Compounds for treating conditions associated with oxidative stress - Patents.com

Abstract

Disclosed herein are compounds, pharma- ceutically acceptable salts, hydrates, solvates, optical isomers, or combinations thereof, for treating conditions associated with oxidative stress / damage, among other causes, comprising the compounds of formula (I) and formula (8); [Formula 1] TIFF2024512388000044.tif43161 (wherein Ri, R2, R3, R4, and R5 are [chemical 2] TIFF2024512388000045.tif32161), where X, Y, and Z are selected from the group consisting of -H, methyl, ethyl, propyl, butyl, substituted or unsubstituted aryl and heteroaryl; n is an integer selected from 0 to 18, and E is a phosphorus or nitrogen atom.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 159,081, filed March 10, 2021, which is incorporated by reference in its entirety.

[0002] The present disclosure relates to compounds comprising one or more quaternary cation moieties chemically linked to one or more antioxidant moieties for treating conditions involving oxidative stress in a mammal. [Background technology]

[0003] Oxidative stress is a factor in many human diseases related to aging, such as Parkinson's disease and Alzheimer's disease, as well as Huntington's chorea, diabetes and Friedreich's ataxia, and is a factor in the non-specific damage that accumulates with aging.It also contributes to inflammation and ischemia-reperfusion tissue damage in stroke and heart attack, and during organ transplantation and surgery.Some antioxidant therapies have been developed to prevent the damage caused by oxidative stress.However, most of these do not target intracellularly, and therefore are not optimally effective.

[0004] Mitochondria are intracellular organelles primarily responsible for energy metabolism. They are also the main source of free radicals and reactive oxygen species (ROS), which cause oxidative stress and / or damage in most cells. Mitochondria are equipped to detoxify ROS due to the presence of antioxidant enzymes (peroxiredoxin, thioredoxin, and GSH-dependent peroxidases). Typically, mitochondrial superoxide (a radical anion generated by one-electron reduction of O) is dismutated by manganese superoxide dismutase (MnSOD), which is localized within the mitochondrial matrix. However, when cellular ROS production exceeds the detoxification capacity of the cell, oxidative damage occurs. This damage disrupts mitochondrial function and oxidative phosphorylation, resulting in significant cellular damage to mitochondrial, cytoplasmic or nuclear cellular proteins, DNA, RNA, and phospholipids, thus inducing cell damage, death, and / or disease. Superoxide can also react with nitric oxide at diffusion-controlled kinetics to form highly potent oxidants and peroxynitrite, which can modify proteins and DNA through oxidation and nitration reactions. In addition to these damaging and pathological roles, ROS also act as redox signaling molecules, promoting cell proliferation, DNA damage repair errors and mutations, leading to inflammatory hyperproliferation, neoplasia, and malignancy.

[0005] Thus, mitochondria are recognized as one of the most important targets for novel drug design in various therapeutic areas, including cancer, inflammation, eye, cardiovascular, and neurological diseases. The most effective way to deliver drugs specifically to mitochondria is by covalently attaching lipophilic cations, such as alkyltriphenylphosphonium moieties, to the pharmacophore of interest. Other delocalized lipophilic cations, such as rhodamine, natural and synthetic mitochondria-targeting peptides, and nanoparticle vehicles, have also been used for mitochondrial delivery of small molecules. Such ions accumulate, unless they are lipophilic enough to shield the positive charge or delocalize over a large surface area, and unless there is an active excretion route, the cations are either not metabolized or are immediately toxic to the cell.

[0006] Oxidative damage also plays an important role in the aging process. In addition, oxidative damage has a role in the pathogenesis of hypoxic injury, cancer, and cardiovascular disease associated with reperfusion, and neurodegenerative disorders (Parkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis). Oxidative stress also contributes to diseases such as Huntington's chorea, diabetes, and Friedreich's ataxia, as well as nonspecific damage that accumulates with age. It also contributes to inflammation and ischemia-reperfusion tissue damage in strokes and heart attacks, and during organ transplants and surgery.

[0007] In the field of ophthalmology, oxidative stress is involved in the pathogenesis of several eye diseases, such as senile cataract, age-related macular degeneration, uveitis, early retinopathy, keratitis and ocular inflammation. It has been reported that oxidized protein levels are elevated during these diseases along with increased inflammatory activity. Oxidative stress plays a major role in the pathogenesis of some forms of dry eye, and treatment with topical / systemic use of antioxidants may be useful.

[0008] The eye requires a high supply of oxygen and nutrients, and this high energy demand of the visual system makes it sensitive to oxidative stress. Retinal cells are vulnerable to ROS due to their synchronous interactions and complex neural networks. Excessive free radicals result in mitochondrial dysfunction, leading to retinal neurodegeneration as an early stage of retinal metabolic disorders. Retinal neurodegeneration is one of the main causes of visual impairment and is highly associated with retinal atrophy or cell death in ocular diseases, such as glaucoma, age-related macular degeneration (AMD), and diabetic retinopathy (DR). Metabolic changes promote both AMD and DR, and ROS-induced damage is central to both of these conditions. Free radical scavengers have shown neuroprotective effects and are known to promote mitochondrial gene expression, which may result in improved retinal cell organization and visual function. SS-31 is a synthetic tetrapeptide known to target the inner mitochondrial membrane and reduce mitochondrial ROS, and has been found to be effective in reversing vision loss in a mouse model of streptozotocin-induced diabetes. Similarly, various other antioxidant therapies have been developed and are found to be effective for treating damage caused by oxidative stress, however, most of these are not targeted intracellularly and therefore are less than optimally effective.

[0009] Therefore, uncontrolled mitochondrial ROS production, resulting in oxidative cell damage-induced diseases and disorders, represents an unsolved problem in the art and represents a compelling target for pharmacological drug therapy using pharmaceutical antioxidant small molecule drug formulations.To prevent cell damage caused by oxidative stress, several prior art antioxidant therapies have been developed to treat various diseases caused by oxidative stress.However, most of these inventions do not target other organelles or mitochondria in the cell, and therefore are not optimally effective.

[0010] The mitochondrial targeting technique developed by Murphy et al. has attracted much interest in recent years, with little success in validating the implications of this concept. According to this concept, a "warhead" group is covalently attached to a bulky and / or lipophilic cationic moiety, such as a quaternary ammonium or phosphonium cationic moiety, via a linker group. These compounds are initially absorbed and accumulate in the cytoplasmic region of the cell in response to a negative cell membrane potential of 30-60 mV. Furthermore, within minutes of drug treatment, lipophilic cations with a positive potential of 30-60 mV permeate through the mitochondrial lipid layer and selectively accumulate within the mitochondria due to the large mitochondrial membrane potential (negative on the inside) of 150-170 mV.

[0011] Depending on the approach used and the membrane potential of cells and mitochondria, mitochondrial concentration can be achieved more than 1000 times higher.Mitochondria targeting has been developed to study mitochondrial physiology and dysfunction and the interaction between mitochondria and other intracellular organelles, and for the treatment of a wide variety of diseases.Various publications also discuss the efforts of targeting small molecule compounds to mitochondria to investigate mitochondrial function as diagnostic tools and potential treatments, all of which are incorporated herein by reference in their entirety.

[0012] In particular, five-membered heterocyclic molecules have been reported to exhibit strong free radical scavenging ability as well as antioxidant properties in various preclinical and clinical studies. They have been effectively exploited in many fields, especially in medicine, because they exhibit bioactivity as analgesics, retinal diseases, antibacterial agents, anti-inflammatory agents, antioxidants, antipsychotics and antitumor agents. However, in some cases, such as the case of edaravone (EDA), the bioactivity is presumed to be due to the transfer of electrons from the anion at physiological pH. It is hypothesized that the EDA radical intermediates generated during this process are less reactive than those of the oxygen free radicals described below. However, their ROS neutralizing ability and their contribution to the therapeutic effect cannot be excluded. Various other modifications of edaravone have been reported, such as the introduction of a C18 hydrocarbon chain at the C-4 position to increase affinity for lipid membranes and derivatives of edaravone lacking pyridoxal scavenging ability, which were found to inhibit glycation that has progressed along with oxidative stress. EDA has also been reported to be useful in certain side effects of cancer chemotherapy, radiotherapy and immunotherapy. This cytoprotective effect of EDA can also be utilized in treating or preventing conditions such as myocardial, retinal, lung, intestinal, liver, pancreatic and kidney injury, all of which are incorporated herein by reference in their entirety. Mitochondrial targeting compounds in this class of agents are shown below, including the mitochondria-targeting ubiquinone (MitoQ) reported by Murphy and coworkers (U.S. Pat. Nos. 6,331,532 and 7,232.809, and European Patent No. 1047701, all of which are incorporated herein by reference in their entirety). MitoQ has shown promise in treating only some, but not all, oxidative stress-induced diseases. MitoQ is currently in Phase 2 clinical trials for the treatment of Parkinson's disease, but it has relatively little activity against other oxidative stress-induced neurodegenerative diseases, such as amyotrophic lateral sclerosis (ALS or Lou Gehrig's disease). This class of compounds is also disclosed in US Patent Application Publication No. 2008 / 0032940, which is incorporated herein by reference in its entirety.

