Haloacetyldrazide, a useful AEP inhibitor.

Abstract

The present invention relates to a compound represented by the general formula (I) 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , W, m and n are as described herein, compositions comprising said compounds, and methods of using said compounds.

Description

Technical Field

[0001] The present invention relates to organic compounds useful for treatment and / or prevention in patients, particularly compounds that inhibit AEP activity.

[0002] The present invention relates to a compound of formula I

Chemical Formula

[0003] Furthermore, the present invention includes all racemic mixtures, all their corresponding enantiomers and / or optical isomers.

Background Art

[0004] Background of the Invention Alzheimer's disease (AD) Alzheimer's disease is the most common form of dementia, primarily affecting older adults and influencing over 5 million people in the United States alone. This number is projected to triple by 2050. The prevalence of AD is age-related, and patients often require institutionalization during the later stages of the disease. Due to its severity, increasing prevalence, long duration, and high treatment costs, AD is likely to continue to be a major public health issue in the coming years.

[0005] Currently approved treatments for Alzheimer's disease (AD) modulate neurotransmission (e.g., Aricept, acetylcholinesterase inhibitors). While these drugs provide short-term relief from some symptoms, they do not alter the underlying pathology or the course of the disease. There is a great unmet medical need for more effective treatments for AD at all stages, particularly novel treatments that slow or delay disease progression.

[0006] The brain of AD patients has two distinct major histopathological lesions: amyloid plaques containing aggregated Aβ peptides and neurofibrillary tangles containing aggregated tau proteins. Extracellular Aβ peptide accumulation is thought to be an early event in a cascade of pathological changes leading to dementia, which includes tau aggregation and neuronal loss (amyloid cascade hypothesis, Selkoe and Hardy, EMBO Mol Med 2016).

[0007] The most advanced drugs currently under development for AD aim to modify disease progression by reducing Aβ. These drugs include Aβ-specific antibodies that promote Aβ clearance, as well as small molecule inhibitors and modulators of the proteases responsible for Aβ production, namely β- and γ-secretases. Strategies to mitigate tau aggregation are needed to complement the Aβ reduction approach. Several companies are pursuing tau-targeted monoclonal antibodies for AD. However, tau antibodies are still in the early stages of clinical development and are unlikely to emerge as standard treatment for AD in the coming years.

[0008] Alzheimer's disease (AD) belongs to a larger group of neurodegenerative dementias known as tauopathies. This group includes frontotemporal dementia (FTD-MAPT), progressive supranuclear palsy (PSP), and corticobasal degeneration (CBD). A common feature of tauopathies is the presence of fibrillary tau aggregates within cells. Tau-targeting therapeutic approaches in AD may be applicable to primary tauopathies. There are no approved drugs for the specific treatment of non-AD tauopathies.

[0009] Tau aggregation and Alzheimer's disease The presence of neurofibrillary tangles within neurons is one of the defining pathological features of Alzheimer's disease (AD). Tau protein, the main component of these tangles, is normally a highly soluble protein that associates with and stabilizes microtubules. In AD and other tauopathies, tau undergoes structural changes that lead to aggregation (Vaquer-Alicea et al., Acta Neuropath 2021). Tau aggregation is central to disease development in tauopathies, as evidenced by multiple studies: (1) Many MAPT mutations associated with frontotemporal dementia promote tau fiber aggregation; (2) Cognitive ability in AD patients is inversely correlated with tau aggregate load; (3) Multimodal imaging in AD patients shows that tau aggregate deposition correlates with focal brain atrophy and functional impairment; (4) Injecting fibrillary tau instead of monomers causes tangling in the mouse brain; (5) Tau added exogenously to cells must aggregate in order to "seed" intracellular tau pathology.

[0010] Therefore, tau aggregation is a toxic function acquisition in disease. Consequently, prevention of tau aggregation is expected to protect against neurodegeneration in Alzheimer's disease (AD) and primary tauopathy.

[0011] Tau cutting by AEP The microtubule-binding region (MBTR) of tau is a crucial part of the molecule nucleating fibril aggregates. Structural studies have shown that the centrally located MBTR is typically covered by the molecule's N-terminal and C-terminal regions, thereby hindering tau-tau interactions in solution. Post-translational modifications, such as phosphorylation and proteolytic cleavage, open up tau molecules, exposing the MBTR and promoting aggregation. PHF tau isolated from AD brains contains the majority of cleaved tau species, suggesting that tau cleavage is essential for the tau aggregation process.

[0012] Lysosomal cysteine ​​proteinase AEP / regmaine has been shown to cleave tau on both sides of the MBTR, thereby exposing the MBTR and promoting aggregation. Stress-induced upregulation of AEP activity in primary mouse neurons promotes tau cleavage. Overexpression of AEP-derived tau fragments 1-368 in neurons is strongly neurotoxic, while overexpression of full-length tau is not. In addition to cleaving tau, AEP may indirectly influence tau aggregation by promoting tau phosphorylation: AEP is known to (indirectly) inhibit protein phosphatase 2A, a key enzyme that modulates tau dephosphorylation.

[0013] AEP in AD Published data show that AEP is upregulated in AD (Zhang et al., Nat Med 2014). Other authors have shown that AEP is hyperactivated in AD (Wang et al., Mol Cell 2017; Basurto-Islas et al., J Biol Chem 2013). Whether due to upregulation or hyperactivation of AEP, AEP-cleaved tau N368 fragments are enriched in AD brain tissue (Zhang et al., Nat Med 2014). The tau N368 / total tau ratio was significantly reduced in CSF from AD patients and was strongly negatively correlated with 18F-GTP1 tau PET signaling (Blennow et al., Brain 2020).

[0014] AEP may contribute to the development of Alzheimer's disease beyond promoting tau aggregation. The AEP cleavage fragment tau N368 has recently been shown to enhance BACE1 expression and Aβ production through binding to the BACE1 transcription factor STAT1 (Zhang et al., Mol Psych 2018). [Overview of the project] [Problems that the invention aims to solve]

[0015] This invention relates to formula I: [ka] (In the formula, R 1 is either halo or alkyl; R 2 is H or halo; R 3 is H or halo; R 4 Is it H or a halo? Alternatively, R 3 and R 4 It forms a 3-membered or 4-membered cycloalkyl ring; R 5 is H, halo, haloalkoxy, alkynyl, alkynylalkoxy, or alkoxyphenyl; R 6 is H; R 7 is H; W is either -C(O)- or -S(O)2-; m is either 0 or 1; n is either 0 or 1. This invention provides novel compounds and pharmaceutically acceptable salts thereof. [Modes for carrying out the invention]

[0016] The term "alkyl" refers to a monovalent linear or branched saturated hydrocarbon group consisting of 1 to 6 carbon atoms. In some embodiments, unless otherwise stated, alkyl refers to a monovalent linear or branched saturated hydrocarbon group consisting of 1 to 6 carbon atoms (C 1~6-alkyl) or 1 to 4 carbon atoms (C 1~4 -Contains alkyl) C 1~6 -Examples of alkyl groups include methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, and pentyl. Specific alkyl groups include methyl and ethyl. The preferred alkyl group is methyl. When an alkyl residue having a specific number of carbon atoms is named, all geometric isomers having that number of carbon atoms may be included. For example, "butyl" may include n-butyl, sec-butyl, isobutyl, and t-butyl, and "propyl" may include n-propyl and isopropyl.

[0017] The term "alkoxy" is derived from the fact that R' is C 1~6 - Represents an alkyl group, the group of formula -O-R'. C 1~6 -Examples of alkoxy groups include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, and tert-butoxy. Specific examples are methoxy and ethoxy. A preferred example is methoxy.

[0018] The term "alkynyl" refers to an unsaturated, unbranched or branched monovalent hydrocarbon chain having at least one acetylene unsaturated portion (i.e., having at least one C≡C part). In some embodiments, unless otherwise specified, an alkynyl contains 2 to 6 carbon atoms or 2 to 4 carbon atoms. Examples of alkynyl groups, but not limited to, include ethynyl (or acetylenyl), propa-1-inyl, propa-2-inyl (or propargyl), buta-1-inyl, buta-2-inyl, and buta-3-inyl. A specific example is ethynyl.

