Cysteine ​​derivatives for the treatment of presbyopia

Abstract

This invention is based on the discovery that N-pivaloylcysteine ​​and its derivatives function to improve the flexibility of the lens and treat presbyopia. N-pivaloylcysteine ​​and N-(2-fluoro-2-methyl-L-oxopropyl)-cysteine ​​showed improvement in reducing lens rigidity and aqueous humor permeability compared to N-acetylcysteine. This invention includes N-pivaloylcysteine ​​and its derivatives, pharmaceutical compositions thereof, and methods of administering them for the prevention of ophthalmic diseases related to lens rigidity, such as presbyopia.

Description

Technical Field

[0001] Technical Field The present invention relates to the therapeutic use of N-pivaloyl-L-cysteine derivatives, particularly for the treatment or prevention of presbyopia or cataract.

Background Art

[0002] Background of the Invention Presbyopia, i.e., age-related farsightedness, is an age-related disorder caused by inappropriate focusing of light on the retina, which gradually reduces the eye's ability to focus on nearby objects and causes problems related to decreased near vision, such as difficulty in reading. Presbyopia usually develops between the ages of 40 and 50 and occurs as a result of various physiological changes in the eye, including atrophy of the ciliary muscle and sclerosis of the lens and lens capsule. These changes综合导致 the loss of the lens's regulatory function and cause the symptoms of presbyopia, but the sclerosis of the lens is presumed to be the most powerful inducer. The rigidity of the lens is due to damage caused by cross-linking of long-lived lens proteins, which contributes to the formation of protein aggregates that can damage the lens at the structural level and inhibit the regulatory function.

[0003] The pathophysiology of cataract also has many characteristics in common with that of presbyopia, which is related to damage to lens proteins associated with aging. Presbyopia is an early symptom of lens aging, while cataract formation is a more advanced and virtually terminal symptom. For example, it has been shown that both diseases are affected by glutathione depletion, and the presence of protective thiol compounds is important for maintaining healthy lens physiology in both cases.

[0004] The standard treatment for presbyopia has traditionally involved the use of corrective lenses such as bifocal lenses, progressive lenses, and multifocal contact lenses. While standard treatment can be an effective treatment strategy, it is cumbersome, does not restore accommodative function, and therefore cannot restore focus to the entire field of vision. Furthermore, its effects are only temporary, as they are only effective while the lenses are being worn. Miotics that restore near vision by inducing the "pinhole effect" have been developed, but their effects are also temporary and often cause side effects such as headaches. Notable miotics include pilocarpine (currently the only eye drop approved for the treatment of presbyopia) and aceline. Drugs that directly reduce the rigidity of the lens and thereby restore accommodative function have a longer-lasting effect due to their mechanism of action and are likely to avoid the side effects associated with other miotics.

[0005] Miotics (including all eye drops currently in clinical development) need to be administered every few hours. Furthermore, contact lenses are designed to be disposable, increasing treatment costs. Eyeglasses solve this problem due to their durability, but both contact lenses and eyeglasses only alleviate symptoms while being worn. Miotics narrow the depth of the field of vision, and corrective lenses do not restore accommodation, generally limiting near vision to specific areas within the field of vision. Monovision laser surgery, which essentially restores near vision in one eye by altering corneal refractive error at the expense of distance vision, can significantly impair stereoscopic vision (and also involves other surgery-related complications and costs).

[0006] The only current treatment, intraocular lens (IOL) implantation, offers a permanent solution to lens damage. However, this surgical option is avoided until cataracts develop or until cataracts do not develop. All current treatments specifically for presbyopia are merely compensatory therapies and do not lead to improvement in lens physiological function over time. Biochemically reducing existing crosslinks may be the only way to improve lens physiological function towards a natural and healthy state.

[0007] Choline lipoate (LACE) has been shown to improve lens flexibility by reducing disulfide. LACE showed initial efficacy in Phase 1 / 2 trials and was being developed for the treatment of presbyopia. However, development was discontinued because LACE failed to meet clinical endpoints. While LACE did not adequately restore near vision, next-generation lens softeners that simultaneously target multiple damage forms may overcome LACE's shortcomings. Ursodeoxycholic acid has been directly compared to LACE. The need for optimized treatments for presbyopia remains.

[0008] N-acetylcysteine ​​(NAC) can simultaneously treat multiple contributing factors to lens damage. For example, NAC acts as a disulfide cleavage agent, AGE inhibitor, antioxidant, and metal chelator, and is also a rate-limiting precursor for glutathione (GSH) synthesis, which in turn acts through these mechanisms.

[0009] S,N-diacetylcysteine, a derivative of NAC modified with a hydrogen sulfide group, suppresses protein aggregation and AGE formation through disulfide reduction and acetylation of the lysine side chain, thereby reducing lens rigidity in ex vivo treated aged mouse lenses.

[0010] While NAC derivatives have been proven to enhance tissue bioavailability and efficacy, acetyl group modification is generally underutilized and has not been explored in the field of presbyopia. [Overview of the project] [Problems that the invention aims to solve]

[0011] Summary of the Invention The objective of the present invention is to provide a sustainable strategy for preventing and treating presbyopia, a common disease that occurs in middle age, without the need for significant disease-modifying treatment strategies, by improving the flexibility of the lens.

[0012] The object of the present invention is to provide a pharmaceutically acceptable ophthalmic composition comprising the N-pivaloyl-L-cysteine ​​derivative of the present invention. The chemical names N-(2,2-dimethyl-1-oxopropyl)-L-cysteine, N-pivaloyl-L-cysteine, and N-pivaloylcysteine ​​are used interchangeably herein.

[0013] The object of the present invention is to provide a rationally designed NAC derivative that is effective, long-acting, and safe as a treatment option for presbyopia by reducing the rigidity of the lens. The present invention is at least in part based on the discovery that N-pivaloylcysteine ​​and its derivatives improve uptake into the lens and function in treating presbyopia. [Means for solving the problem]

[0014] The inventors have discovered that NAC and its derivatives reduce the rigidity of the lens, and that N-pivaloylcysteine ​​and its derivatives show a significant improvement in the effect of NAC.