[0013] Other classes of mitochondria targeted compounds include mitochondria targeted nitric oxides, which have been used in methods for treating neurodegenerative disorders (see U.S. Patent Application Publication No. 2007 / 0066572, which is incorporated herein by reference in its entirety). Mitochondria targeted antioxidants have been used in methods for treating cancer (see U.S. Patent Application No. 11 / 834,799, entitled "Methods for Reducing Anthracycline-Induced Toxicity," filed August 7, 2007, and U.S. Patent Application No. 12 / 554,476, entitled "mitochondria targeted anti-oxidant compounds for prevention, therapy or treatment of hyper-proliferative diseases, neoplasia and cancers," which are incorporated herein by reference in their entirety). Mitochondrial targeted antioxidants have been used in methods for treating degenerative bone disorders (see WO 2006 / 35914, 2006, A1, entitled "Method and composition of treating degenerative bone disorders," which is incorporated by reference in its entirety).

[0014] Notwithstanding the above prior art, there remains a need for compositions and methods for effectively treating conditions caused by excessive mitochondrial production of reactive oxygen species (ROS) at the mitochondrial membrane.

[0015] Therefore, there remains a need for mitochondrial-targeted free radical scavengers and antioxidants with improved pharmacodynamic and pharmacokinetic profiles.To address such a need, the inventors of the present application provide compounds that can utilize the ability of mitochondria to concentrate certain lipophilic cations that take up linked antioxidants to target them to the main source of free radicals and reactive oxygen species that cause oxidative stress. Summary of the Invention

[0016] The present application relates to a compound of formula (I) [ka]

[0017] (In the formula: [ka]

[0018] R1 and R2 are each independently a C group mono-, di- or tri-substituted with hydrogen, optionally halogen, substituted and unsubstituted aryl (monocyclic and bicyclic), substituted and unsubstituted heteroaryl (monocyclic and bicyclic). 1~20 Alkyl, C 2~20 Alkenyl, and C 2~20 alkynyl; R3 is hydrogen, C 1~20 Alkyl, C 2~20 Alkenyl, and C 2~20 alkynyl, wherein R3 is optionally substituted by one or more A; R4 is hydrogen, C 1~20 Alkyl, C 2~20 Alkenyl, and C 2~20 alkynyl, wherein R4 may be substituted by one or more A; and R5 is aryl, heteroaryl, C 1~2 Alkyl-aryl, and C 1~2alkyl-heteroaryl; where A is aryl or heteroaryl. The present invention relates to a compound of the formula:

[0019] In one embodiment disclosed herein, the present application relates to a compound as described above, wherein the aryl and heteroaryl are optionally selected from halogen, C 1~6 Alkyl, -CF3, -OCF3, C 1~6 Alkoxy, C 3~6 Cycloalkoxy, halo-C 1~6 Alkyl, aryl, heteroaryl, heterocyclo, C 3~6 Cycloalkyl, and C 3~6 It is mono-, di- or trisubstituted with substituents selected from the group consisting of cycloalkenyl.

[0020] In one aspect of the embodiments disclosed herein, the present application relates to a compound as described above, wherein at least one of R1, R2, R3, R4, and R5 is [ka]

[0021] (wherein X, Y, and Z are selected from the group consisting of hydrogen, methyl, ethyl, propyl, butyl, substituted or unsubstituted aryl, and heteroaryl; n is an integer selected from 0 to 18, E is a phosphorus or nitrogen atom, and X1 is a sulfur, nitrogen, or oxygen atom).

[0022] In another aspect of the embodiments disclosed herein, the present application relates to a compound as described above, wherein R1, R2, R3 and R4 are each independently hydrogen, C 1~20 Alkyl, C 2~20 Alkenyl, and C 2~20 alkynyl.

[0023] In another aspect of the embodiments disclosed herein, the present application relates to a compound as described above, wherein R5 is aryl, heteroaryl, C 1~2 Alkyl-aryl, and C 1~2 alkyl-heteroaryl.

[0024] In another aspect of the embodiments disclosed herein, the present application relates to a compound as described above, wherein R1, R2 and R4 can be hydrogen; R3 can be -CH3; and R5 is, [ka]

[0025] is; However, at least one of R2, R3, and R4 must be [ka]

[0026] It is.

[0027] In another embodiment disclosed herein, the application relates to pharma- ceutically acceptable salts, hydrates, solvates, optical isomers, esters, and combinations thereof of the compounds as described above.

[0028] The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate several aspects described below, where like numerals represent like elements throughout the drawings. [Brief description of the drawings]

[0029] [Figure 1] FIG. 1 shows dose-response curves (nonlinear regression using 4 parameters) for NWD-19, NWD-14 and NWD-15 using the DPPH assay. [Diagram 2]FIG. 2 shows the viability observed for CT-26 cells following treatment with various concentrations of NWD-19. [Diagram 3] FIG. 3 shows AAPH-induced oxidative cell damage and % survival in RPE cells with or without treatment with NWD-19. [Figure 4] FIG. 4 shows the estimated malonaldehyde equivalents in RPE cells treated with NWD-19 at various concentrations following induction of oxidative stress using AAPH. [Diagram 5] Figure 5 shows disease progression in the eyes of 0.2% BAC treated mice: A) mouse eyes before treatment; B) day 7 after BAC treatment; C) day 14 after BAC treatment (day 7 after NWD-19 treatment); and D) day 21 after BAC treatment (day 14 after NWD-19 treatment). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0030] Terms used in the claims and in this application are defined as follows.

[0031] As used herein, C1-C 20 The alkyl group or moiety may be a straight or branched alkyl group or moiety containing from 1 to 20 carbon atoms. 20 Examples of alkyl groups and moieties include methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, t-butyl, 3-methyl-butyl, and the like.

[0032] As used herein, C2-C 20 Alkenyl groups or moieties are straight-chain or branched alkenyl groups or moieties. Examples include methylene, ethylene and n-propylene groups and moieties.

[0033] As used herein, a halogen group may be selected from the group consisting of chlorine, fluorine, bromine and iodine.

[0034] As used herein, an aryl group is a functional group derived from a simple aromatic ring compound where one hydrogen atom has been removed from the ring; typically, an aryl group or moiety is phenyl, benzyl or naphthyl.

[0035] As used herein, a heteroaryl group is a heterocyclyl group derived from a heteroarene by removing a hydrogen atom from any ring atom and replacing it with a moiety selected from N, O, S, S(O), and S(O). Examples include pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, furanyl, thienyl, pyrazolidinyl, pyrrolyl, oxadiazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, thiadiazolyl, imidazolyl, pyridazolyl, and pyrazolyl groups. Furanyl, thienyl, pyridazolyl, pyrazolyl, pyrimidinyl, and thiazolyl groups are preferred.

[0036] As used herein, a C1-C6 alkoxy group is typically a C1-C6 alkyl group as defined above attached to an oxygen atom.

[0037] As used herein, C 3~6 A cycloalkyl group is a saturated hydrocarbon ring having 3 to 6 carbon atoms; 3~6 A cycloalkenyl group is a hydrocarbon ring having 3 to 6 carbon atoms and 1 or 2 double bonds; and 3~8 Cycloalkoxy is a C linked via an oxygen atom. 3~8 It means one of the cycloalkyl groups.

[0038] The term "straight-chain" as used herein means a chemical structure in the form of an unbranched chain of atoms in a molecule with no side chains attached. Preferably, said (unbranched) chain is open-chained. In contrast, a "branched" structure contains one or more side chains attached to the chain of atoms in a molecule.

[0039] The term "substituted," as used herein, means that any one or more hydrogens in the corresponding group are replaced with another atom or group; i.e., a "substituted alkyl" refers to an alkyl group in which one or more hydrogens are replaced with, for example, halogen, hydroxy, or other atoms or groups.

[0040] The term "alkyl" refers to a hydrocarbon group (straight or branched) of 1 to 20 carbon atoms, including 1 to 6 carbon atoms. In general, the terms C1, C2, C6, C7, C8, C9, C10, C11, C12, C13, C14, C15, C16, C17, C18, C19, C20, C21, C22, C23, C24, C25, C26, C27, C28, C29, C30, C31, C32, C33, C34, C35, C36, C37, C38, C39, C40, C41, C42, C43, C44, C45, C46, ​​C47, C48, C49, C50, C51, C52, C53, C54, C55, C56, C57, C58 20 etc. refer to the number of C atoms (carbon atoms) present in the corresponding group. Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl (e.g., n-propyl and isopropyl), butyl (e.g., n-butyl, isobutyl, t-butyl), and pentyl (e.g., n-pentyl, isopentyl, neopentyl). The term "cycloalkyl" refers to a saturated cyclic hydrocarbon ring system. The terms "alkylaryl", "arylalkyl", "cycloalkylaryl", and "arylcycloalkyl" refer to an alkyl bonded to an aryl, an aryl bonded to an alkyl, a cycloalkyl bonded to an aryl, and an aryl bonded to a cycloalkyl, respectively.

[0041] The term "alkoxy" refers to an alkyl group bonded to an oxygen atom. Alkoxy includes, but is not limited to, methoxy, ethoxy, and the like. The term "alkoxyalkyl" refers to an alkoxy group having an (another) alkyl group bonded to the oxygen atom of the alkoxy group. Alkoxyalkyl includes, but is not limited to, methoxymethyl, ethoxyethyl, and the like. The term "alkenyl" refers to a hydrocarbon group having at least one double bond. The definition of the compounds described herein includes all possible "stereoisomers" and mixtures thereof. In particular, racemates and their isolated optical isomers having the specified activity are included. The racemates can be resolved by physical methods, such as, for example, fractional crystallization, separation or recrystallization of diastereomeric derivatives or separation by chiral column chromatography. The individual optical isomers can be obtained from the racemates by conventional methods, such as, for example, salt formation with an optically active acid followed by crystallization.

