JP2006306877A - Dosage form, pharmaceutical composition and subtenon delivery method - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a pharmaceutical composition for sub Tenon administration to a postocular part close to macula lutea in mammal having age-related macular degeneration. <P>SOLUTION: The pharmaceutical composition comprises an dosage form of a compound represented by formula I, II or III or its pharmaceutically acceptable salt or solvate. The method for treating age-related macular degeneration comprises administering the dosage form. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to drug delivery, and in particular to pharmaceutical compositions, dosage forms, and methods of delivering pharmaceutically active substances to the back of the eye. More particularly, the present invention relates to dosage forms of pharmaceutical compositions and methods of subtenon delivery to the posterior segment of the eye adjacent to the macula of the pharmaceutically active substance.

  In recent years, great progress has been made in optimizing drug delivery to target tissues in the eye and maintaining effective drug doses in those tissues. However, many pharmacological managements of eye diseases continue to use topical application of solutions in the form of water drops to the ocular surface. It has been estimated that typically less than 5% of topically applied drug penetrates the cornea and reaches intraocular tissue. Despite the relatively small portion of locally administered drug dose eventually reaching the pre-segmentation of ocular tissue, many are still effective, thanks to the very high concentration of drug administered . However, delivery of therapeutic amounts of drugs to the tissue in the posterior segment remains a major challenge.

  Currently, treatment of the posterior segment of the disease is severely limited due to the difficulty of delivering an effective amount of the drug to the target tissue in the posterior segment of the eye. Four means can be used for delivery to the posterior segment of the drug: local, systemic, intraocular and periocular (eg subtenon, subconjunctival and postocular) delivery methods. Topically administered drugs can enter the eye by penetrating the conjunctiva and diffusing through the sclera, but this measure usually results in therapeutic drug concentrations in the posterior vitreous, retina, or choroid Absent. Systemic administration can deliver the drug to the back of the eye, but the large systemic doses required are often accompanied by serious side effects. Intravitreal injection provides the most direct means to deliver drugs to the tissue in the posterior segment and can achieve therapeutic tissue drug concentrations, but inherent in retinal detachment, bleeding, endophthalmitis and cataracts The potential for side effects remains and the repeated doses often required are not always well tolerated by patients. Furthermore, direct drug administration to the vitreous is rapidly excreted. Intravitreal sustained release devices have been used to avoid repeated doses. However, these devices require intraocular surgery, must be replaced regularly, and can have side effects similar to those associated with intravitreal injection.

  Drug delivery around the eye by subtenon, subconjunctival or post-ocular administration and placement of a sustained release device provides an alternative route for drug delivery to posterior tissue of the eye. This means of drug delivery is safer and less invasive than intravitreal injection, and also provides the possibility of local, sustained release drug delivery.

Drug delivery by this vehicle is ideally via the sclera, thereby taking advantage of the large surface area of the sclera. The average surface area of the sclera of 17-cm ² occupies 95% of the total surface area of the eyeball, and provides a significantly wider passage for drug diffusion to the inside of the eye compared to the 1-cm ² surface area of the cornea. provide. Also, the region-specific differences in scleral thickness can be used to further optimize drug diffusion through the sclera if a sustained release delivery device or system can be installed in the region with the highest scleral permeability. Will be able to. The sclera is, for example, 1.0 mm thick near the optic nerve, 0.53 mm thick on the corneal scleral annulus, and thinned to 0.39 mm on the eye equator, up to 0.1 mm in many eyes May also be thin. In addition, increasing evidence accumulation suggests that the sclera is very permeable to a wide range of solutes and has enormous potential for post-segment drug delivery. .

  However, due to the high sensitivity of the eye, drug delivery to the posterior eye requires a non-irritating vehicle for the posterior tissue of the eye. Therefore, there is a need in the field of ophthalmology for safe and effective improved pharmaceutical compositions and methods for their use for delivering pharmaceutically active substances to the posterior segment of the eye.

  The present invention relates to indazole compounds, pharmaceutical compositions containing such compounds, and cell growth using such compounds, including angiogenesis and / or angiogenic eye diseases such as age-related macular degeneration. It relates to methods of using such compounds that mediate and / or inhibit specific protein kinase activity for the treatment of disease states. More particularly, the invention relates to dosage forms for the subtenon delivery of such compounds and pharmaceutical compositions to the posterior segment of the eye adjacent to the macula.

  Indazole compounds useful in the compositions, dosage forms, and methods described herein are described in US Pat. Nos. 6,531,491; 6,534,524; and 6,869,962; US Patent Application Publication Nos. 10 / 737,655 and 10 And US Provisional Application No. 60 / 620,394, the disclosures of which are hereby incorporated by reference in their entirety for all purposes.

を有する化合物、およびそれらの製薬的に受容可能な塩または溶媒和物を開示している。 In particular, these patents and applications are of formula I, II, or III:

And pharmaceutically acceptable salts or solvates thereof are disclosed.

  The compounds of formula I, II and III are tyrosine kinase inhibitors intended for the treatment of age-related macular degeneration. Typically, these compounds are formulated as sterile suspension compositions and administered to the back of the retro-ocular segment adjacent to the macula via subtenon administration. Currently, formulations of these compounds are in development for Phase III trials and subsequent commercialization.

  In order to explain the circumstances of the present invention, a discussion of the background of the present invention is included in this application. This was all that was referred to was published, known or part of common sense on the priority date for all claims in the United States or in any other country It should not be understood as an admission.

  Dosage form of a compound of formula I, II or III, or a pharmaceutically acceptable salt or solvate thereof, and in the mammal having an eye disease such as age-related macular degeneration, It is an object of the present invention to provide a pharmaceutical composition thereof for subtenon administration to a section. It is another object of the present invention to provide a method for the treatment of mammalian ocular diseases that administers these dosage forms.

  A therapeutically effective amount of a compound of formula I, II or III can be administered to a mammal via subtenon administration, intraocular, systemic or topical administration. A therapeutically effective amount of a compound of formula I, II or III can be administered to one or both eyes simultaneously or at different intervals.

  Accordingly, in another aspect, the invention includes a compound of formula I, II, or III, or a pharmaceutically acceptable salt or solvate thereof, wherein the compound of formula I, II, or III is from about 1 μg to about 1,500. A dosage form is provided that is μg.

  In another aspect, the present invention provides a dosage form, wherein the dosage form further comprises one or more pharmaceutically acceptable excipients.

In another aspect, the present invention provides a dosage form wherein the compound of formula I, II or III is from about 50 μg to about 1,000 μg.
In another aspect, the invention provides a dosage form wherein the compound of formula I, II or III is from about 100 μg to about 600 μg.
In another aspect, the invention provides a dosage form wherein the compound of formula I, II or III is about 1 μg.
In another aspect, the invention provides a dosage form wherein the compound of formula I, II or III is about 50 μg.
In another aspect, the invention provides a dosage form wherein the compound of formula I, II or III is about 100 μg.
In another aspect, the invention provides a dosage form wherein the compound of formula I, II, or III is about 150 μg.
In another aspect, the invention provides a dosage form wherein the compound of formula I, II, or III is about 300 μg.
In another aspect, the invention provides a dosage form wherein the compound of formula I, II, or III is about 600 μg.
In another aspect, the invention provides a dosage form wherein the compound of formula I, II, or III is about 1,000 μg.
In another aspect, the present invention provides a dosage form wherein the compound of formula I, II or III is about 1,500 μg.

In another aspect, the invention provides for the treatment of a mammalian eye disease comprising administering a therapeutically effective amount of a compound of formula I, II and III, or a pharmaceutically acceptable salt or solvate thereof. A method is provided wherein the therapeutically effective amount is from about 1 μg to about 1,500 μg.
In another aspect, the present invention provides a method for treating a mammalian eye disease, wherein the composition further comprises one or more pharmaceutically acceptable excipients.
In another aspect, the present invention provides a method of treating a mammalian eye disease, wherein the composition is administered by subtenon delivery to the eye.
In another aspect, the invention provides a method for treating a mammalian ocular disease, wherein the ocular disease is age-related macular degeneration (AMD) such as non-exudative (dry AMD) and exudative (wet AMD). ), Choroidal neovascularization, retinopathy such as diabetic retinopathy and retinopathy of prematurity, diabetic macular edema, retinitis, uveitis, cystic macular edema, glaucoma and other posterior segment diseases or symptoms Selected.

In another aspect, the invention provides a method for treating a mammalian eye disease, wherein the therapeutically effective amount is from about 50 μg to about 1,000 μg.
In another aspect, the invention provides a method for treating a mammalian eye disease, wherein the therapeutically effective amount is from about 100 μg to about 600 μg.
In another aspect, the present invention provides a method for treating a mammalian eye disease, wherein the therapeutically effective amount is about 1 μg.
In another aspect, the invention provides a method for treating a mammalian eye disease, wherein the therapeutically effective amount is about 50 μg.
In another aspect, the invention provides a method for treating a mammalian ocular disease, wherein the therapeutically effective amount is about 100 μg.
In another aspect, the invention provides a method for treating a mammalian eye disease, wherein the therapeutically effective amount is about 150 μg.
In another aspect, the invention provides a method for treating a mammalian eye disease, wherein the therapeutically effective amount is about 300 μg.
In another aspect, the invention provides a method for treating a mammalian eye disease, wherein the therapeutically effective amount is about 600 μg.
In another aspect, the invention provides a method for treating a mammalian eye disease, wherein the therapeutically effective amount is about 1,000 μg.
In another aspect, the invention provides a method for treating a mammalian ocular disease, wherein the therapeutically effective amount is about 1,500 μg.

In another aspect, the invention provides a method of treating a mammalian eye disease, wherein the therapeutically effective amount is from about 50 μg / eye to about 600 μg / eye.
In another aspect, the invention provides a method of treating a mammalian eye disease, wherein the therapeutically effective amount is about 150 μg / eye.

In another aspect, the present invention provides a method for treating a mammalian eye disease, wherein the composition is administered twelve times a year.
In another aspect, the present invention provides a method of treating a mammalian eye disease, wherein the composition is administered four times a year.
In another aspect, the present invention provides a method of treating a mammalian eye disease, wherein the composition is administered three times a year.
In another aspect, the present invention provides a method of treating a mammalian eye disease, wherein the composition is administered twice a year.
In another aspect, the present invention provides a method for treating a mammalian eye disease, wherein the composition is administered annually.

In another aspect, the present invention provides a method for stabilizing vision in a mammal comprising administering a dosage form of the present invention.
In another aspect, the present invention provides a method for improving vision of a mammal comprising administering a dosage form of the present invention.

In another aspect, the present invention provides a method for preventing ocular diseases comprising administering a dosage form of the present invention.
In another aspect, the present invention provides a method for preventing ocular disease, wherein the ocular disease is ocular neovascularization.

In another aspect, the present invention provides a method for the treatment of mammalian cancer comprising administering a therapeutically effective amount of a compound of formula I, II or III, or a pharmaceutically acceptable salt or solvate thereof. Wherein the therapeutically effective amount is from about 1 μg to about 1,500 μg.
In another aspect, the present invention provides a method for treating mammalian cancer, wherein the composition further comprises one or more pharmaceutically acceptable excipients.

