Sustained-release biodegradable intratubular insert containing hydrogel and active agent

Abstract

Provided herein are sustained-release biodegradable intracanalicular inserts comprising a hydrogel and an active agent, methods for treating or preventing ocular diseases by administering such inserts, and methods for making such inserts.

Description

Technical Field

[0001] The present invention relates to the treatment of eye diseases, such as diseases affecting the eye surface (e.g., dry eye and dry eye disease "DED"). According to the present invention, eye diseases are treated by administering an insert that is biodegradable and that releases an active agent slowly, for example, into the canaliculus.

Background Art

[0002] Eye diseases and disorders, particularly those affecting the eye surface, are widespread. For example, dry eye disease (DED), also known as keratoconjunctivitis sicca, is one of the most common ophthalmic disorders. Patients presenting to an ophthalmic clinic frequently report symptoms of dry eye, which is becoming an increasingly significant public health problem and one of the most common conditions seen by ophthalmologists. The prevalence is significantly increased in the elderly and in women. In the United States, over 16 million adults are estimated to be diagnosed with this disorder, of whom 9 million are classified as moderate to severe.

[0003] DED is a multifactorial disease of the tear film and ocular surface, and as a result, unpleasant symptoms of the eye such as dryness, burning, itching, redness, stinging, blurring, grittiness, pain, foreign body sensation, visual disturbances, tear film instability, eye fatigue, and often damage to the ocular surface may occur. Also, DED can make it difficult or impossible for patients to wear contact lenses, read, perform computer work, or drive at night.

[0004] Inflammation of both the lacrimal gland and the ocular surface has been shown to be involved in dry eye. Factors that adversely affect the stability and osmotic pressure of the tear film may induce ocular surface damage and trigger an inflammatory cascade that activates innate and adaptive immune responses. These immune-inflammatory responses can lead to further damage to the ocular surface and the development of a persistent inflammatory cycle. For example, inflammation of the ocular surface can result in reduced tear production, which can further worsen the condition and lead to inflammation of the ocular surface and damage to epithelial cells. In animal models, T cell-mediated inflammation has actually been both a cause and a consequence of dry eye. In humans, dry eye has been associated with the presence of conjunctival T cells and elevated levels of inflammatory cytokines in tears compared to the control group, supporting the idea that inflammation is the driving force behind the disorder.

[0005] DED is classified into acute, episodic, and chronic. In some cases, it may be classified as chronic with acute flare. Chronic DED may require year-round attention. Several pharmacological therapies have been investigated for DED, including a stepwise approach starting with commercially available lubricants and artificial tear substitutes (delivered as eye drops), progressing to topical anti-inflammatory therapy, and punctal occlusion using punctal plugs to block tear drainage.

[0006] Artificial tears increase tear volume, but frequent administration is necessary because tear drainage and loss can revert to the original state due to evaporation or absorption via the ocular epithelium, for example. Adding viscosity enhancers may increase retention time, but this can cause blurred vision due to the replacement of tears with higher viscosity ones. While punctal plugs have been shown to be effective in DED patients, they can be lost (due to insufficient retention) and, rarely, can migrate to the nasolacrimal duct, potentially leading to inflammation or other serious conditions. In some cases, surgical closure of the puncta by high-temperature cauterization may be performed as part of an attempt to treat DED. Further treatments approved in the United States for DED patients include Restasis® (cyclosporine) (increases tear production), Cequa® (cyclosporine) (increases tear production), and Xiidra® (Lifitegrast) (for the signs and symptoms of DED). More recently, Eysuvis® (lotepredonol) has been approved for the acute treatment of DED.

[0007] Topical administration of cyclosporine A has been shown to increase tear secretion, likely due to its promotion of the local release of parasympathetic neurotransmitters. Cyclosporine eye drops are currently marketed for topical use in multiple products across several jurisdictions, as shown below. [Table 1]

[0008] However, the application of topical solutions is subject to limitations that affect patient management. These limitations include the difficulty of handling the bottles, limitations in the precision of instillation, the potential for discontinuation of the solution, and limitations in the bioavailability of topical eye drops (Aldrich et al, 2013, Ophthalmic preparations, USP, 39(5), pp.1-21). Specific challenges with currently available cyclosporine eye drop formulations include tolerability issues such as burning and stinging sensations, as well as a slow onset of action that can take several weeks to several months. Furthermore, the high frequency of administration (e.g., several times a day) and the significant impact on patients' daily lives are also issues. In humans, the bioavailability of topical eye drops to reach ocular tissue is less than 5%. Other limitations include a slow onset of action (several weeks to several months) and high drug doses in the eye drops, which may cause adverse reactions (e.g., burning sensation in the eye associated with topical cyclosporine eye drops (Restasis® NDA#021023)).

[0009] Therefore, there is an unmet need for forms of ophthalmic treatment that overcome the shortcomings of current commercially available topical formulations, particularly for dosage forms that enable sustained release of drugs (e.g., cyclosporine A) and associated reductions in administration frequency, thereby improving quality of life and patient compliance, while also reducing the risk of infection and adverse effects (e.g., burning and stinging of the eye).

[0010] Drug delivery via punctal plugs is more beneficial than topical eye drops in that it allows for sustained drug release over time by forming a depot from which the drug is slowly released. Administration consists of a single plug injection, which addresses the aforementioned limitations inherent in long-term administration of topical eye drops.

[0011] However, intratubular plugs also present challenges. Intratubular administration routes have certain anatomically implied constraints (they need to be small enough to enter the lacrimal punctum), and developing an ophthalmic intratubular plug that is easy to administer and, if necessary, remove when the drug depot is depleted, fits well, i.e., is held properly so as not to be lost unnoticed, while at the same time causing no discomfort or unintended administration site reactions (e.g., inflammation) is difficult.

[0012] In addition, drug release needs to be appropriate and consistent over a sustained period. Given the small size of the plug, it is difficult to formulate a drug that includes sufficient drug filling and sustained-release properties.

[0013] Given this background, it is clear that there is a demand for alternative forms of administration that are effective for eye diseases such as DED.

[0014] All references disclosed herein are incorporated herein by reference in their entirety for any purpose. [Overview of the project]

[0015] An objective of some embodiments of the present invention is to provide an intratubular implant containing an active agent effective for treating ocular diseases in patients, particularly DED, over a long period of time.

[0016] Furthermore, an object of some embodiments of the present invention is to provide intratubular implants containing active agents effective for treating patients' eye diseases, particularly blepharitis, over a long period of time.

[0017] Furthermore, an object of some embodiments of the present invention is to provide an intratubular implant containing an active agent effective for treating patients' eye diseases, particularly blepharitis, allergic conjunctivitis, particularly atopic keratoconjunctivitis and vernal keratoconjunctivitis, over a long period of time.

[0018] As outlined above, the main drawback of currently commercially available ophthalmic preparations (e.g., topical eye drops) is the need for frequent administration. The present invention aims to address this by enabling effective and long-term treatment by administering only once an insert that continuously releases an active substance over a long period (e.g., several weeks).

[0019] Thus, another object of some embodiments of the present invention is to provide an intrascleral insert comprising an active agent that provides sustained release of the active agent onto the ocular surface.

[0020] To ensure effective treatment during the implantation period of the insert, the release of the active agent should remain at a substantially constant level (e.g., within the therapeutic window) that provides a therapeutic effect.

[0021] Thus, another object of some embodiments of the present invention is to provide an intrascleral insert comprising an active agent that provides sustained release of the active agent onto the ocular surface, particularly a constant release.

[0022] Another object of some embodiments of the present invention is also to provide an intrascleral insert comprising an active agent that acts more rapidly (e.g., within days or even within hours).

[0023] In addition, the present invention also aims to improve patient compliance, which still has room for improvement in current commercial products.

[0024] Therefore, another object of some embodiments of the present invention is to provide an intrascleral insert comprising an active agent that is highly tolerable and does not cause discomfort that cannot be tolerated during or after insertion into the sclera.

[0025] Also, another object of some embodiments of the present invention is to provide an intrascleral insert comprising an active agent that is easy to administer, i.e., can be easily inserted into the sclera.

[0026] Another objective of some embodiments of the present invention is to provide an intravascular insert containing an active agent that is easy to handle and does not overflow or break easily like eye drops.

[0027] Another objective of some embodiments of the present invention is to provide an intravascular insert containing an active agent that avoids overdosage and underdosage because accidental administration by the patient is unlikely to occur.

[0028] Another objective of some embodiments of the present invention is to provide an intravascular insert containing an active agent that fits well into the blood vessel once inserted and is not easily lost or accidentally discharged through the lacrimal duct.

[0029] Another objective of some embodiments of the present invention is to provide an intravascular insert containing an active agent that is easy to remove or replace, or does not need to be removed.

[0030] Therefore, another objective of some embodiments of the present invention is to provide an intravascular insert containing an active agent that has reduced side effects (such as a burning sensation, stinging, or itching in the eye) compared to general treatments such as eye drops.

[0031] Therefore, another objective of some embodiments of the present invention is to provide an intravascular insert containing an active agent that has reduced associated risks (such as eye infection or systemic toxicity) compared to general treatment methods such as eye drops.

[0032] Therefore, another objective of some embodiments of the present invention is to provide an intravascular insert containing an active agent that does not worsen or reduces the deterioration of the quality of life due to treatment-related restrictions (such as wearing contact lenses, or being unable or limited in reading, computer work, or night-time driving).

[0033] Another object of some embodiments of the present invention is to provide an intratubular insert containing an active agent that is easy to manufacture.

[0034] Another object of some embodiments of the present invention is to provide an intratubular insert containing an active agent that is easy to store and stable during storage.

[0035] Another object of some embodiments of the present invention is to provide an intratubular insert containing an active agent that increases tear production and maintains a patient's tear level at an appropriate level over a long period of time.

[0036] Another object of some embodiments of the present invention is to provide an intraocular implant containing an active agent that relieves or reduces, over a long period of time, symptoms of ocular surface diseases, particularly DED, such as dry eyes, burning sensation, itching, redness, stinging, gritty feeling, pain, foreign body sensation, visual impairment, tear film instability, eye fatigue, and ocular surface disorders in patients.

[0037] Another object of some embodiments of the present invention is to provide a method for treating or preventing ocular diseases, particularly DED, in patients over a long period of time, which can address one or more of the problems mentioned in the above-mentioned object.

[0038] Furthermore, an object of some embodiments of the present invention is to provide a method for treating ocular diseases, particularly blepharitis, in patients over a long period of time, which can address one or more of the problems mentioned in the above-mentioned object.

[0039] Furthermore, an object of some embodiments of the present invention is to provide a method for treating ocular diseases in patients, particularly blepharitis, allergic conjunctivitis, particularly atopic keratoconjunctivitis and vernal keratoconjunctivitis, over a long period of time, which can address one or more of the problems mentioned in the above-mentioned object.

[0040] Another object of some embodiments of the present invention is to provide a method for treating or preventing DED, comprising inserting a biodegradable implant comprising a hydrogel and an active agent into a patient's tubule, which can address one or more of the problems mentioned above with respect to the purpose of providing tubule implants.

[0041] Another object of some embodiments of the present invention is to provide a method for treating or preventing DED, comprising inserting a biodegradable implant containing a hydrogel and an active agent into a patient's tubule, and inserting a second biodegradable implant containing a hydrogel and an active agent into the same tubule after a long period of time (e.g., at least about two months), which can address one or more of the problems mentioned above with respect to the purpose of providing tubular implants.

[0042] Another object of some embodiments of the present invention is to provide a method for treating or preventing ocular diseases, particularly DED, in patients over a long period of time, which provides a therapeutic effect that surpasses that achieved by a treatment consisting of a combination of punctal occlusion with drug-free punctal plugs and administration of eye drops.

[0043] Another object of some embodiments of the present invention is to provide a method for manufacturing tube inserts that can address one or more of the problems mentioned above in relation to tube inserts.

[0044] One or more of these objectives of the present invention, and others, are addressed by one or more embodiments disclosed and claimed herein.

[0045] Individual aspects of the present invention are disclosed herein and claimed in independent claims. Dependent claims, on the other hand, claim specific embodiments and variations of these aspects of the present invention. Details of various aspects of the present invention are shown in the following embodiments for carrying out the invention. [Brief explanation of the drawing]

[0046] [Figure 1] This is a schematic diagram of the eye and lacrimal gland system. [Figure 1.2A] The dried and hydrated insert fibers prepared in Example 1.2 (Execution 1 and Execution 2) are shown. [Figure 1.2B] The dried and hydrated insert fibers prepared in Example 1.2 (Execution 3) are shown. [Figure 1.2C] The dried and hydrated insert fibers prepared in Example 1.2 are shown (low, medium, and high doses). The main portion of the prepared dried insert fibers exhibited a particulate, cylindrical shape with no visible surface defects, regardless of dose (left). In addition, the insert with the most pronounced surface deformation for each dose is also shown (right). [Figure 1.3] The dried and hydrated insert fibers prepared in Example 1.3 are shown. [Figure 1.4A] The dry density and drug filling amount of each insert are shown. [Figure 1.4B] This shows the dry diameter of each insert. [Figure 1.4C] This shows the dry density of each insert. [Figure 1.4D] Microscopic images of each insert are shown. [Figure 1.4E] This graph shows the dry diameter of the insert fibers prepared in Example 1.4, dependent on the hydration time. [Figure 2] This chromatogram shows the conversion from NHS-fluorescein to the fluorescein-trillidine conjugate. [Figure 3.1] The results of the Schirmer tear test in Example 3.4, which tracked tear production over time, are shown. [Figure 3.2] A and B show the cyclosporine A (CsA) concentrations over time measured in the tears of the beagle in Example 3.5. [Figure 3.3] This shows the cyclosporine A (CsA) concentration over time measured in the tear fluid of a beagle in Example 3.6. [Figure 4A] This diagram shows a general overview of the human clinical trial for Example 4. [Figure 4B] This diagram shows a general schematic example of implant placement within the canal of the human eye. [Figure 5] Figures A through D show schematic diagrams illustrating the steps required to insert an implant into the canal of the human eye. [Figure 6] The results of tracking tear production over time using the Schirmer tear test described in Example 4.1 are shown for the test eye (A) and the non-test eye (B). The solid black line represents the average Schirmer score for all individuals analyzed, and the dashed line represents the Schirmer score for a single individual. [Figure 7] A and B track Example 4.1 over time, showing the total corneal fluorescein staining (tCFS) values ​​(mean values ​​across all eyes) in absolute terms and as changes over time from baseline. [Figure 8A] The results of the visual analog scale (VAS) for the severity of eye dryness are shown in absolute values, following Example 4.1 over time. [Figure 8B] Example 4.1 was tracked over time, and the results of the visual analog scale (VAS) for the severity of eye dryness are shown as changes from baseline. [Figure 9] The results of the dry eye frequency score on the Visual Analog Scale (VAS) for Example 4.1 were tracked over time, and the absolute values ​​are shown. [Figure 10] The OSDI results for Example 4.1 are shown as mean and absolute values, tracked over time. [Figure 11] The SPEED score results for Example 4.1 are shown as mean and absolute values, tracked over time.

[0047] definition As used herein, the term “intratubular implant” refers to an object containing an active agent, i.e., administered to, i.e., inserted into, the lacrimal canaliculi, and remaining there for a certain period of time while releasing the active agent into the surrounding environment. The implant may have any predetermined shape before insertion, often a rod-like shape, and this shape can be maintained to some extent once the implant is placed in the desired position, although the dimensions of the implant (e.g., length and / or diameter) may change after administration due to hydration, as further disclosed herein. In other words, what is inserted into the eye is not a solution or suspension, but a pre-formed, tightly fitting object. Thus the implant is fully formed before administration, for example, according to the method disclosed herein. Intratubular implants can be designed to be biodegradable over time (as disclosed below), so that they soften, change shape, and / or decrease in size, and are eventually removed by either dissolution or disintegration, in which case the remainder of the implant can be discharged into the lacrimal duct. In the present invention, the term “insertion” is used to refer to both the insertion in a hydrated state (for example, after the insertion has been administered to the eye or immersed in an aqueous environment by another means and (re)hydrated) and the insertion in a dry (dried / dehydrated) state (for example, after it has been dried to a low moisture content (e.g., 1% by weight or less)).

[0048] In the present invention, the term "eye" refers to the eye as a whole, or any part or section of the eye (since "intraocular implants" can, in principle, be administered to any part or section of the eye). In some embodiments, the present invention focuses on intratubular injection of intraocular implants and the treatment of dry eye disease (DED), as further disclosed below.

[0049] The term "biodegradable" refers to a material or object (e.g., an intraductal insert according to the present invention) that decomposes in vivo, i.e., when placed in the body of a human or animal. In the context of some embodiments of the present invention, as disclosed in detail below herein, an insert comprising a hydrogel containing an active agent slowly biodegrades over time when administered into the intraductal canaliculus of the eye. In some embodiments, biodegradation occurs, at least in part, by ester hydrolysis in the aqueous environment of the canaliculus. The insert slowly softens and disintegrates, resulting in clearance through the nasolacrimal duct.

[0050] A "hydrogel" is a three-dimensional network of one or more hydrophilic natural or synthetic polymers (as disclosed herein) that swells in water and holds a certain amount of water, while maintaining or substantially maintaining its structure, for example, by chemical or physical crosslinking of individual polymer chains. Hydrogels are soft and flexible due to their high water content and are therefore very similar to natural tissues. In the present invention, the term "hydrogel" is used to refer to both a hydrated hydrogel (for example, after it has been formed in an aqueous solution, or after it has been inserted into an eye or immersed in an aqueous environment by another means to be hydrated or (re)hydrated) and a dried (dried / dehydrated) hydrogel (for example, after it has been dried to a low water content (e.g., 1% by weight or less)). In the present invention, if the active ingredient is contained within the hydrogel (e.g., dispersed within it), the hydrogel may also be referred to as a "matrix".

[0051] The term "polymer network" refers to a structure formed from polymer chains (with the same or different molecular structures and the same or different molecular weights) that are crosslinked with each other. Types of polymers suitable for the purposes of the present invention are disclosed below in this specification.

[0052] The term "amorphous" refers to polymers or polymer networks that do not exhibit a crystalline structure in X-ray or electron scattering experiments.

[0053] The term "semi-crystalline" refers to a polymer or polymer network that possesses some crystalline properties, that is, one that exhibits some crystalline properties in X-ray or electron scattering experiments.

[0054] In this specification, the term “precursor” refers to a molecule or compound that reacts with each other, thereby bonding via crosslinking to form a polymer network, and thus a hydrogel matrix. Other materials, such as active agents or buffers, may be present within the hydrogel, but these are not referred to as “precursors.”

[0055] The precursor molecular portions that still exist in the final polymer network are also referred to herein as “units.” Thus, “units” are the building blocks or constituents of the polymer network that forms the hydrogel. For example, a polymer network suitable for use in the present invention may contain identical or different polyethylene glycol units, as further disclosed herein.

[0056] For the purposes of this invention, the term “sustained-release” is intended to characterize a product that is formulated to make the active agent available over a long period of time, thereby enabling a reduction in the frequency of administration compared to immediate-release formulations (e.g., a solution of the active agent applied topically to the eye (i.e., eye drops)). Other terms that may be used interchangeably with “sustained-release” herein include “long-term release” or “controlled release.” In the sense of this invention, the term “sustained-release” includes constant release of the active agent, gradually decreasing release of the active agent, and any combination thereof (e.g., gradually decreasing release of the active agent after a constant release). In the sense of this invention, the terms “gradually decreasing” or “gradually decreasing” refer to a decrease in the release of the active agent over time.

[0057] As used herein, the term “long duration” means any period of time that a person skilled in the art would consider to be an extended treatment of a disease, in particular a period of at least about one week, or at least about one month, for example up to about 12 months, or any intermediate period, for example, about 1 to about 6 months, about 2 to about 4 months, about 2 to about 3 months, or about 3 to about 4 months.

[0058] As used herein, the term “insertion time” refers to the period during which an intratubular implant is present in the tube, i.e., the period from administration of the implant until removal of the implant from the tube. In some embodiments, removal of the implant can be achieved by removal of the implant (which may be intentional or unintentional, but is not expected to occur spontaneously unless an external force is applied), or by spontaneous clearance after a long period of time, once the implant has completely biodegraded, completely disintegrated, or substantially disintegrated and the remaining portion(s) of the implant have been discharged. In some embodiments, the insertion time is at least about one week, or at least about one month or more, for example, up to about 12 months, or any intermediate period, for example, about 1 to about 6 months, about 2 to about 4 months, about 2 to about 3 months, or about 3 to about 4 months.

[0059] As used herein, the term “visualizer” refers to a molecule or composition contained within the hydrogel of an implant that facilitates the visualization of the implant when it is inserted into the canaliculus of the eye. The visualizer may be a fluorophore, such as fluorescein, rhodamine, coumarin, and cyanine. In some embodiments, the visualizer is fluorescein or comprises a fluorescein portion.