[0042] The term "tautomers" refers to constitutional isomers of the compounds described herein that are readily interconverted by a chemical reaction called tautomerization. This reaction usually results in the formal migration of a hydrogen atom or proton, accompanied by the interchange of a single bond with an adjacent double bond. As used herein, a salt of any one of the compounds disclosed in this application is a pharmacologically acceptable salt, examples of which include salts of inorganic bases such as alkali metal salts (e.g., sodium salts, potassium salts, etc.), alkaline earth metal salts (e.g., calcium salts, magnesium salts, etc.), ammonium salts, alkali metal carbonates (e.g., lithium carbonate, potassium carbonate, sodium carbonate, cesium carbonate, etc.), alkali metal hydrogen carbonates (e.g., lithium hydrogen carbonate, sodium hydrogen carbonate, potassium bicarbonate, etc.), alkali metal hydroxides (e.g., lithium hydroxide, sodium hydroxide, potassium hydroxide, cesium hydroxide, etc.); for example, tri(lower)alkylamines (e.g., trimethylamine, triethylamine, N-ethyldiisopropylamine), pyridine, quinoline, etc. , piperidine, imidazole, picoline, dimethylaminopyridine, dimethylaniline, N-(lower) alkylmorpholines (e.g., N-methylmorpholine, 1,5-diazabicyclo[4.3.0]nonene-5 (DBN), 1,8-diazabicyclo[5.4.0]undecene-7 (DBU), 1,4-diazabicyclo[2.2.2]octane (DABCO); salts of inorganic acids such as hydrochloride, hydrobromide, hydroiodide, sulfate, nitrate, phosphate; salts of organic acids such as formate, acetate, propionate, oxalate, malonate, succinate, fumarate, maleate, lactate, malate, citrate, tartrate, carbonate, picrate, methanesulfonate, ethanesulfonate, p-toluenesulfonate, glutamate.

[0043] The compounds described herein may also have the form of a "prodrug". Since prodrugs are known to enhance pharmaceutical qualities (e.g., solubility, manufacturing, etc.), the compounds of the present invention may be delivered in the form of a prodrug. "Prodrugs" are intended to include any covalently bonded carriers that release the active parent drug immediately in vivo when such prodrugs are administered to a mammalian subject.

[0044] Prodrugs include compounds described herein where, for example, a hydroxy, amino or other group is bonded to any group that, upon administration of the prodrug, cleaves to form the free hydroxyl, free amino, etc., respectively. Examples of prodrugs include, but are not limited to, acetate, formate, and benzoate derivatives of alcohol and amine functional groups present in the compounds of the invention.

[0045] As used herein, "pharmaceutically acceptable salts" refers to compounds that are modified by making salts of their acids or bases. Pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines; alkali or organic salts of acidic residues such as carboxylic acids; and the like. In another embodiment, pharmaceutically acceptable salts include the conventional non-toxic salts or quaternary ammonium salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. Preferably, conventional non-toxic salts include those derived from inorganic acids, such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, nitric acid, and the like; as well as salts prepared from organic acids, such as acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pamoic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, sulfamic acid, 2-acetoxybenzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid, isethionic acid, and the like.

[0046] As used herein, the term "pharmaceutically acceptable carrier" refers to a carrier or adjuvant that may be administered to a patient together with a compound disclosed herein, or a pharmaceutically acceptable salt thereof, and that does not destroy the pharmacological activity of the compound and is non-toxic when administered in a dosage sufficient to deliver a therapeutic amount of the compound.

[0047] As used herein, the term "composition" encompasses a product that contains a particular ingredient in a particular amount, as well as any product that results, directly or indirectly, from the combination of the specified amounts of the specified ingredients. Such terms related to pharmaceutical compositions are intended to encompass products that contain the active ingredient and the inactive ingredients that constitute the carrier, as well as any product that results, directly or indirectly, from the combination, complex or aggregate of any two or more ingredients, or from the dissociation of one or more ingredients, or from other types of reactions or interactions of one or more ingredients. Thus, the pharmaceutical compositions described in this application encompass any composition made by mixing a compound described herein, or a pharma-ceutically acceptable salt thereof, and a pharma-ceutically acceptable carrier. "Pharmaceutically acceptable" means that the carrier, diluent or excipient must be compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.

[0048] The phrase "therapeutically effective amount" is intended to include the amount of the compounds described herein that are effective when administered alone or in combination. The phrase is also intended to include the amount of the combination of compounds claimed that is effective in stimulating the endogenous regeneration of terminally differentiated cells in a mammal. Preferably, said combination of compounds is a synergistic combination. Such a synergistic effect occurs when the effect of the compounds when administered in combination is greater than the additive effect of the compounds when administered alone as a single agent.

[0049] The terms "treat" or "treatment", as used herein, include treatment of a disorder condition in a mammal, particularly a human, and include preventing the disorder condition from occurring in a mammal, e.g., said mammal is susceptible to a disorder but has not been diagnosed with the disorder, inhibiting the disorder condition, i.e., arresting further occurrence, and / or alleviating the disorder condition, i.e., ameliorating the symptoms of the disorder.

[0050] Compound: The present application relates to compounds of formula (I), including pharma- ceutically acceptable salts, stereoisomers, and prodrugs thereof. [ka]

[0051] (In the formula: [ka]

[0052] - R1 and R2 are -H, optionally mono-, di- or trisubstituted with halogen, substituted and unsubstituted aryl (monocyclic and bicyclic), substituted and unsubstituted heteroaryl (monocyclic and bicyclic), -C 1~20 Alkyl, C 2~20 Alkenyl, and C 2~20 alkynyl, - R3 is -H, -C 1~20 Alkyl, C 2~20 Alkenyl, and C 2~20 alkynyl, where R3 is optionally substituted by one or more A; aryl and heteroaryl; - R4 is -H, -C 1~20 Alkyl, C 2~20 Alkenyl, and C 2~20 alkynyl, where R4 is optionally substituted by one or more A; aryl and heteroaryl; - R5 is (a) aryl, (b) heteroaryl, (c) -C1~2 Alkyl-aryl and (d)-C 1~2 alkyl-heteroaryl; where A is an aryl or heteroaryl group; wherein the aryl of options (a) and (c) and the heteroaryl of options (b) and (d) are optionally selected from halogen, -C 1~6 Alkyl, -CF3, -OCF3, -C 1~6 Alkoxy, -C 3~6 Cycloalkoxy, halo-C 1~6 Alkyl, aryl, heteroaryl, heterocyclo, -C 3~6 Cycloalkyl, and -C 3~6 cycloalkenyl; However, at least one of R1, R2, R3, R4, and R5 must be [ka]

[0053] (wherein X, Y, and Z are selected from the group consisting of hydrogen, methyl, ethyl, propyl, butyl, substituted or unsubstituted aryl and heteroaryl; n is an integer selected from 0 to 18, and E is a phosphorus or nitrogen atom). The present invention discloses a compound for targeting mitochondria having the structure:

[0054] In one embodiment disclosed herein, the compound of formula (I) has the following structure: [ka]

[0055] and one of their pharma- ceutically acceptable salts; wherein X, Y, and Z are as defined above.

[0056] In another embodiment, the compound of formula (I) has the following structure: [ka]

[0057] as well as one of their pharma- ceutically acceptable salts.

[0058] In another embodiment, the compound of formula (I) has the following structure, where n=6: [ka]

[0059] as well as one of their pharma- ceutically acceptable salts.

[0060] In a preferred embodiment disclosed herein, the compound of formula (I) has the following structure: [ka]

[0061] as well as one of their pharma- ceutically acceptable salts.

[0062] In one embodiment, the compounds of formula (I) contain one or more asymmetric centers and thus occur as racemates and racemic mixtures, enantiomerically enriched mixtures, single enantiomers, individual diastereomers and diastereomeric mixtures.

[0063] In one aspect of one embodiment disclosed herein, the compounds of formula (I) include cis, trans, syn, anti, entgegen (E), zusammen (Z) isomers or mixtures thereof.

[0064] In another embodiment, the compounds described herein may also be represented in multiple tautomeric forms. In such cases, the present application expressly includes all tautomeric forms of the compounds described herein, even if only a single tautomeric form may be represented. All such isomeric forms of such compounds are expressly included in the present invention.

[0065] In another embodiment disclosed herein, the compounds of the present invention include salts and prodrugs. Salts can be formed, for example, between anions and positively charged substituents (e.g., amino) on the compounds described herein. Suitable anions include chloride, bromide, iodide, sulfate, nitrate, phosphate, citrate, methanesulfonate, trifluoroacetate, and acetate. Similarly, salts can also be formed between cations and negatively charged substituents (e.g., carboxylate) on the compounds described herein. Suitable cations include sodium, potassium, magnesium, calcium, and ammonium cations, such as tetramethylammonium.

[0066] In one embodiment where the compound of the present invention is basic, salts may be prepared from pharma- ceutically acceptable non-toxic acids, including inorganic and organic acids. Such acids include acetic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, ethanesulfonic acid, fumaric acid, gluconic acid, glutamic acid, hydrobromic acid, hydrochloric acid, isethionic acid, lactic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, mucic acid, nitric acid, pamoic acid, pantothenic acid, phosphoric acid, succinic acid, sulfuric acid, tartaric acid, p-toluenesulfonic acid, and the like. In one aspect of the present invention, the salts are citric acid, hydrobromic acid, hydrochloric acid, maleic acid, phosphoric acid, sulfuric acid, fumaric acid, and tartaric acid.