It is another object of the present invention to provide compositions and methods for subtenon delivery to the posterior segment of the eye adjacent to the macular of the pharmaceutically active substance.
Accordingly, in one aspect, the invention provides a pharmaceutically active substance having Formula I, II or III, or a pharmaceutically acceptable salt or solvate thereof; one or more excipients; and 1 or more There is provided a pharmaceutical composition comprising the above buffer, wherein the tonicity and pH of the composition is adjusted to physiological conditions; and the composition is adapted for subtenon delivery to the posterior segment of the eye adjacent to the macula. Is for.

In another aspect, the present invention provides a pharmaceutical composition, wherein one or more excipients are flocculants.
In another aspect, the present invention provides a pharmaceutical composition, wherein the flocculant is about 0.001% to about 10% benzalkonium chloride.
In another aspect, the present invention provides a pharmaceutical composition, wherein the flocculant is about 0.02% benzalkonium chloride and the tonicity of the composition is adjusted to physiological conditions with about 4.5% mannitol. .

In another aspect, the present invention provides a pharmaceutical composition, wherein the buffer is about 10 mM PBS buffer.
In another aspect, the present invention provides a pharmaceutical composition comprising about 0.01 mg / mL to about 2 mg / mL of a pharmaceutically active substance having formula I, II or III.
In other aspects, the invention comprises from about 0.1 mg / mL to about 1.2 mg / mL of a pharmaceutically active substance having formula I, II or III; about 0.02% benzalkonium chloride; about 10 mM PBS buffer; and A pharmaceutical composition is provided wherein the tonicity of the composition is adjusted to physiological conditions with about 4.5% mannitol.

In another aspect, the present invention provides a pharmaceutical composition, wherein the flocculant is about 0.001% to about 10% docusate sodium.
In another aspect, the present invention provides a pharmaceutical composition, wherein the flocculant comprises about 0.005% docusate sodium and the tonicity of the composition is adjusted to physiological conditions with about 0.9% sodium chloride. .
In another aspect, the present invention provides a pharmaceutical composition, wherein the buffer is about 10 mM PBS buffer.

In another aspect, the present invention provides a pharmaceutical composition comprising about 0.01 mg / mL to about 2 mg / mL of a pharmaceutically active substance having formula I, II or III.
In other aspects, the invention comprises from about 0.1 mg / mL to about 1.2 mg / mL of a pharmaceutically active substance having formula I, II or III; about 0.005% docusate sodium; about 10 mM PBS buffer; and A pharmaceutical composition is provided wherein the tonicity of the composition is adjusted to physiological conditions with about 0.9% sodium chloride.

In another aspect, the present invention provides a pharmaceutical composition, wherein the flocculant is about 0.001% to about 10% sodium lauryl sulfate.
In another aspect, the present invention provides a pharmaceutical composition, wherein the flocculant comprises about 0.01% sodium lauryl sulfate and the tonicity of the composition is about 4% mannitol and about 0.2% magnesium chloride at physiological conditions. Has been adjusted.
In another aspect, the present invention provides a pharmaceutical composition, wherein the buffer is about 10 mM PBS buffer.
In another aspect, the present invention provides a pharmaceutical composition comprising about 0.01 mg / mL to about 2 mg / mL of a pharmaceutically active substance having formula I, II or III.

  In other aspects, the invention comprises from about 0.1 mg / mL to about 1.2 mg / mL of a pharmaceutically active substance having formula I, II or III; about 0.01% sodium lauryl sulfate; about 10 mM PBS buffer; and A pharmaceutical composition is provided wherein the tonicity of the composition is adjusted to physiological conditions with about 4% mannitol and about 0.2% magnesium chloride.

In another aspect, the present invention provides a pharmaceutical composition, wherein the pharmaceutically active substance is an angiogenesis inhibitor.
In another aspect, the present invention provides a pharmaceutical composition, wherein the angiogenesis inhibitor is an inhibitor of a protein kinase receptor.
In another aspect, the present invention provides a pharmaceutical composition, wherein the protein kinase receptor is a VEGF receptor.

In other aspects, the invention provides a pharmaceutically active substance having Formula I, II or III, or a pharmaceutically acceptable salt or solvate thereof; one or more excipients; and one or more excipients Provided is a method for subtenon delivery to the posterior segment of the pharmaceutical composition adjacent to the macula comprising a buffer, wherein the tonicity and pH of the composition are adjusted to physiological conditions.
In another aspect, the present invention provides a method for subtenon delivery of a pharmaceutical composition, wherein one or more excipients is an aggregating agent.
In another aspect, the invention provides a method for subtenon delivery of a pharmaceutical composition, wherein the flocculant is about 0.001% to about 10% benzalkonium chloride.

In another aspect, the invention comprises about 0.02% benzalkonium chloride and the tonicity of the composition is adjusted to physiological conditions with about 4.5% mannitol for subtenon delivery of a pharmaceutical composition Provide a way.
In another aspect, the invention provides a method for subtenon delivery of a pharmaceutical composition, wherein the buffer is about 10 mM PBS buffer.
In another aspect, the present invention provides a method for subtenon delivery of a pharmaceutical composition comprising about 0.01 mg / mL to about 2 mg / mL of a pharmaceutically active substance having formula I, II or III.
In other aspects, the invention comprises from about 0.1 mg / mL to about 1.2 mg / mL of a pharmaceutically active substance having formula I, II or III; about 0.02% benzalkonium chloride; about 10 mM PBS buffer; and Provided is a method for subtenon delivery of a pharmaceutical composition, wherein the tonicity of the composition is adjusted to physiological conditions with about 4.5% mannitol.

In another aspect, the invention provides a method for subtenon delivery of a pharmaceutical composition, wherein the flocculant is about 0.001% to about 10% docusate sodium.
In another aspect, the invention provides a method for subtenon delivery of a pharmaceutical composition, wherein the flocculant comprises about 0.005% docusate sodium and the tonicity of the composition is about 0.9% sodium chloride. Is adjusted to physiological conditions.
In another aspect, the present invention provides a method for subtenon delivery of a pharmaceutical composition, wherein the buffer is about 10 mM PBS buffer.
In another aspect, the present invention provides a method for subtenon delivery of a pharmaceutical composition comprising about 0.01 mg / mL to about 2 mg / mL of a pharmaceutically active substance having formula I, II or III.

In other aspects, the invention comprises from about 0.1 mg / mL to about 1.2 mg / mL of a pharmaceutically active substance having formula I, II or III; about 0.005% docusate sodium; about 10 mM PBS buffer; and A method for subtenon delivery of a pharmaceutical composition is provided wherein the tonicity of the composition is adjusted to physiological conditions with about 0.9% sodium chloride.
In another aspect, the invention provides a method for subtenon delivery of a pharmaceutical composition, wherein the flocculant is about 0.001% to about 10% sodium lauryl sulfate.
In another aspect, the present invention provides a method for subtenon delivery of a pharmaceutical composition, wherein the flocculant comprises about 0.01% sodium lauryl sulfate and the tonicity of the composition is about 4% mannitol and It is adjusted to physiological conditions with about 0.2% magnesium chloride.
In another aspect, the present invention provides a method for subtenon delivery of a pharmaceutical composition, wherein the buffer is about 10 mM PBS buffer.
In another aspect, the present invention provides a method for subtenon delivery of a pharmaceutical composition comprising about 0.01 mg / mL to about 2 mg / mL of a pharmaceutically active substance having formula I, II or III.

In other aspects, the invention comprises from about 0.1 mg / mL to about 1.2 mg / mL of a pharmaceutically active substance having formula I, II or III; about 0.01% sodium lauryl sulfate; about 10 mM PBS buffer; and Provided is a method for subtenon delivery of a pharmaceutical composition wherein the tonicity of the composition is adjusted to physiological conditions with about 4% mannitol and about 0.2% magnesium chloride.
In another aspect, the present invention provides a method for subtenon delivery of a pharmaceutical composition, wherein the pharmaceutically active agent is an angiogenesis inhibitor.
In another aspect, the present invention provides a method for subtenon delivery of a pharmaceutical composition, wherein the angiogenesis inhibitor is an inhibitor of a protein kinase receptor.
In another aspect, the invention provides a method for subtenon delivery of a pharmaceutical composition, wherein the protein kinase receptor is a VEGF receptor.

Definitions As used in this application, the expressions “stabilization of vision” or “stabilization of vision” are used in early treatment diabetic retinopathy tests at 1 year or more unless otherwise indicated. This means that the loss of visual acuity from the baseline is less than 15 characters, or alternatively, the loss is less than 3 lines when measured using a visual acuity test chart (ETDRS chart). ETDRS charts are discussed by Ferris, FL et al. In the American Journal of Ophthalmology 94: 91-96, 1982, the disclosures of which are incorporated herein by reference in their entirety for all purposes.

  The expressions “improving visual acuity” or “improving visual acuity” as used in the present application indicate that the acquisition of visual acuity from baseline is 15 within a year, unless otherwise indicated, by using an ETDRS chart. Characters or more, or alternatively, means that the acquisition is 3 lines or more.

  As used herein, the expression “preventing ocular neovascularization” or “preventing ocular neovascularization” is non-exudative when measured by techniques well known in the art such as fluorescent angiography and / or colored fundus photography. It means to stop the progression from sexual age-related macular degeneration (dry AMD) to exudative age-related macular degeneration (wet AMD).

  As used herein, the phrase “treat”, unless indicated otherwise, restores, alleviates, and inhibits progression of the disease or condition in which such expression is used, or one or more such disease or condition. Mean to prevent or prevent. As used herein, the term “treatment” means the act of treating unless otherwise indicated (here “treat” is defined immediately above).

  As used herein, the expression “pharmaceutically acceptable salt” includes salts of acidic or basic groups that may be present in the compound unless otherwise indicated. Compounds that are basic in nature are capable of forming many types of salts with various inorganic and organic acids. The acids used to produce pharmaceutically acceptable acid addition salts of such base compounds are those that produce non-toxic acid addition salts, that is, salts that contain a pharmaceutically acceptable anion. For example, acetate, benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, bromide, calcium edetate, camsylate, carbonate, chloride, clavulan Acid salt, citrate salt, dihydrochloride salt, edetate salt, edislyate, estolate, esylate, ethyl succinate, fumarate, gluceptate, gluconate, glutamic acid Salt, glycolylarsanylate, hexyl resorcinate, hydrabamine, hydrobromide, hydrochloride, iodide, isothionate, lactate, Tobionate (lactobionate), laurate, malate, maleate, mandelate, mesylate, methyl sulfate, mucate, napsylate, nitrate, oleate, oxalate, pamoate (Embonate), palmitate, pantothenate, phosphate / diphosphate, polygalacturonate, salicylate, stearate, basic acetate, succinate, tannate, tartaric acid Salts, teoclate, tosylate, triethiodode and valerate.