[0060] As used herein, “surface of the eyeball” includes the conjunctiva and / or cornea, as well as elements such as the lacrimal apparatus (including the puncta and lacrimal canaliculi) and associated eyelid structures.

[0061] As used herein, the terms “tears” or “tears” refer to the fluid secreted by the lacrimal glands that lubricates the eye and forms the tear film. Tears are composed of water, electrolytes, proteins, lipids, and mucin.

[0062] As used herein, the terms “administer,” “insert,” “administer,” and “insert” are used synonymously in the context of the present invention and refer to the placement of an implant in the lacrimal canaliculi, particularly in the vertical portion of the canaliculi, following a procedure such as that described in Example 4.II (Insertion Placement).

[0063] As used herein, the terms “bilateral” or “bilateral” refer to the administration of the implant (in the context of the administration of the implant of the present invention) to both eyes of the patient. The implant may be inserted independently in each eye into the vertical portion of the superior canaliculus or the vertical portion of the inferior canaliculus, or into both the vertical portions of the superior and inferior canaliculus.

[0064] The term "plug" refers to a device that can prevent or reduce tear drainage by causing obstruction, substantial obstruction, or partial obstruction ("lacrimal duct obstruction") of the lacrimal duct, thereby helping to keep the eye moist. Plugs can be classified into "punctal plugs" and "intratubular plugs." Intratubular plugs are also referred to as "tubular plugs" in the literature. Both plug classifications are inserted through the superior and / or inferior lacrimal punctum of the eye. Because punctal plugs remain in the punctal opening, they are easily visible and therefore can be removed without great difficulty. However, the retention rate of punctal plugs is insufficient, and because of their exposed structure, they can be contaminated with microorganisms, which can rarely lead to infection. In contrast, intratubular plugs are not visible and are placed in both the vertical and horizontal parts of the tubercle, resulting in a better retention rate than punctal plugs. However, intratubular plugs are not easily removed and carry a higher risk of migration. Commercially available plugs are often made of collagen, acrylic, or silicone.

[0065] The terms “canaliculus” (plural: canaliculi) or, alternatively, “lacrimal duct” as used herein, refer to the lacrimal duct, i.e., the small channels in each eyelid that drain tears from the punctum to the nasolacrimal duct (see also Figure 1). Thus, the canaliculus form part of the lacrimal apparatus that drains tears from the surface of the eye into the nasal cavity. The canaliculus of the upper eyelid is called the “supra canaliculus” or “upper canaliculus,” and the canaliculus of the lower eyelid is called the “inferior canaliculus” or “lower canaliculus.” Each canaliculus includes a vertical region that continues to the punctum (referred to as the “vertical canaliculus”) and a horizontal region that continues to the vertical canaliculus (referred to as the “horizontal canaliculus”), the horizontal canaliculus which joins the nasolacrimal duct.

[0066] The term "punctum" (plural: puncta) refers to the lacrimal punctum, a tiny opening at the edge of the eyelid, which is the entrance to a small canalicle. When tears are produced, some of the fluid evaporates during blinking, and some is drained through the punctum. Since both the upper and lower eyelids have punctums, these are called the "upper punctum" or "supralapical punctum" and the "lower punctum" or "subcutaneous punctum," respectively (see also Figure 1).

[0067] The term “intratubular implant” refers to an implant that can be administered to the superior or inferior canaliculi of the eye, or both, through the superior or inferior punctum, or through both the superior and inferior punctum, particularly to the vertical portion of the superior or inferior canaliculi, or both. Because the implant is positioned within the canaliculi, it blocks tear drainage by obstructing the lacrimal gland, as is also observed with intratubular plugs. In some embodiments of the present invention, the intratubular implant is inserted bilaterally into the vertical portion of the inferior canaliculi. According to some embodiments of the present invention, the intratubular implant is a sustained-release biodegradable implant.

[0068] The terms “API,” “active(pharmaceutical) ingredient,” “active(pharmaceutical) agent,” “effective(pharmaceutical) ingredient,” “(active) therapeutic agent,” “active,” and “drug” are used interchangeably herein and refer to substances used in a final pharmaceutical product (FPP) and substances used in the preparation of such a final pharmaceutical product, which are intended to impart pharmacological activity or otherwise have a direct effect on the diagnosis, cure, alleviation, treatment, or prevention of a disease or on the restoration, correction, or modification of a patient’s physiological function.

[0069] The active agents used in accordance with the present invention may be active agents for the treatment and / or prevention of diseases or disorders of the eyeball, particularly those affecting the surface of the eyeball. In some embodiments of the present invention, the active agent is used for the treatment and / or prevention of dry eye disease, blepharitis, allergic conjunctivitis, particularly atopic keratoconjunctivitis and vernal keratoconjunctivitis. In certain embodiments of the present invention, the active agent is a low-solubility active agent (i.e., having a solubility in water of less than about 1000 μg / mL or less than about 100 μg / mL).

[0070] In some embodiments, the active agent is selected from immunomodulators / immunosuppressants, particularly calcineurin inhibitors, steroids, specifically synthetic glucocorticoids, nonsteroidal anti-inflammatory drugs (NSAIDs), antibiotics, antihistamines, as well as essential fatty acids such as omega-3 fatty acids, and other agents such as rifitegrast.

[0071] Examples of immunomodulatory / immunosuppressant agents include cyclosporine, everolimus, tacrolimus, sirolimus, and pimecrolimus. Examples of NSAIDs include ibuprofen, mefanamic acid, diclofenac, nepafenac, flurbiprofen, and flurbiprofen sodium. Examples of antibiotics include fusidic acid, becifloxacin (base), clarithromycin, and azithromycin. Examples of antihistamines include ketotifen (base), azelastine (base), and azelastine emponate. Examples of essential fatty acids include linoleic acid and alpha-linolenic acid. Examples of synthetic glucocorticoids include prednisone, prednisolone, prednisolone acetate, methylprednisolone, dexamethasone, dexamethasone acetate, sodium betamethasone phosphate, budesonide, flunisolide, fluticasone propionate, triamcinolone, triamcinolone acetonide, triamcinolone diacetate, fluocinolone acetonide, fludrocortisone acetate, loteprednol, loteprednol etavonate, difluprednate, fluorometholone, mometasone fluate, deoxycorticosterone acetate, aldosterone, rimexolone, beclomethasone, and beclomethasone dipropionate.

[0072] In some embodiments, the API used according to the present invention is cyclosporine A. In some other embodiments, the active agent used herein is not cyclosporine A.

[0073] As used herein, the term "cyclosporine" refers to cyclosporine A, and not, in particular, to cyclosporine B, C, D, E, H, and L, which are metabolites of cyclosporine A, nor to cyclosporine U, G, dihydrocyclosporine A, or isocyclosporine A, which may be present in cyclosporine A as impurities. In some embodiments, cyclosporine A may contain cyclosporine B, C, D, E, G, H, L, and U, as well as dihydrocyclosporine A and isocyclosporine A, as impurities at concentrations of 1.0% or less, or 0.7% or less, respectively, and may also contain unknown impurities at concentrations of 0.3% or less, or 0.1% or less, respectively, with impurities totaling 2.5% or less, or 1.5% or less in total.

[0074] The molecular formula for cyclosporine A is C 62 H 111 N 11 O 12 Its IUPAC name is 30-ethyl-33-(1-hydroxy-2-methylhexa-4-enyl)-1,4,7,10,12,15,19,25,28-nonamethyl-6,9,18,24-tetrakis(2-methylpropyl)-3,21-di(propan-2-yl)-1,4,7,10,13,16,19,22,25,28,31-undecaazacyclotriacontane-2,5,8,11,14,17,20,23,26,29,32-undecone (CAS number 59865-13-3). Its molecular weight is 1203 daltons and it has the following chemical structure. [ka]

[0075] Cyclosporine is a white to substantially white powder that is soluble in various organic solvents (e.g., acetone, methanol, ethanol (96% v / v)) but nearly insoluble in water. In some embodiments, cyclosporine is micronized.

[0076] For the purposes of the present invention, the active agent can be used in all possible forms, including polymorphs or any pharmaceutically acceptable salts, anhydrides, hydrates, other solvates, or derivatives. In this description or claims, whenever the active agent is mentioned without further specification, it also refers to the active agent in any such polymorph, pharmaceutically acceptable salt, anhydride, solvate (including hydrate), or derivative form, even if not explicitly stated. With respect to the active agent, preferred solid forms include, without limitation, pure substance forms in any physical form known to those skilled in the art. For example, the active agent may take the form of particles. The particles may be amorphous or crystalline, or a mixture of these two forms, and can be made of any size which can be classified without limitation as coarse particles, fine particles, or ultrafine particles, the dimensions which can be seen particularly with the naked eye or under a microscope, and can have shapes such as single grains and aggregates. The particles can also be micronized. As used herein, the term “micronization” refers, without limitation, to small-sized particles, particularly microscopic-scale particles, whose particle size has been reduced by, for example, jet milling, jaw crushing, hammer milling, wet milling, precipitation in a non-solvent, cryomilling (milling with liquid nitrogen or dry ice), and ball milling. The active agent may also exist in a dissolved or dispersed state, for example, in a solvent or aqueous medium, or in the form of particles dispersed in, for example, an aqueous suspension (which may optionally contain excipients such as surfactants).

[0077] As used herein, the term “therapeutically effective” refers to the amount of active agent required to produce the desired therapeutic outcome after administration. For example, in the context of the present invention, one desired therapeutic outcome could be a reduction in symptoms associated with DED (measured, for example, by in vivo tests known to those skilled in the art), such as an increase in Schirmer tear test scores, a decrease in staining values ​​measured by conjunctival lysamine green staining or corneal fluorescein staining, a decrease in scores for dry eye severity and / or dry eye frequency on a visual analog scale (VAS), a decrease in the ocular surface disease index and / or standard patient-assessed score for dry eye, and a decrease in best corrected visual acuity. In one embodiment, “therapeutically effective” refers to a concentration of the active agent in a sustained-release biodegradable intratubular implant that, in terms of therapeutic effect, can achieve a tear film concentration equivalent to a cyclosporine concentration of 0.236 μg / mL (considered necessary for immunomodulation (Tang-Liu and Acheampong, Clin. Pharmacokinet. 44(3), pp. 247-261)), and once this tear film concentration is achieved, can be maintained over a long period, particularly substantially throughout the entire remaining period of implantation.

[0078] As used herein, the values ​​"d10", "d50", "d90", and "d100" refer to values ​​that characterize the amount of particles in a particle size distribution that satisfy a particular particle size. In a given particle size distribution, 10% of the particles have a particle size of d10 or less, 50% of the particles have a particle size of d50 or less, 90% of the particles have a particle size of d90 or less, and substantially all particles have a particle size of d100 or less. The percentages can be expressed by different parameters known to those skilled in the art, for example, the percentages can be based on volume, weight, or the number of particles. Thus, d50 may exemplary be a volume-based, weight-based, or number-based median particle size. For example, if volume-based d90 is 43 μm, it means that 90 volume percent of the particles have a particle size of 43 μm or less. In some embodiments, d10, d50, and d90 are volume-based values. The particle size distribution (PSD) can generally be measured by methods known to those skilled in the art, including sieving and laser diffraction. In some embodiments, the PSD is a USP <429> Particle size is measured by optical diffraction, specifically by laser diffraction. In some embodiments, PSD is measured by laser diffraction using a Beckman Coulter LS 13 320, with an obscuration value in the range of 7–9% based on the optical model Fraunhofer rf780z.

[0079] As used herein, the term “approximately” in relation to a measured quantity refers to the normal variation in that measured quantity that can be expected by a person skilled in the art when performing a measurement and exercising a level of care appropriate to the purpose of the measurement and the precision of the measuring instrument.

[0080] The term "at least about" in relation to a measured quantity refers to the normal variation in the measured quantity and amounts higher than the measured quantity that can be expected by a person skilled in the art when performing a measurement and exercising a level of care appropriate to the purpose of the measurement and the precision of the measuring instrument.

[0081] As used herein, unless otherwise explicitly indicated by the context, the singular forms "a," "an," and "the" refer to multiple objects.

[0082] In this specification, the term "and / or" as used in expressions such as "A and / or B" is intended to include both "A and B" and "A or B".

[0083] Open terms such as "include," "including," "contain," and "containing" all mean "comprising." These unrestricted transition clauses are used to introduce an unrestricted list of elements, method steps, etc., that do not exclude additional unlisted elements or method steps. [Modes for carrying out the invention]

[0084] I. Inserts into small tubes The intratubular inserts of the present invention, according to some embodiments, provide sustained release, are biodegradable, and contain a hydrogel and an active agent.

[0085] As outlined in the definition section above, providing sustained release means, in the context of this invention, that the implant can make the active agent available over an extended period. The implant is administered intraocularly and slowly releases the active agent into the tear film. The latter is slowly renewed as new tears are produced from the lacrimal gland and replaced with existing tears on the surface of the eyeball as they are drained through the lacrimal ducts, so the active agent present in the implant is slowly released into the tear film each time the eye blinks, without requiring any action from the patient. Thus, this implant provides a convenient hands-free alternative to conventional eye drops.

[0086] Typically, sustained release can dramatically reduce the frequency of multiple daily administrations of existing products (as the product is in the form of a topical eye drop that is easily rinsed away), as it is maintained for several weeks, for example. This means that patients can benefit from the therapeutic effects of the insert without having to remember to self-administer eye drops multiple times a day, which is a significant advantage in itself, as well as reducing the risk of misadministration due to improper instillation or mishandling / administration, and the risk of infection from repeated use of eye drop bottles.

[0087] Furthermore, as outlined in the definition section above, in some embodiments of the present invention, once administered, the implants are designed to slowly biodegrade over a predetermined period of time. This means that the implant can remain in the duct and does not need to be explanted. Normally, there is no need to remove the implant, and the patient simply needs to leave it in until it is removed. On the other hand, if an unexpected event occurs, such as an allergic reaction, discomfort during insertion, or other adverse event (e.g., irritation), the (partially biodegraded) implant can be removed by applying slight pressure to expel it from the punctum or by moving it further down the duct and removing it through the nasolacrimal duct. The ability and / or ease of removal is also advantageous if the active agent in the implant is depleted at the end of the insertion period and therefore needs to be replaced with a new implant to maintain the therapeutic effect.

[0088] Furthermore, the tube-insertions of the present invention also include hydrogels. Hydrogels, as described in detail in the definition section, can absorb water and transition from a dry state to a hydrated state. In some embodiments, the hydration of the hydrogel results in a change in the shape of the insert. In certain embodiments, the insert expands in diameter and shrinks in length, so that a thin, rod-shaped insert in its dry state can be easily inserted into the tube, and once administered and properly positioned, it expands in diameter within the tube, resulting in a secure fit and reducing the risk of the insert moving or being lost. Because the hydrated insert is soft, it is comfortable to wear even though it is securely fixed in place.

[0089] In some embodiments, the hydrogel includes a polymer network. Further details of the polymer network are provided below.

[0090] The principle by which such highly expandable hydrogel plugs are securely positioned is disclosed, for example, in US8,409,606 (incorporated herein by reference in all purposes, and in case of any conflict, this specification shall prevail).

[0091] Active ingredients: Details of the active agent are provided in the definition section above. As outlined there, in some embodiments the active agent has low solubility in water, and therefore, although we do not wish to be bound by theory, it is hypothesized that contact with tear fluid regulates the drug release rate due to the combination of low drug solubility under physiological conditions (approximately 10 μg / mL) and the limited cross-sectional area of ​​the insert in contact with tear fluid and the volume of tear fluid.

[0092] On the other hand, the form and amount of the active drug embedded in the hydrogel can still affect the dissolution of the active drug and the attainment of the therapeutically effective release rate.

[0093] One aspect of some embodiments of the present invention is a sustained-release biodegradable intratubular insert comprising a hydrogel and an active agent, wherein the active agent is in the form of particles and the particles of the active agent are dispersed within the hydrogel.

[0094] In some embodiments of the present invention, the active agent particles are uniformly dispersed within a hydrogel. As used herein, dispersed within a hydrogel means the active agent particles exist in a substantially pure form embedded within a matrix, but without excluding small amounts of the active agent found on the surface of the matrix. In some embodiments, the active agent particles contain no further excipients other than the active agent and any impurities that may be present in the active material used, and more specifically, encapsulate the drug and form a separate hydrophobic phase, which does not refer to any microspheres, microparticles, or hydrophobic microdomains containing further materials such as the oils, fats, fatty acids, waxes, carbon fluoride, or other water-immiscible phases suggested earlier. The active agent existing in a substantially pure form has the advantage of being easy to manufacture, as it does not require further processing of the active substance to prepare, for example, microspheres, microparticles, or hydrophobic microdomains.

[0095] In some embodiments, the active agent may have a drying loss of 1.5% w / w or less per 100 mg in a capillary stoppered bottle under vacuum at a pressure not exceeding 5 mm of mercury column at 60°C, a heavy metal content of 0.002% or less, organic impurities as defined in the product specifications section of Example 2, specifically the total of all impurities quantified by HPLC of 1.5% or less, an active agent content of 97.0% to 101.5% as quantified by HPLC, residual acetone of 4500 ppm or less as measured by GC headspace, and residual ethyl acetate of 2000 ppm or less.

[0096] In some embodiments and as shown in the examples (see Example 2 in particular), the inserts of the present invention are stable during storage, and the content of the active agent does not substantially change during long-term storage.

[0097] Therefore, in some embodiments, the active agent content measured by HPLC after at least 3 months, at least 6 months, or at least 12 months at a temperature of 2–8°C, and the initial active agent content measured directly by HPLC before storage, are approximately 300–410 μg.

[0098] In some embodiments, the active agent content measured by HPLC after at least 3 months, at least 6 months, or at least 12 months at a temperature of 2–8°C is within 90–110% by weight, or 95–105% by weight, or 98–102% by weight, of the initial active agent content measured directly by HPLC before storage.

[0099] In some embodiments, the amount of impurities measured by HPLC after at least 3 months, at least 6 months, or at least 12 months at a temperature of 2-8°C is 3.0% or less.

[0100] In some embodiments of the present invention, the active agent takes the form of particles. In some embodiments, the particles are micronized particles. While we do not wish to be bound by theory, it is thought that dispersed particles of the active agent, particularly small particles of the active agent, allow for sufficiently rapid dissolution of the active agent and thus enable rapid onset of action.

[0101] In some embodiments of the present invention, the active agent particles have a d50 value of less than about 50 μm, a d90 value of less than about 43 μm, or a d100 value of less than about 45 μm, as measured by laser diffraction. As shown in the examples (see Example 1.4), larger particles are thought to affect the mechanical properties of the tube insert. In addition, larger particles tend to block the tube, making it difficult or impossible to cast the precursor mixture (which will be further outlined below).

[0102] In some embodiments of the present invention, the particles of the active agent have a d50 value in the range of about 3 to about 17 μm, or about 4 to about 12 μm, or about 5 to about 8 μm. As shown in the examples (see Example 1.4), particle size has a considerable effect on the density, swelling behavior, and surface quality of the insert, and the high density and smoother insert surface that can be achieved by smaller particles should be evaluated in comparison to the superior hydration and swelling behavior of larger particles.

[0103] In some embodiments, the d50, d90, and d100 values ​​refer to the values ​​of active agent particles used in the manufacture of the insert, or the values ​​of active agent particles present in the insert.

[0104] Regarding the amount of active agent contained in the implant, a high concentration of the active agent in the implant is desirable in some embodiments. This is because it allows for a higher dose of the active agent in the same implant size (resulting in a longer and / or sustained release at a constant rate, as will be discussed further below), or a smaller product size for the same dose, the latter being preferable in terms of ease of administration and comfort of insertion. On the other hand, as shown in Example 1.1, the concentration has a significant impact on the quality of the implant. That is, both concentrations that are too high and too low tend to result in implants that are "straw-like" with a large dry diameter and hollow structure. In addition, lower drug concentrations appear to lead to improved swelling / hydration behavior.

[0105] In some embodiments of the present invention, the insert contains, in a dry state, an active agent in an amount of about 15% to about 80% by weight, or about 30% to about 65% by weight, or about 45% to about 55% by weight, based on the total weight of the insert.

[0106] One aspect of the present invention is a sustained-release biodegradable intratubular insert comprising a hydrogel and an active agent, wherein the insert, in a dry state, contains about 40% to about 80% by weight of the active agent based on the total weight of the insert.

[0107] In terms of the absolute amount of activity, dose is an important factor in achieving sustained release.

[0108] In some embodiments of the present invention, the insert contains an amount of active agent ranging from about 100 μg to about 800 μg.

[0109] The active agent is included in the insert of the present invention in doses ranging, for example, about 100 μg to about 800 μg, about 100 μg to about 300 μg, about 300 μg to about 450 μg, or about 500 μg to about 800 μg. Any amount within these ranges, for example, about 250 μg, about 360 μg, about 600 μg, or about 670 μg (all values ​​also include dispersions of +25% and -20%, or + / -15%, or + / -10%), may be used.