[0019] The term "alkynylalkoxy" is C 1~6 At least one hydrogen atom of the alkoxy group is C 2~6 C replaced by an alkynyl group 1~6This represents an alkoxy group. Specific examples include ethynylmethoxy, ethynylethoxy, propa-2-inoxy, propynylmethoxy, and propynylethoxy. A specific example is propa-2-inoxy.

[0020] The term "alkoxyphenyl" refers to a phenyl compound substituted with an alkoxy group defined above at the ortho, meta, or para position. A specific example is 4-methoxyphenyl.

[0021] The term "cycloalkyl" refers to monocyclic or polycyclic saturated or partially unsaturated non-aromatic hydrocarbons. In some embodiments, unless otherwise stated, cycloalkyls contain 3 to 8 carbon atoms, 3 to 6 carbon atoms, or 3 to 5 carbon atoms. In some embodiments, cycloalkyls are saturated monocyclic or polycyclic hydrocarbons. In other embodiments, cycloalkyls contain one or more double bonds (e.g., cycloalkyls fused to an aryl or heteroaryl ring, or non-aromatic monocyclic hydrocarbons containing one or two double bonds). Polycyclic cycloalkyls may contain spiro, condensed, or bridging polycyclic moieties, each ring being a saturated or partially unsaturated non-aromatic hydrocarbon. Examples of cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, octahydropentalenyl, and spiro[3.3]heptanyl. A bicyclic structure refers to a ring system consisting of two saturated carbon rings sharing two carbon atoms. Examples of monocyclic cycloalkyl compounds include cyclopropyl, cyclobutanyl, cyclopentyl, cyclohexyl, or cycloheptyl. Specific examples are cyclopropyl and cyclobutanyl. A preferred example is cyclopropyl.

[0022] The terms “halogen,” “halide,” and “halo” are used interchangeably herein and refer to fluoro, chloro, bromo, or iodine. Specific halogens are fluoro and chloro.

[0023] The term "haloalkoxy" is C 1~6- At least one hydrogen atom of the alkoxy group is replaced by the same or a different halogen atom C 1~6 - Represents an alkoxy group. Examples of haloalkoxys are difluoromethoxy, trifluoromethoxy, difluoroethoxy, and trifluoroethoxy. A specific example is trifluoromethoxy.

[0024] The term "pharmaceutically acceptable salt" refers to a salt that retains the biological efficacy and properties of a free base or free acid, and is not biologically or otherwise undesirable. Salts are formed using inorganic acids, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, especially hydrochloric acid, and organic acids, such as formic acid, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and N-acetylcysteine. In addition, these salts can be prepared by adding inorganic or organic bases to free acids. Salts derived from inorganic bases include, but are not limited to, sodium salts, potassium salts, lithium salts, ammonium salts, calcium salts, and magnesium salts. Salts derived from organic bases include, but are not limited to, primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, lysine, arginine, N-ethylpiperidine, piperidine, and polyamine resins. Compounds of formula I may also exist in zwitterionic form. Particularly preferred pharmaceutically acceptable salts of compounds of formula I are salts formed with formic acid, and salts formed with hydrochloric acid that yield hydrochloride, dihydrochloride, or trihydrochloride.

[0025] The abbreviation uM stands for micromoles and is equivalent to the symbol μM.

[0026] The abbreviation uL stands for microliter and is equivalent to the symbol μL.

[0027] The abbreviation ug stands for microgram and is equivalent to the symbol μg.

[0028] Compounds of formula I may contain several chiral centers and may exist in the form of optically pure enantiomers, mixtures of enantiomers, such as racemates, optically pure diastereoisomers, mixtures of diastereoisomers, diastereoisomer racemates, or mixtures of diastereoisomer racemates.

[0029] According to the Kahn-Ingold-Prelogue priority rule, an asymmetric carbon atom can have either an "R" or "S" stereoconfiguration.

[0030] Furthermore, certain embodiments of the present invention provide compounds of formula I described herein and pharmaceutically acceptable salts or esters thereof, in particular compounds of formula I described herein and pharmaceutically acceptable salts thereof, more specifically, compounds of formula I described herein.

[0031] One embodiment of the present invention is R 1 However, it is halo or alkyl; R 2 However, it is H or halo; R 3 However, it is H or halo; R 4 Is it H or a halo? Alternatively, R 3 and R 4 However, it forms a 3-membered or 4-membered cycloalkyl ring; R 5 However, it is H, halo, haloalkoxy, alkynyl, alkynylalkoxy, or alkoxyphenyl; R 6 H is; R 7 H is; W is -C(O)- or -S(O)2-; m is 0 or 1; This specification provides compounds of formula I described herein, and pharmaceutically acceptable salts, wherein n is 0 or 1.

[0032] One embodiment of the present invention is R 1 This specification provides compounds of formula I, wherein the compound is a halo.

[0033] One embodiment of the present invention is R 5 The present specification provides compounds of formula I, wherein the compound is H, halo, or haloalkoxy.

[0034] One embodiment of the present invention provides a compound of formula I described herein, wherein W is -C(O)-.

[0035] One embodiment of the present invention provides a compound of formula I described herein, wherein m is 1 and n is 0.

[0036] One embodiment of the present invention is R 3 and R 4 The present invention provides compounds of formula I described herein that form a three-membered or four-membered cycloalkyl ring.

[0037] One embodiment of the present invention is R 1 That is a halo; R 2 However, it is H or halo; R 3 That is a halo; R 4 Is it a halo? Alternatively, R 3 and R 4 However, it forms a 3-membered or 4-membered cycloalkyl ring; R 5 However, it is H, halo, or haloalkoxyl; W is -C(O)-; m is 1; This specification provides compounds of formula I described herein, and pharmaceutically acceptable salts, where n is 0.

[0038] One embodiment of the present invention is R 1 That is a halo; R 2 However, it is H or halo; R 3 and R 4 However, it forms a 3-membered or 4-membered cycloalkyl ring; R 5 However, it is a halo or haloalkoxyl; W is -C(O)-; m is 1; This specification provides compounds of formula I described herein, and pharmaceutically acceptable salts, where n is 0.

[0039] One embodiment of the present invention is R 1 However, it is either F or Cl; R 2 However, it is either H or F; R 3 and R 4 This forms a three-membered cycloalkyl ring; R 5 However, it is chloro or trifluoromethoxy; W is -C(O)-; m is 1; This specification provides compounds of formula I described herein, and pharmaceutically acceptable salts, where n is 0.

[0040] Specific examples of compounds of formula I described herein are: 2-[(2-fluoroacetyl)-[[(2S)-1-(2,2-difluoro-2-phenyl-acetyl)pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-fluoroacetyl)-[[(2S)-1-(3-phenylpropanoyl)pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-fluoroacetyl)-[[(2S)-1-(1-phenylcyclobutanecarbonyl)pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-fluoroacetyl)-[[(2S)-1-[1-(4-chlorophenyl)cyclobutanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-fluoroacetyl)-[[(2S)-1-(2-phenylacetyl)pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-fluoroacetyl)-[[(2S)-1-[1-(4-chlorophenyl)cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-fluoroacetyl)-[[(2S)-1-(benzenesulfonyl)pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-fluoroacetyl)-[[(2S)-1-(1-phenylcyclopropanecarbonyl)pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-fluoroacetyl)-[[(2S)-1-benzylsulfonylpyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-fluoroacetyl)-[[(2S)-1-(2-phenylethylsulfonyl)pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-fluoroacetyl)-[[(2S)-1-[1-(4-ethynylphenyl)cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-fluoroacetyl)-[[(2S)-1-[1-[4-(trifluoromethoxy)phenyl]cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-fluoroacetyl)-[[(2S)-1-[4-(4-methoxyphenyl)phenyl]sulfonylpyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-fluoroacetyl)-[[(2S)-1-[1-(4-prop-2-inoxyphenyl)cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[((2R)-2-chloro-2-fluoro-acetyl)-[[(2S)-1-[4-(4-methoxyphenyl)phenyl]sulfonylpyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[((2S)-2-chloro-2-fluoro-acetyl)-[[(2S)-1-[4-(4-methoxyphenyl)phenyl]sulfonylpyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[((2R)-2-chloro-2-fluoroacetyl)-[[(2S)-1-[1-[4-(trifluoromethoxy)phenyl]cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[((2S)-2-chloro-2-fluoro-acetyl)-[[(2S)-1-[1-[4-(trifluoromethoxy)phenyl]cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-chloroacetyl)-[[(2S)-1-[4-(4-methoxyphenyl)phenyl]sulfonylpyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-chloroacetyl)-[[(2S)-1-[1-[4-(trifluoromethoxy)phenyl]cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2,2-dichloroacetyl)-[[(2S)-1-[1-[4-(trifluoromethoxy)phenyl]cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2R)-2-chloropropanoyl-[[(2S)-1-[1-[4-(trifluoromethoxy)phenyl]cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2S)-2-chloropropanoyl-[[(2S)-1-[1-[4-(trifluoromethoxy)phenyl]cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; Selected from the and pharmaceutically acceptable salts thereof.