The present invention is also identical to the compounds of formulas I, II and III, but one or more atoms are replaced by an atom having an atomic number or mass number that is different from the normally occurring atomic number or mass number. Isotope-labeled compounds. Examples of isotopes that can be incorporated into the compounds of the present invention include hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine and chlorine isotopes such as ² H, ³ H, ¹³ C, ¹⁴ C, respectively. , ¹⁵ N, ¹⁸ O, ¹⁷ O, ³¹ P, ³² P, ³⁵ S, ¹⁸ F and ³⁶ Cl. Compounds of the present invention, prodrugs thereof, and pharmaceutically acceptable salts of the compounds or prodrugs that contain the aforementioned isotopes and / or isotopes of other atoms are within the scope of the present invention. Certain isotopically-labelled compounds of the present invention, for example those into which radioactive isotopes such as ³ H and ¹⁴ C are incorporated, are useful in drug and / or substrate tissue distribution assays. Tritiated, ie, ³ H, and carbon-14, ie, ¹⁴ C, isotopes are particularly preferred for their ease of manufacture and detectability. In addition, heavier isotopes, such as deuterium, i.e. ² H, have certain therapeutic benefits that result from high metabolic stability, such as increased in vivo half-life or reduced dosage requirements. Can be provided and therefore preferred in certain circumstances. Isotopically labeled compounds of formula I, II or III of the present invention generally replace the methods described for unlabeled compounds with non-isotopically labeled reagents with readily available isotope-labeled reagents. And can be manufactured.

  Embodiments of the present invention and their advantages are best understood by referring to FIG. 1, which schematically illustrates a human eye 10. The eye 10 has a cornea 12, a lens 14, a sclera 16, a choroid 18, a retina 20, and an optic nerve 22. The anterior segment 24 of the eye 10 generally includes the anterior portion 22 of the line 25 of the eye 10, while the posterior segment 26 of the eye 10 generally includes the posterior portion 25 of the line 10 of the eye 10. The retina 20 is physically adhered to the choroid 18 in a circumferential manner adjacent to a pars plana 28. The retina 20 has a macula 30 that is located slightly to the side with respect to the optic nerve 22. The macular 30 is mainly composed of a retinal cone, and is an area showing the maximum visual acuity in the retina 20. A Tenon sac or tenon membrane 34 is located on the sclera 20. The conjunctiva 36 covers a narrow area of the sphere of the eye 10 behind the corneal annulus 32 (bulb conjunctiva) and is folded up (upper cecum) or down (lower sac), respectively, in the upper eyelid 35 And the inner region of the lower eyelid 37. The conjunctiva 36 is located at the top of the Tenon sac 34. The sclera 16 and the Tenon capsule 34 define the outer surface of the eyeball 10. Age-related macular degeneration (AMD), including non-exudative (dry AMD) and exudative (wet AMD), choroidal neovascularization, retinopathy, retinitis, uveitis, cystic macular edema (CME), glaucoma and For the treatment of other posterior segment 26 diseases or conditions, a specific amount of pharmaceutically active pharmaceutically active substance is delivered directly onto the outer surface of the sclera 16 and under the Tenon sac 34. (Depot) 38 is formed. In addition, for age-related macular degeneration (AMD), including non-exudative (dry AMD) and exudative (wet AMD), and CME, depot 38 is directly on the outer surface of sclera 16 under Tenon's capsule 34 It is inserted above and usually above the macular 30. It is well known to those skilled in the art to deliver substances periocularly to the posterior tissue of the eye. For example, US Pat. No. 6,413,245 describes devices useful for subtenon delivery of drugs, the disclosures of which are hereby incorporated by reference in their entirety for all purposes.

  The present invention provides dosage forms of compounds of Formulas I, II and III, pharmaceutical compositions thereof, and methods of subtenon delivery where the compounds and / or compositions are administered to the posterior segment of the eye adjacent to the macula . The compositions described herein further include various excipients and buffers that are safe and non-irritating to delicate ocular tissues.

  The pharmaceutically active substances useful in the compositions of the present invention are severely affected by age-related macular degeneration and other diseases affecting the posterior segment, such as choroidal neovascularization, diabetic retinopathy, glaucoma and retinitis pigmentosa. Useful for the treatment of vision loss.

  For example, the compositions of the present invention used to form drug depot 38 can include one or more pharmaceutically active agents in addition to one or more excipients described herein. Examples of pharmaceutically active substances useful in the compositions of the present invention include anti-infective agents such as, but not limited to, antibiotics, antiviral agents and antifungal agents; antiallergic agents and mast cell stabilizers; steroidal And non-steroidal anti-inflammatory agents (eg nepafenac); cyclooxygenase inhibitors such as, but not limited to, Cox I and Cox II inhibitors; combinations of anti-infective and anti-inflammatory agents; anti-inflammatory agents; anti-glaucoma agents such as , But not limited to, adrenergic agents, β-adrenergic blockers, α-adrenergic agents, parasypathomimetic agents, cholinesterase inhibitors, carbonic dehydrogenase inhibitors and prostaglandins; Combinations; antioxidants; nutritional supplements; therapeutic agents for cystic macular edema, including but not limited to Teloidal anti-inflammatory agents; therapeutic agents for age-related macular degeneration (AMD) including non-exudative (dry AMD) and exudative (wet AMD), such as, but not limited to, angiogenesis inhibitors such as VEGF receptor Angiogenesis inhibitors that inhibit protein kinase receptors, such as the body protein kinase receptor; and nutritional supplements; therapeutic agents for herpes zoster and CMV eye infections; therapeutic agents for proliferative vitreoretinopathy, such as Non-limiting antimetabolic and fibrinolytic agents; wound conditioning agents such as, but not limited to, growth factors; antimetabolic agents; neuroprotective agents such as, but not limited to, eliprodil; Diseases or symptoms such as, but not limited to, age-related macular degeneration (AMD), choroidal neovascularization, including non-exudative (dry AMD) and exudative (wet AMD) Retinopathy, retinitis, uveitis, angiostatic steroids for the treatment of cystoid macular edema and glaucoma, and the like. Such angiogenesis-inhibiting steroids are more fully disclosed in US Pat. Nos. 5,679,666 and 5,770,592. A non-steroidal anti-inflammatory agent for the treatment of cystoid macular edema is nepafenac.

  For administration to the eye, the compounds of the present invention may comprise a compound of which the compound is Delivered in a pharmaceutically acceptable ophthalmic vehicle such that the compound remains on the surface of the eye for a sufficient time to penetrate the rod, lens, choroid / retina and sclera. Pharmaceutically acceptable ophthalmic vehicles can be ointments, vegetable oils, or encapsulating materials. The compounds of the present invention may also be administered directly into the vitreous humor or aqueous humor.

  In addition, the compounds can be administered by well-known acceptable methods, such as subtenon and / or subconjunctival administration. As is well known in the ophthalmologic region, the macula is mainly composed of retinal cones, and is a region that exhibits maximum visual acuity on the retina. The Tenon capsule or Tenon membrane is located on the sclera. The conjunctiva covers a narrow area of the eyeball behind the corneal limbus (ball conjunctiva) and can be folded up (upper cecum) or folded down (lower sac), respectively, in the inner region of the upper and lower eyelids, respectively Covering. The conjunctiva is located at the top of the Tenon sac. The sclera and Tenon's capsule define the outer surface of the eyeball. Age-related macular degeneration (AMD), including non-exudative (dry AMD) and exudative (wet AMD), choroidal neovascularization, retinopathy (retinopathy such as diabetic retinopathy and retinopathy of prematurity), diabetes Specific amount of pharmaceutically active substance that is ophthalmologically acceptable for the treatment of macular edema, retinitis, uveitis, cystic macular edema (CME), glaucoma and other posterior segment diseases or conditions Is preferably inserted directly on the outer surface of the sclera and under the Tenon's capsule. In addition, in the case of age-related macular degeneration (AMD) and CME, including non-exudative (dry AMD) and exudative (wet AMD) and CME, the depot directly on the outer surface of the sclera under the Tenon capsule, and Usually, it is most preferable to be inserted above the macula.

  The compound can be formulated as a depot preparation. Such long acting formulations may be administered by implantation (for example subcutaneously or intramuscularly), intramuscular administration or by the above-mentioned Tenon's capsule or intravitreal injection. Alternatively, the active ingredient may be in powder form for constitution just before use with a suitable vehicle, eg, sterile pyrogen-free water.

  In a particularly preferred embodiment of the invention, the compound is prepared for topical administration in saline (in combination with any of the preservatives and antimicrobial agents commonly used in ophthalmic formulations) and in tear-like form Be administered. Solutions or suspensions are prepared in their pure form and are administered many times a day. Alternatively, the compositions of the present invention produced as described above may also be administered directly to the cornea.

  In a preferred embodiment, the composition can be made with a mucoadhesive polymer that binds to the cornea. Thus, for example, the compound can be formulated with a suitable polymerizable or hydrophobic material (eg, as an emulsion in an acceptable oil) or ion exchange resin, or as a poorly soluble derivative, eg, a poorly soluble salt.

  A pharmaceutical carrier for hydrophobic compounds is a cosolvent system comprising benzyl alcohol, a nonpolar surfactant, a water-miscible organic polymer, and an aqueous phase. This co-solvent system may be a VPD co-solvent system. VPD is a solution of 3% w / v benzyl alcohol, 8% w / v nonpolar surfactant polysorbate 80, and 65% w / v polyethylene glycol 300, and add absolute ethanol to adjust the volume. Yes. This VPD co-solvent system (VPD: 5W) contains VPD diluted 1: 1 with a 5% dextrose aqueous solution. This co-solvent system also dissolves hydrophobic compounds well and as such has low toxicity upon systemic administration. Of course, the ratio of the co-solvent system can be varied within a considerable range without destroying its solubility and toxicity properties. In addition, the identity of the co-solvent components may vary: for example, other low toxicity nonpolar surfactants can be used in place of polysorbate 80; the proportion of polyethylene glycol may vary in size; Of biocompatible polymers such as polyvinylpyrrolidone may be replaced with polyethylene glycol; and other sugars or polysaccharides may be replaced with dextrose.

  Alternatively, other delivery systems for hydrophobic pharmaceutical compounds may be used. Liposomes and emulsions are known examples of delivery vehicles or carriers for hydrophobic drugs. Certain organic solvents such as dimethyl sulfoxide can also be used, but usually have the disadvantage of increased toxicity. In addition, the compounds can be delivered using sustained release systems, such as semipermeable matrices of solid hydrophobic polymers containing therapeutic agents. Various sustained-release materials have been established and are known by those skilled in the art. Sustained release capsules can release the compound for a period of several weeks up to more than 100 days, depending on their chemical properties. Depending on the chemical properties and biological stability of the therapeutic agent, additional contrivances for protein stabilization will be used.

  The pharmaceutical compositions can also contain suitable solid or gel phase carriers or excipients. Examples of such carriers or excipients include calcium carbonate, calcium phosphate, saccharides, starch, cellulose derivatives, ceratin, and polymers such as polyethylene glycol.

  Some of the compounds of the invention are provided as salts with pharmaceutically compatible counterions. Pharmaceutically compatible salts can be formed with many acids, such as hydrochloric acid, sulfuric acid, acetic acid, lactic acid, tartaric acid, malic acid, succinic acid and the like. Salts that are more soluble in aqueous solutions or other protic solvents are the corresponding free base forms.

  The preparation of preferred compounds of the present invention is described in detail in the following examples, but one of ordinary skill in the art will understand that the described chemical reactions can be readily adapted to produce many other compounds of the present invention. Will. For example, the synthesis of compounds not described in the examples of the present invention can be accomplished by modification methods known to those skilled in the art, for example by substituting other suitable reagents known in the art by appropriate protective interfering groups. Or by routine modification of the reaction conditions. Instead, it will be appreciated that other reactions disclosed in the present application or known in the art have applicability for preparing other compounds of the present invention.