[0067] In another embodiment, when the compound of the present invention is acidic, salts may be prepared from pharma- ceutically acceptable non-toxic bases, including inorganic and organic acids. In one embodiment, the salts prepared include lithium salts, sodium salts, potassium salts, magnesium salts, calcium salts, dicyclohexylamine salts, N-methyl-D-glucamine salts, tris(hydroxymethyl)methylamine salts, arginine salts, lysine salts, etc.

[0068] In one embodiment, the compounds of the present invention may be radiolabeled with radioactive isotopes, such as, for example, tritium, iodine-125, or carbon-14. In a preferred embodiment, all isotopic variations of the compounds of the present invention, whether radioactive or not, are intended to be encompassed within the scope of the present invention. In some embodiments, hydrogen atoms of the compounds described herein may be replaced with deuterium atoms.

[0069] In some embodiments, the compounds of Formula I can be prepared as prodrugs, which are drug precursors that, following administration to a subject and subsequent absorption, are converted through some process, such as conversion by a metabolic pathway, to an active or more active species.

[0070] The present application also provides a compound of formula 8 [ka]

[0071] The present invention discloses a process for preparing a pyrazolone core compound of the formula: During the ceremony: [ka]

[0072] - R1 and R2 are mono-, di- or trisubstituted with hydrogen, optionally halogen, substituted and unsubstituted aryl (monocyclic and bicyclic), substituted and unsubstituted heteroaryl (monocyclic and bicyclic), -C 1~20 Alkyl, C 2~20Alkenyl, and C 2~20 alkynyl, - R3 is hydrogen, -C 1~20 Alkyl, C 2~20 Alkenyl, and C 2~20 alkynyl, where R3 is optionally substituted by one or more A; aryl and heteroaryl; - R4 is hydrogen, -C 1~20 Alkyl, C 2~20 Alkenyl, and C 2~20 alkynyl, where R4 is optionally substituted by one or more A; aryl and heteroaryl; - R5 is (a) hydrogen, (b) aryl, (c) heteroaryl, (d) -C 1~2 Alkyl-aryl, and (e)-C 1~2 In formula 8, R5 can also be selected from groups such as halogen, -C1-6 alkyl, -CF3, -SO2, -SON2NH2, -OCF3, -C1-6 alkoxy, -C3-6 cycloalkoxy, halo-C1-6 alkyl, aryl, heteroaryl, heterocyclo, -C3-6 cycloalkyl, and -C3-6 cycloalkenyl; where A is an aryl or heteroaryl group; wherein the aryl of options (a) and (c), and the heteroaryl of options (b) and (d) are as described above; However, at least one of R1, R2, R3, R4, and R5 must be [ka]

[0073] (wherein n=1 to 20, and X, Y, and Z are as defined above).

[0074] In one embodiment, the process for preparing the compounds of formula I and the pyrazolone core compounds of formula 8 is illustrated in the following scheme: [ka] [ka] [ka]

[0075] In one embodiment, the process for preparing rhodamine derivatives of compounds having general formula I is illustrated in the following scheme: [ka]

[0076] In one embodiment, the pyrazolone core of formula (8) found in the compounds of formula I or exemplified compounds can be conveniently accessed, for example, from the corresponding carboxylic acid (1) via an initial non-decarboxylative Claisen condensation reaction with a suitable monoester of malonic acid (2) in the presence of a suitable activating reagent, such as carbonyldiimidazole (i.e., CDI), a suitable Lewis acid, such as magnesium chloride, and a suitable base, such as triethylamine (i.e., TEA).

[0077] In another embodiment, a suitable intermediate (5) for obtaining the desired molecule can be generated from the corresponding suitable ethyl acetoacetate or its derivative (4) and a halogenated alkene or alkane (3) having a lipophilic cation exemplified by bromobutyltriphenylphosphine. This reaction between compounds of formula (4) and (3) can be carried out in an alcoholic solvent such as ethanol or methanol, in the presence of a suitable base such as sodium ethoxide, to generate anions of ethyl acetoacetate or its derivatives, which can then be reacted with bromobutyltriphenylphosphine (3) in THF or a suitable solvent to give the desired intermediate of formula (5).

[0078] In another embodiment, the reaction can be further carried out by heating either the hydrazide of formula (6) or (6') and intermediate (5) in an alcoholic solvent such as ethanol or methanol or optionally a less reactive neat solvent in an enclosed reaction vessel, which then provides the desired compound of formula (7) or (7').

[0079] These intermediates of formula (7) or (7') can then be functionalized, if desired, via either a simple nucleophilic substitution reaction, when R4 is an alkyl or heteroalkyl group, or a copper catalyzed N-arylation reaction, when R4 is an aryl or heteroaryl group.

[0080] In the former simple nucleophilic substitution reaction, a base such as, but not limited to, NaH, CS2CO3 or K2CO3 is added to the pyrazolone intermediate (7) or (7') along with R4-X in a polar aprotic solvent.

[0081] For the latter copper-catalyzed N-arylation reaction, a convenient copper(I) source such as copper iodide, a suitable ligand such as praline, and a suitable base such as Cs2CO3 are heated with R4-X in a polar aprotic solvent to intermediate (7) or (7') to afford the pyrazolone core compound of formula (8) or the compound of formula (I).

[0082] In another embodiment, the reagents and conditions used in the above schematic reaction are as follows: (a) Potassium 3-methoxy-3-oxopropanoate, CDI, MgCl2, TEA; (b) NaOEt, EtOH, rt, then bromobutyltriphenylphosphine in THF, 80 °C; (c) phenylhydrazine, 80-140°C; (d) R4-X, CsCO3, ACN / DMF, 60 °C, 2–16 h.

[0083] In another embodiment, the compound of formula (8) has the following structure: [ka]

[0084] or one of their pharma- ceutically acceptable salts, wherein X, Y, and Z are as defined above.

[0085] In one embodiment disclosed herein, the starting materials used for synthesis are synthesized or obtained from commercial sources such as, but not limited to, Sigma-Aldrich, Fluka, Acros Organics, Alpha Acer, VWR Scientific, etc. General methods for the preparation of compounds can be modified by the use of appropriate reagents and conditions for the introduction of the various moieties found in the structures as provided herein. While preferred embodiments of the invention have been shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, modifications, and substitutions will occur to those skilled in the art herein without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein can be used in carrying out the invention.

[0086] Pharmaceutical Compositions: In one embodiment, the present application provides a pharmaceutical composition comprising a compound of Formula I and Formula 8 described herein, or a salt thereof, and a pharma- ceutically acceptable carrier.

[0087] In another embodiment, the present application provides a pharmaceutical composition and a process for the preparation thereof, the process comprising mixing a mitochondrial targeted compound of Formula I and Formula 8 or a pharma- ceutically acceptable salt thereof with a pharma- ceutically acceptable carrier.

[0088] In another embodiment, the present application includes within its scope prodrugs of the compounds of the present invention. In general, such prodrugs are functional derivatives of the compounds of the present invention that can be easily converted in vivo into the required compound. Thus, in the methods of treatment described herein, the term "administration" of a compound or "administering" a compound will encompass treatment of the various conditions described with a specifically disclosed compound or with a compound that may not be specifically disclosed but that converts to a particular compound in vivo after administration to a patient.

[0089] Conventional procedures for the selection and preparation of suitable prodrug derivatives are described, for example, in "Design of Prodrugs", edited by H. Bundgaard, Elsevier, 1985. Metabolites of these compounds include active species produced upon introduction of the compounds described herein into the biological milieu.

[0090] In one embodiment described herein, the amount to be administered depends on the compound formulation, route of administration, and is generally empirically determined by routine trials. Preferably, the amount of active compound in a unit dose of the formulation may be varied or adjusted by about 0.0001 to 1 mg, 3 mg, 10 mg, or 30 to about 30 mg, 100 mg, 300 mg, or 1000 mg.

[0091] The compositions described herein may be administered in dosage forms as described above, once or in 1-10 divided doses per day. Patients may be advised to start with a low dose combination and gradually increase to a high dose combination. More preferably, the compounds described herein or pharma- ceutically acceptable salts thereof, or compositions thereof, may be administered about 1-6 times per day, or alternatively, as a continuous infusion. Such administration may be used as chronic or acute treatment. Lower or higher doses than those listed above may be required. In another embodiment, the total daily dose may be divided and administered in portions throughout the day as desired.

[0092] In another embodiment, the compound or pharma- ceutically acceptable salt thereof described herein or a composition thereof may be administered orally, parenterally (e.g., by subcutaneous, intradermal, intravenous, intramuscular, intra-articular, intra-articular, intra-synovial, intrasternal, intrathecal, intralesional and intracranial injection or infusion techniques), by inhalation spray, topically, rectally, nasally, buccally, intravaginally, via an implanted reservoir, by injection, subcutaneously, intraperitoneally, transdermally, or as eye drops, for example, at a dosage ranging from about 0.001 mg / kg to 1000 mg / kg (e.g., about 0.001 to about 100 mg / kg, about 0.1 to about 100 mg / kg, about 1 to about 100 mg / kg) every 2 to 168 hours, or depending on the requirements of the particular drug. Preferably, the composition is administered orally or by a parenteral route via injection. The methods described herein contemplate the administration of an effective amount of a compound of the invention or a pharma- ceutically acceptable salt thereof, or a composition of a compound of the invention, to achieve the desired or described effect.