[0015] The compound of the present invention has excellent persistence, and its therapeutic effect is expected to last for a long period even after treatment, making it possible to administer it at a lower frequency than existing miotic drugs.

[0016] The compounds of this invention also address the shortcomings of LACE, which failed in late-stage clinical development. While lipoic acid functions as a disulfide cleavage agent, its beneficial effects on AGE or GSH levels have not been clearly demonstrated. NAC derivatives have the potential for diverse effects, including disulfide reduction and inhibition of AGE.

[0017] The compounds of the present invention are administered as eye drops and, unlike other treatment options such as miotics and corrective lenses, improve the functionality and accommodative ability of the entire lens, restoring the entire field of vision.

[0018] This specification describes the following specific aspects of the invention.

[0019] N-pivaloylcysteine ​​derivatives, halogen-substituted derivatives (more specifically, fluorine derivatives, etc.), and pharmaceutically acceptable salts thereof for all therapeutic applications.

[0020] Halogen-substituted N-pivaloylcysteine ​​derivatives containing additional modifications to the C-terminus or side chain of a cysteine ​​molecule, and pharmaceutically acceptable salts thereof.

[0021] N-pivaloylcysteine ​​derivatives as described herein for the prevention or treatment of presbyopia and related diseases.

[0022] A method for preventing or treating an ophthalmic disease, disorder, or condition characterized by a decrease in the flexibility and / or elasticity of the lens, comprising contacting the eye of a subject (including a human) with an N-pivaloylcysteine ​​derivative as described herein.

[0023] A method for preventing or treating an ophthalmic disease, disorder, or condition characterized by a decrease in the flexibility of the lens, comprising contacting the eye of a subject (including a human) with the N-pivaloylcysteine ​​derivative described herein, wherein the ophthalmic disease is accompanied by a decrease in the accommodative function of the eye.

[0024] A method for preventing or treating presbyopia, comprising contacting the eye of a subject (including a human) with an N-pivaloylcysteine ​​derivative described herein.

[0025] A method for preventing or treating an ophthalmic disease, disorder, or condition, comprising contacting the eye of a subject (including a human) with an N-pivaloylcysteine ​​derivative described herein, wherein the selected route of administration is via an ophthalmic route.

[0026] A composition comprising the N-pivaloylcysteine ​​derivative described herein, which is formulated as eye drops, on contact lenses, or as an eye cream or gel.

[0027] A composition comprising an N-pivaloylcysteine ​​derivative as described herein, wherein the concentration of N-pivaloylcysteine, its derivative, or a pharmaceutically acceptable salt thereof contained in the formulation is 0.000005 to 15% (w / v).

[0028] All aspects of the present invention described above can be individually selected and / or combined. [Modes for carrying out the invention]

[0029] Detailed description of the invention The following detailed description is provided to assist those skilled in the art in carrying out the invention. Illustrative embodiments are described in detail below. However, these embodiments are illustrative only, and this disclosure is not limited thereto, but is defined by the appended claims. Those skilled in the art can modify and alter the embodiments described herein without departing from the spirit or scope of this disclosure.

[0030] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art to which this disclosure pertains. Terms used herein are for the purpose of describing specific embodiments and are not intended to be limiting. Furthermore, terms defined in commonly used dictionaries, etc., should be interpreted in a sense consistent with their meaning in the relevant technical field and in the context of this disclosure, and not in an idealized or overly formal sense unless expressly defined herein.

[0031] The articles "a" and "an" refer to one or more (i.e., at least one) that are the grammatical objects of the article, unless the context clearly indicates otherwise. For example, "one element" means one or more elements.

[0032] Therefore, the following embodiments are merely described to illustrate aspects of this specification by referring to structures and schemes. As used herein, the term "and / or" encompasses any combination of one or more of the associated listed items. The term "or" means "and / or". Expressions such as "at least one", when placed before a list of elements, modify the entire list of elements and not individual elements of the list.

[0033] The methods, compounds, and systems are not limited to the specific methods, compounds, components, or compositions described or exemplified herein. It should also be understood that the terms used herein are for purposes of description and not intended to be limiting.

[0034] Novel substance compositions are described as active ingredients for therapeutic purposes, i.e., the treatment of human diseases. Using these agents, the flexibility of the lens can be restored and / or the visual accommodation function can be restored.

[0035] One aspect of the present invention is the following formula:

Chemical formula

[0036] Another aspect of the present invention is the following formula: [ka] (In the formula, Each R 1 These are independently replaced by C of any choice. 1~6 Alkyl, C 1~6 It is an aryl, halogen, halogen-containing group, or ammonium group; R 2 is OR 4 , SR 4 ,SeR 4 , or N(R 4 )2; R 3 , SR 4 ,SeR 4 , or SC(O)-R 4 and; Here, R 4 C is a hydrogen atom or optionally substituted C 1~6 (It is alkyl.) It is an N-pivaloylcysteine ​​derivative.

[0037] Any substituents, but not limited to, can be selected from alkyl, cycloalkyl, aryl, heteroaryl, hydroxy, alkoxy, halogen, and conjugates, as well as functional groups, solubilizers, lipid groups, etc., to optimize pharmacokinetic and other properties.

[0038] Where used herein, unless otherwise specified, “alkyl” refers to a linear and branched carbon chain having 1 to 20 carbon atoms, or any number within this range, for example, 1 to 6 or 1 to 4 carbon atoms, whether used alone or as part of a substituent. 1~6) independently refers to the number of carbon atoms in the alkyl moiety, or the number of carbon atoms relative to the alkyl moiety of a larger alkyl-containing substituent. Non-limiting examples of alkyl groups include methyl, ethyl, n-propyl, cyclopropyl, isopropyl, n-butyl, sec-butyl, isobutyl, and tert-butyl.