  Compounds of formula I, II and III are tyrosine kinase inhibitors intended for the treatment of age-related macular degeneration. Typically, these compounds are formulated as sterile suspension compositions and are administered by subtenon administration. Currently, suspension formulations of these compounds are under development for commercialization following Phase III trials. However, if these suspensions are filled in glass vials and stored in the reverse orientation, the compound particles will settle in the annulus between the glass and stopper, making it very difficult to resuspend. . This is due in part to the dense packing of micron-sized compound particles on their surfaces. This precipitation results in a decrease in the pharmacological activity of the product when resuspended by subsequent hand shaking. No decrease in pharmacological activity is observed when the vial is stored facing up. Another approach to promote particle resuspension from the annulus of the glass stopper is a loosely structured flocculent precipitate (agglomeration) that minimizes the secondary energy and maximizes the attraction between particles as described in the DLVO theory. To form particles).

  Formulation-based approaches that guide particles to minimize secondary energy balance either steric or electrostatic repulsion, or a combination of the two and van der Waals attraction. In general, the efficiency of flocculation increases with increasing particle concentration. Controlled flocculent aggregation by steric repulsion is achieved by adsorbing large molecules (aqueous or liquid) to the particle surface. The flocculant can be a neutral polymer such as methylcellulose or a surfactant such as polyvinyl alcohol. Such drugs also increase repulsion by increasing the molecular weight of the molecule. Conversely, flocculent aggregation controlled by electrostatic repulsion is achieved by the adsorption of ionic molecules to the particle surface. Such flocculants are cationic surfactants such as benzalkonium chloride, docusate sodium, sodium lauryl sulfate, and the like. Increasing the concentration of these ionic surfactants increases repulsion. If the surfactant adsorption on the particle surface is maximized, then further increase in the surfactant concentration has no effect on repulsion.

  A flocculent formulation is also more susceptible to resuspension because the flocculent flocculent is not packed as closely as the individual particles. For example, the formulations can be made administrable by shaking the vials by hand to break the flocculent aggregates. In these formulations, achieving the optimum conditions for the formation of loosely structured flocculents is facilitated by a clear mechanical understanding of flocculent flocculence and a comprehensive flocculent assessment method. Various techniques using various flocculants have been developed to create and optimize the flocculent agglomeration process. Many of the drugs described have been shown to be minimally toxic at the planned concentration when administered to rabbit eyes.

  In the compositions and methods of the present invention, the excipient can act as a flocculant or suspending agent; or as a surfactant or wetting agent. The expression “suspending agent” means a two-phase system consisting of finely divided solids that have been dispersed (suspended) in a liquid (diffusion medium). Suspensions have certain advantages over other dosage forms. Some pharmaceutically active substances are insoluble in all acceptable media and must therefore be administered as a suspension. Due to its liquid properties, suspension represents an ideal dosage form for subtenon administration. In addition, such drugs are chemically more stable in suspension than in solution. Over time, the suspended drug tends to settle, resulting in a loss of dose uniformity. However, this can be minimized by careful formulation and shaking of the suspension prior to administration.

  Aggregating agents are useful, for example, in suspension compositions, especially when suspended drug particles are desired. The choice and combination of excipients can provide suspension compositions that exhibit improved performance with respect to drug concentration, physical stability and efficiency. The physical stability of the suspension is controlled by the flocculant. The flocculant is an electrolyte that bears a charge opposite to the net zeta potential of the suspended particles. The addition of a critical concentration of flocculant counteracts the surface charge on the suspended particles and allows the formation of flocculent agglomerations or particles that are held together loosely by weak van der Waals attraction. Since the particles are loosely bound together, they do not become cake and can be easily re-diffused by shaking the suspension. In general, flocculent aggregates have particles of approximately the same size, and therefore a clear boundary is seen when the particles are allowed to settle. Non-limiting examples of suitable flocculants include benzalkonium chloride, sodium docusate, sodium lauryl sulfate, cetylpyridinium chloride, polysorbate 80, sorbitan monolaurate, sodium carboxymethylcellulose, xanthan gum, taragacanth, methylcellulose, PEG , Magnesium aluminum silicate, attapulgite, bentonite, potassium dihydrogen phosphate, aluminum chloride, sodium chloride and mixtures thereof.

（式中、Ｒはアルキルの混合で、全てまたはいくつかの基はn-C₈H₁₇から始まっており、より高級の同族体を通して、主要部分を含むn-C₁₂H₂₅、n-C₁₄H₂₉およびn-C₁₆H₃₃と共に伸長している）を有する塩化アルキルベンジルジメチルアンモニウムの混合物である。塩化ベンザルコニウムの平均分子量は360ｇ/moleであり、そして抗微生物性保存剤、防腐剤、殺菌剤、可溶化剤、および湿潤剤として作用することが知られている。塩化ベンザルコニウムは白色または黄白色の非晶質の粉末、粘度の高いゲル、またはゲル状フレークとして存在する。これは吸湿性であり、触ると石鹸状で、そしてかすかな芳香性香と強い苦味を有している。眼科用製剤では、塩化ベンザルコニウムは0.01〜0.02％ w/vの濃度で最も広く使用されている保存剤の１つである。これは、しばしば、シュードモナス株に対するその抗微生物作用を増大させる、他の保存剤または賦形剤と組み合わせて使用される。 Benzalkonium chloride has the general formula:

(Wherein R is a mixture of alkyls, all or some of the groups start with nC ₈ H ₁₇ , and through higher homologues, nC ₁₂ H ₂₅ , nC ₁₄ H ₂₉ and nC ₁₆ containing the main part. A mixture of alkylbenzyldimethylammonium chloride with H ₃₃ ). Benzalkonium chloride has an average molecular weight of 360 g / mole and is known to act as an antimicrobial preservative, preservative, disinfectant, solubilizer, and wetting agent. Benzalkonium chloride exists as a white or yellowish white amorphous powder, a highly viscous gel, or gel-like flakes. It is hygroscopic, soapy to the touch and has a faint fragrance and a strong bitter taste. In ophthalmic formulations, benzalkonium chloride is one of the most widely used preservatives at concentrations of 0.01-0.02% w / v. It is often used in combination with other preservatives or excipients that increase its antimicrobial activity against Pseudomonas strains.

を有している。 Docusate sodium or sodium 1,4-bis (2-ethylhexyl) sulfosuccinate has the empirical formula C ₂₀ H ₃₇ NaO ₇ S and the structural formula:

have.

  Docusate sodium is a white or almost white, waxy, bitter, plastic solid with a characteristic octanol-like scent. It is hygroscopic and is usually available in the form of pellets, flakes or flake roll materials. Docusate sodium and docusate salts are widely used as anionic surfactants in pharmaceutical formulations. Docusate sodium is used primarily as a capsule and direct compression tablet formulation to aid wetting and dissolution. Docusate salts are also used in oral formulations as laxatives and laxatives.

を有するアルキル硫酸ナトリウムの混合物である。 Sodium lauryl sulfate is mainly composed of C ₁₂ H ₂₅ NaO ₄ S and has the structural formula:

A mixture of sodium alkyl sulfate with

  Sodium lauryl sulfate consists of white or cream to pale yellow crystals, flakes or powder, has a smooth feel, is soapy, bitter and has a faint aroma of fatty substances. Sodium lauryl sulfate is an anionic surfactant that is widely used in oral pharmaceutical formulations and cosmetics. It is also a detergent; emulsifier; skin penetrant; tablet and capsule lubricant; and humectant effective in alkaline and acidic conditions.

Surfactants or wetting agents are either hydrophilic or hydrophobic and have the effect of reducing the surface tension of the compositions of the invention containing them. As is well known in the art, the expressions “hydrophilic” and “hydrophobic” are relative expressions. In order to function as a surfactant, the compound must necessarily contain a hydrophobic (lipophilic) moiety (ie, the surfactant compound must be amphiphilic) as well as a polar or charged hydrophilic moiety. I must. An experimental parameter commonly used to characterize the relative hydrophilicity and hydrophobicity of nonionic amphiphilic compounds is the hydrophilic-lipophilic balance (“HLB” value). Surfactants with lower HLB values are more hydrophobic and have greater solubility in oil, while surfactants with higher HLB values are more hydrophilic and in aqueous media Has greater solubility. By using the HLB value as a simple guideline, hydrophilic surfactants generally have an HLB value greater than about 10, and anionic, cationic, or zwitterionic compounds to which the HLB value is not normally applicable It is considered to be. Similarly, a hydrophobic surfactant is a compound having an HLB value of less than about 10.

Surfactants that can be used in the compositions of the present invention include, but are not limited to, polyoxyethylene sorbitan esters (Tween ^™ ) and block copolymers of ethylene oxide and propylene oxide (Poloxamer ^™ ). A wide variety of polyethylene sorbitan esters are commercially available and are suitable for use as surfactants in the compositions of the present invention. In general, these esters are hydrophilic and act as surfactants, although some hydrophobic moieties of this class can also be used. Polyethylene sorbitan esters useful in the compositions of the present invention include polyethylene sorbitan monolaurate (Tween ^™ 20), polyethylene sorbitan monopalmitate (Tween ^™ 40), polyethylene sorbitan monostearate (Tween ^™ 60), and polyethylene sorbitan. And monooleate (Tween ^TM 80).

Poloxamer ^™ is a block copolymer of ethylene oxide and propylene oxide and has the structure: HO (C ₂ H ₄ O) _x (C ₃ H ₆ O) _y (C ₂ H ₄ O) _x H (Pluronic ^™ ); or HO ( C ₃ H ₆ O) _x (C ₂ H ₄ O) _y (C ₃ H ₆ O) _x H (Pluronic ^™ R), where x is about 2 to about 150 and y is about 15 to Or a block copolymer as described above wherein x is about 80 and y is about 27. Block copolymers of ethylene oxide and propylene oxide that meet the above description are available from BASF under the trademark “Pluronic and Lutrol F Block Copolymers”. (For detailed specifications of such polymers, see the BASF pluronic polyol technical data sheet, the disclosure of which is incorporated herein by reference).

Poloxamer ^™ useful in the compositions of the present invention includes block copolymers that are water soluble, in cream or ointment form, can be stored for long periods under anhydrous conditions, and act as surfactants and / or flocculants To do. The poloxamer ^TM described in this application has a hydrophilic-lipophilic balance (HLB) value in the range of 8 to 30 and a molecular weight in the range of 1,000 to 16,000 g / mole. Poloxamer ^™ 188 (Pluronic ^™ F68) is a polyethylene having the structural formula: HO (C ₂ H ₄ O) _x (C ₃ H ₆ O) _y (C ₂ H ₄ O) _x H and an average molecular weight of 8,400 g / mole -Polypropylene glycol copolymer. With Poloxamer ^™ encoding a BASF label, suitable Poloxamers ^™ for use in the compositions of the present invention include, but are not limited to, Pluronic ^™ / Lutrol ^™ F44 (Poloxamer ^™ 124), Pluronic ^™ / Lutrol ^TM
F68 (Poloxamer ^™ 188), Pluronic ^™ / Lutrol ^™ F87 (Poloxamer ^™ 237), Pluronic ^™ / Lutrol ^™ F108 (Poloxamer ^™ 388), and Pluronic ^™ / Lutrol ^™ F127 (Poloxamer ^™ 407).