[0093] In the alternative embodiments described herein, the specific dosage and treatment regimen for any patient will depend on a wide variety of factors, including the activity of the particular compound used, age, body weight, general health, sex, diet, time of administration, excretion rate, drug combination, severity and course of the disease, condition or symptoms, the patient's predisposition to the disease, and the judgment of the treating physician.

[0094] In one embodiment, the compounds of formula I and formula 8 or pharma- ceutically acceptable salts thereof may be administered at a dose of about 0.00005 milligrams to about 2,000 milligrams. In another embodiment, the compounds of formula I and formula 8 or pharma- ceutically acceptable salts thereof may be administered at a dose of about 0.1 milligrams to about 1,000 milligrams. In yet another embodiment, the compounds of formula I and formula 8 or pharma- ceutically acceptable salts thereof may be administered at a dose of about 0.1 milligrams to about 500 milligrams. In yet another embodiment, the compounds of formula I and formula 8 or pharma- ceutically acceptable salts thereof may be administered at a dose of about 0.1 milligrams to about 250 milligrams. Preferably, the compounds of formula I and formula 8 or pharma- ceutically acceptable salts thereof may be administered at a dose of about 0.1 milligrams to about 100 milligrams. More preferably, the compounds of formula I and formula 8 or pharma- ceutically acceptable salts thereof may be administered at a dose of about 0.0001 milligrams to about 50 milligrams. Even more preferably, the compounds of Formula I and Formula 8 or their pharma- ceutically acceptable salts may be administered in a dose of about 0.01 milligrams to about 25 milligrams. The dosage form may further comprise a pharma- ceutically acceptable carrier and / or an additional therapeutic agent.

[0095] In another embodiment disclosed herein, the compound of the present invention may be co-administered with one or more additional therapeutic agents. In one aspect of the embodiment, the additional agents may be administered separately from the compound of the present invention (e.g., sequentially; on different overlapping schedules) as part of a multiple dosing regimen. In some embodiments, these agents may be part of a single dosage form, mixed together with the compound of the present invention in a single composition. In yet another embodiment, these agents may be given as separate doses administered at about the same time that the compound of the present invention is administered. In another embodiment, the composition of the present invention includes a combination of the compound of the present invention and one or more additional therapeutic or prophylactic agents, both the compound and the additional agent may be present at dosage levels ranging from about 0.1% to about 100%; more preferably, the compound and the additional agent may be present at dosage levels ranging from about 0.5% to about 98% of the dosage normally administered in a monotherapy regimen.

[0096] In another embodiment disclosed herein, the composition may contain any conventional non-toxic pharmaceutically acceptable carrier, adjuvant or vehicle.In some cases, the pH of the formulation may be adjusted with pharmaceutically acceptable acid, base or buffer to enhance the stability of the formulated compound or its delivery form.

[0097] In one embodiment, the compositions described herein may be orally administered in any orally acceptable dosage form, including but not limited to capsules, tablets, emulsions, and aqueous suspensions, dispersions, and solutions.For tablets for oral use, commonly used carriers include lactose and cornstarch.Lubricants such as magnesium stearate are also typically added.For oral administration in the form of capsules, useful diluents include lactose and dried cornstarch.For aqueous suspensions and / or emulsions to be orally administered, the active ingredient may be suspended or dissolved in an oil phase, and then combined with emulsifiers and / or suspending agents.If desired, certain sweeteners and / or flavorings and / or colorings may be added.

[0098] In one embodiment, the pharmaceutical preparation is prepared by using a diluent or excipient such as a filler, extender, binder, moisturizer, disintegrant, surfactant, and lubricant. Various forms of the pharmaceutical preparation can be selected according to the purpose of treatment, and typical examples include tablets, pills, powders, solutions, suspensions, emulsions, granules, capsules, suppositories, and injections (solutions, suspensions).

[0099] In one embodiment, the present application relates to tablet formulation, and conventionally known carriers may be widely used. Examples include excipients such as lactose, urea, starch, sucrose, glucose, urea, starch, calcium carbonate, kaolin, sodium chloride, crystalline cellulose, and silicate; binders such as water, ethanol, propanol, gelatin solution, potassium phosphate, glucose solution, shellac, starch solution, carboxymethylcellulose, methylcellulose, and polyvinylpyrrolidone; sodium bicarbonate, agar powder, dry starch, calcium carbonate, sodium alginate, laminaran powder, polyoxyethylene, and the like. Disintegrants such as ethylene sorbitan fatty acid esters, sodium lauryl sulfate, starch, lactose, monoglyceride stearic acid, etc.; anti-disintegrants such as saccharose, stearin, etc.; absorption promoters such as quaternary ammonium bases, cocoa butter, hardened oils, etc.; sodium lauryl sulfate; humectants such as glycerin, starch, etc.; absorbents such as starch, lactose, kaolin, bentonite, colloidal silica, etc.; lubricants such as polyethylene glycol, purified talc, stearates, borate powder, etc. may be used. Furthermore, tablets may be tablets that have been given conventional coatings as necessary, for example, sugar-coated tablets, gelatin capsule tablets, enteric-coated tablets, film-coated tablets, or double tablets, multi-layer tablets.

[0100] In another embodiment, the present application relates to a pill dosage form, and conventionally known carriers may be widely used. Excipients such as glucose, hardened vegetable oil, lactose, starch, cacao butter, kaolin, talc, etc.; binders such as powdered gum arabic, powdered tragacanth, gelatin, ethanol, etc.; disintegrants such as laminarin, agar, etc. may be used.

[0101] In another embodiment, the present application relates to a suppository form, where conventionally known carriers are used, including polyethylene glycol, cacao butter, higher alcohols, esters of higher alcohols, gelatin, and semi-synthetic glycerides.

[0102] In one aspect of the present invention, the present application relates to a capsule dosage form, which can be prepared according to a conventional method by mixing the active ingredient compounds with various carriers as exemplified above and filling them into hard gelatin capsules, soft capsules, etc.

[0103] In another embodiment, the present application relates to an injectable liquid dosage form.Preferably, the solution, emulsion and suspension are sterile and isotonic with blood, and any of those conventionally used in the art as diluents can be used to form these modes, for example, water, ethyl alcohol, macrogol, propylene glycol, ethoxylated isostearyl alcohol, polyoxylated isostearyl alcohol, polyoxyethylene sorbitan fatty acid ester, etc.

[0104] In another embodiment, the present application relates to the formulation of eye drops.Preferably, the solution, emulsion and suspension are sterile and isotonic with blood, and any of the diluents conventionally used in the art can be used to form these modes, such as acetylcysteine, cysteine ​​hydrochloride hydrate, glycerin, PEG and PEG derivatives, sodium chloride, sodium bisulfite, sodium hydroxide, phosphoric acid, benzalkonium chloride, sodium dihydrogen phosphate dihydrate, disodium hydrogen phosphate dodecahydrate, boric acid, citric acid, hypromellose, N-acetyl-carnosine, potassium bicarbonate, purified benzyl alcohol, sterile water, etc.

[0105] There is no particular limitation in the method of administration of the pharmaceutical preparation of the present application, and it may be administered by a method according to the specific form of the preparation, the age, sex and other conditions of the patient, the severity of the disease, etc. For example, in the case of tablets, pills, solutions, suspensions, emulsions, granules and capsules, it is administered orally. In the case of injection, it is administered intravenously alone or in combination with conventional replacement fluids, such as glucose and amino acids, if necessary, and the preparation alone may also be administered intramuscularly, intradermally, subcutaneously, intraperitoneally or intraocularly, if necessary. In the case of suppositories, it is administered rectally. The applied dose of the pharmaceutical preparation of the present invention is appropriately selected according to the administration schedule, the age, sex and other conditions of the patient, the severity of the disease, etc.

[0106] Method of use: In one embodiment disclosed herein, the therapeutically effective amount of the compound of formula I and formula 8, their pharma- ceutically acceptable salts or their compositions is used in antioxidant therapy.In a preferred embodiment, the selected antioxidant therapy for humans or animals is used to prevent mitochondrial damage.This can be done by normalizing or preventing the elevated mitochondrial oxidative stress associated with certain diseases, such as cancer, Parkinson's disease, diabetes, dry eye disease, age-related macular degeneration or diseases associated with mitochondrial DNA mutations.

[0107] In another embodiment, the compounds of formula I and formula 8, their pharma- ceutically acceptable salts or compositions thereof may also be used in combination with cell transplantation therapy for neurodegenerative diseases to increase the survival rate of transplanted cells.

[0108] The compounds described herein, their pharmaceutically acceptable salts or their pharmaceutical compositions can be used in therapeutically effective amounts to treat conditions involving oxidative stress in mammals, where oxidative damage plays an important role in the aging process.By far the greatest risk factor for neurodegenerative diseases, such as Alzheimer's disease (AD), Parkinson's disease (PD) and amyotrophic lateral sclerosis (ALS), is aging.Mitochondria are believed to contribute to aging through the accumulation of mitochondrial DNA mutations and the net production of reactive oxygen species (ROS).

[0109] In addition, oxidative damage plays a role in the pathogenesis of hypoxic injury, cancer, and cardiovascular disease associated with reperfusion, and neurodegenerative diseases. Oxidative stress also contributes to diseases such as Huntington's chorea, diabetes, and Friedreich's ataxia, and to the nonspecific damage that accumulates with age. It also contributes to inflammation and ischemia-reperfusion tissue damage in stroke and heart attack, and during organ transplantation and surgery.