[0039] As used herein, unless otherwise specifically provided, the term “substituted” means: deuterium, halogens (-F, -Cl, -Br, -I), hydroxyl groups (-OH), amino groups (-NH2), carboxyl groups (-CO2H), substituted or unsubstituted C 1~10 Amine group, nitro group (-NO2), C 1~10 Alkyl alkyl group, C 3~10 Cycloalkyl groups, C 6~12 Aryl group, C 1~10 C such as alkoxy groups and trifluoromethyl groups (-CF3) 1~10 This refers to a trifluoroalkyl group or a group substituted with a cyano group (-CN). Examples of substituents include: alkyl, alkyldinyl, alkylcarboxy, alkoxy, alkenyl, alkenylcarboxy, alkenyloxy, aryl, aryloxy, alkylaryl, alkylaryloxy, -OH, amide, carboxysamide, carboxy, sulfonyl, =O, =S, -NO2, halogen, haloalkyl, optionally fused saturated or unsaturated ring, -S(O)R, -SO3R, -SR, -NRR', -OH, -CN, -C(O)R, -OC(O)R, -NHC(O)R, -(CH2) n CO2R or -(CH2) n CONRR'(where n is 0-4, and R and R' are independently H, alkyl, aryl, or alkylaryl). Substitutions also include substitutions of carbon atoms and one or more associated hydrogen atoms.

[0040] As used herein, “biological equivalent” and “biological equivalent” mean chemical substituents on a functional group that are interchangeable and are intended to have substantially similar chemical or physical properties and produce substantially similar biological effects.

[0041] The "halogen-containing group" is not limited to haloalkyl groups, such as halomethyl, dihalomethyl, trihalomethyl, preferably fluoromethyl, chloromethyl, bromomethyl, iodomethyl, difluoromethyl, trifluoromethyl, dichloromethyl, trichloromethyl, dibromomethyl, tribromomethyl, diiodomethyl, triiodomethyl, fluoroethyl, fluoropropyl, fluoroisopropyl, fluoroisobutyl, and fluoro(tert)butyl.

[0042] One embodiment of the present invention includes at least one R 1 R is a halogen, preferably fluorine. In another embodiment of the present invention, one or more R 1 The group is fluorine, preferably R 1 One of the groups is fluorine, and the other R 1 The base is C 1~6 It is alkyl. In another embodiment of the present invention, R 1 One or more of the groups are halogens. In another embodiment of the present invention, R 1 One or more of the groups are halogen-containing groups. In another embodiment of the present invention, R 1 One of the groups is a halogen or halogen-containing group, and the other R 1 The group is a methyl group (CH3). In another embodiment of the present invention, each R 1 The group is a halogen or optionally substituted C 1~6 It is an alkyl group, and this compound is used to treat presbyopia or other ophthalmic diseases.

[0043] In another embodiment of the present invention, R 2 In another embodiment of the present invention, R 2 is OR 4 And here, R 4C is replaced by an optional substitution. 1~6 It is an alkyl group. In another embodiment of the present invention, R 2 is N(R 4 )2. In another embodiment of the present invention, R 2 is SR 4 or SeR 4 In another embodiment of the present invention, R 2 Furthermore, the adjacent carbonyl groups are substituted with bioequivalent groups such as imidazole, oxazole, thiazole, or serenazole, or any salt thereof.

[0044] In one embodiment of the present invention, R 3 In another embodiment of the present invention, R 3 In another embodiment of the present invention, R is SeH, as in the case of selencysteine ​​derivatives. 3 This is serenoether, or SeR 4 And here, R 4 This is C, which has been replaced by an optional substitution. 1~6 It is an alkyl group.

[0045] A preferred compound of the present invention is N-(2-fluoro-2-methyl-1-oxopropyl)-cysteine, as shown below. [ka]

[0046] Specific advantages of the present invention The compounds of the present invention alleviate the underlying causes of presbyopia and have several advantages compared to standard treatments, and include the following compounds:

[0047] Derivatives of N-acetylcysteine ​​include, but are not limited to, N-acetophenyl-cysteine, N-cyclopropylcarbonylcysteine, N-(2,2-dimethyl-1-oxopropyl)-cysteine ​​("N-pivaloylcysteine"), N-(3-methyl-2-butanyl)-cysteine, N-(2-ethyl-2-methyl-1-oxopropyl)-cysteine, N-(2-phenyl-2-methyl-1-oxopropyl)-cysteine, N-(2-methoxy-2-methyl-1-oxopropyl)-cysteine, N-(2-ammonium-2-methyl-1-oxopropyl)-cysteine, and N-(2-fluoro-2-methyl-1-oxopropyl)-cysteine.

[0048] Furthermore, two derivatives, N-pivaloylcysteine ​​and N-(2-fluoro-2-methyl-1-oxopropyl)-cysteine, showed improvement compared to N-acetylcysteine.

[0049] In embodiments described herein, the N-pivaloylcysteine ​​derivatives of the present invention are used in compositions in amounts ranging from 1% to 10% w / v, 1% to 5% w / v, 1% to 2% w / v, 1% to 1.5% w / v, more preferably greater than 1% w / v and less than 1.5% w / v, for example, in the range of 1.16% w / v to 1.32% w / v, or 1.1875% w / v to 1.3125% w / v. Yet another range of N-pivaloylcysteine ​​derivatives that can be used includes 0.95% w / v to 1.2% w / v, 1.1% w / v to 1.4% w / v, and 1.2% w / v to 1.3% w / v. A preferred amount of N-pivaloylcysteine ​​derivative is 1.25% w / v. Other amounts of N-pivaloylcysteine ​​derivatives that can be used include, for example, but are not limited to, 0.5% w / v, 0.6% w / v, 0.7% w / v, 0.8% w / v, 0.9% w / v, 0.95% w / v, 0.99% w / v, 1% w / v, 1.01% w / v, 1.05% w / v, 1.08% w / v, 1.1% w / v, 1.15% w / v, 1.2% w / v, 1.21% w / v, 1.22% w / Examples include v, 1.23% w / v, 1.24% w / v, 1.26% w / v, 1.27% w / v, 1.28% w / v, 1.29% w / v, 1.3% w / v, 1.31% w / v, 1.32% w / v, 1.35% w / v, 1.4% w / v, 1.45% w / v, 1.49% w / v, and 1.5% w / v, as well as any range and amount between these selected amounts of N-pivaloylcysteine ​​derivatives.