[0110] The disclosed amount of the active ingredient (including any variations mentioned) refers to both the final content of the active ingredient in the insert and the amount of the active ingredient used as the starting component of the insert when manufacturing the insert.

[0111] Polymer network: As described above, in some embodiments, the hydrogel comprises a polymer network. The hydrogel can be formed from a precursor having functional groups that form crosslinks to create such a polymer network. These crosslinks between polymer strands or arms can be substantially chemical (i.e., covalent) and / or physical (e.g., ionic, hydrophobic association, hydrogen crosslinking, etc.).

[0112] Polymer networks can be prepared from one type of precursor or from two or more types of reactable precursors. The precursors are selected considering the desired properties of the resulting hydrogel. A variety of suitable precursors exist for use in the preparation of hydrogels. Generally, any pharmaceutically acceptable, crosslinkable polymer that forms a hydrogel can be used for the purposes of this invention. The hydrogel and its constituent elements (including the polymers used to prepare the polymer network) should be physiologically safe, for example, not to induce an immune response or other adverse effects. Hydrogels can be formed from natural polymers, synthetic polymers, or biosynthetic polymers.

[0113] Examples of natural polymers include glycosaminoglycans, polysaccharides (e.g., dextran), polyamino acids, proteins, or mixtures or combinations thereof.

[0114] Synthetic polymers can generally be any polymer produced synthetically from various raw materials by different types of polymerization (including free radical polymerization, anionic or cationic polymerization, chain growth or addition polymerization, condensation polymerization, ring-opening polymerization, etc.). Polymerization can be initiated by some initiator, light and / or heat, and can also be mediated by a catalyst.

[0115] In general, for the purposes of the present invention, one or more synthetic polymers from the group comprising one or more units of polyalkylene glycol may be used, and include polyethylene glycol (PEG), polyethylene oxide, polypropylene oxide, polyvinyl alcohol, poly(vinylpyrrolidinone), polylactic acid, polylactic acid-co-glycolic acid, random or block copolymers, or any combination / mixture thereof (however, this list is not intended to be limiting).

[0116] Precursors can be covalently crosslinked to each other to form a covalently crosslinked polymer network. In some embodiments, a precursor having at least two reaction centers (for example, in free radical polymerization) can function as a crosslinking agent because each reactive group can participate in the formation of different grown polymer chains.

[0117] The precursor may have a biologically inert and hydrophilic portion, such as a core. In the case of branched polymers, the core refers to a continuous portion of molecules linked to arms extending from the core, where the arms often have functional groups at the ends of the arms or branches. Multi-armed PEG precursors are examples of such precursors, which are further disclosed below herein.

[0118] Therefore, a hydrogel for use in the present invention can be prepared, for example, from one multi-arm precursor having a first functional group(s) (or set) and another multi-arm precursor having a second functional group(s) (or set). For example, the multi-arm precursor may have hydrophilic arms terminated with a primary amine (e.g., polyethylene glycol units) (nucleophilic) or activated ester-terminated groups (electrophilic). The polymer network according to the present invention may contain identical or different polymer units that are crosslinked with each other. The precursors may be high molecular weight components (e.g., polymers with functional groups) or low molecular weight components (e.g., low molecular weight amines, thiols, esters, etc.).

[0119] Some functional groups can be made more reactive by using activating groups. Examples of such activating groups include (but are not limited to) carbonyl diimidazole, sulfonyl chloride, aryl halides, sulfosuccinimidyl esters, N-hydroxysuccinimidyl (NHS) esters, succinimidyl esters, epoxides, aldehydes, maleimides, imide esters, and acrylates. NHS esters are useful groups for crosslinking nucleophilic polymers, such as primary amine-terminated or thiol-terminated polyethylene glycols. NHS-amine crosslinking reactions can be carried out in aqueous solution in the presence of a buffer (e.g., phosphate buffer (pH 5.0-7.5), triethanolamine buffer (pH 7.5-9.0), borate buffer (pH 9.0-12), or sodium bicarbonate buffer (pH 9.0-10.0)).

[0120] In some embodiments, each precursor may contain only nucleophilic functional groups or only electrophilic functional groups, insofar as both the nucleophilic and electrophilic precursors are used in the crosslinking reaction. For example, if the crosslinking agent has only nucleophilic functional groups (e.g., amines), the precursor polymer may have electrophilic functional groups (e.g., N-hydroxysuccinimide). On the other hand, if the crosslinking agent has electrophilic functional groups such as sulfosuccinimide, the functional polymer may have nucleophilic functional groups such as amines or thiols. Therefore, the polymer network of the present invention can also be prepared using functional polymers (e.g., proteins, poly(allylamines), or amine-terminated bifunctional or polyfunctional poly(ethylene glycols)).

[0121] In one embodiment of the present invention, precursors for polymer networks forming hydrogels in which an active agent is dispersed to form inserts according to the present invention each have about 2 to about 16 nucleophilic functional groups (referred to as functionality), and in another embodiment, precursors each have about 2 to about 16 electrophilic functional groups (referred to as functionality). Reactive precursors having a number of reactive (nucleophilic or electrophilic) groups that is a multiple of 4, so for example, 4, 8, and 16 reactive groups, are particularly suitable for the present invention. Any number of functional groups (e.g., including any of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16 groups) is possible in order to ensure that the precursor is used according to the present invention and that the functionality is sufficient to form a suitable crosslinked network.

[0122] PEG hydrogel: In some embodiments of the present invention, the polymer network forming the hydrogel comprises polyethylene glycol (PEG) units. PEG is known in the art to form hydrogels when crosslinked, and these PEG hydrogels are suitable for pharmaceutical applications, for example, as a matrix for drugs intended to be administered to any part of the human or animal body.

[0123] The polymer network of the hydrogel insert of the present invention may include 2 to 10 arms, or 4 to 8 arms, or one or more multi-arm PEG units having 4, 5, 6, 7, or 8 arms. In some embodiments, the PEG units used in the hydrogel of the present invention have 4 and / or 8 arms. In some specific embodiments, a 4-arm PEG is utilized.

[0124] The number of arms of the PEG used contributes to controlling the flexibility or softness of the resulting hydrogel. For example, a hydrogel formed by crosslinking 4-arm PEGs is generally softer and more flexible than one formed from 8-arm PEGs of the same molecular weight. More specifically, if it is desired to stretch the hydrogel before (or after) drying, as will be disclosed later in this specification in the section on the manufacture of inserts, more flexible PEG units, such as 4-arm PEGs, may be used in combination with other multi-arm PEGs (e.g., 8-arm PEGs as disclosed above, or other (different) 4-arm PEGs).

[0125] In some embodiments of the present invention, the polyethylene glycol units used as a precursor have an average molecular weight in the range of about 2,000 to about 100,000 daltons, or in the range of about 10,000 to about 60,000 daltons. In some specific embodiments, the polyethylene glycol units have an average molecular weight in the range of about 10,000 to about 40,000 daltons. In a specific embodiment, the polyethylene glycol units used to produce a hydrogel according to the present invention have a molecular weight of about 20,000 daltons.

[0126] The molecular weight of polyethylene glycol and polyethylene glycol derivatives can be quantified by several methods, including gel electrophoresis (e.g., SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis)), gel permeation chromatography (GPC), GPC by dynamic light scattering (DLS), and matrix-assisted laser desorption / ionization time-of-flight (MALDI-TOF) spectroscopy. The molecular weight of the polyethylene glycol precursors disclosed herein can be quantified by any method known to those skilled in the art, including SDS-PAGE, GPC, and MALDI-TOF, and more specifically, by GPC using PEG standards with known molecular weight (e.g., quantified by MALDI-TOF) and polydispersity (e.g., quantified by GPC). MALDI-TOF can be used when high precision is required.

[0127] The molecular weight of polyethylene glycol refers to the average molecular weight, which can be selected from various average values ​​known to those skilled in the art (including number-average molecular weight (Mn), weight-average molecular weight (Mw), and peak-average molecular weight). Any of these average values, particularly the three average molecular weights mentioned above, can be used in the context of the present invention. In some embodiments, the average molecular weight of polyethylene glycol units and precursors disclosed herein is indicated as the number-average molecular weight.

[0128] As used herein, a multi-armed PEG unit having a specified molecular weight may be abbreviated in the form of, for example, 4a20kPEG, which refers to a 4-armed PEG unit with a molecular weight of 20,000.

[0129] In 4-arm PEG, each arm can have an average arm length (or molecular weight) obtained by dividing the total molecular weight of PEG by 4. Therefore, the 4a20k PEG precursor, which is particularly suitable for use in the present invention, has four arms, each with an average molecular weight of approximately 5,000 Daltons. The 8a20k PEG precursor, which can also be used in combination with or as an alternative to the 4a20k PEG precursor, therefore has eight arms, each with an average molecular weight of 2,500 Daltons. Longer arms may be more flexible than shorter arms. PEGs with longer arms may swell more than PEGs with shorter arms. Also, PEGs with fewer arms may swell more and be more flexible than PEGs with more arms. In some specific embodiments, only 4-arm PEG precursors are used in the present invention. In some specific embodiments, two different 4-arm PEG precursors are used in the present invention. In some other embodiments, a combination of 4-arm PEG precursors and 8-arm precursors is used in the present invention. In addition, longer PEG arms result in a higher melting point when dry, which may improve dimensional stability during storage.

[0130] In some embodiments, the electrophilic end groups for use with the PEG precursor for the preparation of the hydrogel of the present invention are N-hydroxysuccinimidyl (NHS) esters, which include, but are not limited to, NHS dicarboxylic acid esters (e.g., succinimidylmalonic acid group, succinimidylmaleic acid group, succinimidyl fumarate group), "SAZ" (referring to a succinimidylazelaic acid end group), "SAP" (referring to a succinimidyladipic acid end group), "SG" (referring to a succinimidylglutaric acid end group), and "SS" (referring to a succinimidosuccinate end group).

[0131] Therefore, in some embodiments, the PEG precursor is an NHS dicarboxylic acid ester-terminated multi-arm PEG precursor that can be represented by the following formula:

[0132] [ka] In the formula, n is determined by the molecular weight of each PEG arm, m is an integer from 0 to 10, specifically 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and x is the number of arms (thus, for example, 2, 4, 8, etc.; see above). When m is 1, each arm is terminated with a succinimidylsuccinate (SS) end group; when m is 2, each arm is terminated with a succinimidylglutaric acid (SG) group; when m is 3, each arm is terminated with a succinimidyladipic acid (SAP) group; and when m is 6, each arm is terminated with a succinimidylazelaic acid (SAZ) group. Using these specific electrophilic end groups, a multi-arm PEG unit can be abbreviated in the form, for example, 4a20k PEG-SAP, which refers to a 4-arm PEG with succinimidyladipic acid end groups and a molecular weight of 20,000 (4 arms, each approximately 5,000 daltons). In the above formula, R is a suitable core structure for providing the desired number of arms. In the case of a 4-arm PEG unit and precursor, R may be a pentaerythritol structure, and in the case of an 8-arm PEG unit and precursor, R may be a hexaglycerol structure.

[0133] In some embodiments, the PEG precursor is 4a20k PEG-SG or 4a20k PEG-SAP.

[0134] In some embodiments, the nucleophilic terminal group for use with the electrophile-containing PEG precursor for the preparation of the hydrogel of the present invention is an amine (indicated as "NH2") terminal group. Thiol (-SH) terminal groups or other nucleophilic terminal groups are also possible.

[0135] In some embodiments, a 4-armed PEG having an average molecular weight of about 20,000 daltons and electrophilic end groups as disclosed above (e.g., SAZ, SAP, SG, and SS end groups) are crosslinked to form a polymer network and, consequently, a hydrogel according to the present invention.

[0136] For example, as a result of a reaction between a nucleophile-containing crosslinking agent and an electrophile-containing PEG unit (for example, a reaction between an amine-containing crosslinking agent and an activated ester-containing PEG unit), multiple PEG units are crosslinked via amide groups by the crosslinking agent.

[0137] In the case of PEG having an NHS-ester terminal group (e.g., succinimidylazelaic acid (SAZ)-, succinimidyladipic acid (SAP)-, or succinimidylglutaric acid (SG)-terminal PEG unit (see above)), as a result of the reaction with an amine-containing crosslinking agent, multiple PEG units are crosslinked by the crosslinking agent, resulting in formula (1) [ka] The linkage is via a hydrolyzable linker having (wherein m is an integer from 0 to 10, specifically 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10). For a SAZ-terminated group, m is 6. For an SAP-terminated group, m is 3; for an SG-terminated group, m is 2; and for an SS-terminated group, m is 1.

[0138] In certain embodiments, the present invention utilizes SG or SAP-terminated groups. SG-terminated groups can shorten the time it takes for the hydrogel to biodegrade compared to the use of other succinimidyl-terminated groups (other than SS groups), such as SAZ-terminated groups. SAZ-terminated groups provide a greater number of carbon atoms in the linker and are therefore more hydrophobic, potentially making them less susceptible to ester hydrolysis than SG-terminated groups.

[0139] In some embodiments, an electrophile-containing multi-arm polymer precursor, particularly a multi-arm PEG precursor having SG or SAP terminal groups (as defined above), is crosslinked with a nucleophile-containing crosslinking agent. The nucleophile can be an amine, particularly a primary amine.

[0140] In some embodiments, the nucleophile-containing crosslinking agent is a nucleophile-containing multi-arm polymer precursor.

[0141] In some other embodiments, the crosslinking agent used is a low molecular weight component containing a nucleophilic terminal group (e.g., an amine or thiol terminal group). In some embodiments, the nucleophilic crosslinking agent is a small molecular weight amine with a molecular weight of 1,000 Da or less containing two or more primary aliphatic amine groups. Specific crosslinking agents for use in the present invention include, for example, dyridine, torilidine, tetralyzine, ethylenediamine, 1,3-diaminopropane, 1,3-diaminopropane, diethylenetriamine, trimethylhexamethylenediamine, their pharmaceutically acceptable salts, hydrates, and derivatives such as conjugates (provided that sufficient nucleophiles for crosslinking are still present), and any mixtures thereof. In some preferred embodiments, torilidine is used as the crosslinking agent. As used herein, torilidine should be understood to refer to any form of torilidine (including torilidine salts such as torilidine acetate, or torilidine derivatives such as labeled torilidine).

[0142] In some embodiments, the nucleophile-containing crosslinking agent is a labeled crosslinking agent, and in particular, labeled trilysine. The crosslinking agent can be labeled with a visualizing agent, for example, to help a physician confirm the presence or absence of an implant during a control test. Fluorophores such as fluorescein, rhodamine, coumarin, and cyanine can be used as visualizing agents to label the crosslinking agent. Labeling can be achieved, for example, by chemical conjugation, and in particular by using the nucleophile of the crosslinking agent for conjugation with the label. Since a sufficient amount of nucleophile (at least more than 1 molar equivalent) is required for crosslinking, the terms "conjugated" or "conjugated" generally include partial conjugation, meaning that only a portion of the nucleophile is used for conjugation with the label. Thus, in some embodiments, the crosslinking agent is trilysine-labeled by partial conjugation with a visualizing agent, and in particular, about 1% to about 20%, or about 5% to about 10%, or about 8%, of the trilysine amine groups are conjugated with the visualizing agent.

[0143] In some embodiments, the nucleophile-containing crosslinking agent is fluorescein-conjugated trilysine. Fluorescein-conjugated trilysine can be obtained by reacting trilysine acetate with N-hydroxysuccinimide (NHS)-fluorescein.

[0144] In some embodiments, the multi-arm polymer unit comprises a 4a20k PEG unit, and the crosslinked unit comprises a fluorescein-conjugated trilysidineamide unit.

[0145] In some embodiments, the polymer network is obtained by reacting 4a20k PEG-SG with fluorescein-conjugated trilysine in a molar ratio ranging from about 1:2 to about 2:1. In some other embodiments, the polymer network is obtained by reacting 4a20k PEG-SS or 4a20k PEG-SAZ with fluorescein-conjugated trilysine in a molar ratio ranging from about 1:2 to about 2:1.

[0146] In some embodiments, the molar ratio of nucleophilic and electrophilic end groups that react with each other is approximately 1:1, i.e., one amine group is provided for each electrophile (e.g., SG or SAP end group). In the case of 4a20k PEG-SG or 4a20k PEG-SAP and fluorescein-linked trilysine as electrophile-containing polymer units, since trilysine has three primary amine groups that can react with the electrophilic SG or SAP ester group, assuming partial conjugation of trilysine using one of the four primary amines on average, the resulting molar ratio of these two components is approximately 1:1. However, an excess of either the electrophilic end group (e.g., NHS end group such as SG) precursor or the nucleophilic (e.g., amine) end group precursor may be used.

[0147] Therefore, in some embodiments, the polymer network can be obtained by reacting 4a20k PEG-SG with fluorescein-conjugated trilysine in a molar ratio ranging from about 1:2 to about 2:1, particularly in a molar ratio of about 1:1.

[0148] In some embodiments, the polymer network can be obtained by reacting 4a20k PEG-SAP with fluorescein-conjugated trilysine in a molar ratio ranging from about 1:2 to about 2:1, particularly in a molar ratio of about 1:1.

[0149] Surfactants: One aspect of the present invention is a sustained-release biodegradable intratubular insert comprising a hydrogel and an active agent, wherein the insert comprises a surfactant.

[0150] In some embodiments, as further outlined above, the active agent is a hydrophobic active agent substantially miscible with the hydrophilic hydrogel material, and in some embodiments, exists in the form of dispersed particles having a specific particle size. Certain particle sizes have been shown to be advantageous in terms of the properties of the insert. However, smaller particles also tend to aggregate. While we do not wish to be bound by theory, the presence of a surfactant is thought to prevent aggregation and improve the uniformity of the hydrogel contents.

[0151] In addition, experimental results show that the presence of surfactants helps prevent undesirable tube adhesion of the cast hydrogel during the manufacturing of the insert (see Example 1.2), thereby improving the quality of the insert.

[0152] Therefore, in some embodiments, the insert contains a surfactant.

[0153] In some embodiments, the insert contains, in a dry state, about 0.01% to about 5% by weight of surfactant, or about 0.01% to about 2% by weight, or about 0.2% to about 2% by weight, or about 0.05% to about 0.5% by weight, based on the total weight of the insert.

[0154] As shown in Example 1.2, the type of surfactant may be important considering its ability to prevent aggregation of active agent particles. In some embodiments, the inserts include nonionic surfactants. Nonionic surfactants may include poly(ethylene glycol) chains. Exemplary nonionic surfactants that can be used herein include poly(ethylene glycol) monolaurate sorbitan (in particular Tween® 20 of PEG-20-monolaurate sorbitan or Tween® 80 of PEG-80-monolaurate sorbitan), commercially available as Tween®, castor oil poly(ethylene glycol) esters (in particular Cremophor 40 of PEG-40-castor oil), and ethoxylated 4-tert-octylphenol / formaldehyde condensation polymers, commercially available as tyloxapol.

[0155] Additional ingredients: The inserts of the present invention may include, in addition to the polymer units, active ingredients, and surfactants that form the polymer network disclosed above, other additional components. Such additional components are, for example, salts resulting from buffers used during the preparation of hydrogels, such as phosphates, borates, bicarbonates, or other buffers such as triethanolamine. In some embodiments of the present invention, sodium phosphate buffers (specifically monobasic sodium phosphate and dibasic sodium phosphate) are used.

[0156] Optionally, preservatives may be used in the implants of the present invention. However, as demonstrated in the examples by the storage stability test data and clinical results indicating the safety of the implants, the implants of the present invention do not require the presence of preservatives, as opposed to, for example, some eye drops. Since preservatives are also thought to cause discomfort to the subject (e.g., stinging and irritation of the eye), in one embodiment of the present invention, the implants are either preservative-free or essentially preservative-free.

[0157] formulation: In some embodiments, the insert according to the present invention comprises an active agent, a polymer network made from one or more polymer precursors disclosed herein in the form of a hydrogel, and an optional additional component (e.g., a surfactant, and salts remaining in the insert from the production process (e.g., phosphates used as buffers, etc.)).

[0158] In some embodiments, the insert according to the present invention, in a dry state, comprises about 15% to about 80% by weight of an active agent and about 20% to about 60% by weight of polymer units based on the total weight of the insert, or about 30% to about 65% by weight of an active agent and about 25% to about 50% by weight of polymer units based on the total weight of the insert, or about 45% to about 55% by weight of an active agent and about 37% to about 47% by weight of polymer units based on the total weight of the insert.

[0159] In one further specific embodiment, on a dry weight basis, the active agent:PEG ratio is approximately 40% by weight of PEG to about 50% to 60% by weight of active agent, based on the total weight of the insert, with the remainder being phosphates and other excipients.

[0160] In some embodiments, the residue of the dry insert (i.e., the remainder of the formulation when the active agent, polymer hydrogel (e.g., PEG hydrogel), and an optional surfactant have already been considered) may be residual salts from the buffer solution as disclosed above. In some embodiments, such salts may be phosphates, borates, or (bi)carbonates. In one embodiment, the buffer salt is sodium phosphate (monobasic and / or dibasic).