Excipients can act as suspending agents in the compositions of the present invention. Suspensions are pharmaceutically inert substances that provide increased stability of the suspension by increasing the viscosity of the composition of the present invention. Suspensions useful in the compositions of the present invention include, but are not limited to, polyethylene glycols (PEG) such as PEG 3350 and monomethoxypolyethylene glycol-distearoylphosphatidyl-ethanolamine (m-PEG-DSPE). PEG-phosphatidylethanolamine derivatives such as; high, medium, and low viscosity (molecular weight) carboxymethylcellulose (CMC), for example, cellulose derivatives such as type, 7H4, 7H3S, 7HOF, 7H, and 9H4; 7M, 7M8S, 7M2, 9M31, 9M8, 12M31, and 12M8; 7L and 7L2, available as Aqualon ^™ from Hercules Inc., and methylcellulose (MC) derivatives.

  PEGs of various molecular weights can be purchased from many different sources, or alternatively they can be synthesized using standard polymerization techniques well known to those skilled in the art and Useful as a surfactant or wetting agent in the composition. PEG is a polyethylene glycol polymer containing about 20 to about 2,000,000 attached monomers. For example, PEG containing various numbers of linked monomers include PEG 20, PEG 30, PEG 40, PEG 60, PEG 80, PEG 100, PEG 115, PEG 200, PEG 300, PEG 400, PEG 500, PEG 600 PEG 1000, PEG 1500, PEG 2000, PEG 3350, PEG 4000, PEG 4600, PEG 5000, PEG 6000, PEG 8000, PEG 11000, PEG 12000, PEG 2,000,000 and all mixtures thereof.

Phosphatidylethanolamines with various acyl chain groups of various chain lengths and saturations are conjugated with polyethylene glycol derivatives to form PEG-lipid conjugates useful as surfactants or wetting agents in the compositions of the present invention. can do. Such phosphatidylethanolamines are commercially available or are isolated or synthesized using methods known to those skilled in the art. Phosphatidylethanolamine preferably contain saturated or unsaturated fatty acids from C ₁₀ has a carbon chain length of C _20. Phosphatidylethanolamine having mono- or di-unsaturated fatty acids and mixtures of saturated and unsaturated fatty acids can also be used. Suitable PEG-phosphatidylethanolamine derivatives include, but are not limited to, monomethoxypolyethylene glycol-distearoylphosphatidyl-ethanolamine (m-PEG-DSPE).

The composition of the present invention may contain one or more suspending agents. Suspensions are pharmaceutically inert substances that provide increased stability of the suspension by increasing the viscosity of the composition of the present invention. These agents also promote the redispersion of particles that have been precipitated by standing for a long time. Various cellulose derivatives are commercially available and are suitable for use as the suspending agent of the present invention. For example, sodium carboxymethylcellulose (CMC) and methylcellulose (MC) are readily available and are suitable as suspending agents in the compositions of the present invention. CMC is a colorless, odorless, non-toxic, water-soluble powder, used in various detergents, soaps, foods, fibers, coatings, paints, cosmetics and medicines, among them water binders, thickeners, Acts as a suspension or emulsion stabilizer. This CMC structure is based on the beta- (1-> 4) -D-glucopyranose polymer of cellulose. Different products of CMC may have different degrees of substitution, but are usually from about 0.6 to about 0.9 derivatives per monomer unit. In general, as the molecular weight and average chain length (ie, degree of polymerization) of CMC increases, the viscosity of these polymers increases correspondingly. High viscosity or high molecular weight CMC has an average molecular weight of about 700,000 g / mole and a degree of polymerization of about 3,200; medium viscosity or medium molecular weight CMC has an average molecular weight of about 250,000 g / mole and a degree of polymerization of about 1,100 The low viscosity or low molecular weight CMC has an average molecular weight of about 90,000 g / mole and a degree of polymerization of about 400; Many different types of CMC are available for purchase. CMCs useful in the compositions of the present invention include, but are not limited to, high viscosity CMCs such as 7H4, 7H3S PH, 7HOF, 7H, and 9H4 type Aqualon ^™ ; 7M, 7M8S, 7M2, 9M31, 9M8, 12M31 And medium viscosity CMC such as 12M8 type; and low viscosity CMC such as 7L and 7L2. Aqualon ^™ is a CMC available from Hercule and its technical information is described in Aqualon ^™ product brochure (Physical and Chemical Properties of Aqualon ^™ , Sodium Carboxymethylcellulose), which is incorporated by reference in its entirety. Included in this application for purposes.

  Methylcellulose (MC) is also useful as a suspending agent in the composition of the present invention. MC is also a semi-synthetic water-soluble polymer derived from cellulose. This MC structure is based on the beta- (1-> 4) -D-glucopyranose polymer of cellulose. MC is a powdery substance produced by methylation of natural cellulose and is used as a food additive, a bulking laxative, an emulsifier, and a thickener that swells in water to form a gel.

  Glycerin is a trihydric alcohol and is a clear, colorless, transparent, viscous and hygroscopic liquid at room temperature. Glycerin has been widely used in the pharmaceutical industry as a solvent and solubilizer in various pharmaceutical vehicles for both inside and outside, and is also useful as a vehicle in the compositions of the present invention.

  The compositions of the present invention can optionally include one or more aqueous buffers, or mixtures thereof. Buffers are commonly used in pharmaceutical compositions and have the effect of stabilizing the pH of the composition when acid or base is added to the solution, minimizing acid or basic fluctuations in the solution. The buffer maintains the pH of the solution by reacting with a small amount of added acid or base. For buffers that can do this, both acid and base must be included; the acid will react with any added base and the base will react with any added acid. However, they need to be able to coexist without reacting to each other. In order to do so, the acid and base must be a conjugated acid-base pair. Examples of buffers include ethanoic acid and sodium ethanoate, or a mixture of ammonia solution and ammonium chloride.

Buffers useful in the compositions of the present invention are commonly known as biological buffers and include, but are not limited to, potassium dihydrogen phosphate (KH ₂ PO ₄ ), potassium hydrogen phosphate (K ₂ HPO ₄ ), potassium phosphate (K ₃ PO ₄ ), sodium dihydrogen phosphate (NaH ₂ PO ₄ ), sodium hydrogen phosphate (Na ₂ HPO ₄ ), and sodium phosphate (Na ₃ PO ₄ ) and their A phosphate buffer, such as a mixture of Other useful buffers include, but are not limited to, citric acid and sodium citrate; citric acid and sodium hydroxide; citric acid and sodium hydrogen phosphate; boric acid-citric acid-potassium dihydrogen phosphate-diethyl barbi Tool acid and sodium hydroxide; acetic acid and sodium acetate; potassium hydrogen phthalate and sodium hydroxide; sodium cacodylate and sodium acetate; potassium dihydrogen phosphate and sodium hydrogen phosphate; sodium dihydrogen phosphate and sodium hydrogen phosphate Sodium tetraborate and boric acid; 2-amino-2-methyl-1,3-propanediol and hydrochloric acid; diethanolamine and hydrochloric acid; potassium chloride-boric acid and sodium hydroxide; boric acid-sodium hydroxide and potassium chloride; Glycine and sodium hydroxide; sodium carbonate and sodium bicarbonate Sodium hydrogen phosphate and sodium hydroxide; potassium chloride and sodium hydroxide; succinic acid, imidazole and hydrochloric acid, phosphoric acid, tris (hydroxymethyl) aminomethane, glycylglycine, and boric acid buffers, and mixtures thereof. Can be mentioned. Other buffers are also useful in the compositions of the present invention and can be readily replaced by those skilled in the art. Ideally, the buffer used in the composition of the present invention is at about physiological pH (7.38) and is present in a range of concentrations from about 1 mM to 100 mM (1 mM = 0.001 M). Such buffers are readily available commercially or are made using laboratory methods well known in the art. For example, 10 mM PBS (phosphate buffered saline) is commercially available (Zymed Cat. No. 00-3000), or KH ₂ PO ₄ 0.26 g, Na ₂ HPO ₄ -7H ₂ 0 2.17g, 8.71g NaCl, 800mL distilled water, and use NaOH or HCl to adjust the pH to 7.4 so that it does not go too far and back titration as it may change the salt concentration Adjust the volume to 1L with water. The compositions of the present invention can also include one or more tonicity modifiers. Tonicity modifiers act to increase the effective osmotic pressure of the compositions, thereby improving their compatibility within the cellular environment. For example, if a cell is placed in a hypotonic solution (ie, a solution that has a low solute concentration and therefore a high water concentration), there will be a net transfer of water into the cell, Will swell and break (dissolve). Conversely, if the cells are placed in a hypertonic solution, the cells will contract. Tonicity modifiers include, but are not limited to, salts such as sodium chloride; sugars such as mannitol, and are present in the compositions of the present invention in an amount sufficient for an approximately isotonic preparation. ing. In the composition of the present invention, the tonicity and pH of the composition are adjusted according to physiological conditions. See, for example, Remington's Pharmaceutical Sciences, Mack Publishing Company, Easter, Pa., 15th Edition (1975).

  The preparation of the compositions of the present invention and methods for subtenon delivery of pharmaceutically active substances or pharmaceutically acceptable salts or solvates thereof are described in detail in the examples below. One skilled in the art will appreciate that the compositions and methods of the present application can be readily adapted to the manufacture of many other inventive compositions for subtenon, intraocular, systemic or topical delivery.

Example 1
Bioanalytical method for the determination of compounds of formula I in human plasma (treated with heparin sodium) by LC-API / MS / MS The compound of formula I and the internal standard substance (compound of formula I-d7) are mixed with ethyl acetate: hexane Extracted from 500 μL of human plasma (heparin sodium treated) by liquid-liquid extraction using (50:50). After evaporating to dryness and reconstitution, the extract was analyzed by LC-API / MS / MS. The operation time was about 2.5 minutes. The lower weigh limit (LLOQ) of the compound of formula I was 0.0100 ng / mL (FIG. 1). The confidence response range was 0.0100 to 10.0 ng / mL.
The bioanalytical method is based on the results of a complete validation test on human plasma. The relevant results and reports are in the ALTA validation report AR139K.

Acceptance criteria Based on the verification results referenced above:
1. An acceptable analytical run should have a standard curve such that at least 75% of the individual standard points meet the method criteria. The back-calculated standard deviation (% DEV) must be within ± 15% of the nominal concentration, with the exception of ± 20% deviation allowed for LLOQ. At least one calculated concentration of the lowest measured double point of the standard curve must meet the criteria for quantification as an LLOQ. If this criterion is not met, the LLOQ is raised to the next level.
2. The calculated concentration of acceptable analytical QC should be within ± 15% of the nominal concentration. At least two-thirds of all analytical QCs must meet the criteria given by the respective QC level displayed.
3. The calculated concentration of acceptable dilution QC should be within ± 15% of the nominal concentration. At least two thirds of the dilution QC must meet the criteria given for each dilution scheme. If the dilution QC for a given dilution scheme does not meet the criteria, then all test samples associated with this dilution scheme must be repeated.
4). If neither standard curve nor analytical QC criteria are met, the analysis result is rejected.

Revision Record Revision 1 provided protection against the carry-over effect of the injector, including assessment, precautions, and addition of a new rinse solvent.

Analytical instrument and equipment
SCIEX API 4000TM mass spectrometer (PE-SCIEX Concord, Ontario, Canada)
Shimadzu LC-10AD HPLC pump (2) Shimadzu SCL-10A controller with mixing tee (or equivalent) or HTC-PAL automatic injector (or equivalent)
IEC Centra GP8R centrifuge (or equivalent)
Turbobop LV evaporator reagents and chemicals Reference materials, compounds of formula I, and internal standards (compounds of formula I-d7) were obtained from Agouron Pharmaceutical Company under Pfizer. When used, acetonitrile (ACN), methanol (MeOH), ethyl acetate, hexane and water are HPLC grade or equivalents.