[0050] In accordance with the present invention, N-pivaloylcysteine ​​derivatives, or pharmaceutically acceptable salts, analogs, metabolites, prodrugs, derivatives, metabolites, cocrystals, modifiers, solvates, hydrates, isotopes, tautomers, esters, polymorphs, or stereoisomers thereof, can be administered in combination with one or more other agents, for example, a miotic agent such as pilocarpine, but is not limited to these.

[0051] In certain embodiments, the method further includes administering to the patient's eye a drug that promotes pupillary constriction or improvement of visual function.

[0052] In certain embodiments, the additional agent is administered to the patient's eye at the same time as the administration of the N-pivaloylcysteine ​​derivative. In certain embodiments, the additional agent is administered to the patient's eye either before or after the administration of the N-pivaloylcysteine ​​derivative to the patient's eye.

[0053] This method can be further characterized depending on what the additional agent is. In certain embodiments, the additional agent improves visual function. In certain embodiments, the additional agent promotes pupillary constriction in the patient. In certain embodiments, the additional agent is selected from the group consisting of muscarinic acetylcholine receptor agonists, α-2 adrenergic receptor agonists, prostaglandins, and miotics.

[0054] A pharmaceutically acceptable ophthalmic composition may contain the N-pivaloylcysteine ​​derivative of the present invention as the sole active ingredient in the pharmaceutically acceptable ophthalmic composition. In some embodiments, the pharmaceutically acceptable ophthalmic composition further comprises boric acid, sodium citrate dihydrate, sodium chloride, hydrochloric acid and / or sodium hydroxide, and water.

[0055] A pharmaceutically acceptable ophthalmic composition can be administered once daily. A pharmaceutically acceptable ophthalmic composition can be administered twice daily.

[0056] A pharmaceutically acceptable ophthalmic composition can be administered to both eyes of a patient. A pharmaceutically acceptable ophthalmic composition can be administered to one eye of a patient. A pharmaceutically acceptable ophthalmic composition can be administered to the patient's non-dominant eye. A pharmaceutically acceptable ophthalmic composition can be administered to the patient's dominant eye.

[0057] Additional pharmaceutically acceptable components can be prepared with the compounds of the present invention, particularly in the form of eye drops. Examples of these additives include tonicity modifiers, buffers, surfactants, thickeners, corneal permeability enhancers, stabilizers, preservatives, antioxidants, solubilizers, suspending agents, pH adjusters, excipients, binders, fluidizers, lubricants, and solvents.

[0058] Examples of tension adjusting agents include nonionic compounds and ionic agents containing salts such as NaCl.

[0059] Examples of surfactants include ionic surfactants (including anionic surfactants and cationic surfactants) and nonionic surfactants.

[0060] Examples of thickening agents include hydroxypropyl methylcellulose.

[0061] Examples of corneal permeability enhancers include detergents, particularly benzalkonium chloride (BAK) and EDTA.

[0062] Examples of buffering agents include sodium dihydrogen phosphate, sodium hydrogen phosphate, other phosphate derivatives, or salts thereof.

[0063] Examples of solubilizing agents include Tween-80, polyethylene glycol, cyclodextrins including hydroxypropyl-β-cyclodextrin, and other α- and β-cyclodextrins.

[0064] The compositions described herein may contain suitable preservatives. Examples of suitable preservatives include benzalkonium chloride ("BAK"), polyquaternium-1 (Polyquad®), chlorobutanol, and stabilized chlorine dioxide. Stabilized chlorine dioxide (also known as Purite®) can be described as an aqueous solution of sodium chlorite (NaClO2). U.S. Patent No. 5,424,078, which is incorporated herein by whole reference, further discusses the use of stabilized chlorine dioxide as a preservative for ophthalmic formulations.

[0065] The additive components listed above (such as tonicity modifiers, buffers, surfactants, thickeners, corneal permeability enhancers, stabilizers, preservatives, antioxidants, solubilizers, suspending agents, pH adjusters, excipients, binders, fluidizers, lubricants, and solvents) may be used at any appropriate and pharmaceutically acceptable concentration.

[0066] Examples of solvents include water and physiological saline.

[0067] In addition to eye drops, the present invention includes compositions comprising the N-pivaloylcysteine ​​derivatives described herein, which are incorporated onto contact lenses or used as eye creams or gels. These compositions are not limited to chitosan, hyaluronic acid, poloxamer, PLGA, PVCL-PVA-PEG, cetaconium chloride, glengham, hydroxyethylcellulose, or gelatin.

[0068] Another aspect of the present invention provides a method for treating presbyopia in a patient according to a monotherapy regimen, which comprises administering a dose of a single therapeutic agent effective for treating presbyopia to the eye of the patient in need, wherein the single therapeutic agent is an N-pivaloylcysteine ​​derivative which is an agonist of the present invention.

[0069] This method may be further characterized by additional features such as dose setting regimens and dosage specification, as will be described in more detail below. The present invention includes all permutations and combinations of these features. Thus, in certain embodiments, the dosage is administered at or near the patient's bedtime. In certain embodiments, the dosage is administered within one hour of the patient's bedtime.

[0070] In certain embodiments, the dosage is administered as a single eye drop. In certain embodiments, the dosage is administered as two or more eye drops.

[0071] In the embodiments described herein, the composition may be administered once daily, twice daily, or more times. Preferably, the composition is administered once daily.

