycin levels in cornea (ng/mg) and retinachoroid (ng/mg) at 21 and 35 days. The retinal choroidal level is labeled "R/choroidal" in FIG. 22 . the

The aqueous humor was homogenized and then analyzed by liquid chromatography and mass spectrometry. Four rabbit eyes were analyzed at each time point. The aqueous humor does not include the injection site, so this measurement indicates the level of rapamycin delivered into the aqueous humor. The mean levels of rapamycin in the aqueous humor were about 0.875, 1.0, 7.0, 0.725, 0.5, 0.525, 0.0, 0.125, 0.014 at 1, 4, 7, 11, 14, 21, 28, 35, 54, and 56 days after injection, respectively and 0.0485ng/ml. the

Corneal homogenates were then analyzed by liquid chromatography and mass spectrometry. The cornea does not include the injection site, so this measurement indicates the level of rapamycin delivered into the cornea. Four rabbit eyes were analyzed at each time point. Mean levels of rapamycin in the cornea were approximately 0.3225, 0.1, 0.0275 and 0.0125 ng/mg at 4, 14, 21 and 35 days post-injection, respectively. the

Retinochoroids were homogenized and analyzed as described in Example 2, and samples were taken as described above for the vitreous. The retinochoroid does not include the site of administration, therefore, this measurement indicates the level of rapamycin delivered into the retinochoroid. Mean levels of rapamycin in the retina and choroid were approximately 11.61, 0.2, 0.0275 and 2.655 ng/mg at 4, 14, 21 and 35 days post-injection, respectively. the

Example 41 - Intravitreal injection of a solution containing rapamycin

1.0 µl of the solution described in Example 17 was injected into the vitreous of New Zealand White rabbit eyes. The injected solution forms a non-dispersed mass relative to the surrounding medium. Table 2 reports the mean levels of rapamycin in the aqueous humor one day after injection. For comparison, Table 2 also reports the results of the studies described in Examples 42-45 below. the

Aqueous humor was homogenized and analyzed as described in Example 40. Analyze two rabbit eyes. The aqueous humor does not include the injection site, so this measurement indicates the level of rapamycin delivered into the aqueous humor. The mean level of rapamycin in the aqueous humor was about 0.438 ng/ml one day after the injection, with a standard deviation of about 0.141 ng/ml. the

Example 42 - Intravitreal injection of a solution containing rapamycin

3.0 µl of the solution described in Example 17 was injected into the vitreous of New Zealand White rabbit eyes. The injected solution forms a non-dispersed mass relative to the surrounding medium. Table 2 reports the mean levels of rapamycin in the aqueous humor one day after injection. the

Aqueous humor was homogenized and analyzed as described in Example 40. Analyze two rabbit eyes. The aqueous humor does not include the injection site, so this measurement indicates the level of rapamycin delivered into the aqueous humor. The mean level of rapamycin in the aqueous humor was about 0.355 ng/ml one day after the injection, with a standard deviation of about 0.234 mg/ml. the

Example 43 - Subconjunctival injection of a solution containing rapamycin

3.0 µl of the solution described in Example 17 was injected between the sclera and conjunctiva of New Zealand white rabbit eyes. The injected solution forms a non-dispersed mass relative to the surrounding medium. Table 2 reports the mean levels of rapamycin in the aqueous humor one day after injection. the

Aqueous humor was homogenized and analyzed as described in Example 40. Analyze two rabbit eyes. The aqueous humor does not include the injection site, so this measurement indicates the level of rapamycin delivered into the aqueous humor. The mean level of rapamycin in the aqueous humor was about 0.338 ng/ml one day after the injection, with a standard deviation of about 0.122 ng/ml. the

Example 44 - Anterior chamber administration of a solution containing rapamycin

5.0 µl of the solution described in Example 17 was injected into the anterior chamber of New Zealand white rabbits by injection into the anterior end of the eye. Aqueous humor is withdrawn using a syringe. Table 2 reports the mean levels of rapamycin in the aqueous humor 14 days after injection. the

Aqueous humor was homogenized and analyzed as described in Example 40. Analyze two rabbit eyes. The aqueous humor does not include the injection site, so this measurement indicates the level of rapamycin delivered into the aqueous humor. The mean level of rapamycin in the aqueous humor 14 days after injection was about 0.166 ng/ml, with a standard deviation of about 0.183 ng/ml. the

Example 45 - Anterior chamber administration of a solution containing rapamycin

10 µl of the solution described in Example 17 was injected into the anterior chamber of New Zealand white rabbits. Table 2 reports the mean levels of rapamycin in the aqueous humor 14 days after injection. the

Aqueous humor was homogenized and analyzed as described in Example 40. Analyze two rabbit eyes. The aqueous humor does not include the injection site, so this measurement indicates the level of rapamycin delivered into the aqueous humor. The mean level of rapamycin in the aqueous humor 14 days after injection was about 0.004 ng/ml, with a standard deviation of about 0.006 ng/ml. the

All references (including patents, patent applications, and publications) cited herein are hereby incorporated by reference in their entirety, whether or not specifically previously cited. the

Table 1 Liquid Formulations

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Table 2 Concentration of rapamycin in aqueous humor

Claims (1)

1. liquid preparation, it comprises by weight 2% rapamycin altogether, 4% ethanol and 94% PEG400, forms nondispersive agglomerate when wherein this liquid preparation is in the vitreous body that is expelled to experimenter's eye.