[0161] The amounts of the active agent and polymer(s) can be varied, and inserts according to the present invention can be prepared using other amounts of the active agent and polymer hydrogel.

[0162] In some embodiments, the amount of drug in the formulation is less than about twice the amount of polymer (e.g., PEG) units, but in some cases it may be higher. However, it is desirable that the mixture containing, for example, the precursor, buffer, and drug (before the hydrogel has completely gelled) can be uniformly cast into the mold or tube.

[0163] In one embodiment of the present invention, the hydrogel contains about 3% to about 20% polyethylene glycol (equivalent to the weight of polyethylene glycol ÷ the weight of the fluid × 100) after it is formed and before it is dried, i.e., in a wet state. In one embodiment, the hydrogel contains about 7.5% to about 15% polyethylene glycol (equivalent to the weight of polyethylene glycol ÷ the weight of the fluid × 100) in a wet state.

[0164] In some embodiments, a solid content of about 10% to about 30% (w / v) (where “solid” means the total weight of the polymer precursor(s), salts, and drugs in the solution) is used to form a hydrogel of the insert according to the present invention.

[0165] In some embodiments, the water content of the hydrogel in its dry (dehydrated / dried) state (e.g., before insertion into the canaliculus of the eye) may be very low (e.g., less than 1% by weight of water). In other words, in some embodiments, the insert contains less than about 1% by weight of water in its dry state. In some embodiments, the water content may be even lower, for example, less than 0.25% by weight or less or less than 0.1% by weight of the total weight of the insert.

[0166] Dimensions of the insert, and dimensional changes during hydration due to stretching: The dried inserts can have various forms depending on the manufacturing method (e.g., the use of a mold or tube to cast a mixture containing a hydrogel precursor with an active agent before complete gelation). In one embodiment, the insert has a cylindrical or essentially cylindrical shape and a circular or essentially circular cross-section. The shape of the insert may also be described as fibrous (because the length of the cylinder greatly exceeds its diameter) or rod.

[0167] Polymer networks (for example, PEG networks in hydrogel inserts according to certain embodiments of the present invention) may be semi-crystalline in a dry state below room temperature and amorphous in a wet state. Even in a stretched form, dry inserts may be dimensionally stable below room temperature, which may be advantageous for inserting the insert into a tube.

[0168] When the implant is hydrated in the eye (which can be simulated by immersing the implant in PBS at 37°C and pH 7.4), the dimensions of the implant according to the present invention may change, generally with the diameter increasing while the length decreases or at least remains the same or essentially the same. The advantage of this dimensional change is that the implant is thin enough to be easily inserted into the tubercle in its dry state, but once placed in the tubercle, it shortens, improving comfort of wear for the short vertical portion of the tubercle and the corresponding limited available space, as well as increasing in diameter, allowing it to fit snugly against the tubercle wall and secure the implant in place, thus preventing unintended movement and loss of the implant. The implant can also soften, allowing for comfortable wear despite the tight fit. In some embodiments, the dimensional change is made possible, at least in part, by a “shape memory” effect introduced by stretching the implant longitudinally during the manufacturing of the implant (also disclosed in the “Manufacturing Method” section below). In some embodiments, stretching can be performed either in a dry or wet state, i.e., after or before drying the hydrogel insert. It should be noted that if the hydrogel insert is simply dried and cut to the desired length without stretching, the dimensions of the insert may not change substantially, or both the diameter and length of the insert may increase during hydration. If this is undesirable, the hydrogel fibers can be dry-stretched or wet-stretched, i.e., stretched before or after drying. In particular, the fibers can be stretched before drying.

[0169] In the case of pre-formed dry hydrogels, some degree of molecular orientation can be imparted by dry-stretching the material, then solidifying it, and fixing the molecular orientation. In some embodiments, this can be achieved by stretching the material (optionally while heating the material to a temperature above the melting point of the crystallizable region of the material), and then crystallizing the crystallizable region. As an alternative, in some embodiments, the glass transition temperature of the dry hydrogel may be used to fix the molecular orientation of a polymer such as PVA having a suitable glass transition temperature. Yet another alternative is to stretch the gel before it is completely dry (also referred to as "wet stretching"), and then dry the material under tension. This molecular orientation results in a mechanism by which it swells anisotropically when introduced into a hydration medium such as a vitreous body. During hydration, in some embodiments, the insert swells only in the radial dimension, while its length decreases, is maintained, or is essentially maintained. The term "anisotropic swelling" refers to a phenomenon where a cylindrical object swells predominantly in one direction, but hardly swells (or even shrinks) in the longitudinal direction, while swelling is predominantly in the other direction.

[0170] The degree of dimensional change during hydration may depend, in particular, on the stretch coefficient. As used herein, the stretch coefficient refers to the coefficient by which the hydrogel is stretched in measurements in the stretching direction, i.e., the change in length (not the change in diameter) immediately before and after stretching, without considering any further incidental dimensional changes (e.g., due to drying or rehydration). For example, stretching with a stretch coefficient of approximately 1.3 (e.g., by wet stretching) may result in little effect or little change in length during hydration. In contrast, stretching with a stretch coefficient of approximately 1.8 (e.g., by wet stretching) may result in a significant decrease in length during hydration. For example, stretching with a stretch coefficient of 4 (e.g., by dry stretching) may result in a much smaller length during hydration (e.g., a reduction in length from 15 mm to 8 mm). Those skilled in the art will understand that other factors besides stretching may also affect the swelling behavior.

[0171] One aspect of the present invention is a sustained-release biodegradable intratubular insert comprising a fibrous hydrogel and an active agent, wherein the fibers are stretched.

[0172] In some embodiments, the fibers are stretched with a longitudinal stretch coefficient of about 1.0 to about 4.0, or about 1.5 to about 3.0, or about 2.7.

[0173] The composition of the polymer network is among the other factors that influence the stretching of the hydrogel and the possibility of changing the dimensions of the implant during hydration. When PEG precursors are used, precursors with fewer arms (e.g., 4-arm PEG precursors) contribute more to the flexibility of the hydrogel than precursors with more arms (e.g., 8-arm PEG precursors). If the hydrogel contains a large amount of less flexible components (e.g., a large amount of PEG precursors with more arms (e.g., 8-arm PEG units)), the hydrogel may become stiffer and more difficult to stretch without breakage. On the other hand, a hydrogel containing more flexible components (e.g., PEG precursors with fewer arms (e.g., 4-arm PEG units)) may be easier to stretch and softer, but may also swell more during hydration. Therefore, the behavior and properties of the implant once it is placed in the eye (i.e., when the hydrogel is (re)hydrated) can be tuned by altering the structural characteristics and by changing the treatment of the implant after it has been initially formed.

[0174] Exemplary dimensions of the inserts used in the following examples of this specification are shown in Table 2.1 of the Examples section. The dimensions of the dried insert depend, in particular, on the amount of active agent incorporated and the active agent:polymer unit ratio, and can also be controlled by the diameter and shape of the mold or tube used to gel the hydrogel. Furthermore, the diameter of the insert is further determined, in particular, by the (wet or dry) stretching of the hydrogel fibers once formed. The dried fibers (after stretching) are cut into segments of the desired length to form the insert. Thus, the diameter and length of the insert can be adapted as desired. On the other hand, the anatomical dimensions of the lacrimal canaliculi impose certain dimensional requirements on the intracanaliculi inserts.

[0175] In some embodiments, the insert takes the form of a fiber. The fiber may have an average length of about 1.5 mm to about 4.0 mm and an average diameter of 0.8 mm or less in its dry state, an average length of about 2.0 mm to about 2.5 mm and an average diameter of 0.62 mm or less in its dry state, or an average length of about 2.5 mm to about 2.9 mm and an average diameter of 0.62 mm or less in its dry state.

[0176] In some embodiments and as shown in the examples (see Example 2 in particular), the inserts of the present invention are stable during storage, and the dimensions of the product do not change or substantially change during long-term storage.

[0177] Therefore, in some embodiments, the insert, after being stored at a temperature of 2-8°C for at least 3 months, at least 6 months, or at least 12 months, takes the form of fibers having an average length of about 2.5 mm to about 2.9 mm and an average diameter of 0.62 mm or less in its dry state.

[0178] The dimensions of the implant can be adjusted, for example, by selecting an appropriate active agent concentration, taking into account the dosage of the active agent incorporated into the implant. The dimensions of the implant can be reduced by increasing the active concentration. On the other hand, the active substance concentration affects not only the properties and quality of the implant, such as swelling behavior or diameter after drying, but also its release behavior (see Example 1.1 in particular). Surprisingly, the inventors have found that in some embodiments, certain combinations of implant dimensions, active concentration, and / or dosage not only result in high-quality implants with appropriate swelling behavior that facilitates manufacturing, administration, and comfort of wear, but also provide effective release over a long period.

[0179] Considering the above, one aspect of the present invention is a sustained-release biodegradable tubular insert comprising about 360 μg of hydrogel and an active agent, which takes the form of a fiber (or cylinder) having an average length of about 2.5 mm to about 2.9 mm and an average diameter of about 0.62 mm or less in its dry state. Another aspect of the present invention is a sustained-release biodegradable tubular insert comprising hydrogel and an active agent in an amount of about 45% to about 55% by weight based on the total weight of the insert, which takes the form of a fiber (or cylinder) having an average length of about 2.5 mm to about 2.9 mm and an average diameter of about 0.62 mm or less in its dry state.

[0180] In addition to such inserts, the inserts of the present invention in some other embodiments may also decrease in length and increase in diameter upon in vivo hydration in the eye, i.e., in the lacrimal canaliculi, or in vitro (where in vitro hydration is measured after 24 hours in phosphate-buffered saline at pH 7.4, 37°C), reaching an average diameter of at least 1.0 mm in an expanded state 10 minutes after in vitro hydration in phosphate-buffered saline at pH 7.4, 37°C, or an average diameter of at least 1.3 mm in an equilibrium state 24 hours after in vitro hydration in phosphate-buffered saline at pH 7.4, 37°C. In one embodiment, this dimensional change can be achieved by dry-stretching the hydrogel fibers with a draw coefficient of about 1 to about 4, or a coefficient of about 1.5 to about 3.0, or a coefficient of about 2.7.

[0181] In certain embodiments, this stretching induces shape memory, meaning that the insert shrinks in length (also called snapback) and expands in diameter until it approaches (approximately) an equilibrium dimension determined by the original molded dimensions and compositional parameters upon hydration when administered into the lacrimal canaliculi. The narrower dry dimensions facilitate insertion of the product into the canaliculi, while the expansion in diameter and reduction in length after administration results in a shorter, thicker insert that is comfortable to wear, yet securely fixed in place, minimizing the risk of unintended movement. Thus, in one embodiment, the present invention also relates to a method for imparting shape memory to a hydrogel mixture fiber containing particles of an active agent, wherein the active agent is dispersed within the hydrogel, and the shape memory is imparted by stretching the hydrogel mixture fiber longitudinally.

[0182] In vitro release: The in vitro release of the active drug from the insert of the present invention can be quantified by various methods, for example, by replacing a volume of PBS (phosphate-buffered saline, pH 7.4) at 37°C daily under non-sink simulated physiological conditions, similar to the volume of human eye tears.

[0183] In vitro release tests can be used to compare different inserts (e.g., from different manufacturing batches, with different compositions, and with different dosage intensities) with each other for purposes such as quality control or other qualitative evaluations.

[0184] In vivo release and persistence: In one embodiment of the present invention, when a dried insert of the present invention is inserted into a tubule, it hydrates as disclosed above, changes in size, and then completely disintegrates, biodegrading and disintegrating over time until any residue is discharged into the tear duct. As the insert biodegrades (e.g., through ester hydrolysis), it may gradually swell and soften. As the inventors have observed from clinical trials presented in the following Examples section of this specification, inserts according to some embodiments of the present invention can last for several months, for example, about 2 to about 4 months or longer, and also allow for the sustained release of the active agent over several months.

[0185] After the implant has completely disintegrated, any remaining undissolved active drug particles can be excreted through the tear drainage system. Therefore, the duration of sustained release can be designed, among other things, by adjusting the disintegration time, provided that the implant contains a sufficient amount of active drug to last for the time required for the implant to completely disintegrate. In some embodiments, if two implants are used to treat one eye, for example, one implant in each of the lower and upper canaliculi, to achieve the desired total dose, these can be designed to disintegrate over the same or substantially the same time.

[0186] Within the lacrimal canaliculi, the implants of the present invention in some embodiments disintegrate within a long period of time, for example, within about 1 to 6 months after insertion, or within about 2 to 4 months after insertion, or within about 2 to 3 months after insertion, or within about 3 to 4 months after insertion. This has been demonstrated in clinical trials (see the Examples section, particularly Example 4).

[0187] In one embodiment, the implant, after being inserted into the tube, releases a therapeutically effective amount of active drug over a period of at least about one month, at least about two months, at least about three months, or at least four months after insertion.

[0188] In one embodiment of the present invention, the active agent is released from the implanted material at an average rate of approximately 0.1 μg / day to approximately 10 μg / day, or approximately 1 μg / day to approximately 5 μg / day, or approximately 2 μg / day to approximately 4 μg / day.

[0189] In one embodiment of the present invention, the active agent is released from the implant after insertion into a human subject at an average rate of approximately 0.1 μg / day to approximately 10 μg / day, or approximately 1 μg / day to approximately 5 μg / day, or approximately 2 μg / day to approximately 4 μg / day.

[0190] As presented in the Examples section of this specification, preclinical studies in animals and clinical studies in humans have shown that, in some embodiments, the inserts of the present invention can continuously release therapeutically effective amounts of the active agent over a long period of time until the insert is completely disintegrated. However, in certain embodiments, the entire amount of the active agent contained in the insert is released from the insert before the insert is completely biodegraded.

[0191] One aspect of the present invention is a sustained-release biodegradable intratubular implant comprising a hydrogel and an active agent dispersed within the hydrogel, wherein the implant, after being inserted into a tube, releases a therapeutically effective amount of the active agent over a period of at least about three months after insertion.

[0192] In some embodiments, the tear fluid concentration of the active drug after insertion into a human subject is in the range of approximately 0.1 μg / mL to approximately 10 μg / mL.

[0193] In some embodiments, the tear film concentration of the active agent after insertion of the implant is in the range of approximately 0.1 μg / mL to approximately 10 μg / mL, or approximately 1 μg / mL to approximately 5 μg / mL.

[0194] In some embodiments, the insert disintegrates within the tube before the active agent particles contained within the insert are completely dissolved.

[0195] In some embodiments, the implants of the present invention can be designed to disintegrate within the lacrimal canaliculi within approximately 1 to 6 months after insertion, or within approximately 2 to 4 months after insertion, or within approximately 2 to 3 months after insertion, or within approximately 3 to 4 months after insertion.

[0196] In one embodiment, when the polymer network of the hydrogel is crosslinked based on linking groups derived from NHS ester-terminated groups (e.g., SG, SAP, or similar groups disclosed above), the persistence of the hydrogel in an aqueous environment and within a small tube depends, in particular, on the hydrophobicity of the carbon chain adjacent to the degradable ester group. In the case of the inserts used herein, this carbon chain contains three or four carbon atoms, as it is derived from the SG and SAP functional groups of the 4a20k PEG precursor. This allows for long-term persistence of about two to three months, or about three to four months, in the human eye. In other embodiments, hydrogel inserts that are biodegradable in the human eye and have similar or different persistence to the inserts exemplified in the examples can be prepared using different precursors and crosslinking agents than 4a20kPEG-SG / SAP.

[0197] In some embodiments, the hydrogel inserts soften over time with degradation, which may depend, in particular, on the structure of the linkers that crosslink the PEG units within the hydrogel. Inserts formed from 4a20k PEG-SAZ and 8a20k PEG-NH2, as used in the embodiments of this application, soften fairly slowly over time.

[0198] Mechanism of release: While we do not wish to be bound by theory, the mechanism by which the active agent is released from the insert of the present invention can be explained as follows.

[0199] As outlined above, in some embodiments, the hydrophobic active agent is virtually miscible with the hydrophilic hydrogel material. Once inserted and positioned within the tube, the insert comes into contact with the tear fluid, which slowly absorbs and permeates the hydrophilic hydrogel. Biodegradation, i.e., hydrolysis of the hydrogel matrix, softens the hydrogel, allowing further permeation by the tear fluid. However, the hydrophobic active substance, particularly in the form of uniformly dispersed active agents, remains encapsulated within the hydrogel matrix and is released by slow distribution into the hydrogel due to its low solubility in aqueous solution.

[0200] Furthermore, while we do not wish to be bound by any particular theory, it is thought that the tear fluid at the top of the tuberotubular insert results in a fluid column that tends to allow the release of the active substance to be limited by the cross-sectional area of ​​the proximal portion of the plug. The tuberotubular wall appears to release the drug at a much slower rate than the depletion of the therapeutic agent through the fluid column, and / or the tuberotubular wall may become saturated with the drug, thus slowing the release from the wall. Therefore, in some embodiments, the tuberotubular inserts of the present invention do not require, and more specifically, do not include, any complex and more difficult-to-manufacture barrier or reservoir systems (e.g., coatings on the side walls of the insert that block the release of the active substance and limit it to release from the cross section) as has been conventionally proposed.

[0201] Drug release from the cross-section of the implant into the tear film may first occur in the outer region of the hydrogel in contact with the liquid environment (i.e., drug particles located in the region of the hydrogel closer to the punctum dissolve and diffuse first, while particles located at the other end of the implant closer to the tear duct diffuse last). As a result, in some embodiments, the region of the hydrogel closer to the punctum becomes devoid of drug particles. This region is therefore also called the "clearance zone," and it is limited to dissolved drug and has a concentration below the drug's solubility.

[0202] In some embodiments, when the drug dissolves and diffuses from the hydrogel to form a clearance zone, this area of ​​the hydrogel becomes void and softer and more brittle. Simultaneously with the diffusion of the drug from the hydrogel, the hydrogel can also be slowly degraded, for example, by ester hydrolysis in the aqueous environment of the eye. This degradation occurs uniformly throughout the entire bulk of the hydrogel. As the degradation stage progresses, distortion and erosion (also referred to herein as disintegration) of the hydrogel begin to occur. When this happens, the hydrogel becomes softer and more liquid (and thus distorts its shape), and eventually the hydrogel completely disintegrates, with any remaining hydrogel fragments and / or active substances being removed by the tear drainage system.

[0203] In some embodiments, if the active agent is a drug with relatively low solubility, undissolved particles of the active agent may remain when the insert has completely disintegrated; that is, particles of the active agent contained in the insert may disintegrate in the tube before they are completely dissolved. However, as outlined above, in such cases, any remaining active particles are removed by the tear drainage system.

[0204] However, in one embodiment, the entire amount of the active agent is released before the hydrogel completely disintegrates. The release of the active agent from the hydrogel can be maintained at a relatively constant rate because the hydrogel can hold the active agent particles in place and prevent aggregation. II. Manufacturing of Inserts

[0205] In some embodiments, the present invention also relates to a method for producing sustained-release biodegradable intratubular inserts comprising a hydrogel and an active agent disclosed herein. In some embodiments, the method is a) A step of preparing a precursor mixture comprising a hydrogel precursor and particles of an active agent dispersed in the precursor mixture, b) A step of molding a precursor mixture, crosslinking the hydrogel precursor to form a polymer network, and obtaining a molded hydrogel mixture containing the polymer network, c) The step of drying the hydrogel mixture to obtain an insert.

[0206] In one embodiment, the active agent particles can be used in a micronized form to prepare the insert, i.e., used in the form of micronized particles and dispersed to prepare a precursor mixture in which the micronized particles are homogeneously dispersed. In another embodiment, the active agent can be used in a non-micronized form to prepare the insert. Further details of the active ingredient are disclosed in detail above and apply to the active substance used in the manufacture in all embodiments.

[0207] Precursors for forming hydrogels in some embodiments are disclosed in detail in the section relating to the inserts themselves.

[0208] In some embodiments, in step a), the precursor mixture is prepared by mixing an electrophile-containing multi-arm polymer precursor with a nucleophile-containing crosslinking agent in a buffered aqueous solution in the presence of particles of a particulating activator.

[0209] In some embodiments, in step a), the buffered aqueous precursor solution is prepared by dissolving a multi-arm polymer precursor in an aqueous buffer solution, and then mixed with a buffered aqueous precursor suspension containing particles of a nucleophilic crosslinking agent and a micronizing agent, for example, within 60 minutes.

[0210] When preparing a crosslinked PEG network using a PEG precursor, a method for producing the insert in some embodiments may include mixing an electrophile-containing multi-arm polyethylene glycol (e.g., 4a20k PEG-SG or 4a20k PEG-SAP) with a nucleophile-containing crosslinking agent (e.g., trilysine) in a buffered aqueous solution in the presence of particles of a micronizing activator.