[0072] When administered, the composition preferably has a sufficient duration of action throughout the day. The duration of action refers to the length of time the administered composition is effective against at least one visual parameter or eye condition (e.g., presbyopia). In some embodiments, the composition may have a duration of action of at least 2 hours, at least 3 hours, preferably at least 4 hours, more preferably at least 6 hours, even more preferably at least 8 hours, even more preferably at least 10 hours, and at any point in between. Some embodiments may provide compositions with a duration of action exceeding 10 hours, for example, 12 hours, or even exceeding 24 hours.

[0073] In certain embodiments, the method provides a therapeutic effect on presbyopia with a duration of at least 12 hours. In certain embodiments, the method provides a therapeutic effect on presbyopia with a duration of at least 16 hours. In certain embodiments, the method provides a therapeutic effect on presbyopia with a duration of at least 18 hours. In certain embodiments, the method provides a therapeutic effect on presbyopia with a duration of at least 20 hours. In certain embodiments, the method provides a therapeutic effect on presbyopia with a duration of at least 24 hours. In certain embodiments, the method provides a therapeutic effect on presbyopia with a duration of at least 36 hours. In certain embodiments, the method provides a therapeutic effect on presbyopia with a duration of at least 48 hours.

[0074] Examples are provided below to illustrate the methods and results obtained in accordance with the disclosed subject matter. These examples are not intended to encompass all aspects of the subject matter disclosed herein, but rather to illustrate representative methods, compositions, and results. These examples are not intended to exclude equivalents and modifications of the invention that would be readily apparent to those skilled in the art. [Examples]

[0075] Examples Example 1. Synthesis Unless otherwise specified, reactions were carried out in anhydrous solvents, under a nitrogen gas (N2) atmosphere, and in oven-dried glassware (140°C). S-trityl L-cysteine ​​methyl ester was prepared starting from commercially available L-cysteine ​​methyl ester hydrochloride according to a known method (Gale, et al., Inorg. Chem., 50: 10460-10471 (2011)). A general reaction scheme is shown below, followed by specific information for each step. [ka]

[0076] N-2,2-dimethylpropionyl-S-trityl-L-cysteine ​​methyl ester: A mixture of S-trityl-L-cysteine ​​methyl ester (1.00 g, 2.64 mmol), hexafluorophosphate azabenzotriazole tetramethyluronium (HATU, 1.05 g, 2.77 mmol), and pivalic acid (283 mg, 2.77 mmol) in dichloromethane (DCM) (26.4 mL) was cooled to 0°C. DIPEA (0.92 mL, 5.28 mmol) was added dropwise to this solution. After all additions, the mixture was stirred at 0°C for 10 minutes and then allowed to return to room temperature. After 1 hour, the reaction mixture was transferred to a separatory funnel and washed with NaHCO3 (20 mL x 2 times), followed by water (20 mL x 2 times), and saline solution (10 mL x 1 time). The organic layer was dried over MgSO4, filtered, and concentrated under reduced pressure. The crude product was purified by flash chromatography (SiO2, 3:17 siRNA / hexane) to obtain the title compound as an off-white solid (1.15 g, 2.49 mmol, 94%). 1 H NMR(600MHz,CDCl3)δ7.38(dt,J=8.5,1.8Hz,6H),7.28(t,6H),7.22(t,3H),6.24(d,J=7.6Hz,1H),4.59(ddd, J=7.6,5.6,4.6Hz,1H),3.71(s,3H),2.64(dd,J=12.1,4.7Hz,1H),2.58(dd,J=12.1,5.7Hz,1H),1.21(s,9H); 13C NMR(150MHz, CDCl3)δ178.2,171.3,144.4,129.6,128.1,127.0,66.8,52.7,51.1,38.9,34.1,27.5.

[0077] To a solution of N-2,2-dimethylpropionyl S-trityl L-cysteine ​​methyl ester (1.14 g, 2.47 mmol) in a 1:1 THF / MeOH solution (7.06 mL), water (1.84 mL) was added, followed by LiOH·H2O (311 mg, 7.41 mmol). After stirring for 1 hour, the reaction mixture was cooled to 0°C, adjusted to pH 2 with 1 M HCl, and transferred to a separatory funnel. The aqueous layer was extracted with DCM (3 × 25 mL). The combined organic layers were washed with water (1 × 25 mL) and saline solution (1 × 25 mL); dried over MgSO4, filtered, and concentrated under reduced pressure to obtain the title compound. The obtained solid was used without further purification (1.04 g, 2.32 mmol, 94%). 1 H NMR(600MHz,CDCl3)δ7.41(d,6H),7.29(t,J=7.7Hz,6H),7.23(t,3H),6.16(d,J=6 .5Hz,1H),4.22(td,J=6.8,4.8Hz,1H),2.72(qd,J=12.9,5.9Hz,2H),1.20(s,9H); 13 C NMR(151MHz, CDCl3)δ180.2,172.9,144.3,129.6,128.3,127.1,67.2,51.8,38.9,32.9,27.5.

[0078] To a mixture of N-2,2-dimethylpropionyl-S-trityl-L-cysteine ​​(211 mg, 0.47 mmol) in N-2,2-dimethylpropionyl-L-cysteine:DCM (0.73 mL), TFA (2.24 mL) and triethylsilane (0.07 mL, 0.47 mmol) were sequentially added. After stirring for 10 minutes, the reaction mixture was concentrated under reduced pressure. The resulting crude product was purified by flash chromatography (SiO2, 3:100 i-PrOH / DCM) to obtain the title compound as a white solid (81 mg, 0.40 mmol, 84%).1 H NMR(400MHz,CDCl3)δ6.67(d,J=6.8Hz,1H),4.82(dt,J=6.9,4.3Hz,1H),3.15(ddd,1H),2.99(ddd,J=14.3,9.6,4.6Hz,1H),1.46(t,1H),1.26(s,9H); 13 C NMR(151MHz,CDCl3)δ179.7,173.3,53.5,39.1,27.5,26.5.