[0041] Further specific examples of Formula I described herein are: 2-[(2-fluoroacetyl)-[[(2S)-1-(2,2-difluoro-2-phenyl-acetyl)pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-fluoroacetyl)-[[(2S)-1-[1-(4-chlorophenyl)cyclobutanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-fluoroacetyl)-[[(2S)-1-[1-(4-chlorophenyl)cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-fluoroacetyl)-[[(2S)-1-[1-[4-(trifluoromethoxy)phenyl]cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[((2R)-2-chloro-2-fluoroacetyl)-[[(2S)-1-[1-[4-(trifluoromethoxy)phenyl]cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[((2S)-2-chloro-2-fluoro-acetyl)-[[(2S)-1-[1-[4-(trifluoromethoxy)phenyl]cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-chloroacetyl)-[[(2S)-1-[1-[4-(trifluoromethoxy)phenyl]cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; Selected from the and pharmaceutically acceptable salts thereof.

[0042] The most specific example of Formula I described herein is: 2-[(2-fluoroacetyl)-[[(2S)-1-[1-(4-chlorophenyl)cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-fluoroacetyl)-[[(2S)-1-[1-[4-(trifluoromethoxy)phenyl]cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[((2R)-2-chloro-2-fluoroacetyl)-[[(2S)-1-[1-[4-(trifluoromethoxy)phenyl]cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[((2S)-2-chloro-2-fluoro-acetyl)-[[(2S)-1-[1-[4-(trifluoromethoxy)phenyl]cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; Selected from the and pharmaceutically acceptable salts thereof.

[0043] The method for producing the compound of formula I described herein is the objective of the present invention.

[0044] General synthesis scheme R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 Compounds of formula I, where W, n, and m are as described above, can be obtained as shown in scheme 1. 1 and R 2 The compound of formula II as described above is R 3 , R 4 , R 5 , R 6 , R 7, where W, n, and m are as described above, the compound of formula III can be reacted with a suitable functional group selected from F, Cl, Br, OH, ON-succinimidyl (OSu) depending on the properties of W. Specifically, when W=-C(O)-, X can be selected from F, Cl, Br, OH, ON-succinimidyl, preferably Cl and OH. When X=OH, the reaction is carried out under standard amide coupling conditions known in the art, for example. When X=F, Cl, Br, or OSu, the reaction can be carried out in a suitable polar aprotic or nonpolar solvent, such as dimethylformamide or dichloromethane, in the presence of a stoichiometric amount of a suitable chemically inert base, such as a tertiary amine such as triethylamine or diisopropylethylamine. When W=-S(O)2-, X can be selected from F, Cl, preferably Cl. The reaction is carried out in a suitable polar aprotic or nonpolar solvent, such as dimethylformamide or dichloromethane, in the presence of a stoichiometric amount of a suitable chemically inert base, such as a tertiary amine such as triethylamine or diisopropylethylamine. [ka] Scheme 1

[0045] Alternatively, R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 Compounds of formula I, where W, n, and m are as described above, can be obtained as shown in scheme 2. 3 , R 4 , R 5 , R 6 , R 7 A compound of formula IV in which W, n, and m are as described above, R 1 and R 2is as described above, and can be reacted with a compound of formula V in which Y is a suitable functional group selected from F, Cl, Br, OH, O-N-succinimidyl (OSu), preferably Cl and OH. When Y = OH, the reaction is carried out, for example, under standard amide coupling conditions known in the art. When Y = F, Cl, Br or OSu, the reaction can be carried out in a suitable polar aprotic or nonpolar solvent, such as dichloromethane, in the presence of a stoichiometric amount of a suitable base, such as pyridine.

Chemical Structure

[0046] R 3 、R 4 、R 5 、R 6 、R 7 The intermediate compounds of formulas III and IV in which W, n, and m are each as described above can be obtained by methods known in the art.

[0047] One embodiment of the present invention is a method for preparing a compound of formula I as defined above, which comprises reacting a compound of formula II with a compound of formula III.

[0048] One embodiment of the present invention is a method for preparing a compound of formula I as defined above, which comprises reacting a compound of formula IV with a compound of formula V.

[0049] Another embodiment of the present invention provides a pharmaceutical composition or pharmaceutically containing the compound of the present invention and a therapeutically inert carrier, diluent, or excipient, as well as a method of using the compound of the present invention to prepare such a composition and pharmaceutically. In one example, the compound of formula I can be formulated into a galenic dosage form by mixing it with a physiologically acceptable carrier, i.e., a carrier that is nontoxic to the recipient at the dose and concentration used, at ambient temperature, at a suitable pH, and to a desired degree of purity. The pH of the formulation depends primarily on the specific application and the concentration of the compound, but is preferably somewhere in the range of about 3 to about 8. In one example, the compound of formula I is formulated in an acetate buffer at pH 5. In another embodiment, the compound of formula I is sterile. The compound can be stored, for example, as a solid or amorphous composition, as a lyophilized formulation, or as an aqueous solution.

[0050] The composition is formulated, administered, and given in a manner consistent with good medical practice. Factors to be considered in this context include the specific disorder being treated, the specific mammal being treated, the clinical symptoms of the individual patient, the cause of the disorder, the site of drug delivery, the method of administration, the administration schedule, and other factors known to medical professionals.

[0051] The compounds of the present invention may be administered by any suitable means, including orally, topically (including buccal and sublingual), rectally, vaginally, percutaneously, parenterally, subcutaneously, intraperitoneally, intrapulmonaryly, intradermally, intrathecally, and epidurally and intranasally, and, if topical treatment is desired, intrafocal administration. Parenteral administration includes intramuscular, intravenous, intra-arterial, intraperitoneal, or subcutaneous administration.

[0052] The compounds of the present invention can be administered in any convenient dosage form, such as tablets, powders, capsules, solutions, dispersions, suspensions, syrups, sprays, suppositories, gels, emulsions, patches, etc. Such compositions may contain conventional components in pharmaceutical preparations, such as diluents, carriers, pH adjusters, sweeteners, fillers, and further activators.

[0053] Typical formulations are prepared by mixing the compound of the present invention with a carrier or excipient. Suitable carriers and excipients are well known to those skilled in the art and are described in detail, for example, Ansel, Howard C. et al., Ansel's Pharmaceutical Dosage Forms and Drug Delivery Systems. Philadelphia: Lippincott, Williams & Wilkins, 2004; Gennaro, Alfonso R. et al., Remington: The Science and Practice of Pharmacy. Philadelphia: Lippincott, Williams & Wilkins, 2000; and Rowe, Raymond C. Handbook of Pharmaceutical Excipients. Chicago, Pharmaceutical Press, 2005. The formulation may also include one or more buffers, stabilizers, surfactants, wetting agents, lubricants, emulsifiers, suspending agents, preservatives, antioxidants, opacifiers, flow enhancers, processing aids, colorants, sweeteners, fragrances, flavorings, diluents, and other known additives to provide a better appearance for the drug (i.e., the compound of the present invention or its pharmaceutical composition) or to assist in the manufacture of the pharmaceutical (i.e., the pharmaceutical).

[0054] Compounds of formula I and pharmaceutically acceptable salts thereof can be processed with pharmaceutically inert inorganic or organic adjuvants for the manufacture of tablets, coated tablets, sugar-coated tablets, hard gelatin capsules, injections, or topical formulations. Lactose, corn starch or derivatives thereof, talc, stearic acid or salts thereof, etc., can be used as such adjuvants for tablets, sugar-coated tablets, and hard gelatin capsules, for example.