[0110] In one embodiment, the compounds described herein, their pharma- ceutically acceptable salts, or their compositions can also be used in ophthalmology, where oxidative stress is involved in the pathogenesis of some eye diseases, such as senile cataract, age-related macular degeneration, uveitis, retinopathy of prematurity, keratitis, and ocular inflammation.Oxidized protein levels increase during these diseases with increased inflammatory activity.Oxidative stress plays a major role in the pathogenesis of some forms of dry eye, and treatment with topical / systemic use of antioxidants would be useful.

[0111] In another aspect, the present application provides a method for transplanting cells or tissues into a subject.In a preferred embodiment, the present application is directed to a method that can be used as a prophylactic agent to protect organs during transplantation or improve ischemia-reperfusion injury that occurs during surgery.The compounds of the present invention, their pharmacologic acceptable salts, or their compositions can also be used to reduce cell damage after stroke and heart attack, or to be given prophylactically to premature infants who are prone to cerebral ischemia.

[0112] In another embodiment, the methods of treatment by administering the compounds described herein include treatment of the various conditions described with compounds that are specifically disclosed or compounds that may not be specifically disclosed but that convert to a particular compound in vivo after administration to a patient.

[0113] In one embodiment disclosed herein, the method has a major advantage over current antioxidant therapies in that it allows antioxidants to selectively accumulate in mitochondria, the part of the cell under greatest oxidative stress. This would greatly enhance the efficacy of antioxidant therapy. Related lipophilic cations have been tested as potential antioxidants (in vitro) and are known to be relatively non-toxic to whole animals, so these mitochondrial-targeted antioxidants may lack adverse side effects.

[0114] The compounds disclosed herein may be prepared, for example, according to the following reaction schemes.

[0115] Example 1: Synthesis of (4-(5-methyl-3-oxo-2-phenyl-2,3-dihydro-1H-pyrazol-1-yl)butyl)triphenylphosphonium bromide (NWD-11) [ka]

[0116] Equimolar amount of ethyl acetoacetate was added to a 100 mL round bottom flask containing phenylhydrazine in ethanol. The mixture was then stirred at room temperature for 5 minutes and then refluxed for 5 hours. After completion of the reaction, the solvent was removed and a yellowish solid (Intermediate-I) was precipitated by adding diethyl ether. The precipitated solid was dried under vacuum and dissolved in 10 mL of DMF to obtain a clear solution. 4-Bromobutyltriphenylphosphine (1.1 equiv.) and CsCO3 (1.1 equiv.) were added to the above solution and the mixture was then heated at 65-70°C for 4 hours. After 4 hours, the reaction mixture was cooled to room temperature and poured into ice-cold water. Finally, the desired compound was extracted with chloroform three times. All the organic phases were combined and dried over sodium sulfate. The solvent was removed under reduced pressure and the resulting crude product was purified by column chromatography (DCM:MeOH, 9:1) to give a white solid as the final compound (45% yield), which was characterized by TLC (Rf=0.45, DCM:MeOH, 9:1) and mass spectrometry (M / Z=491.3) as (4-(5-methyl-3-oxo-2-phenyl-2,3-dihydro-1H-pyrazol-1-yl)butyl)triphenylphosphonium bromide.

[0117] Example 2: (4-(5-oxo-1-phenyl-2,5-dihydro-1H-pyrazol-3-yl)butyl)triphenylphosphonium bromide (NWD-14) [ka]

[0118] To a solution of (4-carboxybutyl)triphenylphosphonium (1 eq.) in acetonitrile (0.05M) was added CDI (1.1 eq.). The resulting yellow solution was then stirred for 2.5 h at RT (the solution turns into a white precipitate) before it was added in portions over a period of 0.5 h to a white suspension of potassium 3-methoxy-3-oxopropanoate (2.1 eq.), magnesium chloride (2.5 eq.) and triethylamine (3.2 eq.). The resulting suspension was then stirred for 2 h at RT and finally heated at reflux for a further 24 h. The thus obtained crude reaction suspension was cooled to RT, diluted with water and the pH was adjusted to 7.9-8.2 with 10% HCl. The desired intermediate was extracted three times with DCM. All organic phases were combined and washed with brine and water. Finally, the organic phase was dried over sodium sulfate and concentrated to give Intermediate-II as a dark oily mass (75% yield), which can be used without further purification. Intermediate-II was dissolved in ethanol and 1.5 equivalents of phenylhydrazine monohydrate was added therein to obtain a solution. The reaction mixture was refluxed at 80° C. for 1 h and then at 140° C. for 4 h. The reaction turned into a sticky mass upon cooling to RT. The desired compound was precipitated with diethyl ether and filtered. Finally, it was purified by silica gel chromatography to give (4-(5-oxo-1-phenyl-2,5-dihydro-1H-pyrazol-3-yl)butyl)triphenylphosphonium bromide as a dark yellow solid (35% yield) as the final compound characterized by TLC (Rf=0.46, DCM:MeOH, 9:1) and mass spectrometry LC-MS: 477.1 (M / Z).

[0119] Example 3: (10-(5-oxo-1-phenyl-2,5-dihydro-1H-pyrazol-3-yl)decyl)triphenylphosphonium bromide (NWD-22) [ka]

[0120] (9-Carboxynonyl)triphenylphosphonium bromide was prepared by refluxing 10-bromodecanoic acid with triphenylphosphine in toluene. (9-Carboxynonyl)triphenylphosphonium bromide was purified and reacted with CDI (1.1 eq.). The resulting yellow solution was then stirred for about 2-2.5 h at RT (the solution turns into a white precipitate) before it was added in portions over a period of 0.5 h to a white suspension of potassium 3-methoxy-3-oxopropanoate (2.1 eq.), magnesium chloride (2.5 eq.) and triethylamine (3.2 eq.). The resulting suspension was then stirred for 2 h at RT and finally heated at reflux for an additional 24 h. The thus obtained crude reaction suspension was cooled to RT, diluted with water and the pH was adjusted to 7.9-8.2 with 10% HCl. The desired intermediate was extracted three times with DCM. All organic phases were combined and washed with brine and water. Finally, the organic phase was dried over sodium sulfate and concentrated to obtain (12-methoxy-10,12-dioxododecyl)triphenylphosphonium bromide (Intermediate-III) as a dark oily mass, which was used without further purification. Intermediate-III was then dissolved in ethanol and 1.5 equivalents of phenylhydrazine monohydrate was added therein to obtain a solution. The reaction mixture was refluxed at 80° C.-110° C. for 4-6 hours. The reaction mass turned into a sticky mass upon cooling to RT. The desired compound was precipitated with diethyl ether and filtered. Finally, it was purified by silica gel chromatography to give (10-(5-oxo-1-phenyl-2,5-dihydro-1H-pyrazol-3-yl)decyl)triphenylphosphonium bromide as a dark yellow solid (30% yield) as the final compound characterized by TLC (Rf=0.51, DCM:MeOH, 9:1) and mass spectrometry LC-MS: 561.5 (M / Z).

[0121] Example 4: (4-(3-methyl-5-oxo-1-phenyl-4,5-dihydro-1H-214-pyrazol-4-yl)butyl)triphenylphosphonium bromide (NWD-15) [ka]

[0122] To a stirred solution of sodium ethoxide (21% wt, 25 mmol) in ethanol was added ethyl acetoacetate (25 mmol) at room temperature. The mixture was heated at 80° C. and after 30 min, 4-bromobutyltriphenylphosphine (25 mmol) in ethanol was added to obtain a yellow solution. After 30 min at RT, the reaction mixture turned into a white suspension, which was then refluxed for 24 h. After 24 h, the reaction mixture was cooled at RT and the pH was adjusted to 6.5 with 1N HCl. The reaction mixture was partitioned with water and ethyl acetate. The organic phases were combined, subjected to washing with water and brine, and dried over MgSO4. The organic phase was concentrated under vacuum to obtain an oily mass, which was confirmed by MS analysis to be the desired intermediate-IV, which can be used without further purification. An equimolar amount of phenylhydrazine monohydrate was added to the vessel containing intermediate-IV, and the mixture thus formed was heated at 140° C. for 4 h. Finally, the desired compound was precipitated using ether and purified by silica gel chromatography to give the desired compound as a sticky solid in 40% yield, characterized by TLC (Rf=0.53, DCM:MeOH, 9:1) and mass spectrometry LC-MS: 491.3 (M / Z).

[0123] Example 5: (10-(3-methyl-5-oxo-1-phenyl-4,5-dihydro-1H-pyrazol-4-yl)decyl)triphenylphosphonium bromide (NWD-19) To a stirred solution of sodium ethoxide (21% wt, 25 mmol) in ethanol was added ethyl acetoacetate (25 mmol) at room temperature. The mixture was heated at 80° C. and after 30 min, (10-bromodecyl)triphenylphosphonium (25 mmol) in ethanol was added to give a yellow solution. After 30 min at RT, the reaction mixture turned into a white suspension, which was then refluxed for 24 h. After 24 h, the reaction mixture was cooled at RT and the pH was adjusted to 6.5-7.0 with 1N HCl. The reaction mixture was partitioned with water and ethyl acetate. The organic phases were combined, subjected to washing with water and brine, and dried over MgSO4. The organic phase was concentrated under vacuum to give an oily mass, which was confirmed by MS analysis to be the desired (11-(ethoxycarbonyl)-12-oxotridecyl)triphenylphosphonium bromide (Intermediate-V), which was used without further purification. An equimolar amount of phenylhydrazine monohydrate was added to the vessel containing intermediate-V and the mixture thus formed was heated at 100-120 °C for 4-5 h. Finally, the desired compound was precipitated using ether and purified by silica gel chromatography (28% yield). The compound thus isolated was characterized by TLC (Rf=0.48, DCM:MeOH, 9:1) and mass spectrometry LC-MS: 575.2 (M / Z).