[0079] Following the procedure from N-2-methyl-2-fluoropropionyl S-trityl-L-cysteinemethyl ester:N-2,2-dimethylpropionyl S-trityl-L-cysteinemethyl ester, S-trityl-L-cysteinemethyl ester (3.28 g, 6.68 mmol), HATU (2.67 g, 7.01 mmol), 2-fluoroisobutyric acid (0.66 mL, 7.01 mmol), and DIPEA (2.33 mL, 13.4 mmol) were reacted in DCM (67 mL). The title compound (2.39 g, 5.13 mmol, 77%) was obtained as a pale yellow solid by purification by flash chromatography (SiO2, 15:100 siRNA / hexane). 1 H NMR(400MHz,CDCl3)δ7.39(d,6H),7.29(t,J=7.5Hz,6H),7.23(t,3H),6.92-6. 84(m,1H),4.52(q,J=6.3Hz,1H),3.71(s,3H),2.70-2.57(m,2H),1.56(dd,6H); 13 C NMR(151MHz,CDCl3)δ170.6,144.4,129.7,128.2,127.1,67.1,52.9,50.9,33.9,25.2,25.1,24.9; 19 F NMR (376 MHz, CDCl3) δ-147.51.

[0080] Following the procedure from N-2-methyl-2-fluoropropionyl-S-trityl-L-cysteine:N-2,2-dimethylpropionyl-S-trityl-L-cysteine, the title compound was prepared from N-2-methyl-2-fluoropropionyl-S-trityl-L-cysteine ​​methyl ester (2.39 g, 5.13 mmol), LiOH·H2O (646 mg, 15.4 mmol), and water (3.83 mL) in 1:1 THF / MeOH (14.7 mL). No further purification was necessary, and the title compound was obtained as a pale yellow solid (2.13 g, 4.72 mmol, 92%). 1 H NMR(600MHz,CDCl3)δ7.42(d,J=7.7Hz,6H),7.31(t,J=7.7Hz,6H),7.25(t,J=7.2Hz,3H),6. 87(t,J=6.6Hz,1H),4.39(d,J=6.6Hz,1H),2.74(qd,J=12.8,5.7Hz,2H),1.69-1.46(dd,6H); 13 C NMR(151MHz,CDCl3)δ173.7,173.5,144.1,129.5,128.1,127.0,96.6,95.4,67.2,50.9,33.1,25.0,24.9,24.8,24.8; 19 F NMR (376 MHz, CDCl3) δ-147.39.

[0081] Following the procedure for N-2-methyl-2-fluoro-propionyl-L-cysteine:N-2,2-dimethyl-propionyl-L-cysteine, the title compound was prepared from N-2-fluoro-2-methyl-propionyl S-trityl L-cysteine ​​(250 mg, 0.56 mmol), TFA (2.64 mL), and triethylsilane (0.06 mL, 0.56 mmol) in DCM (0.87 mL). The resulting crude product was dissolved in water (10 mL) and Et2O (10 mL) and then transferred to a separatory funnel. The aqueous layer was washed with Et2O (3 × 10 mL) and freeze-dried to obtain the title compound as a white solid (104 mg, 0.50 mmol, 89%). 1H NMR(600MHz,CDCl3)δ7.23(t,1H),4.91-4.85(m,1H),3.14(ddd,J=14.2,8.7,4.2Hz ,1H),3.03(ddd,J=14.2,9.4,4.6Hz,1H),1.67-1.53(dd,6H),1.48(t,J=9.0Hz,1H); 13 C NMR(151MHz,CDCl3)δ174.1,173.9,173.4,96.7,95.5,53.3,26.5,25.2,25.1,24.9,24.8; 19 F NMR (376 MHz, CDCl3) δ-147.48.

[0082] Example 2. Lens rigidity in exvivo To evaluate the effects of multiple compounds on lens rigidity, ex vivo screening was performed using a more efficient variant of the coverslip method (Cheng, et al., J.Vis.Exp., 111: 53986 (2016)). Our variant uses a single measurement point that enables multi-drug screening, employing a weight determined to be optimal based on the coverslip amount-response curve. Pre-dissected and frozen eyeballs were thawed from 8-month-old C57BL / 6 mice, and the lenses were removed by microscopic dissection. The intact lenses were then immersed in solutions of various test compounds and incubated at 37°C for 12 hours. Each washed lens was placed in a recess of a plexiglass measurement chamber filled with the solution, and its axial diameter in the uncompressed state was measured. Subsequently, three glass coverslips, bonded together, with a total weight of 588.0 mg, were placed on top of the lens, and the axial diameter was remeasured. The change in the rigidity of the lens is calculated as the ratio (compression / decompression) between the axial diameter of the compressed lens and the axial diameter of the lens without added weight. A higher ratio indicates greater flexibility.

[0083] NAC(20mM) significantly improved lens rigidity compared to untreated (medium) control lenses in screening.

[0084] To address a significant limitation of ex vivo lens treatment (the stiffness of lenses increases over time when extracted from postmortem eyeballs), we compared newly dissected 8-month-old lenses with NAC treatment. We observed spontaneous hardening during a 12-hour incubation period (mean compressibility: 10.34%, n=10). Therefore, to consider the possibility that NAC primarily slows the standard hardening rate rather than severing existing cross-linking bonds, we examined the stiffness after 1 hour incubation with NAC and found a significant improvement of 2.07% compared to the vehicle.

[0085] Example 3 Initial in vivo studies of NAC were conducted using 7-month-old C57BL / 6 mice. These mice were then administered 100 mM and 20 mM NAC eye drops twice daily for one month (artificial tears were used as the vehicle). On the final day, the animals were sacrificed, their eyeballs were extracted, and the lenses were removed. Lens rigidity was measured using the same assay as in the ex vivo study, within 5–10 minutes after lens extraction. Cover glass weights of 588 mg and 1047 mg were used in the assay. NAC 100 mM showed a significant improvement in lens rigidity compared to the vehicle at both cover glass weights, while NAC 20 mM showed a mild and insignificant improvement. Subsequent studies showed that N-acetyl-D-cysteine, an enantiomer of NAC, also showed a significant improvement compared to the vehicle, but with lower efficacy than NAC.