[0055] Suitable adjuvants for soft gelatin capsules include, for example, vegetable oils, waxes, fats, semi-solid substances, and liquid polyols.

[0056] Suitable adjuvants for the production of solutions and syrups include, for example, water, polyols, sucrose, invert sugar, and glucose.

[0057] Suitable adjuvants for injection solutions include, for example, water, alcohol, polyol, glycerol, and vegetable oil.

[0058] Suitable adjuvants for suppositories include, for example, natural or hydrogenated oils, waxes, fats, semi-solid or liquid polyols.

[0059] Suitable adjuvants for topical ophthalmic formulations include, for example, cyclodextrin, mannitol, or many other carriers and excipients known in the art.

[0060] Furthermore, pharmaceutical preparations may contain preservatives, solubilizers, viscosity-enhancing substances, stabilizers, humectants, emulsifiers, sweeteners, colorants, flavoring agents, salts for altering osmotic pressure, buffers, masking agents, or antioxidants. These may also contain other substances of therapeutic value.

[0061] Dosages can vary over a wide range and, of course, are tailored to the individual requirements of each specific case. Generally, for oral administration, a daily dose of approximately 0.1 mg to 20 mg per kg of body weight, preferably approximately 0.5 mg to 4 mg per kg of body weight (e.g., approximately 300 mg per person), is preferably divided into 1 to 3 individual doses, which, if appropriate, may consist of equal amounts, for example. For topical administration, the formulation may contain 0.001% to 15% by weight of the drug, and the required dose, which may be between 0.1 and 25 mg, can be administered by a single dose per day or per week, by multiple doses per day (2 to 4 times), or by multiple doses per week. However, where indicated, it will be clear that the upper or lower limits given herein may be exceeded.

[0062] The present invention also relates in particular to the following: Compounds of formula I for use as therapeutically active substances; A compound of formula I for use in the treatment of diseases regulated by AEP.

[0063] One embodiment of the present invention involves the use of compounds of formula I for the treatment or prevention of Alzheimer's disease, primary age-related tauopathy (PART) dementia, frontotemporal dementia (FTD-MAPT), chronic traumatic encephalopathy (CTE), progressive supranuclear palsy (PSP), corticobasal degeneration (CBD), frontotemporal dementia and parkinsonism associated with chromosome 17 (FTDP-17), Ritico-Bodig disease (Guam Parkinson's dementia complex), ganglioglioma and gangliocytoma, meningeal hemangioma, post-encephalitis parkinsonism, subacute sclerosing panencephalitis (SSPE), Pick's disease, or corticobasal degeneration.

[0064] One embodiment of the present invention is the use of a compound of formula I for the treatment or prevention of Alzheimer's disease, primary age-related tauopathy (PART) dementia, frontotemporal dementia (FTD-MAPT), chronic traumatic encephalopathy (CTE), progressive supranuclear palsy (PSP), or Pick's disease.

[0065] One embodiment of the present invention is the use of a compound of formula I for treating or preventing Alzheimer's disease.

[0066] One embodiment of the present invention involves the use of a compound of formula I for preparing a medicament for treating or preventing Alzheimer's disease, primary age-related tauopathy (PART) dementia, frontotemporal dementia (FTD-MAPT), chronic traumatic encephalopathy (CTE), progressive supranuclear palsy (PSP), corticobasal degeneration (CBD), frontotemporal dementia and parkinsonism associated with chromosome 17 (FTDP-17), Ritico-Bodig disease (Guam Parkinson's dementia complex), ganglioglioma and gangliocytoma, meningeal hemangioma, post-encephalitis parkinsonism, subacute sclerosing panencephalitis (SSPE), Pick's disease, or corticobasal degeneration.

[0067] One embodiment of the present invention is the use of a compound of formula I for preparing a medicament for treating or preventing Alzheimer's disease, primary age-related tauopathy (PART) dementia, frontotemporal dementia (FTD-MAPT), chronic traumatic encephalopathy (CTE), progressive supranuclear palsy (PSP), or Pick's disease.

[0068] One embodiment of the present invention is the use of a compound of formula I for preparing a medicament for treating or preventing Alzheimer's disease.

[0069] One embodiment of the present invention is a compound of formula I for treating or preventing Alzheimer's disease, primary age-related tauopathy (PART) dementia, frontotemporal dementia (FTD-MAPT), chronic traumatic encephalopathy (CTE), progressive supranuclear palsy (PSP), corticobasal degeneration (CBD), frontotemporal dementia and parkinsonism associated with chromosome 17 (FTDP-17), Ritico-Bodig disease (Guam Parkinson's dementia complex), ganglioglioma and gangliocytoma, meningeal hemangioma, post-encephalitis parkinsonism, subacute sclerosing panencephalitis (SSPE), Pick's disease, or corticobasal degeneration.

[0070] One embodiment of the present invention is a compound of formula I for treating or preventing Alzheimer's disease, primary age-related tauopathy (PART) dementia, frontotemporal dementia (FTD-MAPT), chronic traumatic encephalopathy (CTE), progressive supranuclear palsy (PSP), or Pick's disease.

[0071] One embodiment of the present invention is a compound of formula I for treating or preventing Alzheimer's disease.

[0072] One embodiment of the present invention is a method for treating or preventing Alzheimer's disease, primary age-related tauopathy (PART) dementia, frontotemporal dementia (FTD-MAPT), chronic traumatic encephalopathy (CTE), progressive supranuclear palsy (PSP), corticobasal degeneration (CBD), frontotemporal dementia and parkinsonism associated with chromosome 17 (FTDP-17), Ritico-Bodig disease (Guam Parkinson's dementia complex), ganglioglioma and gangliocytoma, meningeal hemangioma, post-encephalitis parkinsonism, subacute sclerosing panencephalitis (SSPE), Pick's disease, or corticobasal degeneration.

[0073] One embodiment of the present invention is a method for treating or preventing Alzheimer's disease, primary age-related tauopathy (PART) dementia, frontotemporal dementia (FTD-MAPT), chronic traumatic encephalopathy (CTE), progressive supranuclear palsy (PSP), or Pick's disease.

[0074] One embodiment of the present invention is a method for treating or preventing Alzheimer's disease.

[0075] One embodiment of the present invention is a method for treating or preventing Alzheimer's disease, primary age-related tauopathy (PART) dementia, frontotemporal dementia (FTD-MAPT), chronic traumatic encephalopathy (CTE), progressive supranuclear palsy (PSP), corticobasal degeneration (CBD), frontotemporal dementia and parkinsonism associated with chromosome 17 (FTDP-17), Ritico-Bodig disease (Guam Parkinson's dementia complex), ganglioglioma and gangliocytoma, meningeal hemangioma, post-encephalitis parkinsonism, subacute sclerosing panencephalitis (SSPE), Pick's disease, or corticobasal degeneration, comprising administering an effective amount of a compound of formula I to a patient in need thereof.

[0076] One embodiment of the present invention is a method for treating or preventing Alzheimer's disease, primary age-related tauopathy (PART) dementia, frontotemporal dementia (FTD-MAPT), chronic traumatic encephalopathy (CTE), progressive supranuclear palsy (PSP), or Pick's disease, comprising administering an effective amount of a compound of formula I to a patient in need thereof.

[0077] One embodiment of the present invention is a method for treating or preventing Alzheimer's disease, comprising administering an effective amount of a compound of formula I to a patient in need thereof.

[0078] Furthermore, one embodiment of the present invention provides a compound of formula I described herein when prepared according to any one of the methods described herein.

[0079] Similarly, an object of the present invention is a pharmaceutical composition comprising a compound of formula I described herein and a therapeutically inactive carrier.

[0080] Herein, the present invention will be explained by the following embodiments, which do not have limiting features.

[0081] If the prepared product is obtained as a mixture of enantiomers, epimers, and / or diastereoisomers, the pure enantiomer can be obtained by methods known to those skilled in the art, such as chiral chromatography or crystallization.