The following methods for the preparation of reagents and solutions are provided for illustration only. Other preparation methods can be substituted as long as the final concentration is the same.
1. Constituent reagents / diluent: MeOH: water (50:50) Mix equal volumes of MeOH and water.
2.1M NH ₄ OAc (ammonium acetate) 77.09 g of NH ₄ OAc is dissolved in 1 L of water.
Mix 5 mL of 3.5 mM NH ₄ OAc 1M NH4OAc with 995 mL of water.
4). Ethyl acetate: hexane (50:50) Mix 500 mL of ethyl acetate and 500 mL of hexane.
5. Reconstitution solvent: MeOH: 5 mM NH ₄ OAc (50:50). Mix 50 mL of MeOH and 50 mL of 5 mM NH ₄ OAc.
6). Mobile phase A: 0.1% formic acid 999 mL of water and 1 mL of formic acid are mixed.
7). Mobile phase B: 999 mL of ACN CAN containing 0.1% formic acid and 1 mL of formic acid are mixed.
8). Autoinjector rinse 1: Mix ACN: water (50:50) with 0.3% polyethyleneimine (PEI): 500 mL of ACN, 500 mL of water and 3 mL of PEI.
9. Autoinjector rinse 2: MeOH: water (10:90) with 0.5% formic acid: Mix 100 mL of MeOH, 900 mL of water and 5 mL of formic acid. Mix the solution well and bring the solution to room temperature before use.

Preparation of Standards and Quality Control Samples The following methods are provided as examples for the preparation of calibration curve (CC) and quality control (QC) solutions. Other weighings and dilutions can be substituted if the calibration range remains the same and the QC concentration is approximately the same. Examples of calibration curve points are shown below.

1. CC and QC stock solutions a) A CC stock solution of a compound of formula I is prepared in MeOH at a concentration of 100 μg / mL.
b) A QC stock solution of the compound of formula I is prepared in MeOH by independent weighing at a concentration of 100 μg / mL.

2. CC substock and spiking solution in CC spiking solution dilution solvent (MeOH: water (50:50)) are prepared as shown below.

3. Internal standard solution: a) reacting a compound of formula I-d7 internal standard 100 [mu] g / mL of ² compounds (IS) stock solution is prepared.
b) The IS spiking solution in dilute solvent is prepared on the day of analysis as shown below:

Pooled matrix QC
QC is prepared by spiking a control matrix (human plasma supplemented with sodium heparin) with a 10 μg / mL QC substock solution of the compound of formula I (prepared by diluting 1 mL of QC stock solution to 10 mL with MeOH). Next, dilute with a control matrix to obtain an appropriate concentration. An example of QC preparation is described below:

c) After fortification, vortex each QC pool thoroughly.
d) Place an appropriate amount (eg 1.2 mL) aliquot of each QC pool into a labeled polypropylene tube (eg 2 mL).
e) Store the pooled QC samples frozen at -20 ° C until use.

4). Test solution for system suitability
Test solutions containing compounds of formula I (0.025 ng / mL) and IS (5 ng / mL) can be prepared at or near the concentration of the final extract of LLOQ, and a low calibration curve point is used for test injection be able to. To prepare the former, 50 μL of 5 ng / mL of a compound of formula I (eg, CC-0.5) and 2.5 mL of 20 ng / mL of a compound of formula I-d7 (eg, IS spiking solution) are mixed. And dilute to a final volume of 10 mL with reconstitution solvent (50:50 MeOH: 5 mM NH ₄ OAc).

Definition of control and blank Blank without IS (BI / BI) : Specimens identified as BI / BI contain a control matrix and no internal standard added.
2. Blank with IS (BI / IS) : Samples identified as BI / IS contain a control matrix and an internal standard.
3. Solvent blank : The appropriate solvent or mobile phase was used to demonstrate background loss or carry-over from the chromatography system.

Special considerations Protect specimens and extracts from light. Additional BI / BI specimens must be extracted to protect the specimen from the carry-over effect of the injector.

Extraction method for each stage Label all tubes (glass tubes with screw caps or equivalent).
2. Thaw the control matrix, QC and test specimen to room temperature.
3. Mix the control matrix, QC and test specimen container.
4). If necessary, centrifuge the specimen to remove particles.
5. In a suitable tube:
a) Pipet 500 μL of control matrix (sodium heparin-treated human plasma) into each tube labeled with BI / BI (N> 1), BI / IS (N> 1) and weighing curve (N = 2) .
b) Pipette 500 μL of QC (N> 2 at low, medium and high levels) and test specimen (N = 1) into each appropriately labeled tube. If a smaller sample aliquot is taken, dilute (QS) to 500 μL in the control matrix or in a separate tube with control matrix and pipette a 500 μL aliquot into the extraction tube.
c) When the test sample is diluted, a set of QC samples (N = 3) must be diluted in the same manner as the test sample.

6). Add 50 μL of each CC spiking solution to the appropriate calibration curve tube (prepared in duplicate).
7). Add 50 μL of IS spiking solution (20 ng / mL compound of formula I-d7 prepared on the day of analysis) to each labeled tube except for all BI / BI.
8). Add makeup reagent (50:50 MeOH: H2O):
a) Add 100 μL of makeup reagent to the BI / BI sample.
b) Add 50 μL of makeup reagent to BI / IS, QC and test sample.
9. Mix by vortexing (about 5-10 seconds).
Ten. Add approximately 8 mL of ethyl acetate: hexane (50:50) to each tube.
11． Cap (polypropylene or Teflon) and mix with an orbital mixer for about 5 minutes at 100 RPM.
12． Centrifuge the sample for 5 minutes at 2500 RPM setting.
13. Carefully freeze the aqueous phase with a dry ice / acetone bath or place the tube in a −70 ° C. freezer (approximately 30 minutes).
14. The supernatant is placed in a glass tube (eg, a 16 × 100 mm glass culture tube or equivalent) for use in a turbo-bop.
15. Evaporate to dryness with nitrogen at 30 ° C.
16． Reconstitute with 200 μL of reconstitution solvent (50:50 MeOH: 5 mM NH4OAc) and stir briefly (about 10-20 seconds).
17． Transfer to a smoked automatic injection vial using polyethylene insert (or equivalent).
18． The specimen extract can be stored in the refrigerator until analysis.

Chromatography Separation is performed using reverse phase chromatography.
HPLC operating conditions:
Column: Keystone Betasil C18, (5μm) 100x2mm (Thermo Hypersil-Keystone)
Guard: Keystone Betasil C18, (5μm) 10 × 2mm (Thermo Hypersil-Keystone)
Column temperature: ambient temperature Injection volume: 30 μL (different injection volumes may be used as long as chromatographic integrity is maintained)
Flow rate: 0.4 mL / min Mobile phase: Isocratic, Mobile phase A: Mobile phase B 55:45 (v / v)
Analyte ~ Retention time (min)
Compound of formula I 1.2
Compound of formula I-d7 1.2

A 0.5 μm stainless steel pre-column filter and / or frit can be used to extend the life of the column. This pre-column filter can be used in various analytical sets if no obvious back pressure is observed.
Suitable reagents for autoinjector rinsing are autoinjector rinses 1 and 2. Other suitable rinses can be used.

Detection by mass spectrometry Detection is performed by tandem mass spectrometry (MS / MS). HPLC system is combined with mass spectrometry using turbo Ron spray ^TM (TurbolonSpray ^TM) atmospheric pressure ionization inlet. Analysis is performed by positive ionization MS / MS. High purity nitrogen gas from high pressure liquid nitrogen Java is used for the nebulizer, Turbolon Spray ^™ and curtain gas. Other nitrogen or air sources can also be used.
Detection is performed in multiple reaction monitoring mode (MRM). The collision gas for generating fragmented ions is high purity nitrogen.
If the appropriate sensitivity is achieved, the column eluate should be split appropriately using a tee with zero dead volume (Valco, Houston, Texas, or equivalent). In ratio, after the column, all other eluates can be directed to the inlet.

The following reactions are monitored:
Analyte transition (± 0.2 amu)
Compound of formula I m / z 468.1? 193.1
Compound of formula I-d7 m / z 475.1? 193.1

The following instrument parameters were used during validation. The measured parameter values may be changed daily to optimize instrument sensitivity. The actual parameter values used in the analysis set must be recorded in the raw data.
Mass spectrometry parameters:
ISV voltage (IS): 4400 V
Curtain gas (CUR): 15
Turbolon spray temperature (TEM): 700 ℃
Impact pressure (CAD): N2 @ 7
Cluster separation potential (DP): 90
Collision energy (CE): 59
GS1: 50
GS2: 75

Quantification method Peak regions are integrated with Applied Biosystems MDS SCIEX Analyst ^™ software. A leveling factor can be used. If used, the same leveling factor must be used for all integrations in the set used. A calibration curve is derived from the peak area ratio (analyte / internal standard) using the ratio of the least squares regression method to the nominal concentration of the standard calibration curve. The deviation from the regression line is calculated using a regression equation that reversely calculates the concentration of each calibration standard level. The concentration of the QC specimen is also calculated from their regression curves using the calculated peak area ratio.
Regression type: linear
Weight coefficient: 1 / × 2 <sup>*
* When</sup> x is the concentration of a given calibration standard level.

Recording of human plasma parameters of compounds of formula I Project: Mode: ISV
Record: Equipment: SX9

Example 2
Achieving effective choroid concentrations with minimal systemic effects of compounds of formula I (VEGFR inhibitors) in cynomolgus monkeys and humans with age-related macular degeneration:
The compounds of formula I can be delivered to the posterior segment of the eye adjacent to the macula by subtenon administration to minimize side effects. The compounds of formula I have been found to be potent inhibitors of VEGER2 tyrosine kinase activity and are selective for VEGER tyrosine kinase. The compound is also a potent, long-lasting and reproducible inhibitor of ocular neovascularization in various preclinical models, including atrophy with reduced preexisting vascular leakage and suppression of angiogenesis I know.

  The drug solubility of the compound of formula I is low, about 0.8 μg / ml in PBS and about 5 μg / ml in the vitreous. This compound binds strongly to proteins, for example, serum binding is about 99.9%. Such high protein binding and low solubility means that the drug forms a depot that slowly disappears. For example, the 14C radioactive equivalent of a compound of formula I is first taken up by pigment cells and remains present for 8 weeks after administration. Thus, drug levels in ocular tissue reach equilibrium and remain constant within a relatively prolonged period. Furthermore, the physicochemical properties of this compound are suitable for subtenon administration.

  Phase I clinical randomized, masked, single-time, safety and efficacy of compounds of formula I in subjects suffering from subcentral choroidal neovascularization with age-related macular degeneration In addition, a multiple dose, continuous dose-dose test was performed. In the implementation of this test type, single and multiple doses of Random Mask Phase 1/2, continuous administration with 52 weeks of repeated posterior subtenon administration followed by 52 weeks of follow-up. -Included a weight gain test. This study included two stages, and stage 2 was conducted after safety was demonstrated at week 13. Participants in these trials included exudative age-related macular degeneration, including non-exudative (dry) and exudative (wet) 55 years and older with all types of subfoveal choroidal neovascularization Includes 114 men and women with AD (AMD). Their visual acuity was measured as 73 to 24 characters (20/40 20/320), and the first PDT was allowed within 3 months prior to screening. The number of groups in this study included: 4 branches of 3 cohorts: 50 mg (q13 and 26 w), 150 mg (q26 w) and 600 mg (q52 w), and the participating study centers were in the Netherlands (4 ), The United States (24), the United Kingdom (6) and Australia (5), as well as a single independent decoding center of the University of Wisconsin FPRC.