[0211] In some embodiments, the molar ratio of electrophiles to nucleophiles in the PEG precursor is approximately 1:1, but it is also possible to use a molar ratio in the range of approximately 1:2 to 2:1, where the nucleophiles (e.g., amine groups) are in excess of the electrophiles, or vice versa. In some embodiments, the method involves reacting 4a20k PEG-SG or 4a20k PEG-SAP with fluorescein-conjugated trilysine in a weight ratio in the range of approximately 30:1 to approximately 50:1.

[0212] As shown in Example 1.4, it was found that treating the precursor mixture by vacuum degassing had a significant impact on the quality of the insert. Specifically, it was possible to prevent aggregation of the active agent particles. Therefore, in some embodiments, in step a), the precursor mixture containing the active agent particles is degassed under vacuum after its components have been mixed.

[0213] In some embodiments, once the precursor mixture is prepared as outlined in step a) above, the mixture can be molded in step b) by casting it into a suitable mold or tube before complete gelation to obtain the final shape of the desired hydrogel, i.e., in step b), the molding of the precursor mixture consists of filling the mold or tube with the precursor mixture before complete crosslinking to obtain the final shape of the desired hydrogel mixture, and crosslinking the hydrogel precursor.

[0214] When the insert should have a fibrous shape, the reactive mixture can be filled into a fine-diameter tube (e.g., a polyurethane (PU) tube) to obtain an elongated cylindrical shape. Depending on the desired final cross-sectional geometric shape of the hydrogel fibers, their initial diameter (which may be further reduced by stretching), and the ability of the reactive mixture to uniformly fill the tube, various tube geometric shapes and diameters can be used.

[0215] Therefore, the inside of the tube may have a circular geometric shape or a non-circular geometric shape (for example, a cross-shaped geometric shape). The tube may have a circular geometric shape with an inner diameter of, for example, about 1 mm to about 3 mm or about 2.0 mm.

[0216] In some embodiments, after forming a hydrogel and curing it until it is completely gelled, the hydrogel can be stretched longitudinally in a wet or dry state, as already disclosed in detail in the section relating to the dimensional change of the insert during hydration above. In some embodiments, the stretch coefficient can be in the range of about 1 to about 4.5, or other ranges as also disclosed above. In some embodiments, when dry stretching is performed, the hydrogel is first dried and then stretched. In some embodiments, when wet stretching is performed, the hydrogel is stretched in a wet (substantially not dry) state and then dried under tension. Optionally, heat may be applied during stretching. Further optionally, the fibers may be further twisted.

[0217] In some embodiments, the insert can be obtained by preparing a precursor mixture containing a hydrogel precursor and an active agent, filling the precursor mixture into a tube, crosslinking the hydrogel precursor within the tube to obtain a hydrogel mixture formed as fibers, and stretching the hydrogel mixture fibers to obtain the insert.

[0218] In some embodiments, the insert maintains its dimensions even after stretching, provided it is stored in a dry state below room temperature.

[0219] In some embodiments, the inserts are packaged separately and sterilized, for example, by gamma irradiation.

[0220] The shape memory effect due to stretching has already been disclosed in detail above regarding the properties of the insert. In some embodiments, the degree of shrinkage during hydration depends, in particular, on the stretching coefficient, as already disclosed above.

[0221] In some embodiments, the present invention thus relates to a method for imparting shape memory to hydrogel mixture fibers containing an active agent dispersed within a hydrogel, wherein the shape memory is imparted by stretching the hydrogel strands longitudinally.

[0222] The elongation coefficients for use in these methods of the present invention can be utilized as already disclosed above. III. Treatment

[0223] In some embodiments, the present invention further relates to a method for treating or preventing an ocular disease in a patient requiring such treatment or prevention, comprising administering to the patient a first sustained-release biodegradable intratubular insert comprising a hydrogel and an active agent as disclosed above.

[0224] In some embodiments, the aforementioned first implant is left in the duct until it completely disintegrates, or is removed before it completely disintegrates. In some embodiments, the aforementioned first implant can be designed to disintegrate over a long period, for example, within about 1 to 6 months, 2 to 4 months, 2 to 3 months, or 3 to 4 months after insertion, although in some cases, disintegration may take longer. Under normal circumstances, the implant decomposes without requiring any action from the patient, so the implant can remain in the duct until it completely disintegrates. This is an advantage because the patient does not need to consult a doctor or optician to have the implant removed. On the other hand, as biodegradation progresses, the implant softens, so if an unexpected event occurs, such as an allergic reaction, discomfort during wear, or other adverse event (e.g., irritation), early removal is facilitated by applying slight pressure to expel the implant from the punctum, or by moving the implant further down the duct and removing it through the nasolacrimal duct, if necessary. Furthermore, because the first implant is soft, it is possible to insert the second implant without having to remove the first implant beforehand. By inserting the second implant, the first implant is further pushed into the canal and remains there without discomfort until disintegration is complete, or is pushed down into the tear drainage system. The second implant can be inserted, for example, immediately after the intended treatment period of the first implant has elapsed, or when the patient feels that the treatment effect has worn off.

[0225] Therefore, in some embodiments, the second insert can be inserted at least one month or at least two months later without prior removal of the aforementioned first insert. In other embodiments, the aforementioned first insert is removed before it completely disintegrates, and the second insert is administered to replace the removed first insert.

[0226] One aspect of the present invention is a method for treating a target dry eye disease, (a) The step of inserting a first biodegradable implant into the first cannula of the first eye of the subject, wherein the implant is (1) Biodegradable hydrogel and (2) The hydrogel contains approximately 100 μg to approximately 800 μg of active agent, (3) The insertion step, wherein the active drug is released from the implant at an average rate of approximately 0.1 μg / day to approximately 10 μg / day over a period of at least approximately 2 months from the day the first implant is inserted into the subject, (b) A method comprising the step of inserting a second implant into a first canaliculus of a first eye of a subject at least two months after the date of insertion of a first implant, wherein the second implant is similar to or substantially similar to the first implant.

[0227] In some embodiments, the aforementioned first implant is designed to disintegrate within the tube within approximately 2 to 3 months after insertion, and the aforementioned first implant is removed within 2 months after administration.

[0228] In some embodiments, the dose of the active agent administered once per eye during a treatment period of at least two months is approximately 300 μg to approximately 400 μg of the active agent. Other suitable doses are further disclosed above.

[0229] In some embodiments, the ocular disease is a disorder of the tear film and the surface of the eyeball.

[0230] In some embodiments, the ocular disease is dry eye disease. In alternative embodiments, the implants and methods of the present invention can be used to treat other ocular surface diseases, such as blepharitis, allergic conjunctivitis, and especially atopic keratoconjunctivitis and vernal keratoconjunctivitis.

[0231] In some embodiments, ocular diseases are associated with one or more conditions selected from the group consisting of burning, itching, redness, stinging, pain, foreign body sensation, visual impairment, inflammation of the lacrimal gland, inflammation of the ocular surface, T cell-mediated inflammation, presence of conjunctival T cells in the tears, and elevated levels of inflammatory cytokines in the tears.

[0232] In some embodiments, sustained-release biodegradable intratubular inserts comprising the hydrogel and active agent of the present invention can be applied to prevent such ocular conditions in subjects at risk of developing dry eye disease or any related condition (e.g., subjects who wear contact lenses).

[0233] In some embodiments, the treatment is effective in improving tear production, as measured by the Schirmer tear test, in patients with a Schirmer score of less than 10 mm prior to administration, and / or in reducing dry eye symptoms, as determined by one or more assessments selected from the group consisting of assessment of the severity of dry eye symptoms on a visual analog scale, assessment of the frequency of dry eye symptoms on a visual analog scale, quantification of tear film breakup time, corneal fluorescein staining, conjunctival lysamine green staining, best corrected visual acuity, quantification of the ocular surface disease index, and standard patient assessments of dry eye.

[0234] In some embodiments, this treatment is effective in improving tear production in patients whose Schirmer score is less than 10 mm prior to administration, as measured by the Schirmer tear test.

[0235] In some embodiments, the dose per eye administered once during the treatment period is contained in one or two implants.

[0236] In some embodiments, the insert is inserted into the lower or upper tube, or one insert is inserted into both the lower and upper tubes, respectively. The insert can be inserted into the vertical portion of the tube.

[0237] The active agent administered once per eye during the treatment period can be contained in one or two implants.

[0238] In some embodiments, the dose per eye administered once during the treatment period is contained, for example, in one implant (e.g., one implant containing a dose of approximately 250 μg or approximately 360 μg of the active agent).

[0239] In some embodiments, the insert can be inserted into the tube using a grasping device selected from the group consisting of forceps, tweezers, and applicators.

[0240] In embodiments in which two inserts are administered, the inserts are inserted simultaneously, as disclosed above herein. The inserts inserted simultaneously may be the same or different.

[0241] In some embodiments, the treatment period is at least one month, at least two months, or at least three months. “Treatment period” according to one embodiment of the present invention means that the therapeutic effect of the implant of the present invention, once inserted, is maintained, or essentially maintained, over that period. In other words, in some embodiments, only a single insertion (of the implant of the present invention) is required to maintain the therapeutic effect over the long period referred to herein as “treatment period.” This represents a significant advantage over currently used eye drops for treating dry eye, which require very frequent administration several times a day, and thus greatly improves the patient’s quality of life.

[0242] One aspect of the present invention is a method for treating a patient who requires treatment for a dry eye disease, the method comprising administering to the patient a sustained-release biodegradable intratubular insert containing a hydrogel and an active agent, wherein punctal occlusion and release of the active agent into the eye result in a synergistic effect.

[0243] This synergistic effect is thought to be due to the higher bioavailability of the active agent compared to administering eye drops containing the active agent designed to release the same amount of active agent daily. This can be quantified, for example, by calculating the amount of active agent released into the tear film over time based on the concentration of the active agent in the tear film.

[0244] In some embodiments, the systemic concentration of the active drug is below a quantifiable level. Because the systemic concentration of the active drug is kept to a minimum, the risk of drug interactions or systemic toxicity is also minimized. Therefore, in one embodiment, the additional medication(s) taken by the patient pose no significant risk. This is particularly beneficial for elderly patients who frequently suffer from eye diseases and are taking other medications in addition.

[0245] One aspect of the present invention is the use of a sustained-release biodegradable intratubular insert comprising a hydrogel and an active agent as disclosed above in the preparation of a pharmaceutical for the treatment of an eye disease as disclosed above in a patient requiring such treatment, or for the treatment of a dry eye disease / keratoconjunctivitis as disclosed above in a patient requiring such treatment.

[0246] One aspect of the present invention is a sustained-release biodegradable intratubular insert comprising a hydrogel and an active agent as disclosed above, for use in the treatment of patients requiring treatment for ophthalmic diseases as disclosed above, or for use in the treatment of patients requiring treatment for dry eye diseases / keratoconjunctivitis sicca as disclosed above.

[0247] One aspect of the present invention is a method for increasing tear production, as measured by the Schirmer tear test, in a patient having a Schirmer score of less than 10 mm prior to administration, the method comprising administering to the patient a sustained-release biodegradable intratubular insert containing a hydrogel and an active agent as disclosed above.

[0248] In some embodiments, in this method, the Schirmer score can be increased by at least 2 mm 6 weeks after insertion of the implant, or at least 3 mm 12 weeks after insertion.

[0249] One aspect of the present invention is a method for reducing symptoms of dry eyes, which is quantified by one or more assessments selected from the group consisting of assessment of the severity of symptoms of dry eyes on a visual analog scale, assessment of the frequency of symptoms of dry eyes on a visual analog scale, quantification of tear film breakup time, corneal fluorescein staining, conjunctival lysamine green staining, best corrected visual acuity, quantification of the ocular surface disease index OSDI, and standard patient assessment of dry eyes SPEED, the method comprising administering to a patient a sustained-release biodegradable intratubule insert containing a hydrogel and an active agent as disclosed above.

[0250] In some embodiments, in such a method, the total corneal fluorescein staining value tCFS can decrease by at least 1.5 six weeks after implantation, or by at least 3 twelve weeks after implantation.

[0251] In some embodiments, in such a method, the assessment of the severity of symptoms of dry eyes on a visual analog scale may decrease by at least 10 two weeks after insertion of the implant, or by at least 15 six weeks after insertion. [Examples]

[0252] The following examples are included to demonstrate certain aspects and embodiments of the present invention as described in the claims. However, those skilled in the art will understand that the following description is illustrative and should not be construed as limiting the invention in any way. Example 1: Preparation of Cyclosporine Inserts

[0253] The cyclosporine insert of this application is essentially cylindrical (formed as a fiber) in order to release cyclosporine slowly based on its low solubility in tear fluid, and the cyclosporine is homogeneously dispersed and encapsulated within a PEG-based hydrogel matrix.

[0254] I. Example 1.1: Evaluation of drug concentration To evaluate the effect of drug concentration, three different formulations were prepared using low, medium, and high doses of cyclosporine. The compositions of the three formulations are shown in Table 1.1.1 below. [Table 2]

[0255] Low-dose, medium-dose, and high-dose inserts were prepared essentially according to the manufacturing process outlined for the test product insert (see Example 1.5 below). However, tyroxapol was not used, and the cyclosporine-containing syringe was first mixed with the multi-arm PEG solution-containing syringe, and then with the trilidine acetate (TLA) / fluorescein solution-containing syringe, without vacuum degassing.

[0256] After casting, the hydrogel was cured, i.e., crosslinked, and then the tube strands were stretched and dried in an incubator under a nitrogen flow. The dried strands were removed from the flexible tube and cut to length. The dimensions and physical properties were as follows: [Table 3]

[0257] In both low-dose and high-dose formulations, undesirable "straw-forming" fibers were obtained, meaning fibers with a large dry diameter and a hollow center. As a result, the density of the straw-forming fibers was low.

[0258] To reverse the "roundness" of the fibers, the cross-sectional diameter was measured on the thick and thin sides, and the smaller diameter value was identified as diameter 1 and the larger as diameter 2, and the aspect ratio was calculated as follows. Aspect ratio = diameter 1 / diameter 2

[0259] In the medium-dose formulation, strawification was not shown, but flat fibers with a very low aspect ratio were produced. The aspect ratio was 0.7, indicating a lack of roundness. Although the strawified fibers in the low-dose and high-dose formulations were rounder, due to their low density, much larger fibers than desired were produced. Also, as summarized in Table 1.1.3 below, the drug concentration also affected the hydration properties of the insert.

Table 4

[0260] As can be seen from this, the hydrated diameter far exceeded the target of 1.45 mm, decreased with an increase in dose, and the shrinkage rate also decreased with an increase in dose.

[0261] II. Example 1.2: Evaluation of Surfactants Presence of Surfactant To evaluate the effect of the surfactant, three different formulations were prepared. One contained a surfactant (control, Run 1), one contained 0.05% Tween® 20 (Run 2), and the third contained no surfactant but was prepared using ethanol (Run 3). The compositions of the three formulations are shown in Table 1.2.1 below.

Table 5

[0262] The inserts in Runs 1-3 were prepared essentially according to the manufacturing process outlined for the test inserts (see Example 1.5 below), namely, one hydrogel suspension precursor syringe containing a suspension of cyclosporine in trilysine acetate (TLA) / fluorescein aqueous solution (Run 1: without tyroxapole; Run 2: containing Tween® 20 instead of tyroxapole; Run 3: without tyroxapole, but dissolved in ethanol instead of water) and a second hydrogel solution precursor syringe containing a multi-arm PEG aqueous solution were prepared. These two syringes were mixed, and the liquid suspension was then injected to cast it onto a subset of flexible tube pieces, after which the material was crosslinked and solidified. Vacuum degassing was not performed. After casting, the hydrogel was cured, i.e., crosslinked, and then the tube strands were stretched and dried in an incubator under a nitrogen stream. The dried strands were removed from the flexible tubes and cut to length. The dimensions and physical properties were as follows: [Table 6]

[0263] To reverse the "curvature" of the fibers, the cross-sectional diameter was measured on both the thicker and thinner sides. The smaller diameter was identified as diameter 1, and the larger diameter as diameter 2. The aspect ratio was then calculated as follows. Aspect ratio = diameter 1 / diameter 2

[0264] As can be seen here, adding Tween® 20 resulted in a higher aspect ratio (indicating rounded fibers). The average diameter was smaller and the density was higher than the control, which is advantageous in terms of ease of insertion into the product. The control fibers in Run 1 adhered to the tube during drying and showed a low aspect ratio, indicating flattened fibers (see Figure 1.2A).

[0265] In experiment 3, which investigated the use of ethanol, a major problem was encountered. The solution began to evaporate within one minute of the start of curing, and bubbles formed within the fibers. As can be seen in Figure 1.2B, when the fibers were removed from the incubator, they were all damaged and deformed. Drug concentration in the presence of surfactant

[0266] To evaluate the effect of drug concentration in the presence of a surfactant, three different formulations were prepared using low, medium, and high doses of cyclosporine in the presence of 0.05% Tween® 20. The compositions of the three formulations are shown in Table 1.2.3 below. [Table 7]

[0267] Three inserts for Runs 1-3 were prepared, essentially following the manufacturing process outlined for the test inserts (see Example 1.5 below), except that Tween® 20 was used instead of thyroxapole. Vacuum degassing was not performed. After casting, the hydrogel was cured, i.e., crosslinked, and then the tube strands were stretched and dried in an incubator under a nitrogen stream. The dried strands were removed from the flexible tubes and cut to length. Sterilization was not performed. The dimensions and physical properties were as follows: [Table 8]

[0268] The aspect ratio was 0.9 or greater in all cases, indicating rounded fibers. Dry fiber diameter increased with increasing dose. However, the increase in dry fiber diameter was slight in all cases, which may be related to the decrease in fiber density with increasing dose.

[0269] Figure 1.2C shows the dried and hydrated inserts produced in this run. As can be seen, aggregation appears to occur more frequently at higher drug concentrations.

[0270] Table 1.2.5 below shows the characteristics of the hydrated inserts 10 minutes and 24 hours after hydration with PBS at 37°C. As can be seen, both the hydrated diameter and the shrinkage coefficient decrease with increasing dose. This indicates that the drug content has a significant adverse effect on the rehydration rate. [Table 9]

[0271] Surfactant type To address aggregation, the effect of various surfactants on the particle size of cyclosporine was quantified. The surfactants were tested at the maximum surfactant concentration according to FDA guidance.

[0272] Stock solutions containing 0.05% Tween® 20 (PEG-20 sorbitan monolaurate), 4% Tween® 80 (PEG-80 sorbitan monolaurate), 0.5% Cremophor RH 40 (PEG-40 hydrogenated castor oil), and 0.3% tyloxapol (ethoxylated 4-tert-octylphenol / formaldehyde condensation polymer) were prepared by adding PBS to known weights of surfactants in glass vials. The surfactant solutions were then vortexed and sonicated. Solutions containing approximately 9-10% (w / w) of cyclosporine were prepared by adding approximately 1 mL of surfactant / buffer stock solution to approximately 100 mg of cyclosporine in the vial. Controls were prepared using distilled water (DIW) or phosphate-buffered saline (PBS) and did not contain surfactants. The solutions were then vortexed and the particle size was measured by laser diffraction using a Beckman Coulter LS 13 320 based on the optical model Fraunhofer rf780z at an obscuration value in the range of 7-9%.

[0273] Table 1.2.6 shows the solution concentration and particle size. As can be seen, the use of surfactants significantly reduced particle size, suggesting a reduction in aggregation. Of the surfactants tested, tyroxapole showed the greatest reduction in particle size. In addition, the particle sizes measured using tyroxapole were closest to those provided by the supplier (D10, D50, and D90 showed 0.5, 2.1, and 7.5 μm, respectively). [Table 10]

[0274] I. Example 1.3: Evaluation of Vacuum Degassing To evaluate the effect of vacuum degassing, a 600 μg dose of cyclosporine insert was prepared using a hydrogel precursor mixture of 0.3% (w / w) tyroxapol and 9% (w / v) 4a20k PEG-SG. The composition of the insert formulation is shown in Table 1.3.1 below. [Table 11]

[0275] Three different insert sets were prepared using the same manufacturing process for hydrogel suspension precursor syringes containing cyclosporine, tyroxapol, and trilysine acetate / fluorescein, with the following details regarding the treatment after mixing the components and before further mixing with the PEG-containing hydrogel solution precursor.

[0276] For the first insert set, (i) the hydrogel suspension in the syringe was processed by priming the syringe, vortexing the suspension for 3 minutes, and manually degassing it (as is customary for all formulations). Manual degassing was performed by pulling the plunger of a capped syringe to create a vacuum, then removing the syringe cap to release the vacuum and collapse any large bubbles. In the next step (ii), the hydrogel suspension precursor was placed in a vacuum chamber, degassed, and then mixed with the hydrogel solution precursor. The hydrogel suspension precursor for the second insert set was processed in essentially the same manner, except that it was further sonicated in step (iii) after step (ii). The hydrogel suspension precursor for the third insert set was prepared in the same manner as the second set, except that degassing in the vacuum chamber in step (ii) was omitted. A summary of the preparation steps is shown in Table 1.3.2 below. [Table 12]

[0277] The first set of inserts obtained by vacuum degassing alone showed almost no aggregates. The second set of inserts obtained by both vacuum degassing and ultrasonic treatment showed numerous large aggregates and was not compatible with the cutting device. The third set of inserts obtained with ultrasonic treatment but without vacuum degassing showed some aggregates, but otherwise had a good appearance (Figure 1.3).