[0082] Abbreviation The following abbreviations were used in the laboratory: THF = tetrahydrofuran; MTBE = methyl tert-butyl ether; DMF = Dimethylformamide; TLC = Thin-layer chromatography; rt=room temperature, 20~25℃; Cbz = benzyloxycarbonyl; BOC = t-butyloxycarbonyl; HPLC = High-Performance Liquid Chromatography; HBTU = Hexafluorophosphate benzotriazole tetramethyluronium; HATU = Hexafluorophosphate azabenzotriazole tetramethyluronium.

[0083] Starting material The basic chemicals and solvents were purchased and used as is without further purification. Intermediates Int-1, Int-13, and Int-14 can be commercially available or synthesized using methods known in the art.

[0084] intermediate Intermediate 7: 2-[(2-fluoroacetyl)-[[(2S)-pyrrolidine-2-carbonyl]amino]amino]acetamide [ka]

[0085] Step 1: Benzyl 2-(2-tert-butoxycarbonylhydrazino)acetate (Int-2)

[0086] 49761 mg, 376 mmol of tert-butylcarbazate and 65 mL, 376 mmol of N,N-diisopropylethylamine were dissolved in toluene (500 mL), and benzyl 2-bromoacetate (Int-1, 75.00 g, 327 mmol) was added. The mixture was stirred at 75°C for 16 hours. After cooling, the mixture was filtered, and the filtrate was concentrated to obtain a yellow oil. The crude product was purified by column chromatography (silica gel, 220 g, petroleum ether / ethyl acetate, gradient 100:1 to 40:1 (v / v)), and the product-containing fractions were combined and concentrated under vacuum to obtain the marked compound as a colorless oil (60.00 g, 214 mmol, yield 65%). MS m / z (ESI): 225.2 [M-CH2=C(CH3)2] + .

[0087] Step 2: Benzyl 2-hydrazinoacetate hydrochloride (Int-3)

[0088] Benzyl 2-(2-tert-butoxycarbonylhydrazino)acetate (Int-2, 58.0 g, 207 mmol) was dissolved in ethyl acetate (200 mL), and a solution of hydrogen chloride in ethyl acetate (4 M, 300 mL, 1200 mmol) was added. A large amount of white precipitate was formed. The mixture was stirred at 25°C for 5 hours. The mixture was then filtered, the solid was washed with ethyl acetate (200 mL), and dried under vacuum to obtain the marked compound as a white solid (42.0 g, 194 mmol, yield 94%). MS m / z(ESI): 181.2 [M+H] + .

[0089] Step 3: Benzyl(2S)-2-[[(2-benzyloxy-2-oxo-ethyl)amino]carbamoyl]pyrrolidine-1-carboxylate(Int-4)

[0090] Benzyl 2-hydrazinoacetate hydrochloride (Int-3, 1.10 g, 5.08 mmol) was dissolved in dichloromethane (15 mL), and Z-PRO-OH (1.27 g, 5.08 mmol), N,N-diisopropylethylamine (2.65 mL, 15.2 mmol), and N-[3-(dimethylamino)propa-1-yl]-N-ethylcarbodiimide hydrochloride (1.261 g, 6.60 mmol) were added at 0°C. The mixture was stirred at 25°C for 12 hours. The reaction product was then diluted with cold water (30 mL) and extracted with dichloromethane (2 × 15 mL). The combined organic layer was washed with brine (20 mL), dried on sodium sulfate, and concentrated under vacuum to obtain a yellow oil. The crude product was purified by column chromatography (silica gel, 25 g, petroleum ether / ethyl acetate, gradient 10:1~1:2 (v / v)) to obtain the marked compound as yellow oil (1800 mg, 4.37 mmol, yield 86%). MS m / z(ESI): 412.3 [M+H] + .

[0091] Step 4: Benzyl(2S)-2-[[(2-amino-2-oxo-ethyl)amino]carbamoyl]pyrrolidine-1-carboxylate(Int-5)

[0092] A solution of benzyl(2S)-2-[[(2-benzyloxy-2-oxoethyl)amino]carbamoyl]pyrrolidine-1-carboxylate (Int-4, 10.0 g, 24.3 mmol) in methanol (7N, 10 mL, 700 mmol) was stirred at 25°C for 14 hours. The mixture was then concentrated to obtain a pale yellow oil. The pale yellow oil was dissolved in water (100 mL) and extracted with methyl tert butyl ether (5 × 70 mL; the entire extraction process needed to be held still for 20 minutes for emulsification, and the addition of a small amount of EtOH was beneficial for separation). The aqueous solution was freeze-dried to obtain the marked compound (15.9 mg, 49.6 mmol, 68% yield) as a pale yellow solid. MS m / z(ESI): 321.3 [M+H] + .

[0093] Step 5: Benzyl(2S)-2-[[(2-amino-2-oxo-ethyl)-(2-fluoroacetyl)amino]carbamoyl]pyrrolidine-1-carboxylate(Int-6)

[0094] Benzyl(2S)-2-[[(2-amino-2-oxo-ethyl)amino]carbamoyl]pyrrolidine-1-carboxylate (Int-5, 4.20 g, 13.1 mmol) was dissolved in dichloromethane (35 mL), and the solution was cooled to -78°C (acetone / dry ice). Then, a solution of fluoroacetyl chloride (1645 mg, 17.0 mmol) in dichloromethane (15 mL) was added dropwise at -78°C. The mixture was stirred at -78°C for 0.5 hours. The mixture was then warmed to 25°C and stirred for 1.5 hours. Afterward, the mixture was concentrated under vacuum to obtain the crude labeled compound as a pale yellow solid (4.90 g, 12.9 mmol, 98% yield), which was used in the next step without further purification. MS m / z(ESI): 403.2 [M+H] + .

[0095] Step 6: 2-[(2-fluoroacetyl)-[[(2S)-pyrrolidine-2-carbonyl]amino]amino]acetamide(Int-7)

[0096] Benzyl (2S)-2-[[(2-amino-2-oxo-ethyl)-(2-fluoroacetyl)amino]carbamoyl]pyrrolidine-1-carboxylate (Int-6, 3.90 g, 10.2 mmol) was dissolved in methanol (50 mL), and palladium carbon (10%, 800 mg) was added. The resulting mixture was stirred at 25 °C for 4 hours under a hydrogen atmosphere (hydrogen balloon). Then, the mixture was filtered through a celite pad, washed with methanol (50 mL), and the filtrate was concentrated in vacuo to obtain the title compound as a pale yellow solid (2.50 g, 10.1 mmol, 99% yield). MS m / z (ESI): 269.1 [M+H] + .

[0097] Intermediate 12: 2-[2-[(2S)-1-[1-[4-(trifluoromethoxy)phenyl]cyclopropanecarbonyl]pyrrolidine-2-carbonyl]hydrazino]acetamide

[0098] Step 1: Benzyl (2S)-2-[[(2-amino-2-oxo-ethyl)-tert-butoxycarbonyl-amino]carbamoyl]pyrrolidine-1-carboxylate (Int-9)

[0099] Benzyl (2S)-2-[[(2-amino-2-oxo-ethyl)amino]carbamoyl]pyrrolidine-1-carboxylate (Int-5, 5.00 g, 15.6 mmol) was dissolved in dichloromethane (100 mL), and di-tert-butyl dicarbonate (4088 mg, 18.7 mmol) and subsequently triethylamine (4.73 g, 46.8 mmol) were added. The mixture was stirred at 25 °C for 16 hours. Then, the reaction mixture was concentrated in vacuo, and the crude product was purified by reverse-phase chromatography. After lyophilization, the title compound was isolated as a white solid (3.00 g, 7.14 mmol, 46% yield). MS m / z (ESI): 421.1 [M+H] + .

[0100] ​Step 2: tert-butyl N-(2-amino-2-oxo-ethyl)-N-[[rac-(2S)-pyrrolidine-2-carbonyl]amino]carbamate (Int-10)

[0101] A mixture of benzyl(2S)-2-[[(2-amino-2-oxo-ethyl)-tert-butoxycarbonyl-amino]carbamoyl]pyrrolidine-1-carboxylate (Int-9, 1.50 g, 3.57 mmol) and palladium carbon (10% m / m, 100 mg) in tetrahydrofuran (60 mL) was stirred at 25°C for 48 hours under a hydrogen atmosphere (balloon). The mixture was then filtered through a celite pad, washed with tetrahydrofuran (50 mL), and the filtrate was concentrated under vacuum to obtain the marked compound as a white solid (1.00 g, 3.49 mmol, 98% yield).