[0124] Example 6: (10-(1-(3-ethylphenyl)-3-methyl-5-oxo-4,5-dihydro-1H-pyrazol-4-yl)decyl)triphenylphosphonium (NWD-36) To a stirred solution of sodium ethoxide (21% wt, 25 mmol) in ethanol was added ethyl acetoacetate (25 mmol) at room temperature. The mixture was heated at 80° C. and after 30 min, (10-bromodecyl)triphenylphosphonium (25 mmol) in ethanol was added to give a yellow solution. After 30 min at RT, the reaction mixture turned into a white suspension, which was then refluxed for 24 h. After 24 h, the reaction mixture was cooled at RT and the pH was adjusted to 6.5-7.0 with 1N HCl. The reaction mixture was partitioned with water and ethyl acetate. The organic phases were combined, subjected to washing with water and brine, and dried over MgSO4. The organic phase was concentrated under vacuum to give an oily mass, which was confirmed by MS analysis to be the desired (11-(ethoxycarbonyl)-12-oxotridecyl)triphenylphosphonium bromide (Intermediate-V), which was used without further purification. An equimolar amount of (3-ethyl)phenylhydrazine monohydrate was added to the vessel containing intermediate-V and the mixture thus formed was heated at 100-120° C. for 4-5 h. Finally, the desired compound was precipitated using ether and purified by silica gel chromatography to give (42% yield) and characterized by TLC (Rf=0.61, DCM:MeOH, 9:1) and mass spectrometry LC-MS: 603.2 (M / Z).

[0125] Example 7: (10-(3-methyl-1-(3-(methylsulfonyl)phenyl)-5-oxo-4,5-dihydro-1H-pyrazol-4-yl)decyl)triphenylphosphonium bromide To a stirred solution of sodium ethoxide (21% wt, 25 mmol) in ethanol was added ethyl acetoacetate (25 mmol) at room temperature. The mixture was heated at 80° C. and after 30 min, (10-bromodecyl)triphenylphosphonium (25 mmol) in ethanol was added to give a yellow solution. After 30 min at RT, the reaction mixture turned into a white suspension, which was then refluxed for 24 h. After 24 h, the reaction mixture was cooled at RT and the pH was adjusted to 6.5-7.0 with 1N HCl. The reaction mixture was partitioned with water and ethyl acetate. The organic phases were combined, subjected to washing with water and brine, and dried over MgSO4. The organic phase was concentrated under vacuum to give an oily mass, which was confirmed by MS analysis to be the desired (11-(ethoxycarbonyl)-12-oxotridecyl)triphenylphosphonium bromide (Intermediate-V) and used without further purification. An equimolar amount of 1-[4-methylsulfonyl]phenyl]hydrazine was added to the vessel containing intermediate-V and the mixture thus formed was heated at 100-120° C. for 4-5 h. Finally, the desired compound was precipitated using ether and purified by silica gel chromatography to give (45% yield) and characterized by TLC (Rf=0.63, DCM:MeOH, 9:1) and mass spectrometry LC-MS: 653.2 (M / Z).

[0126] Example 8: 2-(10-(3-methyl-5-oxo-1-phenyl-4,5-dihydro-1H-pyrazol-4-yl)decyl)guanidine, bromide salt (NWD-30) Guanidine (2.2 g, 0.023 mol) was dissolved in 25 mL of dichloromethane and 10 mL of 25% NaOH (aq) was added to it to obtain a clear solution. The above clear solution was cooled on an ice bath and 5 mL of 1,10 dibromodecane was added thereto, followed by 1 mg of NaI. The ice bath was warmed to rt and the reaction was continued for 48 h. After 48 h, the reaction was stopped and the phases were separated. The aqueous phase was washed with 2×25 mL of dichloromethane.

[0127] All organic phases were combined and dried over Na2SO4. The organic phase was concentrated under reduced pressure at 40°C water bath temperature to obtain a low melting product characterized as 2-(10-bromodecyl)guanidine by MS. This was then reacted with activated ethyl acetoacetate at 85°C for 24 hours to obtain ethyl 2-acetyl-12-((diaminomethylene)amino)dodecanoate. After acid workup and purification, this was reacted with an equimolar amount of phenylhydrazine by using the conditions mentioned in Example 7. After hexane precipitation and column, the desired compound was obtained as a pale yellow semi-solid (12% yield), characterized as 2-(10-(3-methyl-5-oxo-1-phenyl-4,5-dihydro-1H-pyrazol-4-yl)decyl)guanidine by LC-MS: 359.3 (M / Z). [ka]

[0128] Example 9: N,N,N-trimethyl-5-(3-methyl-5-oxo-1-phenyl-4,5-dihydro-1H-2λ 4 -pyrazol-4-yl)pentan-1-aminium bromide (NWD-31) To a stirred solution of sodium ethoxide (21% wt, 25 mmol) in ethanol was added ethyl acetoacetate (25 mmol) at room temperature. The mixture was heated at 80° C. and after 30 min, (5-bromopentyl)trimethylammonium bromide (25 mmol) in ethanol was added to give a yellow solution. [ka]

[0129] After 30 min at RT, the reaction mixture turned into a white suspension, which was then refluxed for 24 h. After 24 h, the reaction mixture was cooled at RT and EtOH was removed from the reaction mass under vacuum at 55° C. water bath temperature. 5 mL of H2O was added into the reaction mixture to obtain a yellow solution. The yellow solution was then extracted with 3×10 mL of EA (EA phase turned yellow, aqueous phase still slightly yellow remail). All organic phases were combined and dried over MgSO4. It was then evaporated to obtain a yellowish sticky mass. 1.8 mL of phenylhydrazine was added into the sticky mass and reacted at 85° C. for 5 h. After workup and column chromatography, the final compound was collected as a yellow crystalline solid (45% yield). The desired compound was characterized by LC-MS: 302.4 (M / Z).

[0130] Example 10: In vitro antioxidant capacity determination of synthesized compounds by DPPH assay NWD-19, NWD-15, and NWD-14 were evaluated for antioxidant activity using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. The DPPH assay was used to predict antioxidant activity by a mechanism whereby antioxidants act to inhibit lipid oxidation, thus scavenging DPPH radicals and thus determining free radical scavenging capacity. A DPPH antioxidant assay kit (Dokindo Molecular Technologies, Inc.) was used and plates were read at 517 nm on a BioTek Synergy HT plate reader (Gen5 software). All compounds were tested in duplicate at five concentrations to determine the antioxidant activity of each compound. Dose response curves were generated for all tested compounds (as shown in Figure 1) and IC50 was determined to be 2.1 μM, 2.3 μM, and 2.8 μM for NWD-19, NWD-15, and NWD-14, respectively.

[0131] Example 11: In vitro anticancer activity evaluation of NWD-19 in CT-26 cancer cell line CT-26 cell lines were seeded in 96-well plates at the appropriate pre-determined seeding density and allowed to attach for approximately 16 hours. After ensuring adequate confluence (approximately 70-80%), the medium was replaced with fresh medium and test compounds were added to the plates in triplicate at six concentrations. Appropriate vehicle and positive controls were also added and the plates were incubated overnight in a CO2 (95%) / O2 (5%) incubator at a temperature of 37°C. After incubation, the plates were assayed for cytotoxicity using the cell titer glo2.0 assay (Promega) on a BioTek Synergy HT plate reader (Gen5 software). Viability was calculated at each NWD-19 treatment concentration and inhibition curves were obtained as shown in Figure 2. The calculated EC50 was 9.6 μM for NWD-19.

[0132] Example 12: Protection of RPE cell lines from AAPH-induced oxidative stress by NWD-19 Primary retinal pigmented endothelial cell lines (RPE) were seeded in appropriate growth medium in 96-well plates overnight (16-24 hours). Upon reaching 80-90% confluence, cells were then treated with NWD-19 at concentrations ranging from 0.039uM to 1.25uM. Appropriate blanks were also added and plates were incubated overnight in a CO2 (95%) / O2 (5%) incubator at a temperature of 37°C. Following incubation, all wells were treated with 10mM AAPH to induce oxidative stress and incubated overnight. After incubation, plates were assayed for viability using the cell titer glo2.0 assay (Promega). As shown in Figure 3, AAPH caused oxidative stress-induced cell death in nearly all RPE cells overnight when compared to untreated cells. However, treatment with NWD-19 helped reduce oxidative cell damage and increased survival in a dose-dependent manner, with a dose of 1.25 μM showing nearly a 50% chance of cell survival.

[0133] Example 13: In vitro TBARS assay in RPE primary cells to evaluate the antioxidant potential of NWD-19 Primary retinal pigmented endothelial cell lines (RPE) were seeded in appropriate growth medium in 96-well plates overnight (16-24 hours). Cells were then treated with NWD-19 at seven concentrations and incubated overnight. Following incubation, all wells were treated with 10 mM AAPH to induce oxidative stress and incubated overnight. TBARS assay was then utilized to evaluate the antioxidant potential of compounds in RPE cells at various concentrations. As shown in Figure 4, NWD-19 showed significant antioxidant activity in the TBARS assay against AAPH-induced oxidative stress in retinal epithelial cell lines at concentrations below 0.078 μM.