  The primary endpoint of this study is: ocular and systemic safety at 13 weeks after a single sub-Tenon dose and 104 weeks after multiple doses. Failure rate:% of subjects lost (ie, less than 15 characters). Secondary endpoints were:% response rate of subjects who acquired 15 characters or more; mean variation in visual acuity from baseline; changes in fundus photography, fluorescein angiography and OCT; formula I after subtenon administration Includes compound drug metabolism; as well as changes in health-related quality of life scores.

  For clinical safety, the longest follow-up period is 12 months (4 injections); treatment-related AEs-8/60 (13%) of subjects (all mild); There were no SAEs associated with; and there were no deaths.

  In humans, the compound of formula I has an assay sensitivity of 10 pg / mL; plasma concentration measured during stage 1: 378 samples collected and 35 (9%) of the samples have a measurable plasma concentration. Of the 35 active drugs, 19 (54%) have at least one measurable plasma concentration; the maximum measurable plasma concentration is 102 pg / mL ( (Day 3); no measurable plasma concentration after 4 weeks. Plasma concentrations were lower than the expected minimum effective tissue concentration estimated at IC90 = 60 ng / g.

  In a masked efficacy study, 47 subjects achieved a 13 week visit in a low dose (50 μg), medium dose (150 μg) and high dose (600 μg) cohort, of which 42 were active drugs and Five were given a placebo. The changes from baseline in visual acuity at the end of the 13-week follow-up period were as follows: 4 subjects gained more than 15 characters; 36 subjects had stable visual acuity (baseline Within ± 14 characters); and 7 subjects lost more than 15 characters.

  The compounds of formula I are potent and effective small molecule VEGFR2 inhibitors. In preclinical animal species, the compound of formula I exceeded the targeted concentration in the choroid for at least 16 weeks. Human plasma concentrations have proven to be significantly lower than therapeutic levels. No significant side effects have been observed in previous Phase 1/2 trials. Thus, the compounds of formula I may be effective drugs for the treatment of angiogenic age-related macular degeneration (AMD), including non-wetting (dry AMD) and wet (wet AMD) .

Example 3
VEGFR tyrosine kinase inhibitors of compounds of formula I when sub-tenon administration of a compound of formula I to cynomolgus monkeys is a tenon for the treatment of choroidal neovascularization associated with age-related macular degeneration Clinical development is underway for subcapsular administration. The ocular and systemic distribution of the compound of formula I was evaluated using cynomolgus monkeys after subtenon administration.

  Single dose drug and toxic metabolism studies were conducted in male and female cynomolgus monkeys. A compound of formula I was administered by subtenon injection in a sterile sucrose suspension at a dose several times higher than the possible clinical dose. Drug deposition was targeted at the posterior part of the macula adjacent to the optic nerve. Plasma was collected up to 48 hours after administration. Ocular tissues including choroid, retina, administration site tissue, eye muscle, sclera, vitreous, iris-ciliary body, lens and aqueous humor were collected at various time points up to 16 weeks in one study. The concentration of compound of formula I in the administration site tissue, choroid, retina and vitreous was measured at week 13 of the subsequent study. Plasma and tissue concentrations were measured by LC-MS / MS.

  The plasma concentration of the compound of formula I was less than 0.2 ng / mL in all examples in all administration groups except one sample. Compounds of formula I are observed in the administration site tissue (conjunctiva and adipose tissue collected close to the sclera at the administration site) up to 16 weeks after a single sub-Tenon administration at all doses tested It was. The concentration of the compound of formula I in the choroid, the target tissue for age-related macular degeneration, is 25-125 times the IC90 of the compound of formula I for inhibition of VEGF-R2 in the rat retinal PK / PD model, Achieved within 3 days and maintained up to 16 weeks. The retina concentration of the compound of formula I is usually below IC90 and is encased in a Tenon membrane, showing mild but varying levels. The concentration of the compound of formula I in the sclera increased up to 8 weeks and decreased at 12 and 16 weeks. Very low concentrations of the compound of formula I were observed in the iris-ciliary body or lens and not in the vitreous or aqueous humor. [Figure 2]

  The compound of formula I (ie AG-013958) is a VEGFR inhibitor that achieves effective choroidal concentrations with minimal systemic effects in cynomolgus monkeys and humans with age-related macular degeneration. Subtenon administration of a suspension of a compound of formula I forms drug deposits on the posterior surface of the sclera, has little effect on peripheral tissues and has no measurable systemic exposure and is directed against the choroid Sustained exposure was given. This data supports a separately reported preclinical safety assessment of the compound of Formula I and also an ongoing clinical assessment for the treatment of age-related macular degeneration.

  The physicochemical properties of the compounds of formula I include low drug solubility: about 0.8 μg / ml in PBS; about 5 μg / ml in the vitreous; strong binding to protein; serum binding about 99.9% High protein binding and low solubility means that the drug forms a depot that slowly disappears, thus the drug level in the eye tissue reaches equilibrium and within an extended period of time Remains relatively constant. These physicochemical properties are suitable for subtenon administration.

  The compounds of formula I are potent and effective small molecule VEGFR2 inhibitors. In preclinical animal species, the compound of formula I exceeded the targeted concentration in the choroid for at least 16 weeks. Human plasma concentrations have proven to be significantly lower than therapeutic levels. No significant side effects have been observed in previous Phase 1/2 trials. The compounds of formula I may be effective drugs for the treatment of angiogenic age-related macular degeneration.

Example 4
VEGF receptor tyrosine kinase inhibitor, compound of formula I, evaluation after ocular administration in rabbits and primates The compound of formula I is currently used for the treatment of angiogenic eye diseases such as age-related macular degeneration It is a vascular endothelial growth factor receptor tyrosine kinase inhibitor that is undergoing Phase 1/2 clinical trials. The safety of the compound of formula I is 1 to 30 times higher than the expected clinical dose (each in the eye), subtenon (ST), intravitreal (IVT) or intravenous (IV) administration. Evaluated in a series of nonclinical trials. Single subtenon administration of the compound of formula I (intended clinical route of administration) provided no gross findings due to the compound until 13 weeks of treatment in primates. Microscopic findings of the eyes and whole body tissues showed the presence of macrophages at the site of subtenon administration at 8 and 13 weeks after treatment. This finding was consistent with a foreign body response to the presence of the compound of formula I and was not accompanied by tissue damage or a fulminant inflammatory response. A major finding in studies using the IVT route of administration in rabbits and primates was the identification of test substances that were visible in the vitreous of both animal species up to 8 weeks after injection. The presence of macrophages in the vitreous of IVT-treated rabbits was considered a foreign body reaction. The dose selected in the IVT test resulted in a choroidal concentration of the compound of formula I that was 2-3 fold higher than that tested with subtenon administration. The compound of formula I given in a single bolus IV dose at the selected maximum IVT dose produced no signs of systemic toxicity by 5 days after treatment. Overall, preclinical safety assessment of the compound of formula I shows that the compound is well tolerated in rabbits and primates, and angiogenesis including the non-wetting form (dry AMD) and the wet form (wet AMD) of this compound Support further development for the treatment of age-related macular degeneration (AMD).

  Ocular vascular diseases such as age-related macular degeneration are the leading cause of vision loss and blindness in the United States (1). Vascular endothelial growth factor (VEGF) is thought to be associated with angiogenesis, and inhibition of the VEGF pathway has proven hope as a pharmaceutical intervention for the treatment of choroidal neovascularization (2) . The compounds of formula I are small molecules that inhibit the activity of the tyrosine kinase (TK) domain of vascular endothelial growth factor receptor 2 (VEGF-R2). The compounds of formula I are currently undergoing phase 1/2 clinical trials as angiogenesis inhibitors intended for the treatment of choroidal neovascularization in patients with age-related macular degeneration. The clinical route of delivery is by subtenon (ST) administration. This study explores preclinical safety assessment by subtenon and intravitreal (IVT) administration in Dutch-belted rabbits and cynomolgus monkeys and by intravenous bolus injection in rabbits.

Materials and Methods Animals: Male and female Dutch belt rabbits (1.5-2 kg) and cynomolgus monkeys (2-4 kg).
All animal-related operations were performed in accordance with animal protection standards and laboratory animal care and use guidelines, under procedures approved by IACUC. This study complies with ARVO recommendations for the use of animals in ophthalmic and visual studies.

Ophthalmic administration Subtenon administration in primates: The animal is anesthetized (xylazine / ketamine), then a small incision (1-2 mm) is made in the temporary lower inlaid eye and exposed to the sclera to expose the space under the Tenon capsule To be able to touch. A bent 23 gauge blunt end cannula was inserted into the subtenon subcapsular space and advanced in the posterior direction. A compound of formula I or vehicle (sucrose-based formulation) was administered as a 0.5 ml bolus injection.

  IVT administration in primates and rabbits: Animals were anesthetized (xylazine / ketamine) and then a 27 gauge needle was inserted approximately 3 mm behind the corneal limbus of the upper / temporary inlay. The tip chamfer of the needle was placed in the forward position and advanced to the center of the vitreous. The compound of formula I or vehicle (formulation based on hyaluronic acid) was administered as a 0.05 ml bolus injection.

Experimental Design Rabbit and primate IVT test and subtenon test plan are shown in FIG.

result:

  The compounds of formula I are well tolerated under Subtenon, IVT and intravenous administration. Electroretinograms and ophthalmic studies were within normal range throughout the duration of the study after subtenon administration. Systemic toxicity measured at the endpoints obtained in this study was not observed after ocular administration of the compound of formula I. Compounds of formula I administered intravenously did not cause systemic toxicity as measured by clinical observation and tissue histopathology.

Example 5
Ocular and systemic drug metabolism of compounds of formula I by subtenon administration to cynomolgus monkeys VEGFR tyrosine kinase inhibitor, a compound of formula I, is a tenone for the treatment of choroidal neovascularization associated with age-related macular degeneration Clinical development is underway for subcapsular administration. The ocular and systemic distribution of the compound of formula I was evaluated using cynomolgus monkeys after subtenon (ST) administration.

  Single dose drug and toxic metabolism studies were conducted in male and female cynomolgus monkeys. A compound of formula I was administered by subtenon injection in a sterile sucrose suspension at a dose several times higher than the possible clinical dose. Drug deposition was targeted at the posterior part of the macula adjacent to the optic nerve. Plasma was collected up to 48 hours after administration. Ocular tissues including choroid, retina, administration site tissue, eye muscle, sclera, vitreous, iris-ciliary body, lens and aqueous humor were collected at various time points up to 16 weeks in one study. The concentration of the compound of formula I in the administration site tissue, choroid, retina and vitreous was measured on the 13th week of the subsequent study. Plasma and tissue concentrations were measured by LC-MS / MS.