[0278] II. Example 1.4: Evaluation of drug particle size Further inserts were prepared to evaluate and determine the optimal particle size of the active pharmaceutical ingredient. Sieving effect of large cyclosporine particles

[0279] To evaluate the effectiveness of sieving to exclude larger particles, 400 μg doses of cyclosporine inserts were prepared using a hydrogel precursor mixture of 0.3% (w / w) tyroxapole and 9% (w / v) 4a20k PEG-SG. The composition of the formulations is shown in Table 1.4.1 below. [Table 13]

[0280] The cyclosporine used was sieved, and the particle size fractions shown in Table 1.4.1 below were used. During manufacturing, some cast strands were damaged and unusable, as summarized in Table 1.4.2 below. The dry density and drug filling rate for each insert are shown in Figure 1.4A. [Table 14]

[0281] Thus, we can conclude that large particle sizes have serious adverse effects. Effect of cyclosporine particle size on hydration, dry density, and mechanical defects

[0282] To evaluate the effects of different particle sizes, inserts were prepared using micronized cyclosporine particles with five different particle size distributions (PSDs). The compositions of the five formulations are shown in Table 1.4.3 below. [Table 15]

[0283] The d10, d50, and d90 values ​​of the cyclosporine used are shown in Table 1.4.4 below. [Table 16]

[0284] The following was observed during manufacturing: Inserts prepared with larger CSI particles tended to have a larger dry diameter (see Figure 1.4B), while density decreased with increasing CSI particle size (see Figure 1.4C). Smaller CSI particles resulted in smoother insert surfaces (see Figure 1.4D, showing microscopic images taken under a stereomicroscope with a camera). Inserts derived from small and small / medium particles appeared to be covered with PEG. Inserts prepared with medium and large particles exhibited rough surfaces with significant irregularities and deformation. Medium particles showed the highest rate of breakage during drying. Larger particles exhibited better hydration and swelling behavior (see Figure 1.4E).

[0285] In conclusion, larger particle sizes improved swelling and snapback, but also increased strand breakage. Additionally, high-dose formulations showed higher breakage rates than low-dose and medium-dose formulations, and the removal of very large particles (over 45 microns) by sieving significantly improved the breakage rate.

[0286] Overall, the "small" to "medium" particle sizes strike a balance between the increased density and fiber smoothness of small particles and the rehydration rate of medium particles, which reduces strand breakage.

[0287] III. Example 1.5: Preparation of clinical trial support inserts The following describes the preparation process for the test inserts used in human clinical trials (Formulations 1, 2A, 2B, and 3, see Example 4) and high-dose beagle dog trials (Formulations 4A and 4B, see Example 3.6). The test inserts are based on a medium-term sustained-release cyclosporine / hydrogel formulation based on PEG-SG (Formulations 1 and 4A, designed to last approximately 2-3 months), a long-term sustained-release cyclosporine / hydrogel formulation based on PEG-SAP (Formulations 2A and 4B, designed to last approximately 3-4 months), and two cyclosporine-free hydrogel vehicle (HV) formulations that function as placebos: a long-term sustained-release formulation based on PEG-SAP (Formulation 2B, designed to last approximately 3-4 months), and a short-term sustained-release formulation based on PEG-SS (Formulation 3, designed to last approximately 1 week).

[0288] The target diameter for the cyclosporine-containing implant was 0.55 mm ± 0.03 mm, and the target length was 2.72 mm ± 0.08 mm. The target diameter for the HV implant was 0.41 mm ± 0.05 mm, and the target length was 2.72 mm ± 0.08 mm. The composition of the formulations is shown in Table 1.5 below. [Table 17]

[0289] I. Overview of the Preparation Process Two precursor syringes were prepared to form a polymer network of cyclosporine-containing inserts. One was a hydrogel suspension precursor syringe containing a cyclosporine suspension in tyroxapole and trillidine acetate (TLA) / fluorescein solution, and the second was a hydrogel solution precursor syringe containing a multi-arm PEG (4-arm 20K PEG based on a pentaerythritol core structure containing amine-reactive NHS groups) solution. The two syringes were mixed and then cast into a subset of flexible tubing by injecting the liquid suspension before the material crosslinked and solidified. After casting, the hydrogels were cured (i.e., crosslinked). The strands of cyclosporine-containing tubing encapsulated within the hydrogel network were then stretched and dried in an incubator under a nitrogen flow. The dried strands were removed from the flexible tubing, cut to length, and stored in vials. The chemicals were then packaged in protective foam carriers and heat-sealed in laminate foil pouches under a nitrogen environment. The cyclosporine-containing preparations were sterilized using electron beam irradiation, and the HV preparations were sterilized using gamma irradiation.

[0290] For the HV placebo inserts, the amount of excipients used was adjusted according to the composition (variations of multi-arm PEG) in Table 1, and they were prepared similarly, except that cyclosporine was not used.

[0291] II. Preparation of Hydrogel Suspension Precursor Syringe Except for the type of multi-arm PEG used and the amount of dibasic sodium phosphate, the same procedures and quantities as for formulations 1 and 2A were used. For formulations 4 and 5, the amount of cyclosporine was also adjusted according to the composition shown in Table 1.5 above.

[0292] The hydrogel suspension precursor syringe consisted of a mixture of two different additional precursor syringes.

[0293] Each syringe contained micronized cyclosporine suspended in tyroxapol solution. This was prepared by weighing 704.8 mg ± 5.0 mg of micronized cyclosporine (d50: 5-8 μm, d100: ≥ 45 μm) and suspending it in a 0.8 wt-% solution of tyroxapol in 2,775.0 mg ± 20.0 mg of sterile water for injection (WFI). For the HV formulations (formulations 2B and 3), the syringes contained only a 0.8 wt-% tyroxapol solution.

[0294] The other syringe contained a dibasic sodium phosphate buffered trilidine-fluorescein conjugate solution, which was prepared by (i) mixing 25.0 mg ± 0.5 mg of NHS-fluorescein with 8,025.0 mg ± 5.0 mg of a solution containing 97.5 mg ± 2.5 mg of trilidine acetate and 243.75 mg ± 2.5 mg (formulations 1 and 3) or 690 mg ± 5.0 mg (formulations 2A and 2B) of dibasic sodium phosphate in 9,750.0 ± 10.0 mg of WFI; (ii) reacting the resulting mixture at room temperature for 1 to 24 hours; (iii) filtering the solution; and (iv) filling each syringe with 1,575.0 mg ± 10.0 mg of the resulting solution. Completion of the reaction in step (ii) was confirmed by reversed-phase (RP) HPLC with UV detection, which allowed identification of unreacted components and product amides by retention time (RT). After step (ii), no quantifiable peaks remained at RTs consistent with NHS-fluorescein (RT approximately 6.6 mins), and a new peak created by the formation of the product amide appeared at a higher RT (RT approximately 8.2 mins). The results of the initial test demonstrated that the product was converted to an amide after 1 hour and that the reaction product was stable in solution for up to 7 days (Figure 2).

[0295] Two additional precursor syringes were connected with a female-to-female Luer lock connector, and their contents were mixed by moving the syringes back and forth a total of 25 times to form a hydrogel suspension precursor syringe. Because cyclosporine is very poorly soluble in water and prone to aggregation, tyroxapole, USP, was added to the solution used to suspend the cyclosporine to help disperse the cyclosporine and suppress any aggregation, as well as to prevent the hydrogel mixture from adhering to the inner wall of the tube, which is thought to be the cause of the insert having a flat shape with a low aspect ratio.

[0296] II. Preparation of Hydrogel Solution Precursor Syringe The hydrogel solution precursor syringe contained a PEG buffer solution, which was prepared by combining 1,565.0 mg ± 10.0 mg of a solution containing 24.5 mg ± 1.0 mg of monobasic sodium phosphate in 7,985.0 mg ± 50.0 mg of WFI with 542.0 mg ± 5.0 mg of either 4a20K PEG-SG (20 kDa PEG with four arms accompanied by N-hydroxysuccinimidylglutaric acid terminal groups, used in formulation 1) or 4a20K PEG-SAP (20 kDa PEG with four arms accompanied by N-hydroxysuccinimidyladipic acid terminal groups, used in formulation 2a).

[0297] III. Casting, stretching, and drying To form the hydrogel / cyclosporine suspension, hydrogel suspension precursor syringes containing cyclosporine, tyroxapole, and trilysine-fluorescein conjugate, as well as hydrogel solution precursor syringes containing 4a20k PEG-SG, 4a20k PEG-SAP, or 4a20k PEG-SS, were first placed in a vacuum chamber, degassed by exposure to a programmed vacuum cycle, and then connected to a female-to-female Luer lock connector. The contents of the precursor syringes were mixed together by moving them back and forth a total of 25 times between each syringe, and the hydrogel / cyclosporine suspension thus produced was transferred to a single syringe.

[0298] Next, the hydrogel / cyclosporine suspension syringe was connected to a barbed fitting of autoclaved polyurethane tubing (2.0 mm inner diameter and 2.8 mm outer diameter), cut to the appropriate length, and the suspension was cast through the prepared tubing before the material crosslinked and solidified.

[0299] Once the tube was full, it was removed from the syringe and the barbed fitting of the tube was capped. Gelation time was confirmed by performing a gel-tap test. For the gel-tap test, a small amount of the remaining hydrogel / cyclosporine suspension was placed on a glass slide and tapped with the tip of a pipette until the suspension gelled and began to form strands (indicating that polymerization had begun), which took approximately 2–8 minutes (i.e., the pipette tip remained attached throughout the complete tapping cycle).

[0300] A filled tube containing a hydrogel / cyclosporine suspension (hereinafter referred to as "cast strand") was placed vertically and stored in a curing chamber (at ambient temperature and humidity) for 3 to 6 hours to allow the gel to harden.

[0301] After the curing time had elapsed, the cast strand was placed in a stretching fixture and secured in place with a dynamic clamp. The cast strand was stretched to the fixing length of the stretching fixture, which was approximately 2.7 times the original tube length. The stretching fixture was then moved and placed vertically into an incubator set to 32.0 ± 2.0 °C and a nitrogen flow rate of 53 ± 3 SCFH (standard cubic feet / hour) for drying. The cast strand was left in the incubator for several days to dry completely.

[0302] IV. Cutting, packaging, sterilization, and inspection The dried cast strands were removed from the tubes and cut to a length of approximately 2.7 mm. A Vision System was used at 10x magnification to perform 100% in-process visual and dimensional inspection of the inserts (acceptability criteria: no particulate, cylindrical shape, no visible surface defects; diameter of cyclosporine-containing inserts: 0.55 mm ± 0.03 mm; diameter of HV- inserts: 0.41 mm ± 0.05 mm; length of all inserts: 2.72 mm ± 0.08 mm).

[0303] Inserts that met all in-process specifications were packaged by placing one insert per foam carrier and sealing them in an aluminum-LDPE foil pouch that could be peeled open by the user. To hold the insert, the bottom of the foam carrier had a V-notch with an opening, into which the insert could be inserted with forceps, allowing a portion of the insert to protrude and be easily removed. The foam carrier containing the insert was placed inside the foil pouch. The unsealed foil pouch was moved to a glove box providing an inert nitrogen environment to reduce residual moisture from the foam and pouch material, where it was stored for a minimum of 16 hours and not exceeding 96 hours. The pouch was then sealed completely and continuously inside the glove box using a pouch sealer. The pouch seal was inspected, and the package was stored at 2-8°C until sterilization.

[0304] The packaged inserts were sterilized by electron beam or gamma ray irradiation and stored at 2-8°C until final quality inspection.

[0305] After sterilizing the inserts in the pouches, a final quality check of the chemicals was performed.

[0306] Example 2: Specifications of the insert The obtained inserts were characterized by visual inspection, microscopic analysis, HPLC analysis, and storage stability testing.

[0307] I. Visibility Test To confirm that the implants could be visually observed through the surrogate test model when illuminated with blue light, the implants were visually inspected.

[0308] II. Microscopic Analysis The inserts were microscopically examined using a Unitron Z850 / NSZ-606 microscope to confirm the dimensions of the product in a dry state and in a hydrated state after hydration in phosphate-buffered saline at pH 7.4 and 37°C for 10 minutes (magnified state) and 24 hours (equilibrium state) (Table 2.1). [Table 18]

[0309] III. Further testing of product specifications Further product specifications will be tested and reported according to Table 2.2 below. [Table 19-1] [Table 19-2]

[0310] IV. Storage Stability Test The stability of cyclosporine-containing inserts according to formulations 1 and 2A was evaluated over 12 months under refrigerated conditions (2-8°C).

[0311] The results of the storage stability tests can be found in Tables 2.3.1 and 2.3.2 (Formulation 1), and Tables 2.3.3 and 2.3.4 (Formulation 2A). [Table 20-1] [Table 20-2] [Table 20-3] [Table 20-4]

[0312] Stability data obtained over 12 months continued to meet the specified stability specifications. According to ICH Q1E, for refrigerated products, the proposed shelf life can be up to twice as long if long-term data shows little or no change over time and little or no variation, but it should not exceed 12 months if the proposal is supported by the analytical results and relevant reference data. The real-time long-term stability data contradicted the upper and lower limits of the stability specifications using 95% confidence intervals (CI). Overall, there was little trend in the quantitative data. Statistical analysis showed that the product fits a shelf life of 23 months, based on degradation analysis of assays that indicate potential interference at 23 months. All other stability parameters indicate potential interference beyond 36 months. Therefore, a shelf life of at least 23 months can be expected.

[0313] Example 3: Evaluation of cyclosporine implants in Beagle dogs To test the pharmacokinetics of cyclosporine-containing inserts, different formulations are being tested in pharmacokinetic studies using beagle dogs.

[0314] Six different Beagle dog studies were conducted, and the amount of drug released from the insert and / or the cyclosporine concentration in the tear fluid were quantified during the study period. A summary of the studies and formulations is shown in Table 3. [Table 21-1] [Table 21-2]

[0315] Example 3.1: Proof-of-Concept PK Test Inserts containing a nominal dose of 0.7 mg of cyclosporine were administered to beagle dogs, as summarized in Table 3.

[0316] The amount of drug released from the implant was evaluated over time, and the cyclosporine concentration in the tear fluid was quantified throughout the test period.

[0317] The median tear concentration of cyclosporine during the study period was 1.1–1.9 μg. See Table 3.1 below. A subset of the implant was removed at week 6, and the amount of residual drug compared to the administered dose was quantified. The average amount of cyclosporine released over 6 weeks was 0.37 mg. Assuming a constant daily release rate throughout the administration period, supported by the relatively constant tear concentration, the estimated daily delivered dose is calculated to be 8.8 μg / day. [Table 22]

[0318] Example 3.2: Proof-of-Concept PK Test Inserts containing a nominal dose of 0.44 mg of cyclosporine were administered to beagle dogs, as summarized in Table 3.

[0319] The amount of drug released from the implant was evaluated over time, and the cyclosporine concentration in the tear fluid was quantified throughout the test period.

[0320] The mean tear concentration of cyclosporine during the study period was 1.1–2.8 μg. See Table 3.2 below. On day 32, the insert was removed and the amount of remaining drug was quantified compared to the administered dose. The mean amount of cyclosporine released over 32 days was 0.2 mg. Assuming a constant daily release rate throughout the administration period, the estimated daily delivery dose is calculated to be 6.3 μg / day. [Table 23]

[0321] Example 3.3: Proof-of-Concept PK Test The initial cyclosporine dose in the implants before administration was 671 ± 13 μg (mean and standard deviation) for all n=10 samples. Four and five implants were removed on days 28 and 50, respectively, and their cyclosporine content was analyzed. Since similar results were obtained from both eyes, the average of all removed implants was calculated, resulting in release amounts (initial dose minus the amount remaining in the removed implants (576 ± 18 μg and 494 ± 32 μg, respectively)) of 3.4 and 3.5 μg, respectively, as shown in Table 3.3. The average daily release rate was quantified by comparing the initial dose with the released dose. The test results indicate that the estimated daily delivery dose was approximately 3.5 μg / day over days 28 and 50. The range of drug release from the implants over the test period was from a low of 2.7 μg / day to a high of 4.4 μg / day. [Table 24]

[0322] Example 3.4: Dry eye model test Inserts containing a nominal dose of 0.36 mg of cyclosporine, with a hydration diameter of 1.5 mm and a hydration length of 2.5 mm, were administered to beagle dogs as summarized in Table 3.

[0323] An artificial dry eye model was created by surgically removing the lacrimal gland from the right eye of a beagle. The left eye remained untreated and served as a healthy control. The Schirmer tear test tracked tear production over time and showed that tear production in the right eye decreased to almost zero, demonstrating proof of concept in the dry eye model (see Figure 3.1). The amount of drug released from the implant was evaluated over time, and the cyclosporine concentration in the tear fluid was quantified throughout the study period.

[0324] The mean tear cyclosporine concentration ranged from 0.8 to 1.4 μg / mL in healthy eyes and from 1.4 to 4.8 μg / mL in dry eyes. See Table 3.4 below. The higher tear cyclosporine concentrations in the Beagle dry eye model (compared to healthy eyes) demonstrate both that cyclosporine can be successfully transported from the implant into the tear film and possibly to the ocular surface under dry eye conditions, and that the cyclosporine concentration on the ocular surface is higher than that observed in healthy eyes because, under dry eye conditions, tear volume decreases (as a result of reduced tear production), thus reducing drug dilution. [Table 25]

[0325] Example 3.5: Pharmacokinetic study Inserts of formulations 1 and 2A (PEG-SG and PEG-SAP hydrogel, 0.36 mg of cyclosporine) were administered bilaterally to 24 beagle dogs, as summarized in Table 3. Tear samples were collected at 1, 3, and 6 hours post-administration, on days 1 and 3, and at weeks 1, 2, 4, 6, 8, 10, and 12. The results are shown in Table 3.5 and Figures 3.2A and 3.2B below. Cyclosporine was released from the inserts into the tears of the beagles over 12 weeks in both formulation 1 (group 1) and formulation 2A (group 2). The results indicate that a similar maximum tear concentration of 2.7 μg / mL occurred at approximately 3 hours between the two formulations. The drug release profile and tear concentration appeared to be similar (within the 95% confidence interval) between the two formulations over the first 6 weeks.

[0326] Between weeks 8 and 12, it has been noted that the cyclosporine tear concentration in Beagle dogs may decrease with SG hydrogel formulations compared to SAP hydrogel formulations. This difference may be due to the faster degradation of SG hydrogel compared to SAP hydrogel. [Table 26]

[0327] Example 3.6: High-dose study As summarized in Table 3, the pharmacokinetics of high-dose (0.7 mg cyclosporine) inserts in two hydrogel formulations 4A and 4B (PEG-SG and PEG-SAP) were evaluated in GLP-compliant toxicity studies.

[0328] Tear samples were collected before administration, and at 1, 2, 4, and 24 hours after administration, as well as on days 7, 30, 60, 90, and 104. Cyclosporine concentrations in the tears are shown in Table 3.6.1 and Figure 3.3. The results indicate that tear concentrations in this Beagle study were comparable (within the 95% confidence interval) between the two formulations over the study period, and that drug levels were below the lower limit of quantification (LLOQ = 30 ng / mL) in recovered animals (test samples were removed two weeks before the 104-day sample collection).

[0329] The pharmacokinetic profiles were comparable between the two formulations, demonstrating continuous steady-state dose exposure throughout the study period. The true peak concentration is not clear because tear film concentration reached an apparent steady state within one hour of dose exposure throughout the study period. [Table 27]

[0330] Example 4: Randomized, multicenter, double-blind, vehicle-controlled human Phase 1 / 2 clinical trial To evaluate the safety, tolerability, and efficacy of cyclosporine implants in the treatment of DED patients, a randomized, multicenter, double-blinded, vehicle-controlled phase 1 / 2 clinical trial was designed, enrolling 145 subjects (290 eyes) into two cohorts. Cohort 1 was an open-label group consisting of approximately 5 subjects (10 eyes) treated with formulation 2A hydrogel / cyclosporine implants, while Cohort 2 was a randomized, double-blind group consisting of approximately 140 subjects treated with two different formulations 1 and 2A hydrogel / cyclosporine implants, as well as two different formulations 2B and 3HV implants.

[0331] Both eyes will be treated with the same medication / formulation. If both eyes are eligible in terms of dry eye symptoms, the eye with the higher total score from corneal fluorescein staining will be designated as the test eye, and the other eye as the non-test eye. If the total scores from corneal fluorescein staining are the same for both eyes, a biostatistician will determine the test eye before analysis. If only one eye is being studied, that eye will be designated as the test eye, but both eyes will still be administered the same medication / formulation.