[0102] Step 3: tert-butyl N-(2-amino-2-oxo-ethyl)-N-[[(2S)-1-[1-[4-(trifluoromethoxy)phenyl]cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]carbamate (Int-11)

[0103] tert-butyl N-(2-amino-2-oxo-ethyl)-N-[[(2S)-pyrrolidine-2-carbonyl]amino]carbamate (Int-10, 120 mg, 0.420 mmol) was dissolved in dimethylformamide (2 mL), and HATU (148 mg, 0.630 mmol), diisopropylethylamine (163 mg, 1.26 mmol), and 1-[4-(trifluoromethoxy)phenyl]cyclopropanecarboxylic acid (103 mg, 0.420 mmol) were added. The mixture was stirred at 25°C for 1 hour. The reaction mixture was then purified by reverse-phase chromatography (Phenomenex luna C18 150*25 mm*10 μm, eluent (water + 0.225% formic acid) / acetonitrile). After lyophilization, the marked compound was obtained as a white solid (100 mg, 0.190 mmol, yield 46%). MS m / z (ESI): 515.1 [M+H] + .

[0104] Step 4: 2-[2-[(2S)-1-[1-[4-(trifluoromethoxy)phenyl]cyclopropanecarbonyl]pyrrolidine-2-carbonyl]hydrazino]acetamide(Int-12)

[0105] tert-butyl N-(2-amino-2-oxo-ethyl)-N-[[(2S)-1-[1-[4-(trifluoromethoxy)phenyl]cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]carbamate (Int-11, 50 mg, 0.10 mmol) was dissolved in dichloromethane (0.3 mL), and trifluoroacetic acid (0.1 mL) was added. The mixture was stirred at 25°C for 1 hour. The reaction mixture was then concentrated under vacuum to obtain the crude labeled compound as a white solid (40 mg, 0.10 mmol, 99% yield), which was used in the next step without further purification. MS m / z(ESI): 415.1 [M+H] + .

[0106] Intermediate 18: 2-[(2-fluoroacetyl)-[[(2S)-pyrrolidine-2-carbonyl]amino]amino]acetamide [ka]

[0107] Step 1: Methyl(2S)-1-[4-(4-methoxyphenyl)phenyl]sulfonylpyrrolidine-2-carboxylate(Int-15)

[0108] Methyl L-proline hydrochloride (1.34 g, 7.93 mmol, Int-13) was dissolved in dichloromethane (25 mL), and the solution was cooled to 0°C (ice bath). Then, pyridine (3.1 g, 3.2 mL, 39.6 mmol) was added, followed by the dropwise addition of a solution of 4'-methoxy-[1,1'-biphenyl]-4-sulfonyl chloride (2.69 g, 9.51 mmol, Int-14) in dichloromethane (25 mL). The yellow reaction mixture was stirred at 0°C to 23°C for 3 hours. After that, the reaction mixture was poured into ice-cold citric acid aqueous solution (5% m / m, 50 mL), and after phase separation, the aqueous layer was extracted with dichloromethane (3 × 50 mL). The combined organic layers were dried over sodium sulfate, filtered, and the filtrate was concentrated under vacuum. The residue was purified by column chromatography (silica gel, 80 g, n-heptane / ethyl acetate, gradient 100:0~20:80 (v / v)). After concentration of the product-containing fraction, the marked compound was obtained as a white solid (2.55 g, 6.79 mmol, yield 86%). MS m / z (ESI): 376.1 [M+H] + .

[0109] Step 2: (2S)-1-[4-(4-methoxyphenyl)phenyl]sulfonylpyrrolidine-2-carboxylic acid (Int-16)

[0110] Methyl((4'-methoxy-[1,1'-biphenyl]-4-yl)sulfonyl)-L-prolinate (2.52 g, 6.71 mmol, Int-15) was dissolved in a mixture of tetrahydrofuran (20 mL), methanol (5 mL), and water (10 mL). Lithium hydroxide monohydrate (321 mg, 7.62 mmol) was added, and the reaction mixture was stirred at room temperature (23 °C) for 2 hours. The mixture was slowly acidified by adding aqueous citric acid solution (5% m / m, pH 3-4), extracted with ethyl acetate (2 × 20 mL), and the combined organic layer was dried over sodium sulfate and concentrated under vacuum to obtain the marked compound as a white solid (2.40 g, 6.64 mmol, 99% yield). No further purification was performed. MS m / z (ESI): 362.1 [M+H] + .

[0111] Step 3: 2-[2-[(2S)-1-[4-(4-methoxyphenyl)phenyl]sulfonylpyrrolidine-2-carbonyl]hydrazino]acetamide(Int-17)

[0112] A suspension of ((4'-methoxy-[1,1'-biphenyl]-4-yl)sulfonyl)-L-proline (100 mg, 277 μmol, Int-16) and 1-hydroxybenzotriazole hydrate (56.8 mg, 360 μmol) in dichloromethane (2 mL) was cooled to 0°C (ice bath). 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (70.4 mg, 360 μmol) was added, and the mixture was stirred at 0°C for 1 hour. Then, ethylaminoglycinate hydrochloride (85.6 mg, 553 μmol) and N,N-diisopropylethylamine (95 mg, 125 μl, 719 μmol) were added. The reaction mixture was heated to room temperature (23°C) and stirred for 30 minutes. After that, it was poured into water (5 mL) and extracted with ethyl acetate (2 × 10 mL). The combined organic layers were washed with saturated sodium bicarbonate aqueous solution (20 mL), dried on sodium sulfate, and concentrated under vacuum. The crude material was purified by column chromatography (silica gel, 12 g, n-heptane / ethyl acetate, gradient 100:0~0:100 (v / v)) to obtain the labeled compound as a colorless gum (115 mg, 249 μmol, 90% yield). MS(ESI) m / z: 462.2[M+H] + .

[0113] Step 4: (S)-2-(2-(((4'-Methoxy-[1,1'-biphenyl]-4-yl)sulfonyl)prolyl)hydrazineyl)acetamide(Int-18)

[0114] A solution of ethyl(S)-(1-((4'-methoxy-[1,1'-biphenyl]-4-yl)sulfonyl)pyrrolidine-2-carboxamide)glycinate (505 mg, 1.09 mmol, Int-17) in dimethylformamide (500 μL) was cooled to 0°C (ice bath). Ammonia (7 M in methanol, 15.6 mL, 109 mmol) was added, followed by sodium cyanide (5.4 mg, 109 μmol). The reaction mixture was stirred at 0°C for 2 hours and at room temperature (23°C) for 15 hours. The reaction mixture was then concentrated under vacuum, poured into water / brine (1:1 v / v, 10 mL), and extracted with dichloromethane / methanol (9:1 v / v, 2 × 20 mL). The combined organic layers were dried over sodium sulfate and concentrated under vacuum to obtain the marked compound as a white solid (454 mg, 1.05 mmol, 96% yield). MS(ESI)m / z:433.2[M+H] + . [Examples]

[0115] Example 1 2-[(2-fluoroacetyl)-[[(2S)-1-(2,2-difluoro-2-phenyl-acetyl)pyrrolidine-2-carbonyl]amino]amino]acetamide [ka]

[0116] 2-[(2-fluoroacetyl)-[[(2S)-pyrrolidine-2-carbonyl]amino]amino]acetamide (Int-7, 206 mg, 0.840 mmol) was dissolved in dimethylformamide (1.5 mL), and 2,2-difluoro-2-phenylacetic acid (80 mg, 0.46 mmol), HBTU (211 mg, 0.56 mmol), and N-methylmorpholine (94 mg, 0.93 mmol) were added. The mixture was stirred at 25°C for 4 hours. Subsequently, it was directly purified by preparative HPLC (Shim-pack C18 150*25*10 μm, eluent (water + 0.225% formic acid) / acetonitrile, gradient 76:24~56:44 within 10 minutes). After lyophilization, the marked compound was obtained as a white solid (30 mg, 0.07 mmol, yield 16%). MS(ESI)m / z:401.2[M+H] + .