[0086] All compounds were tested at a 20 mM concentration, and if the solubility in the medium was insufficient to achieve this concentration, 0.1% tween-80 and 0.4% PEG-400 were added. Initially, several acetyl derivatives modified at the R2 site (N-acetophenylcysteine, N-acetobenzylcysteine, N-cyclopropylcarbonylcysteine, N-(N,N-dimethylformamidoylcysteine, N-(2,2-dimethyl-1-oxopropyl)-cysteine ​​("N-pivaloylcysteine"), and N-pentanylcysteine) were generated, of which N-pivaloylcysteine ​​showed the greatest improvement. Subsequently, several N-pivaloyl variants with modifications at the R1 site (N-(3-methyl-2-butanyl)-cysteine, N-(2-ethyl-2-methyl-1-oxopropyl)-cysteine, N-(2-phenyl-2-methyl-1-oxopropyl) N-(2-methoxy-2-methyl-1-oxopropyl)-cysteine, N-(2-ammonium-2-methyl-1-oxopropyl)-cysteine, and N-(2-fluoro-2-methyl-1-oxopropyl)-cysteine) were tested ex vivo. Two compounds, N-pivaloylcysteine ​​and N-(2-fluoro-2-methyl-1-oxopropyl)-cysteine, showed significant improvement over NAC, while several other compounds showed improvement over the control and not significant improvement over NAC (Table 1). N-(2-fluoro-2-methyl-1-oxopropyl)-cysteine ​​had the greatest mean significance and the highest significance compared to the vehicle.

[0087] [Table 1]

[0088] [Table 2]

[0089] *The vehicle consisted of Epilife (60 μM CaCl) treated lenses and combinations of Epilife + 0.1% Tween-80 and 0.4% PEG-400. No significant or evaluable difference was observed between the two, so they were aggregated here. Phenyl-substituted, benzyl-substituted, and cyclobutene-substituted variants were treated with Epilife; all others were treated with Epilife + PEG / Tween as the solvent to improve the solubility of the test reagents.

[0090] Example 4. In vivo efficacy Rationale and Objective: To evaluate the effects of N-acetylcysteine ​​(NAC) and its variants on the reduction of lens rigidity in vivo. The following specific variables differ significantly between in vivo and ex vivo studies: in particular the presence of the cornea as a barrier during in vivo treatment, the 12-hour duration of ex vivo treatment versus the 28-day duration of in vivo treatment, and the effect of postmortem lens hardening in ex vivo.

[0091] The following test groups were evaluated in 7-month-old C57BL / 6 mice: vehicle (artificial tears) (40 mice), N-acetylcysteine ​​20 mM (20 mice), N-(2-fluoro-2-methyl-1-propylcarbonyl)-cysteine ​​200 mM (10 mice), LACE (choline lipoate) 1.5% (PBS vehicle) (10 mice), and 2-month-old mice (untreated) (7 mice).

[0092] Mice treated with N-(2-fluoro-2-methyl-1-propylcarbonyl)-cysteine ​​showed significantly reduced lens rigidity compared to both LACE-treated and vehicle-treated mice. Vehicle-treated mice had an average total compression value of 199.34 micrometers (standard deviation 32.16). Mice treated with N-(2-fluoro-2-methyl-1-propylcarbonyl)-cysteine ​​had an average total compression value of 281.05 micrometers (standard deviation 40.70). Mice treated with LACE had an average total compression value of 209.68 micrometers (standard deviation 28.16). Younger control mice aged 2 months had an average total compression value of 319.71 micrometers (standard deviation 40.05). N-(2-fluoro-2-methyl-1-propylcarbonyl)-cysteine ​​was superior to the 8-month vehicle (p<.0001) and LACE (p<.0001), and was able to restore 67.9% of the lens rigidity lost due to aging over 2-8 months. N-(2-fluoro-2-methyl-1-propylcarbonyl)-cysteine ​​showed a significant improvement over NAC. LACE itself did not result in a significant improvement compared to vehicle-treated lenses.

[0093] Example 5. Toxicity testing and dose optimization in mice The maximum tolerable dose of N-(2-fluoro-2-methyl-1-propylcarbonyl)-cysteine ​​(and its S-acetylated counterpart, N-(2-fluoro-2-methyl-1-propylcarbonyl)-S-acetylcysteine) for reducing lens rigidity was evaluated, and tests were conducted to histologically assess whether any safety issues arise in the lens and cornea after one week of treatment with these compounds.

[0094] Two-month-old C57BL / 6 mice were treated with the therapeutic compound 2x / day (see sample groups (n=5) and concentrations in the table below). All animals were evaluated on a scale of 1–4 for post-administration pruritus and scratching / blinking, conjunctivitis, eyelid swelling, discharge, and strabismus. Evaluators were blinded to the treatment groups. After euthanasia, the eyeballs were fixed, stained with H+E, and the anatomical structure of the cornea and lens was evaluated in the two highest concentration groups deemed tolerable.

[0095] [Table 3]

[0096] N-(2-fluoro-2-methyl-1-propylcarbonyl)-cysteine ​​and its S-acetylated counterpart, N-(2-fluoro-2-methyl-1-propylcarbonyl)-S-acetylcysteine, had maximum solubility concentrations of 200 mM and 300 mM, respectively, in artificial tears. Post-administration blink rate increased in all treatment groups but returned to normal levels before administration. With 200 mM N-(2-fluoro-2-methyl-1-propylcarbonyl)-cysteine, conjunctivitis increased slightly, but there were no changes in strabismus, eyelid swelling, or eye discharge. Histological examination of N-(2-fluoro-2-methyl-1-propylcarbonyl)-cysteine-treated lenses was evaluated as completely normal. 200 mM N-(2-fluoro-2-methyl-1-propylcarbonyl)-cysteine ​​was evaluated as tolerable.