The plasma concentration of the compound of formula I was less than 0.2 ng / mL in all examples in all administration groups except one sample. Compounds of formula I are observed in the administration site tissue (conjunctiva and adipose tissue collected close to the sclera at the administration site) up to 16 weeks after a single sub-Tenon administration at all doses tested It was done. The concentration of the compound of formula I in the choroid, the target tissue for age-related macular degeneration, is 25-125 times the IC ₉₀ of the compound of formula I for inhibition of VEGF-R2 in the rat retinal PK / PD model Achieved within 3 days and maintained up to 16 weeks. The retina concentration of the compound of formula I was usually below IC ₉₀ . The eye muscles were also encased in a tenon membrane, showing mild but fluctuating levels. Concentrations in the scleral specimen increased up to 8 weeks and decreased at 12 and 16 weeks. Very low concentrations of the compound of formula I were observed in the iris-ciliary body or lens and were only low or not observed in the vitreous or aqueous humor.

  The compound of formula I (ie AG-013958) is a VEGFR inhibitor that achieves effective choroidal concentrations with minimal systemic effects in cynomolgus monkeys and humans with age-related macular degeneration. Subtenon administration of a suspension of a compound of formula I forms a drug deposit on the posterior surface of the sclera, has little effect on peripheral tissues and has no measurable systemic exposure and persists to the choroid Exposure was given. This data supports a separately reported preclinical safety assessment of the compound of Formula I and also an ongoing clinical assessment for the treatment of age-related macular degeneration.

  The following tests were conducted to determine the eye and systemic distribution of compounds of formula I in cynomolgus monkeys. The compound was prepared as a sterile sucrose suspension.

  In the 16 week study, male and female monkeys were lightly anesthetized and then administered 0.5 mL / eye of a compound of formula I. Two groups received 50 or 1500 μg / eye subtenon injection. Blood samples were collected at 1, 2, 4, 8, 12, 24, 48, 72 hours and 4 and 8 weeks after administration. The animals were euthanized and the endpoints were on day 3 and at weeks 4, 8, 12, and 16. The eyes were removed, dissected, and the collected ocular tissues were the administration site, choroid, retina, vitreous humor, aqueous humor, lens, sclera, iris-ciliary body and eye muscle. Test specimens were analyzed by LC-MS / MS.

  In the 13 week study, three groups of male monkeys were lightly anesthetized and then dosed with 50, 250 or 1500 μg / eye of a compound of formula I. Administration was performed on both sides by subtenon injection near the optic nerve. Blood samples were collected at 1, 6 and 24 hours after administration. After incision at week 13, the collected ocular tissues were the administration site, choroid, retina and vitreous humor. Eye tissue and plasma samples were analyzed by LC-MS / MS.

Tissue analyzed: site of administration ^* ; retina; iris-ciliary body; sclera; eye muscle; vitreous humor; aqueous humor; lens;
^{* The} administration site tissue consists of conjunctiva and adipose tissue in the subtenon space taken from the drug deposition area behind the macula and adjacent to the optic nerve.

Analysis Administration site tissue, sclera, eye muscle and lens-CAN (acetonitrile) with 0.1% TFA (trifluoroacetic acid).
Retina and choroid—CAN: Protein precipitation with MeOH.
LOQ:
Retina-0.2 ng / g (concentration corrected by tissue mass)
Choroid-0.5 ng / g (concentration corrected with tissue mass)
Plasma-0.2 ng / mL
Vitreous-0.5 ng / mL
Administration site tissue, sclera, eye muscle and lens-0.5 ng / mL

Results The results of these studies showed that the compound of formula I was detected in the dosing site tissue by 16 weeks at both doses. In the 50 μg / eye dose group, 1 μg or more was observed at 13 weeks.

By day 3, a compound of formula I at a concentration of at least 1,100 ng / g was detected in choroidal tissue and maintained until week 16. The concentrations detected in both tests were 125 times higher than the target IC ₉₀ .

The concentration of the compound of formula I in the retina was below the target IC ₉₀ and fluctuated. The compound of formula I was moderate in ocular muscles and iris-ciliary body and low in a few samples of lens and vitreous humor. The compound of formula I was not detectable in aqueous humor or plasma, and no noticeable systemic or ocular toxicity was observed.

The compound of formula I was 25-125 times higher than the target effective concentration (IC ₉₀ ) as early as 3 days in the choroid and was maintained until 16 weeks. The concentration of Formula I compound in the retina was significantly lower and more variable than the choroid and administration site. The compound of formula I did not cause any ocular or systemic toxicity.

  Accumulation in the administration site tissue and choroid of the compound of Formula I demonstrates slow release of the drug and suggests a long interval of clinical administration.

Example 6
Subtenon injection into female Dutch belt rabbits Female Dutch belt rabbits (1.5-2 kg) were anesthetized (ketamine / xylazine mixture) and both eyes were prepared for sterile subtenon injection. Each eye was administered a 0.5 mL bolus injection of the test vehicle or formulation using a curved 23G needle. Animals were observed for clinical signs of toxicity / irritation for 1 week after dosing. The eyes were also removed and fixed in Davidson's solution for histopathology.

As shown in Table 1, vehicle excipients for the composition and methods described herein include flocculants such as benzalkonium chloride, docusate sodium and sodium lauryl sulfate, carboxymethylcellulose (CMC; low Molecular weight (90 kDa), medium molecular weight (250 kDa) and high molecular weight (700 kDa), 0.25-1.0%), polysorbate ^TM 80 (0.02 and 0.2%), polyethylene glycol 3350 (PEG 3350; 0.2 and 1.0%), poloxamer ^TM 188 (0.01 And 0.25%), nano-edges (Poloxamer ^™ 188 and PEG3350 combination) and methylcellulose (MC; 0.25%); isotonic agents such as mannitol and salts (eg, NaCl and MgCl), and PBS buffer Contains various buffer solutions.

  After subtenon injection, moderate redness was observed in the conjunctiva around the eyelids and around the injection entry site in most treated eyes, most often disappearing within 2-4 days after treatment. This effect was not related to the administered test substance and was considered to be related to the subtenon administration procedure. Overall, the vehicle vehicle tested was well tolerated in ocular tissues after subtenon administration.

Example 7
Subtenon formulation A suitable formulation for subtenon administration of compounds of Formula I, II and III has been developed. These compounds are in a concentration range of about 0.05 to about 10 mg / mL; with sodium dihydrogen phosphate buffer at physiological pH; and either sodium chloride or mannitol in an amount sufficient for a nearly isotonic formulation And was formulated.
1. Tween ^TM 80, 0.001% to 0.2% by weight;
2. CMC, 0.01% to 1.0% by weight;
3. Poloxamer ^TM 188, 0.01% to 0.25% by weight;
4). 4. methylcellulose (MC), 0.1% to 0.25% by weight; and Polyethylene glycol 3350 (PEG 3350), 0.01% to 1.0% by weight.
All these formulations can be diluted to achieve a drug concentration of at least 0.1 mg / ml upon dilution with an appropriate vehicle.

The compound of formula II was diluted with sodium phosphate buffer and either sodium chloride or mannitol in an amount sufficient for a nearly isotonic formulation:
1.2mg / ml 0.25% CMC, 0.01% Tween ^TM 80 autoclaved
10 mg / ml filtered 0.1% MC ^* + 0.05% Tween ^™ 80 Irradiated Compound of formula III with sodium phosphate buffer and either sodium chloride or mannitol in sufficient quantity for a nearly isotonic formulation Diluted with:
1.2mg / ml 0.01% Tween ^TM 80 autoclaved
10mg / ml filtered 0.5% MC ^* + 0.05% Tween ^TM 80 Autoclaved

Schemes for the preparation of compounds of formulas I, II and III:
All surfactant or suspension solutions were prepared as 1 to 20-fold concentrates and sufficient volume was added to achieve the desired concentration. Provided below is a scheme for the preparation of a sub-Tenon capsule composition comprising compounds of Formulas I, II and III:
Prepare 10 mM pH 7.4 phosphate buffer / 0.9% saline solution.
Prepare a surfactant solution using PBS. Filter through a 0.2 μm filter into a clean bottle.
If necessary, prepare a suspension solution with PBS. Filter through a 0.22 or 5 μm filter into a clean bottle.
Measure drug substance and place in appropriate dispersion container.

Add the required volume of surfactant / suspension solution. Place on magnetic stirrer until drug is completely wet.
The suspended drug is transferred to a clean mixing container and then the dispersing container is washed with PBS and placed in this container.
If used, add the required amount of suspending agent solution to the mixing vessel.
Bring the final volume to 80-90% with PBS.
Mix for 15 minutes using a homogenizer.
Transfer to weighing cylinder; wash mixing vessel and mixer head with PBS into bulk.
Adjust the volume after defoaming.
Transfer to a clean bottle and place on a magnetic stirrer. The mixing speed is set so that the suspension is homogeneous during filling.
Fill a 2 mL vial with 1 mL suspension using calibrated 1-5 mL.
Cap and cap all vials.
If necessary, sterilize the vial in an autoclave (conditions: 121 ° C. for 15 minutes).

  While the invention has been described with reference to many specific compositions, those skilled in the art will appreciate that the invention can be embodied in other specific forms without departing from the spirit of the invention. Accordingly, those skilled in the art will appreciate that the invention is not limited by the foregoing detailed description, but rather is defined by the appended claims.

1 is a schematic view of an eye. Figure 2 shows the change in concentration after administration of a compound of formula I in the choroid, the target tissue for age-related macular degeneration. Shown are the IVT and sub-Tenon sac test plans in rabbits and primates.

Claims (15)

Formula I, II or III:

Or a pharmaceutically acceptable salt or solvate thereof, wherein the compound of formula I, II or III is from about 1 μg to about 1,500 μg.   2. The dosage form of claim 1, wherein the dosage form further comprises one or more pharmaceutically acceptable excipients.   The dosage form of claim 1, wherein the compound of formula I, II or III is about 50 μg to about 1,000 μg.   2. The dosage form of claim 1, wherein the compound of formula I, II or III is about 100 [mu] g to about 600 [mu] g. Formula I, II or III:

A method of treating an ophthalmic disease in a mammal, comprising administering to the mammal a composition comprising a therapeutically effective amount of a compound of the above, or a pharmaceutically acceptable salt or solvate thereof, The above method of about 1 μg to about 1,500 μg.   6. The method of claim 5, wherein the composition further comprises one or more pharmaceutically acceptable excipients.   6. The method of claim 5, wherein the composition is administered by subtenon delivery to the eye.   Eye diseases include non-exudative (dry AMD) and exudative (wet AMD) age-related macular degeneration (AMD), choroidal neovascularization, retinopathy such as diabetic retinopathy and retinopathy of prematurity, diabetic 6. The method of claim 5, wherein the method is selected from macular edema, retinitis, uveitis, cystic macular edema, glaucoma and other diseases or symptoms of the posterior segment of the eye.   6. The method of claim 5, wherein the therapeutically effective amount is from about 50 μg to about 1,000 μg.   6. The method of claim 5, wherein the therapeutically effective amount is from about 100 [mu] g to about 600 [mu] g.   6. The method of claim 5, wherein the composition is administered 1-12 times a year.   A method for stabilizing vision of a mammal comprising administering the dosage form of claim 1 to the mammal.   A method for improving vision in a mammal comprising administering the dosage form of claim 1 to the mammal.   A method for preventing ocular diseases comprising administering the dosage form of claim 1 to a mammal.   15. The method of claim 14, wherein the eye disease is ocular neovascularization.

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