[0117] Example 2 2-[(2-fluoroacetyl)-[[(2S)-1-(3-phenylpropanoyl)pyrrolidine-2-carbonyl]amino]amino]acetamide [ka]

[0118] 2-[(2-fluoroacetyl)-[[(2S)-pyrrolidine-2-carbonyl]amino]amino]acetamide (Int-7, 50 mg, 0.20 mmol) was dissolved in dimethylformamide (1.5 mL), and 3-phenylpropionic acid (30.5 mg, 0.20 mmol), HBTU (92 mg, 0.24 mmol), and N-methylmorpholine (41 mg, 0.41 mmol) were added. The mixture was stirred at 25°C for 4 hours. Subsequently, it was directly purified by preparative HPLC (Phenomenex luna C18 150*25 mm*10 μm, eluent (water + 0.225% formic acid) / acetonitrile, gradient 81:19~51:49 within 11 minutes). After lyophilization, the marked compound was isolated as a white solid (13 mg, 0.03 mmol, yield 17%). MS(ESI)m / z:379.1[M+H] + .

[0119] Example 3 2-[(2-fluoroacetyl)-[[(2S)-1-(1-phenylcyclobutanecarbonyl)pyrrolidine-2-carbonyl]amino]amino]acetamide [ka]

[0120] 2-[(2-fluoroacetyl)-[[(2S)-pyrrolidine-2-carbonyl]amino]amino]acetamide (Int-7, 84 mg, 0.34 mmol) was dissolved in dimethylformamide (1.5 mL), and 1-phenylcyclobutanecarboxylic acid (50 mg, 0.28 mmol), HATU (129 mg, 0.34 mmol), and N,N-diisopropylethylamine (0.1 mL, 0.57 mmol) were added. The mixture was stirred at 25°C for 12 hours. It was then directly purified by preparative HPLC (Phenomenex luna C18 150*25 mm*10 μm, eluent (water + 0.225% formic acid) / acetonitrile). After lyophilization, the marked compound was obtained as a white solid (22 mg, 0.05 mmol, yield 18%). MS(ESI) m / z: 405.2[M+H] + .

[0121] Example 4 2-[(2-fluoroacetyl)-[[(2S)-1-[1-(4-chlorophenyl)cyclobutanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide [ka]

[0122] 2-[(2-fluoroacetyl)-[[(2S)-pyrrolidine-2-carbonyl]amino]amino]acetamide (Int-7, 70 mg, 0.28 mmol) was dissolved in dimethylformamide (1.5 mL), and 1-(4-chlorophenyl)-1-cyclobutanecarboxylic acid (50 mg, 0.24 mmol), HATU (108 mg, 0.28 mmol), and N,N-diisopropylethylamine (0.04 mL, 0.24 mmol) were added. The mixture was stirred at 25°C for 12 hours. Subsequently, it was purified directly by preparative HPLC (Phenomenex luna C18 150*25 mm*10 μm, eluent (water + 0.225% formic acid) / acetonitrile). After lyophilization, the marked compound was obtained as a white solid (23 mg, 0.05 mmol, yield 21%). MS(ESI)m / z:461.2 [M+Na] + .

[0123] Example 5 2-[(2-fluoroacetyl)-[[(2S)-1-(2-phenylacetyl)pyrrolidine-2-carbonyl]amino]amino]acetamide [ka]

[0124] 2-[(2-fluoroacetyl)-[[(2S)-pyrrolidine-2-carbonyl]amino]amino]acetamide (Int-7, 50 mg, 0.20 mmol) was dissolved in dimethylformamide (1.5 mL), and phenylacetic acid (28 mg, 0.20 mmol), HBTU (92 mg, 0.24 mmol), and N-methylmorpholine (41 mg, 0.41 mmol) were added. The mixture was stirred at 25°C for 4 hours. This was then purified directly by preparative HPLC (Phenomenex luna C18 150*25 mm*10 μm, eluent (water + 0.225% formic acid) / acetonitrile, gradient 88:12~58:42). After lyophilization, the marked compound was isolated as a white solid (22 mg, 0.06 mmol, yield 30%). MS(ESI) m / z: 365.1[M+H] + .

[0125] Example 6 2-[(2-fluoroacetyl)-[[(2S)-1-[1-(4-chlorophenyl)cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide [ka]

[0126] 2-[(2-fluoroacetyl)-[[(2S)-pyrrolidine-2-carbonyl]amino]amino]acetamide (Int-7, 50 mg, 0.20 mmol) was dissolved in dimethylformamide (1.5 mL), and 1-(4-chlorophenyl)cyclopropanecarboxylic acid (48 mg, 0.24 mmol), HBTU (92 mg, 0.24 mmol), and N-methylmorpholine (41 mg, 0.41 mmol) were added. The mixture was stirred at 25°C for 4 hours. Subsequently, it was directly purified by preparative HPLC (Phenomenex luna C18 150*25 mm*10 μm, eluent (water + 0.225% formic acid) / acetonitrile, gradient 73:27~43:57 within 10 minutes). After lyophilization, the marked compound was obtained as a white solid (18 mg, 0.04 mmol, yield 21%). MS(ESI)m / z:425.0[M+H] + .

[0127] Example 7 2-[(2-fluoroacetyl)-[[(2S)-1-(benzenesulfonyl)pyrrolidine-2-carbonyl]amino]amino]acetamide [ka]

[0128] 2-[(2-fluoroacetyl)-[[(2S)-pyrrolidine-2-carbonyl]amino]amino]acetamide (Int-7, 50 mg, 0.20 mmol) was dissolved in dimethylformamide (1.5 mL), and benzenesulfone chloride (0.03 mL, 0.22 mmol) and N,N-diisopropylethylamine (0.07 mL, 0.41 mmol) were added. The mixture was stirred at 25°C for 4 hours. It was then purified directly by preparative HPLC (Phenomenex luna C18 150*25 mm*10 μm, eluent (water + 0.225% formic acid) / acetonitrile). After lyophilization, the marked compound was obtained as a white solid (29 mg, 0.08 mmol, yield 37%). MS(ESI) m / z: 409.2 [M+Na] + .

[0129] Example 8 2-[(2-fluoroacetyl)-[[(2S)-1-(1-phenylcyclopropanecarbonyl)pyrrolidine-2-carbonyl]amino]amino]acetamide [ka]

[0130] 2-[(2-fluoroacetyl)-[[(2S)-pyrrolidine-2-carbonyl]amino]amino]acetamide (Int-7, 50 mg, 0.20 mmol) was dissolved in dimethylformamide (1.5 mL), and 1-phenyl-1-cyclopropanecarboxylic acid (33 mg, 0.20 mmol), HBTU (92 mg, 0.24 mmol), and N-methylmorpholine (41 mg, 0.41 mmol) were added. The mixture was stirred at 25°C for 4 hours. It was then directly purified by preparative HPLC (Phenomenex luna C18 150*25 mm*10 μm, eluent (water + 0.225% formic acid) / acetonitrile). After lyophilization, the marked compound was isolated as a white solid (23 mg, 0.06 mmol, yield 29%). MS(ESI) m / z: 413.3 [M+Na] + .

[0131] Example 9 2-[(2-fluoroacetyl)-[[(2S)-1-benzylsulfonylpyrrolidine-2-carbonyl]amino]amino]acetamide [ka]

[0132] 2-[(2-fluoroacetyl)-[[(2S)-pyrrolidine-2-carbonyl]amino]amino]acetamide (Int-7, 50 mg, 0.20 mmol) was dissolved in dimethylformamide (1.5 mL), and α-toluenesulfonyl chloride (0.03 mL, 0.22 mmol) and N,N-diisopropylethylamine (0.07 mL, 0.41 mmol) were added. The mixture was stirred at 25°C for 4 hours. It was then directly purified by preparative HPLC (Phenomenex luna C18 150*25 mm*10 μm, eluent (water + 0.225% formic acid) / acetonitrile). After lyophilization, the marked compound was obtained as a white solid (32 mg, 0.08 mmol, yield 39%). MS(ESI) m / z: 423.2 [M+Na] + .