[0097] Example 6. Comparison of aqueous humor LC / MS concentrations Tests were conducted to evaluate the extent to which N-(2-fluoro-2-methyl-1-propylcarbonyl)-cysteine ​​penetrates the mouse cornea compared to NAC and N-pivaloylcysteine.

[0098] Eight-month-old C57BL / 6 mice were administered 100 mM N-(2-fluoro-2-methyl-1-propylcarbonyl)-cysteine, 100 mM N-pivaloylcysteine, or 100 mM NAC as 3 μm eye drops twice daily for one month. Ten mice were used per group. No significant increase in tolerability issues such as blinking or conjunctivitis was observed over time. Histological examination revealed good safety.

[0099] On the final day, aqueous humor was collected from the eyeball one hour after the last dose (average 1-5 μL extracted), and the concentration of each compound was quantified by LC / MS (column: Atlantis T3, 100 × 2.1 mm, 5 μm; mobile phase: A: water / formic acid (1000 / 5, v / v), B: acetonitrile / methanol / formic acid (50 / 50 / 0.3, v / v / v).

[0100] N-(2-fluoro-2-methyl-1-propylcarbonyl)-cysteine ​​showed the highest concentration in aqueous humor (47 μm), followed by N-pivaloylcysteine ​​(38 μm). NAC was much lower, with 17 out of 20 samples below the detection limit.

[0101] N-(2-fluoro-2-methyl-1-propylcarbonyl)-cysteine ​​improved corneal permeability because its concentration in aqueous humor was higher than that of NAC and N-pivaloylcysteine. N-(2-fluoro-2-methyl-1-propylcarbonyl)-cysteine ​​also showed the additional benefit of a superior reduction in lens rigidity in the ex vivo environment where the cornea is absent, which may also be due to improved lens permeability.

Claims

1. formula: 【Chemistry 1】 (In the formula, Each R １ These are independently replaced by C of any choice. １～６ Alkyl, C １～６ It is an aryl, halogen, halogen-containing group, or ammonium group; Y is -C- or -CO- or -N-; R ２ is OR ４ , SR ４ , SeR ４ , or N(R ４ ) ２ and; R ３ , SR ４ ,SeR ４ , or SC(O)-R ４ And; Here R ４ C is a hydrogen atom or optionally substituted C １～６ (It is alkyl.) Compounds thereof, and pharmaceutically acceptable salts thereof.

2. The following formula: 【Chemistry 2】 (In the formula, R １ , R ２ and R ３ (As defined in claim 1) The compound according to claim 1, having the following characteristics.

3. R ２ The compound according to claim 1 or 2, wherein the compound is an OH group.

4. R ２ However, OR ４ And here, R ４ However, C was replaced by an arbitrary choice. １～６ A compound according to any one of claims 1 to 3, wherein it is alkyl.

5. R ２ However, N(R ４ ) ２ The compound according to any one of claims 1 to 4.

6. R ２ However, SR ４ Or SeR ４ The compound according to any one of claims 1 to 4.

7. R ２ The compound according to any one of claims 1 to 4, wherein the adjacent carbonyl group together forms a biological equivalent group selected from the group consisting of imidazole, oxazole, thiazole, and serenazole.

8. R ３ The compound according to any one of claims 1 to 7, wherein SH is present.

9. R ３ The compound according to any one of claims 1 to 7, wherein the compound is SeH.

10. R ３ However, it is SeH, and R ４ However, C was replaced by an arbitrary choice. １～６ A compound according to any one of claims 1 to 7, wherein it is alkyl.

11. at least one R １ The compound according to any one of claims 1 to 10, wherein the compound is a halogen.

12. at least one R １ The compound according to claim 11, wherein the compound is fluorine.

13. The aforementioned R １ One or more of the groups are fluorine, and the other R １ The base is replaced by C of any choice. １～６ The compound according to claim 2, wherein it is alkyl.

14. The aforementioned R １ The compound according to claim 13, wherein one of the groups is fluorine.

15. R １ The compound according to any one of claims 1 to 12, wherein one or more of the members are halogen-containing groups.

16. The aforementioned R １ One of the groups is a halogen-containing group, and the other R １ The base is CH ３ The compound according to claim 15.

17. The aforementioned R １ The compound according to any one of claims 1 to 12, wherein one of the groups is an ammonium group.

18. N-(2,2-dimethyl-1-oxopropyl)-cysteine ​​or a pharmaceutically acceptable salt or ester thereof.

19. N-(2-fluoro-2-methyl-1-oxopropyl)-cysteine ​​or a pharmaceutically acceptable salt or ester thereof.

20. A pharmaceutically acceptable ophthalmic composition comprising a compound according to any one of claims 1 to 19, and one or more pharmaceutically acceptable excipients selected from the group consisting of tonicity modifiers, buffers, surfactants, thickeners, corneal permeability enhancers, stabilizers, preservatives, antioxidants, solubilizers, suspending agents, pH adjusters, excipients, binders, fluidizers, lubricants, and solvents.

21. The ophthalmic composition according to claim 20, which is formulated as an eye drop.

22. The ophthalmic composition according to claim 21, wherein the one or more excipients are boric acid, sodium citrate dihydrate, sodium chloride, hydrochloric acid, or sodium hydroxide.

23. A method for reducing the rigidity of the lens, comprising administering a therapeutically effective amount of a compound described in any one of claims 1 to 19 or a pharmaceutically acceptable salt, analog, metabolite, prodrug, metabolite, solvate, hydrate, isotope, or ester thereof to a subject in need thereof.

24. The method according to claim 23, further comprising administering a pupil-constricting agent.

25. The method according to claim 24, wherein the miotic agent is pilocarpine.

26. A method for treating presbyopia, comprising administering a therapeutically effective amount of a compound described in any one of claims 1 to 19 or a pharmaceutically acceptable salt, analog, metabolite, prodrug, metabolite, solvate, hydrate, isotope, or ester thereof to a subject in need thereof.

27. The method according to claim 26, further comprising administering a pupil-constricting agent.

28. The method according to claim 27, wherein the miotic agent is pilocarpine.

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