[0134] Example 14: In vivo efficacy evaluation of benzalkonium chloride-induced dry eye in C57BL / 6 mice Benzalkonium chloride (BAC) is a preservative commonly used in eye drops. However, it has been reported to increase the risk of dry eye disease due to its toxicity to the ocular surface, conjunctiva, cornea, and deeper ocular structures. 5uL of 0.2% BAC in mouse eyes has been reported to cause dry eye symptoms without affecting tear production, inflammation, or corneal smoothness (Richard Zhang,et al.Dose-dependent benzalkonium chloride toxicity imparts ocular surface epithelial changes with features of dry eye disease,The Ocular Surface,Volume 18,Issue 1,2020,Pages 158-169).

[0135] Symptoms of dry eye disease were induced in 24 male and female C57BL / 6 mice for 21 days using daily eye drops of 0.2% BAC. Daily topical application of NWD-19 at a concentration of 10 μM began 8 days after BAC treatment and continued until day 21, while control mouse eyes were treated with PBS during this period. The results of disease progression in mouse eyes treated with 0.2% BAC are shown in FIG. 5: A) mouse eyes before treatment, B) day 7 after BAC treatment, C) day 14 after BAC treatment (7 days after NWD-19 treatment), D) day 21 after BAC treatment (14 days after NWD-19 treatment). The eyes of the mice were examined by a veterinary ophthalmologist for corneal lesions before dosing and then 7, 14 and 21 days after dosing, and dry eye was also assessed using a drop of 0.1% fluorescein solution in each treated eye and observation and scoring using a cobalt blue filter. BAC (0.2%) caused severe dryness and corneal lesions in the eyes of the mice by the 7th day of treatment. Treatment with NWD-19 drops provided a masked improvement in eye condition compared to vehicle control eyes, as can be seen in Figure 5.

[0136] While specific embodiments of the subject invention have been discussed, the above specification is illustrative and not limiting. Many variations of the embodiments will become apparent to those of skill in the art upon review of this specification and the claims that follow. The full scope of the invention should be determined by reference to the claims, along with their full scope of equivalents, and the specification, along with such variations.

Claims

1. Formula (I) 【Chemistry 1】 (In the formula: 【Chemistry 2】 represents a single or double bond, - R １ and R ２ is -C, optionally mono-, di- or trisubstituted with -H, halogen, substituted and unsubstituted aryl (monocyclic and bicyclic), substituted and unsubstituted heteroaryl (monocyclic and bicyclic), １～２０ Alkyl, C ２～２０ Alkenyl, and C ２～２０ alkynyl, - R ３ is -H, -C １～２０ Alkyl, C ２～２０ Alkenyl, and C ２～２０ alkynyl, where R ３ is optionally substituted by one or more substituents selected from A, aryl and heteroaryl; - R ４ is -H, -C １～２０ Alkyl, C ２～２０ Alkenyl, and C ２～２０ alkynyl, where R ４ is optionally substituted with one or more substituents selected from A, aryl and heteroaryl; - R ５ is (a) aryl, (b) heteroaryl, (c) -C １～２ Alkyl-aryl, and (d) -C １～２ alkyl-heteroaryl; where A is aryl or heteroaryl; Said aryl in options (a) and (c), and said heteroaryl in options (b) and (d) are optionally selected from halogen, —C １～６ Alkyl, -CF ３ , -OCF ３ , -C １～６ Alkoxy, -C ３～６ Cycloalkoxy, halo-C １～６ Alkyl, aryl, heteroaryl, heterocyclo, -C ３～６ Cycloalkyl, and -C ３～６ Cycloalkenyl is mono-, di- or trisubstituted with a substituent selected from the group consisting of: However, the above R １ , R ２ , R ３ , R ４ and R ５ At least one of 【Chemistry 3】 wherein X, Y, and Z are each independently selected from the group consisting of -H, methyl, ethyl, propyl, butyl, substituted or unsubstituted aryl and heteroaryl; n is an integer selected from 0 to 18, and E is a phosphorus or nitrogen atom. or a pharma- ceutically acceptable salt, hydrate, solvate, optical isomer, or combination thereof.

2. R １ , R ２ , R ３ , and R ４ are each independently -H, -C １～２０ Alkyl, C ２～２０ Alkenyl, and C ２～２０ The compound of claim 1 , wherein the aryl group is selected from alkynyl.

3. R ５ is aryl, heteroaryl, -C １～２ Alkyl-aryl, and -C １～２ The compound of claim 1, which is selected from the group consisting of alkyl-heteroaryl.

4. The R １ , R ２ , R ３ , R ４ and R ５ At least one of the formula 【Chemistry 4】 2. The compound of claim 1, wherein X, Y, Z are selected from the group consisting of -H, methyl, ethyl, propyl, butyl, substituted or unsubstituted aryl and heteroaryl; n is an integer selected from 0 to 18, and E is a phosphorus or nitrogen atom.

5. - R １ , R ２ and R4 is H; - R ３ But -CH ３ and - R ５ but 【Chemistry 5】 and However, the above R ２ , R ３ and R ４ At least one of 【Chemistry 6】 wherein n is an integer selected from 0 to 18. Selected from the group consisting of: The compound of claim 1.

6. 2. The compound of claim 1, wherein the compound of formula (I) comprises cis, trans, syn, anti, entgegen (E), zusammen (Z) isomers or mixtures thereof.

7. The compound of claim 1, wherein the compound of formula (I) contains one or more asymmetric centers and therefore exists as racemates and racemic mixtures, enantiomerically enriched mixtures, single enantiomers, individual diastereomers and diastereomeric mixtures.

8. 13. A pharmaceutical composition comprising a therapeutically effective amount of a compound of claim 1 and a pharma- ceutically acceptable carrier or diluent.

9. 9. The pharmaceutical composition of claim 8, which is in a dosage form selected from the group consisting of tablets, pills, powders, solutions, suspensions, emulsions, granules, capsules, suppositories and suspensions.

10. Use of a compound described in claim 1 in the manufacture of a medicine for treating or preventing oxidative stress-related diseases, including but not limited to neurodegenerative diseases, cardiovascular diseases, radiation-induced damage, metabolic disorders, and eye diseases.

11. Use of a compound according to claim 1 in the manufacture of a medicament for restoring and / or improving mitochondrial function.

12. 2. Use of a compound according to claim 1 in the manufacture of a medicament for the treatment or prevention of acute radiation syndrome, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS), cancer, cardiovascular disease associated with reperfusion, neurodegenerative diseases, Huntington's disease, diabetes and its complications, Friedreich's ataxia, and eye diseases such as senile cataract, age-related macular degeneration, uveitis, dry eye disease, retinopathy of prematurity, diabetic retinopathy, keratitis, and ocular inflammation.

13. A process for preparing a compound of formula (I), or a pharma- ceutically acceptable salt thereof, comprising the steps of: 【Chemistry 7】 (a) reacting a carboxylic acid of formula (1) with a malonic acid monoester of formula (2) in the presence of an activating agent selected from carbonyldiimidazole (CDI), a Lewis acid selected from magnesium chloride (MgCl 2 ), and a base selected from N-methylmorpholine (NMM), pyridine, N-ethylmorpholine (NEM), triethylamine (TEA) and / or the like; (b) reacting ethyl acetoacetate or its derivative of formula (4) with a halogenated alkene or alkane having a lipophilic cation of formula (3) in the presence of a base selected from sodium ethoxide (NaOEt) in an alcoholic solvent selected from the group of protic polar solvents, followed by reaction in a suitable solvent such as tetrahydrofuran (THF) at a temperature of about 80° C. to obtain an intermediate compound of formula (5); 【Chemistry 8】 (c) reacting the intermediate of formula (5) with a hydrazine derivative of formula (6) or (6') in an alcoholic solvent such as ethanol or methanol or under solvent-free conditions at a temperature between 80°C and 140°C in a sealed reaction vessel to obtain an intermediate compound of formula (7) or (7'); (d) reacting an intermediate of formula (7) or (7') with (i) when R 4 is an alkyl or heteroalkyl group, carrying out a nucleophilic substitution reaction with R 4 -X in the presence of a base selected from sodium hydride (NaH), cesium carbonate (Cs 2 CO 3 ), or potassium carbonate (K 2 CO 3 ) in a polar aprotic solvent; or (ii) when R 4 is an aryl or heteroaryl group, carrying out a copper catalyzed N-arylation reaction with R 4 -X in the presence of a copper(I) source selected from copper iodide (CuI), a ligand such as proline, and a base selected from Cs 2 CO 3 in a polar aprotic solvent; to obtain the desired pyrazolone core compound of formula (8) or compound of formula (I); 【Chemistry 9】 (e) optionally purifying the product obtained in step (d) using a purification method selected from chromatography or crystallization; (f) optionally converting the resulting compound into a pharma- ceutically acceptable salt, hydrate, or solvate by treatment with an acid or base selected from hydrochloric acid (HCl), sulfuric acid (H2SO4), sodium hydroxide (NaOH), or a suitable organic acid or base in a suitable solvent; Including, A process wherein the compound is selected as defined in claim 1.

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