[0332] The treatment details are summarized in Table 4 below. [Table 28]

[0333] I. Exam Schedule Both cohort groups followed essentially the same trial schedule. Participants were eligible if: ● The individual has self-reported a history of dry eye disease in both eyes for more than 6 months, or has a clinically confirmed diagnosis by an ophthalmologist. ●At the time of the screening visit, the test eye must have an ongoing dry eye disease defined by a Visual Analog Scale (VAS) dry eye severity score of ≥30, and ● In the same eligible eye or both eyes, the corneal fluorescein staining (tCFS) score at 5 minutes is between 6 and 15 (NEI scale, see the "Assessment" section below), and the Schirmer score (unanesthetized) is wetted to more than 0 mm and less than or equal to 10 mm. ●Further selection and exclusion criteria will apply.

[0334] Participants will be screened 14 days before insertion day 1 (visit 2), and eligibility will be confirmed at visit 2 (insertion day 1). Participants in Cohort 2 will be randomly assigned to one of four treatment groups in a ratio of 2:2:2:1 at visit 2. From week 2 to week 16, all participants will be scheduled for further treatment follow-up visits at regular intervals (visits 3-8), particularly to determine the presence or absence of implants. The presence or absence of implants will be assessed non-invasively by irradiating the corresponding area with blue light and using a yellow filter.

[0335] In all cohorts, if the implant is visible at week 16 (visit 8), the subject will visit the clinic 30 days later (±10 days; visit 9) and will continue to visit the clinic every 30 days as needed until the implant is no longer visible and the physician determines there is no evidence of biological activity. If the implant is not visible at week 16 (visit 8) and the physician determines there is no evidence of biological activity, the subject will discontinue the study.

[0336] A general schematic diagram of this examination is shown in Figure 4A.

[0337] Various assessments will be performed at each visit. In particular, all ophthalmic assessments outlined in Point III below will be performed at visits 1-8 and visit 9 (if a visit 9 is available for an individual patient), with the exception of the TBUT assessment (performed only at visits 1, 2, 5, and 7).

[0338] If any adverse events occur, they should be evaluated similarly at each follow-up visit from the day of insertion / day 1 onward, according to point III below (any signs, symptoms, and conditions that occurred before insertion on day 1 should be recorded in the patient's medical history).

[0339] II. Insertion of Inserts The ophthalmic implant is placed in the vertical portion of the duct (see Figure 5B). In the clinical trials disclosed herein, the implant was placed in the inferior lacrimal punctum.

[0340] To position the insert within the ductus, lateral pressure was applied to stretch the ductal system, and the skin was temporarily pulled down near the punctum (Figure 5A). The inferior punctum was dilated towards the nose using a punctal dilator to confirm that the system was stretching, and the depth and width of the ductus were expanded through the punctum. If necessary, the dilator was rotated with a spinning motion to assist the dilation process (Figure 5B).

[0341] The surface around the punctal opening was dried using an ophthalmic sponge (Figure 5C).

[0342] The hydrogel / cyclosporine or HV implant was inserted at a slight angle towards the nose using forceps (Figure 5D), with the goal of inserting 70% in the initial movement. The remaining portion of the implant was then tapped or pressed using forceps, while avoiding excessive pressure on the implant to prevent deformation. The implant was confirmed to be positioned slightly below the lacrimal punctal opening.

[0343] If the insert hydrated before it was positioned slightly below the lacrimal punctum opening (resembling a trumpet shape), or before it was in ideal position, or if part of the insert protruded and could not be inserted, the insert was discarded and a new one was used.

[0344] The ease of insertion of ophthalmic implants was graded as "easy" (1), "moderate" (2), or "difficult" (3).

[0345] III. Evaluation This study used the following evaluation methods. Schirmer tear film test

[0346] The Schirmer test quantifies tear production and works through capillary action, which allows tears to move along the length of a paper test strip. The speed at which tears move along the strip is proportional to the rate of tear production. The subject is instructed to look upward, and the test strip is applied so that its curved tip rests between the lower eyelid's conjunctiva and the bulbar conjunctiva of the eye. After 5 minutes, the patient is instructed to open both eyes and look upward, and the test strip is removed. The Schirmer test score is quantified by the length of the moist area on the test strip. The test is performed on both eyes simultaneously. If anesthesia is used, only basal tear secretion is measured.

[0347] A Schirmer score of 10 mm or higher is considered normal, while a score of less than 5 mm indicates tear deficiency.

[0348] Lacrimal membrane rupture time (TBUT) and corneal fluorescein staining (tCFS) The time it takes for the tear film to break down after blinking is called TBUT (Tear Film Tolerance Time). This is a quantitative test to measure the stability of the tear film. A normal tear film break-down time is over 15 seconds. To evaluate TBUT, a saline-moistened fluorescein strip is applied to the inferior conjunctival sac. After several blinks, the tear film is examined for the appearance of the first dry spot on the cornea using a broad-beam slit lamp with a blue filter.

[0349] A TBUT value of less than 5-10 seconds indicates tear instability, which is observed in patients with mild to moderate dry eye.

[0350] The tCFS (total corneal fluorescein staining) value is measured to evaluate the condition of the cornea. Damage to the corneal surface, such as abrasions, can be caused by dry eye, for example, and this can be visualized using fluorescein dye.

[0351] To assess tCFS, wet a fluorescein strip with saline / eyewash solution, instruct the subject to look upward, and apply the wet strip to the lower palpebral conjunctiva without touching the bulbar conjunctiva. Since TBUT is also assessed by applying fluorescein, if tCFS measurement is performed immediately after TBUT, there is no need to apply additional fluorescein dye. Instruct the subject to blink several times to distribute the fluorescein dye, and after a waiting period of 2-3 minutes, use cobalt blue illumination and a Wratten yellow filter to assess corneal staining in each of the five corneal regions (central, inferior, nasal, temporal, and superior) using the NEI (National Eye Institute) 0-3 scale (0=no staining, 1=mild staining, 2=moderate staining, 3=severe staining), and the total score of the five regions (0-15 points) is taken as the total CFS score.

[0352] A higher tCFS score indicates greater corneal surface damage.

[0353] Conjunctival lysamine green staining (LGS) The LGS value is measured to assess the condition of the conjunctiva. A lysamine strip is moistened with saline / eyewash solution, and the subject is instructed to look upward. The moistened strip is applied to the lower palpebral conjunctiva without touching the bulbar conjunctiva. The subject is instructed to blink several times to distribute the lysamine dye. After a waiting period of 1-4 minutes, using appropriate lighting, conjunctival staining is assessed for each of the six regions of the conjunctiva (temporal, upper temporal, lower temporal, and upper nasal, lower nasal, and nasal) using the NEI (National Eye Institute) scale of 0-3. The sum of the six regions (0-18) is taken as the total LGS score.

[0354] Staining caused by pinguecula elevation may not improve. Staining associated with pinguecula can be consistently excluded from the total lysamine score.

[0355] The higher the LGS score, the greater the damage to the conjunctival surface.

[0356] Best corrected visual acuity BCVA Visual acuity testing should be performed before any tests requiring eye contact or eye drops in the test eye. LogMAR visual acuity must be assessed using the Early Treatment Diabetic Retinopathy Test (ETDRS) or the Modified ETDRS visual acuity chart. This chart consists of five letters per row, each row representing 0.1 log units (logMAR) of the minimum separation angle at a given test distance.

[0357] Visual acuity testing is performed using the Early Treatment Diabetic Retinopathy Study (ETDRS) or the modified ETDRS visual acuity chart, with the subject's own corrective lenses (glasses only) or pinhole refraction used to achieve the best corrected visual acuity. The ETDRS or modified ETDRS visual acuity chart consists of rows of five letters, each row representing the minimum separation angle at a given test distance in 0.1 log units (logMAR).

[0358] Visual acuity (VA) is scored as a logMAR value. The base logMAR value is taken from the last line in which the characters were read correctly, and N × 0.02 is added to this value (where N is the total number of incorrect characters up to the last line read). This sum (base logMAR + N × 0.02) corresponds to the BCVA of that eye.

[0359] The lower the BCVA score, the better the visual acuity.

[0360] Eye dryness score / Visual Analog Scale (VAS) To assess the dry eye score, participants are instructed to rate the severity and frequency of their dry eye symptoms as a percentage by marking a vertical line on a horizontal line (representing a value from 0 to 100%) that indicates the level of eye discomfort and frequency of dry eye currently experienced in both eyes (0% corresponds to "not uncomfortable," and 100% corresponds to "most uncomfortable").

[0361] Ocular Surface Disease Index (OSDI) (Copyright) The OSDI is a 12-item questionnaire that assesses the impact of dry eye symptoms and visual function on a subject's lifestyle over the past week, allowing for rapid assessment of eye irritation symptoms in dry eye disease (see, for example, Arch Ophthalmol. 2000;118(5):615-621 by RMSchiffman et al., incorporated herein by reference).

[0362] The higher the final score, the greater the obstacle.

[0363] Standard Patient Evaluation of Eye Dryness (SPEED) The SPEED questionnaire (Korb and Blackie, Ocular Surgery News Europe Edition, 2012 (as incorporated herein by reference)) is another assessment for monitoring dry eye symptoms over time, consisting of eight items with a score of 0 to 28 that evaluate the frequency and severity of symptoms including dryness, grittiness, scratching, irritation, burning, tearing, pain, and eye fatigue.

[0364] A higher score indicates a greater disability.

[0365] IV. Adverse Events An adverse event (AE) is any undesirable medical event in a patient receiving a drug or in a clinical trial where a causal relationship with the treatment is not necessarily clear. Therefore, an AE can be any undesirable, unintended sign (including abnormal clinical laboratory findings), symptom, or disease that is temporarily related to the use of a drug (investigation drug), regardless of whether it is directly related to the drug (investigation drug).

[0366] A serious adverse event (SAE) refers to an undesirable medical event that occurs at any dose, including the following: ● It can lead to death. ● Life-threatening (This refers to events in which the subject was at risk of death at the time of the event, and does not include events in which death would have occurred if the event had been more serious). ● Hospitalization is required, or an extension of the current hospitalization is required (hospitalization for elective surgery does not qualify as SAE). ● Causes persistent or significant impairment / inability. ● This is a congenital anomaly / birth defect.

[0367] At each patient visit, questions about adverse events will be asked using an open-ended questioning method, taking care not to influence the patient's responses.

[0368] Record any adverse events (AEs) and serious adverse events (SAEs) experienced by the subject from visit 2 (insertion / day 1) to visit 9 (follow-up 30 days later), regardless of the severity of the event or its association with the investigational treatment.

[0369] Exacerbations (AEs) that were previously documented and designated as ongoing in previous assessments will be re-examined at subsequent visits as needed, and any resolutions will be documented.

[0370] Changes in the intensity or frequency of AEs are recorded as separate events (i.e., a new recording is initiated).

[0371] Any ongoing SAEs when a subject completes or discontinues the trial will be followed up until the event is resolved, stabilized, or returns to the baseline state.

[0372] All events are evaluated to determine whether they meet the criteria for SAE, their severity, and their relevance to the investigational treatment.

[0373] Example 4.1: Cohort 1: Open-label, single-center phase 1 trial Cohort 1 was an open-label Phase 1 trial intended to evaluate the safety, tolerability, durability, and biological activity of a hydrogel / cyclosporine implant. Treatment allocation was communicated to the sponsor, principal investigator, and subjects, and followed the trial schedule outlined in Section I. Study Schedule above. After eligibility confirmation, all 5 enrolled patients (10 eyes) in Cohort 1 received implantation of Formulation 2A during visit 2, following the procedure outlined in Section II. Implant Placement above.

[0374] I. Safety and Tolerability All subjects completed the 16-week study period, and there were no dropouts. No serious adverse events were reported. The implants were observed to be well-tolerated, and no adverse events such as stinging, irritation, blurred vision, or lacrimation were reported or observed. Replacement implants were not required. The ease of insertion of the implants was evaluated as follows: ●8 eyes: easy ●1 eye: moderate ●1 eye: difficult

[0375] The assessments of moderate and difficult difficulty were for the right and left eyes of the same subject. This indicates that the overall dimensions and swelling behavior of the implant are excellently tuned for easy implant administration.

[0376] II.Durability All implants were visualized during the last visit (Clinic 8).

[0377] III. Biological activity / efficacy The effectiveness of the implants was evaluated based on ophthalmological assessments performed at each visit.

[0378] Tear production, as measured by the Schirmer test (without anesthesia), improved from a mean of 4.2 mm at baseline to 8.2 mm at week 12, and in 1 out of 5 patients (20%), the Schirmer score increased by 10 mm or more from baseline at week 12 (see Figures 6A and 6B).

[0379] The subjects demonstrated improvement in signs of dry eye disease (DED) as measured by tCFS. tCFS improved from a mean 6.7 at baseline to a mean 2.7 at week 12 (see Figure 7A), resulting in a change from baseline (CFB) of 4.0 at week 12 (see Figure 7B).

[0380] Symptoms of DED, as measured by the Visual Analog Scale (VAS) dry eye severity score, also improved. (i) The mean value improved from a baseline of 51 to a mean value of 33 at week 12 (see Figures 8A and 8B), and (ii) The VAS dry eye frequency score improved from a mean of 51 at baseline to a mean of 31 at week 12 (see Figure 9).

[0381] The effects of the implant were observed as early as two weeks for both signs and symptoms of DED (measured by VAS ocular dryness severity and frequency scores) and continued throughout the 16-week study period.

[0382] In addition, the Ocular Surface Disease Index (OSDI) and the Standard Patient Assessment of Dry Eyes (SPEED) scores also decreased over 16 weeks (see Figures 10 and 11).

[0383] Example 4.2: Cohort 2: Randomized, multicenter, double-blinded, vehicle-controlled phase 2 trial Cohort 2 is an ongoing randomized, double-blind, vehicle-controlled phase 2 trial intended to evaluate the safety, tolerability, and efficacy of hydrogel / cyclosporine implants. Treatment allocation was blinded to subjects, the principal investigators and their staff, and the sponsor's representatives, and followed the trial schedule outlined in Section I. Trial Schedule above.

[0384] The randomization schedule was computer-generated by a qualified biostatistician unrelated to the trial implementer or project team, and the hydrogel / cyclosporine and HV inserts administered to subjects at randomization during the double-blind treatment phase were made identical in appearance. Information regarding the treatment was shielded from subjects, the principal investigator, and their staff until the final database was locked, and sponsor staff involved in conducting and monitoring the trial remained shielded until the trial was completed and the database was locked.

[0385] In visit 2, after eligibility confirmation, each of the 140 patients enrolled in Cohort 2 received one of the implants (formulations 1, 2A, 2B, and 3) according to the randomization schedule for visit 2. The administration of the implants followed the procedure outlined in Section II. Implant Placement above.

[0386] If unlocking is required, access to the unlocked data will be limited to two individuals not involved in the trial (the sponsor, the medical monitor, and the sponsor's statistical officer), or, in emergency situations, the principal investigator will be allowed to access it directly, thereby maintaining the integrity of the trial evaluation and objectives.

[0387] IV. Safety and Tolerability Adverse events, including stinging, irritation, blurred vision, or watery eyes, were continuously monitored.

[0388] V.Durability The durability of an implant is evaluated by monitoring its presence or absence.

[0389] VI. Biological activity / efficacy The effectiveness of the implant will be evaluated based on ophthalmological assessments performed at each visit. The effectiveness endpoints are defined as follows: Primary endpoints ● Change from baseline (CFB) and absolute value of Schirmer test (unanesthetized) at 12 weeks Secondary endpoints Symptoms: ●Percentage of subjects whose Schirmer score increased by 10mm or more in week 12 ●Corneal fluorescein staining (tCFS) using the NEI scale at each visit after baseline: CFB and absolute value ●CFB and absolute values ​​of corneal fluorescein-stained subregions using the NEI scale at each visit after baseline. ●CFB and absolute value of conjunctival lysamine green staining using the NEI scale at each visit after baseline Symptoms (reported by subject): ●CFB and absolute values ​​of the visual axis score (VAS) at each trial visit after baseline. ●CFB and absolute values ​​of the Ocular Surface Disease Index (OSDI (copyright) total score, each of the three domains, and individual questions) at each visit after baseline. ● CFB of the SPEED questionnaire (overall score and individual questions) at each visit after baseline. Exploratory items: ● CFB of tear membrane permeability time (TBUT) at 12 weeks ● Presence or absence of implants at all follow-up examinations after baseline ● Ease of insertion as assessed by the principal investigator. ● Ease of visualization through evaluation by the principal investigator.

[0390] While several embodiments of the present invention have been described, it is clear that the inventors' basic embodiments can be modified to provide other embodiments utilizing the compounds and methods of the present disclosure. Therefore, it will be understood that the scope of the present disclosure should be defined by the appended claims rather than by the specific embodiments shown as examples.

[0391] Throughout this application, various references are cited. The disclosures of these references are incorporated herein by reference. The present invention also includes the following embodiments. <1> A sustained-release biodegradable intratubular insert comprising a hydrogel and an active agent, wherein the active agent takes the form of particles and the particles of the active agent are dispersed within the hydrogel. <2> The sustained-release biodegradable intratubular insert according to claim 1, wherein the particles of the active agent have a d50 value of less than approximately 50 μm. <3> The sustained-release biodegradable tube insert according to item 2, wherein the particles of the active agent have a d50 value in the range of 3 to 17 μm, preferably 4 to 12 μm, and more preferably 5 to 8 μm. <4> The active agent is a low-solubility active agent, and is preferably used to treat ocular diseases or disorders affecting the surface of the eye, including dry eye diseases, blepharitis, allergic conjunctivitis, and especially atopic keratoconjunctivitis and vernal keratoconjunctivitis. The active agent is more preferably selected from the group consisting of immunosuppressants, nonsteroidal anti-inflammatory drugs (NSAIDs), antibiotics, antihistamines, essential fatty acids, and rifitegrass. The active agent is most preferably cyclosporine, everolimus, tacrolimus, sirolimus, pimecrolimus, ibuprofen, mefanamic acid, diclofenac, nepafenac, flubiprofen, flubiprofen sodium, fusidic acid, becifloxacin (base), clarithromycin, azithromycin, ketotifen (base), azelastine (base), azelastine emponate, linoleic acid, α-linoleic acid, prednisone, prednisolone, prednisolone acetate, methylprednisolone, dexamethasone, dexamethasone acetate, or betamethasone phosphate sodium. A sustained-release biodegradable intratubular insert according to any one of the above 1 to 3, selected from the group consisting of lium, budesonide, flunisolide, fluticasone propionate, triamcinolone, triamcinolone acetonide, triamcinolone hexacetonide, triamcinolone diacetate, fluocinolone acetonide, fludrocortisone acetate, loteprednol, loteprednol etavonate, difluprednate, fluorometholone, mometasone furoate, deoxycorticosterone acetate, aldosterone, rimexolone, beclomethasone, beclomethasone dipropionate, and rifiteglast. <5> The sustained-release biodegradable tube insert according to any one of 1 to 4 above, wherein the hydrogel comprises a polymer network, preferably the polymer network comprises the same or different crosslinked polymer units, and more preferably the crosslinked polymer units are one or more crosslinked polyethylene glycol units. <6> The sustained-release biodegradable tube insert according to 5, wherein the polymer network comprises polyethylene glycol units having an average molecular weight in the range of about 2,000 to about 100,000 daltons, preferably about 10,000 to about 60,000 daltons, and more preferably about 20,000 to about 40,000 daltons. <7> The sustained-release biodegradable tube insert according to 5 or 6 above, wherein the polymer network comprises one or more crosslinked multi-arm polymer units, preferably the multi-arm polymer unit comprising one or more 2-10 arm polyethylene glycol units, more preferably one or more 4-8 arm polyethylene glycol units, and most preferably one 4-arm polyethylene glycol unit. <8> The sustained-release biodegradable tube insert described in 7 above, wherein the four arms of the four-arm polyethylene glycol unit are connected to a pentaerythritol core molecule. <9> The sustained-release biodegradable intratubular insert according to any one of 5 to 8 above, wherein the polymer network is formed by reacting an electrophile-containing multi-arm polymer precursor with a nucleophile-containing crosslinking agent. <10> The sustained-release biodegradable intratubular insert described in 9 above, wherein the electrophile is an activated ester group, more preferably an N-hydroxysuccinimidyl (NHS) ester group, and even more preferably selected from the group consisting of succinimidylmalonic acid group, succinimidylsuccinic acid (SS) group, succinimidylmaleic acid group, succinimidyl fumarate group, succinimidylglutaric acid (SG) group, succinimidyladipic acid (SAP) group, succinimidylpimelic acid group, succinimidylsuberic acid group, and succinimidylazelaic acid (SAZ) group. <11> The sustained-release biodegradable intratubular insert according to 9 or 10 above, wherein the nucleophilic group-containing crosslinking agent is an amine, preferably a low molecular weight amine with a molecular weight of less than 1,000 Da containing two or more primary aliphatic amine groups, more preferably a low molecular weight amine selected from the group consisting of dyridine, trilidine, tetralysine, ethylenediamine, 1,3-diaminopropane, 1,3-diaminopropane, diethylenetriamine, and trimethylhexamethylenediamine, even more preferably trilidine, and most preferably trilidine acetate. <12> The sustained-release biodegradable intratubular insert according to 11, wherein the nucleophilic group-containing crosslinking agent is labeled trillidine, preferably labeled trillidine labeled with a visualizing agent, more preferably labeled trillidine labeled with a visualizing agent selected from the group consisting of fluorophores (e.g., fluorescein), rhodamine, coumarin, and cyanine, even more preferably fluorescein-conjugated trillidine, and most preferably fluorescein-conjugated trillidine obtained by reacting trillidine acetate with N-hydroxysuccinimide (NHS)-fluorescein. <13> A sustained-release biodegradable intratubular insert according to any one of 7 to 13 above, wherein the multi-arm polymer unit comprises a 4a20k PEG unit and the crosslinking unit comprises a fluorescein-conjugated trilysidineamide unit. <14> The polymer network is obtained by reacting 4a20k PEG-SG or 4a20k PEG-SG with fluorescein-conjugated trilysine in a molar ratio in the range of about 1:2 to about 2:1, preferably in a molar ratio of about 1:1, as described in any of 5 to 13 above. <15> A sustained-release biodegradable tube insert according to any one of 1 to 14 above, wherein the insert, in a dry state, contains, based on the total weight of the insert, about 15% to about 80% by weight of the active agent and about 20% to about 60% by weight of polymer units based on the total weight of the insert, preferably, 30% to 65% by weight of the active agent and 25% to 50% by weight of polymer units based on the total weight of the insert, more preferably, 45% to 55% by weight of the active agent and 37% to 47% by weight of polymer units based on the total weight of the insert. <16> A sustained-release biodegradable intratubular insert comprising a hydrogel and an active agent, wherein the insert, in a dry state, contains approximately 40% to approximately 80% by weight of the active agent based on the total weight of the insert. <17> A sustained-release biodegradable intratubular insert according to any one of 1 to 16 above, wherein the insert contains, in a dry state, 45% to 55% by weight of the active agent based on the total weight of the insert. <18> The sustained-release biodegradable tube insert according to any one of items 1 to 17 above, wherein the insert contains a surfactant. <19> A sustained-release biodegradable intratubular insert comprising a hydrogel and an active agent, wherein the insert comprises a surfactant. <20> The sustained-release biodegradable tube insert according to 18 or 19, wherein the insert contains, in a dry state, about 0.01% to about 5% by weight, preferably 0.2% to 2% by weight, of a surfactant based on the total weight of the insert. <21> The sustained-release biodegradable tube insert according to any one of the above 18 to 20, wherein the insert comprises a nonionic surfactant, preferably a nonionic surfactant containing a poly(ethylene glycol) chain, and the surfactant is more preferably selected from the group consisting of poly(ethylene glycol) monolaurate sorbitan, castor oil poly(ethylene glycol) ester, and ethoxylated 4-tert-octylphenol / formaldehyde condensation polymer, and even more preferably selected from the group consisting of poly(ethylene glycol)-20-monolaurate sorbitan, poly(ethylene glycol)-80-monolaurate sorbitan, castor oil poly(ethylene glycol)-35 ester, and ethoxylated 4-tert-octylphenol / formaldehyde condensation polymer, and most preferably ethoxylated 4-tert-octylphenol / formaldehyde condensation polymer. <22> A sustained-release biodegradable intratubular insert according to any one of items 1 to 21 above, wherein the content of the active agent, measured by HPLC after storage at a temperature of 2 to 8°C for at least 3 months, preferably at least 6 months, and more preferably at least 12 months, is about 300 to about 410 μg or about 90 to about 110% by weight. <23> A sustained-release biodegradable intratubular insert according to any one of items 1 to 22 above, wherein the amount of impurities measured by HPLC after storage at a temperature of 2 to 8°C for at least 3 months, preferably at least 6 months, and more preferably at least 12 months, is 3.0% or less. <24> The sustained-release biodegradable tube insert according to any one of the above 1 to 23, wherein the insert takes the form of fibers, and the fibers preferably have an average length of about 1.5 mm to about 4.0 mm and an average diameter of 0.8 mm or less in a dry state, more preferably an average length of 2.0 mm to 2.5 mm and an average diameter of 0.62 mm or less in a dry state, and even more preferably an average length of 2.5 mm to 2.9 mm and an average diameter of 0.62 mm or less in a dry state. <25> A sustained-release biodegradable tube insert according to any one of the above 1 to 24, wherein the insert, after being stored at a temperature of 2 to 8°C for at least 3 months, preferably at least 6 months, and more preferably at least 12 months, takes the form of fibers having an average length of about 2.5 mm to about 2.9 mm and an average diameter of 0.62 mm or less in its dry state. <26> The sustained-release biodegradable intratubular insert according to any one of 1 to 25 above, wherein the insert takes the form of fibers having an average diameter of at least 1.0 mm in a swollen state 10 minutes after hydration, or an average diameter of at least 1.3 mm in an equilibrium state 24 hours after in vitro hydration in phosphate-buffered saline at 37°C and pH 7.4. <27> The sustained-release biodegradable tube insert according to any one of the above 1 to 26, wherein the insert disintegrates within the tube within approximately 1 to 6 months, preferably within 2 to 4 months, more preferably within 2 to 3 months or 3 to 4 months after insertion. <28> A sustained-release biodegradable intratubular insert according to any one of the above 1 to 27, wherein the insert, after being inserted into the tube, releases a therapeutically effective amount of active drug over a period of at least about one month, preferably at least two months, and more preferably at least three months after insertion. <29> A sustained-release biodegradable intratubular implant according to any one of the above 1 to 28, wherein the active agent is released from the implant at an average rate of about 0.1 μg / day to about 10 μg / day, preferably at an average rate of 1 μg / day to 5 μg / day, more preferably at an average rate of 2 μg / day to 4 μg / day after insertion into a human subject. <30> A sustained-release biodegradable intratubular implant according to any one of the above 1 to 29, wherein the concentration of the active drug in the tear film after insertion into a human subject is in the range of approximately 0.1 μg / mL to approximately 10 μg / mL, preferably in the range of approximately 1 μg / mL to approximately 5 μg / mL. <31> A sustained-release biodegradable tube insert according to any one of the above 1 to 30, wherein the insert disintegrates within the tube before the particles of the active agent contained in the insert are completely dissolved. <32> The sustained-release biodegradable tube insert according to any one of items 1 to 31, wherein the fibers are stretched before or after drying, preferably before drying. <33> A sustained-release biodegradable intratubular insert comprising a fibrous hydrogel and an active agent, wherein the fibers are stretched. <34> The sustained-release biodegradable tube insert according to 32 or 33, wherein the fibers are stretched in the longitudinal direction with a stretch coefficient of about 1.0 to about 4.0, preferably about 1.5 to about 3.0, and more preferably about 2.7. <35> A sustained-release biodegradable intratubular insert, A hydrogel containing a polymer network obtained by reacting 4a20k PEG-SG with fluorescein-conjugated trilysine in a molar ratio of approximately 1:1, It contains approximately 360 μg of the active agent, The sustained-release biodegradable tube insert takes the form of fibers having an average length of approximately 2.5 mm to approximately 2.9 mm and an average diameter of 0.62 mm or less in its dry state. <36> A sustained-release biodegradable intratubular insert, A hydrogel containing a polymer network obtained by reacting 4a20k PEG-SAP with fluorescein-conjugated trilysine in a molar ratio of approximately 1:1, It contains approximately 360 μg of the active agent, The sustained-release biodegradable tube insert takes the form of fibers having an average length of approximately 2.5 mm to approximately 2.9 mm and an average diameter of 0.62 mm or less in its dry state. <37> A method for performing the treatment or prevention of an eye disease in a human subject requiring treatment or prevention, comprising inserting a first sustained-release biodegradable intratubular insert containing the hydrogel and active agent described in any of items 1 to 36 above into a tube in the human subject. <38> The method according to 37, wherein the first insert is designed to remain in the tube until it completely disintegrates, and / or the first insert is designed to disintegrate within about 3 to 4 months after insertion. <39> A method for treating dry eye disease in a subject, wherein the method is (a) The step of inserting a first biodegradable insert into the first cannula of the first eye of the object, wherein the insert is (1) Hydrogel and, (2) comprising approximately 100 μg to approximately 800 μg of active agent dispersed within the hydrogel, (3) The insertion step, wherein the active agent is released from the insert at an average rate of about 0.1 μg / day to about 10 μg / day for a period of at least about two months from the day the first insert is inserted into the target, (b) The method comprising the step of inserting a second implant into the first canal of the first eye of the subject at least about two months after the date of insertion of the first implant, wherein the second implant is substantially similar to the first implant. <40> The first insert is removed before it completely collapses, preferably A second insert is inserted to replace the removed first insert, and / or the first insert is designed to disintegrate within the tube within approximately 2 to 3 months after insertion, and / or The method according to 39, wherein the first insert remains in the small tube until it completely disintegrates. <41> The method according to any one of the above 37 to 40, wherein a second insert is inserted at least two months later without prior removal of the first insert, or the first insert is removed before it completely disintegrates, and the second insert is inserted to replace the removed first insert, preferably the first insert is designed to disintegrate within the tube within about two to three months after insertion, and the first insert is removed within two months after insertion. <42> The method according to any one of 37 to 41 above, wherein the eye disease is a disorder of the tear film and the surface of the eye, preferably a dry eye disease, and more preferably the eye disease is related to one or more conditions selected from the group consisting of burning sensation, itching, redness, stinging, pain, foreign body sensation, visual impairment, inflammation of the lacrimal gland, inflammation of the surface of the eye, T cell-mediated inflammation, presence of conjunctival T cells in the tears, and elevated levels of inflammatory cytokines in the tears. <43> The treatment is effective in improving tear production, as measured by the Schirmer tear test, in subjects with a Schirmer score of less than 10 mm before insertion of the implant, preferably the Schirmer score increases by at least 2 mm 6 weeks after insertion and / or increases by at least 3 mm 12 weeks after insertion and / or The treatment is effective in reducing symptoms of dry eyes as determined by one or more assessments selected from the group consisting of assessment of the severity of dry eye symptoms on a visual analog scale, assessment of the frequency of dry eye symptoms on a visual analog scale, quantification of tear film breakup time, corneal fluorescein staining, conjunctival lysamine green staining, best corrected visual acuity, quantification of the ocular surface disease index, and a standard patient assessment of dry eyes, preferably the method according to any one of 37 to 42 above, wherein the total tCFS of corneal fluorescein staining values ​​decreases by at least 1.5 six weeks after insertion and / or at least 3 twelve weeks after insertion, and preferably the assessment of the severity of dry eye symptoms on a visual analog scale decreases by at least 10 two weeks after insertion and / or at least 15 six weeks after insertion. <44> The method according to any of the above 37-43, wherein the treatment period is at least one month, at least two months, or at least three months. <45> A method for producing a sustained-release biodegradable intratubular insert containing a hydrogel and an active agent as described in any of items 1 to 36 above, a) A step of preparing a precursor mixture comprising a hydrogel precursor and particles of an active agent dispersed in the precursor mixture, b) The step of molding the precursor mixture, crosslinking the hydrogel precursor to form a polymer network, and obtaining a molded hydrogel mixture containing the polymer network, c) The method comprising the step of drying the hydrogel mixture to obtain the insert. <46> The method according to 45, wherein in step a), the precursor mixture is prepared by mixing an electrophile-containing multi-arm polymer precursor with a nucleophile-containing crosslinking agent in a buffered aqueous solution in the presence of micronized active agent particles, preferably the electrophile-containing multi-arm polymer precursor is provided in a buffered aqueous precursor solution and the nucleophile-containing crosslinking agent is provided in a buffered aqueous precursor suspension containing the micronized active agent particles. <47> The method according to 45 or 46, wherein in step a), the precursor mixture containing particles of the active agent is degassed under vacuum after the mixing of its components. <48> The method according to any one of the above 45 to 47, further comprising stretching the hydrogel mixture fibers, wherein the stretching is performed before or after drying the hydrogel mixture, and preferably the fibers are stretched with a draw coefficient of about 1 to about 4.5. <49> A method for imparting shape memory to a hydrogel mixture fiber, wherein the hydrogel mixture fiber contains particles of an active agent dispersed within the hydrogel, and the shape memory is imparted by stretching the hydrogel mixture fiber in the longitudinal direction. <50> Use of a sustained-release biodegradable intratubular insert containing the hydrogel and active agent described in any of items 1 to 36 above in the preparation of a pharmaceutical for the treatment of an eye disease in a subject requiring treatment, as described in any of items 37 to 44 above. <51> A sustained-release biodegradable intratubular implant comprising the hydrogel and active agent described in any of items 1 to 36 above, for use in the treatment of an eye disease in a subject requiring treatment as described in any of items 37 to 44 above.

Claims

1. A sustained-release biodegradable intratubular insert containing hydrogel and cyclosporine, The cyclosporine takes the form of particles, and the cyclosporine particles are dispersed within the hydrogel. The hydrogel comprises a polymer network comprising one or more crosslinked multi-arm polymer units, and the multi-arm polymer unit comprises one or more 2- to 10-arm polyethylene glycol units. The cyclosporine particles have a d50 value of less than 50 μm when measured by laser diffraction using USP<429>. The sustained-release biodegradable tube insert wherein the insert, in a dry state, contains 40% to 80% by weight of cyclosporine and 0.05% to 5% by weight of a surfactant based on the total weight of the insert.

2. The sustained-release biodegradable tube insert according to claim 1, wherein the cyclosporine particles have a d50 value in the range of 3 to 17 μm.

3. The sustained-release biodegradable tube insert according to claim 1 or 2, wherein the polymer network comprises the same or different cross-linked polymer units.

4. The sustained-release biodegradable tube insert according to any one of claims 1 to 3, wherein the polymer network comprises polyethylene glycol units having an average molecular weight in the range of 2,000 to 100,000 daltons.

5. The sustained-release biodegradable tube insert according to any one of claims 1 to 4, wherein the multi-arm polymer unit comprises one or more 4- to 8-arm polyethylene glycol units.

6. The sustained-release biodegradable intratubular insert according to any one of claims 1 to 5, wherein the polymer network is formed by reacting an electrophile-containing multi-arm polymer precursor with a nucleophile-containing crosslinking agent.

7. The electrophile is an N-hydroxysuccinimidyl (NHS) ester group selected from the group consisting of succinimidylmalonic acid group, succinimidylsuccinic acid (SS) group, succinimidylmaleic acid group, succinimidyl fumarate group, succinimidylglutaric acid (SG) group, succinimidyladipic acid (SAP) group, succinimidylpimelic acid group, succinimidylsuberic acid group, and succinimidylazelaic acid (SAZ) group, and / or The sustained-release biodegradable tube insert according to claim 6, wherein the nucleophilic group-containing crosslinking agent is selected from the group consisting of dyridine, trilysine, tetralysine, ethylenediamine, 1,2-diaminopropane, 1,3-diaminopropane, diethylenetriamine, and trimethylhexamethylenediamine.

8. The sustained-release biodegradable intratubular insert according to claim 6 or 7, wherein the nucleophilic group-containing crosslinking agent is labeled trilysine.

9. The sustained-release biodegradable tube insert according to any one of claims 1 to 8, wherein the insert, in a dry state, contains 20% to 60% by weight of polymer units based on the total weight of the insert.

10. The sustained-release biodegradable intratubular insert according to any one of claims 1 to 9, wherein the surfactant is selected from the group consisting of poly(ethylene glycol) monolaurate sorbitan, castor oil poly(ethylene glycol) ester, and ethoxylated 4-tert-octylphenol / formaldehyde condensation polymer.

11. The sustained-release biodegradable tube insert according to any one of claims 1 to 10, wherein the insert takes the form of fibers.

12. The aforementioned fibers have an average length of 1.5 mm to 4.0 mm and an average diameter of 0.8 mm or less in their dry state, and / or The sustained-release biodegradable tube insert according to claim 11, wherein the fibers have an average diameter of at least 1.0 mm in a swollen state 10 minutes after hydration, or an average diameter of at least 1.3 mm in an equilibrium state 24 hours after in vitro hydration in phosphate-buffered saline at 37°C and pH 7.

4.

13. The aforementioned insert disintegrates within the aforementioned tube within 1 to 6 months after insertion, and / or The sustained-release biodegradable tube insert according to any one of claims 1 to 12, wherein the insert disintegrates within the tube before the cyclosporine particles contained in the insert are completely dissolved.

14. A sustained-release biodegradable intratubular implant according to any one of claims 1 to 13, wherein the cyclosporine concentration in the tear film after insertion into a human subject is in the range of 0.1 μg / mL to 10 μg / mL.

15. A sustained-release biodegradable tube insert according to any one of claims 1 to 14, wherein the insert is in the form of fibers and the fibers are stretched.

16. The sustained-release biodegradable tube insert according to claim 15, wherein the fibers are stretched before or after drying.

17. The sustained-release biodegradable tube insert according to claim 16, wherein the fibers are stretched in the longitudinal direction with a stretching coefficient of 1.0 to 4.

0.

18. A sustained-release biodegradable tube insert according to any one of claims 1 to 17, comprising a hydrogel containing a polymer network obtained by reacting 4a20k PEG-SG or 4a20k PEG-SAP with fluorescein-conjugated trilysine in a 1:1 molar ratio, and 360 μg of cyclosporine, wherein the fibrous material has an average length of 2.5 mm to 2.9 mm and an average diameter of 0.62 mm or less in its dry state.

19. A method for performing the treatment or prevention of an eye disease in a human subject requiring treatment or prevention, comprising inserting a first sustained-release biodegradable intratubular insert containing the hydrogel and cyclosporine described in any one of claims 1 to 18 into a tube in the human subject, for use in the method, the sustained-release biodegradable intratubular insert according to any one of claims 1 to 18.

20. The sustained-release biodegradable tube insert according to claim 19, wherein the first insert is designed to remain in the tube until it completely disintegrates, and / or the first insert is designed to disintegrate in the tube within 3 to 4 months after insertion.

21. A method for treating dry eye disease in a subject, wherein the method is (a) A step of inserting a first biodegradable insert into the first tubule of the first eye of the object, wherein the insert is (1) Hydrogel and, (2) comprising 100 μg to 800 μg of cyclosporine dispersed in the hydrogel, (3) The insertion step, wherein the cyclosporine is released from the insert at an average rate of 0.1 μg / day to 10 μg / day for a period of at least two months from the day the first insert is inserted into the target, (b) A sustained-release biodegradable intratubular implant according to any one of claims 1 to 18, for use in the method, comprising the step of inserting a second implant into the first tube of the first eye of the subject at least two months after the date of insertion of the first implant, wherein the second implant is substantially similar to the first implant.

22. The first insert is removed before it completely collapses, and / or The first insert is designed to disintegrate within the tube within two to three months after insertion, and / or The sustained-release biodegradable tube insert according to claim 21, wherein the first insert remains inside the tube until it completely disintegrates.

23. A sustained-release biodegradable tube insert according to any one of claims 19 to 22, wherein a second insert is inserted at least two months later without prior removal of the first insert, or the first insert is removed before it completely disintegrates and the second insert is inserted to replace the removed first insert.

24. The aforementioned eye disease is a disorder of the tear film and the surface of the eyeball, and / or The treatment is effective in improving tear production, as measured by the Schirmer tear test, in subjects with a Schirmer score of less than 10 mm before insertion of the implant, and / or The treatment is effective in reducing symptoms of dry eyes as determined by one or more assessments selected from the group consisting of assessment of the severity of dry eye symptoms on a visual analog scale, assessment of the frequency of dry eye symptoms on a visual analog scale, quantification of tear film breakup time, corneal fluorescein staining, conjunctival lysamine green staining, best corrected visual acuity, quantification of the ocular surface disease index, and standard patient assessment of dry eyes, and / or A sustained-release biodegradable intratubular implant according to any one of claims 19 to 23, wherein the treatment period is at least one month, at least two months, or at least three months.

25. A method for producing a sustained-release biodegradable tube insert according to any one of claims 1 to 18, a) A step of preparing a precursor mixture comprising a hydrogel precursor and cyclosporine particles dispersed in the precursor mixture, b) The steps of molding the precursor mixture, crosslinking the hydrogel precursor to form a polymer network, and obtaining a molded hydrogel mixture containing the polymer network, c) The method comprising the step of drying the hydrogel mixture to obtain the insert.

26. In step a), the precursor mixture is prepared by mixing an electrophile-containing multi-arm polymer precursor with a nucleophile-containing crosslinking agent in a buffered aqueous solution in the presence of finely atomized cyclosporine particles, and in step a), the precursor mixture containing cyclosporine particles is degassed under vacuum after mixing of its components, and / or The method according to claim 25, further comprising stretching the hydrogel mixture fibers, wherein the stretching is performed before or after drying of the hydrogel mixture.

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