[0133] Example 10 2-[(2-fluoroacetyl)-[[(2S)-1-(2-phenylethylsulfonyl)pyrrolidine-2-carbonyl]amino]amino]acetamide [ka]

[0134] 2-[(2-fluoroacetyl)-[[(2S)-pyrrolidine-2-carbonyl]amino]amino]acetamide (Int-7, 100 mg, 0.41 mmol) was dissolved in dimethylformamide (1.0 mL), and 2-phenylethanesulfonyl chloride (83 mg, 0.41 mmol) and N,N-diisopropylethylamine (0.14 mL, 0.81 mmol) were added. The mixture was stirred at 25°C for 4 hours. It was then directly purified by preparative HPLC (Phenomenex luna C18 150*25 mm*10 μm, eluent (water + 0.225% formic acid) / acetonitrile). After lyophilization, the marked compound was isolated as a white solid (40 mg, 0.10 mmol, yield 24%). MS(ESI) m / z: 415.2[M+H] + .

[0135] Example 11 2-[(2-fluoroacetyl)-[[(2S)-1-[1-(4-ethynylphenyl)cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide [ka]

[0136] 2-[(2-fluoroacetyl)-[[(2S)-pyrrolidine-2-carbonyl]amino]amino]acetamide (Int-7, 100 mg, 0.41 mmol) was dissolved in dimethylformamide (2 mL), and 1-(4-ethynylphenyl)cyclopropanecarboxylic acid (75.6 mg, 0.41 mmol), HBTU (185 mg, 0.49 mmol), and N,N-diisopropylethylamine (0.21 mL, 1.22 mmol) were added. The mixture was stirred at 25°C for 4 hours. Subsequently, it was directly purified by preparative HPLC (Phenomenex luna C18 150*25 mm*10 μm, eluent (water + 0.225% formic acid) / acetonitrile). After lyophilization, the marked compound was obtained as a white solid (100 mg, 0.24 mmol, yield 59%). MS(ESI)m / z:415.2[M+H] + .

[0137] Example 12 2-[(2-fluoroacetyl)-[[(2S)-1-[1-[4-(trifluoromethoxy)phenyl]cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide [ka]

[0138] 2-(2-((1-(4-(trifluoromethoxy)phenyl)cyclopropane-1-carbonyl)-L-prolyl)hydrazinyl)acetamide (Int-12, 35 mg, 84 μmol) was dissolved in dichloromethane (1 mL), and pyridine (8.7 mg, 8.8 μl, 110 μmol) was added. The solution was cooled to 0°C (ice bath), and 2-fluoroacetyl chloride (11 mg, 7.8 μl, 110 μmol) was added. The reaction mixture was stirred at 0°C for 1 hour. Thereafter, additional amounts of pyridine (8.7 mg, 8.8 μl, 110 μmol) and 2-fluoroacetyl chloride (11 mg, 7.8 μl, 110 μmol) were added, and stirring was continued at 0°C for 1 hour. Next, the mixture was concentrated under vacuum, and the residue was purified by silica gel flash chromatography (12 g, ethyl acetate / n-heptane, gradient 0:100~100:0, then methanol / ethyl acetate, gradient 0:100~10:90) to obtain the marked compound as a white solid (34 mg, yield 85%). MS(ESI)m / z:475.2[M+H] + .

[0139] Example 13 2-[(2-fluoroacetyl)-[[(2S)-1-[4-(4-methoxyphenyl)phenyl]sulfonylpyrrolidine-2-carbonyl]amino]amino]acetamide [ka]

[0140] (S)-2-(2-(((4'-methoxy-[1,1'-biphenyl]-4-yl)sulfonyl)prolyl)hydrazineyl)acetamide (Int-18, 50 mg, 116 μmol) was suspended in dichloromethane (1.5 mL), and pyridine (10.1 mg, 10.2 μl, 127 μmol) was added. The solution was cooled to 0°C (ice bath), and a solution of 2-fluoroacetyl chloride (12.5 mg, 9.1 μl, 127 μmol) in dichloromethane (100 μl) was added. The reaction mixture was stirred at 0°C for 30 minutes. Thereafter, further portions of pyridine (10.1 mg, 10.2 μl, 127 μmol) and 2-fluoroacetyl chloride (12.5 mg, 9.1 μl, 127 μmol) in dichloromethane (100 μl) were added, and stirring was continued at 0°C for 30 minutes. Next, the mixture was diluted with dichloromethane (5 mL), washed with water (5 mL), and the organic layer was dried on sodium sulfate and concentrated under vacuum. The residue was purified by silica gel flash chromatography (12 g, ethyl acetate / n-heptane, gradient 0:100~100:0, then methanol / ethyl acetate, gradient 0:100~10:90). After concentration of the product-containing fraction, the marked compound was obtained as a white solid (54 mg, yield 95%). MS(ESI)m / z:493.2[M+H] + .

[0141] Example 14 2-[(2-fluoroacetyl)-[[(2S)-1-[1-(4-prop-2-inoxyphenyl)cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide [ka]

[0142] 2-[(2-fluoroacetyl)-[[(2S)-pyrrolidine-2-carbonyl]amino]amino]acetamide (Int-7, 228 mg, 0.92 mmol) was dissolved in dimethylformamide (2 mL), and 1-(4-prop-2-inoxyphenyl)cyclopropanecarboxylic acid (100 mg, 0.46 mmol), HBTU (351 mg, 0.92 mmol), and N,N-diisopropylethylamine (0.24 mL, 1.39 mmol) were added. The mixture was stirred at 20°C for 2 hours. Subsequently, it was purified directly by preparative HPLC (Phenomenex luna C18 150*25 mm*10 μm, eluent (water + 0.225% formic acid) / acetonitrile, gradient 83:17~50:50 within 11 minutes). After freeze-drying, the marked compound was obtained as a white solid (64 mg, 0.13 mmol, 30% yield). MS(ESI)m / z:445.1[M+H] + .

[0143] Example 15 2-[((2R)&(2S)-2-chloro-2-fluoroacetyl)-[[(2S)-1-[4-(4-methoxyphenyl)phenyl]sulfonylpyrrolidine-2-carbonyl]amino]amino]acetamide [ka]

[0144] (S)-2-(2-(((4'-methoxy-[1,1'-biphenyl]-4-yl)sulfonyl)prolyl)hydrazineyl)acetamide (Int-18, 45 mg, 104 μmol) was suspended in dichloromethane (1.5 mL), and pyridine (9.0 mg, 9.2 μl, 114 μmol) was added. The solution was cooled to 0°C (ice bath), and a solution of 2-chloro-2-fluoroacetyl chloride (15.3 mg, 10.2 μl, 114 μmol) in dichloromethane (100 μl) was added. The reaction mixture was stirred at 0°C for 30 minutes. Subsequently, further fractions of pyridine (9.0 mg, 9.2 μl, 114 μmol) and 2-chloro-2-fluoroacetyl chloride (15.3 mg, 10.2 μl, 114 μmol) in dichloromethane (100 μl) were added, and the mixture was stirred at 0°C for 15 minutes. This was then diluted with dichloromethane (5 mL), washed with water (5 mL), and the organic layer was dried over sodium sulfate and concentrated under vacuum. The residue was purified by silica gel flash chromatography (12 g, ethyl acetate / n-heptane, gradient 0:100~100:0, then methanol / ethyl acetate, gradient 0:100~10:90). After concentration of the product-containing fraction, the marked compound was obtained as a white solid (35 mg, yield 64%). MS(ESI)m / z:527.1[M+H] + .

[0145] Example 16 2-[((2R)&(2S)-2-chloro-2-fluoroacetyl)-[[(2S)-1-[1-[4-(trifluoromethoxy)phenyl]cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide [ka]

[0146] 2-(2-((1-(4-(trifluoromethoxy)phenyl)cyclopropane-1-carbonyl)-L-prolyl)hydrazinyl)acetamide (Int-12, 100 mg, 241 μmol) was dissolved in dichloromethane (2 mL), and pyridine (24.8 mg, 25.2 μl, 314 μmol) was added. The solution was cooled to 0°C (ice bath), and 2-chloro-2-fluoroacetyl chloride (41.9 mg, 27.9 μl, 314 μmol) was added. The reaction mixture was stirred at 0°C for 1 hour. Thereafter, additional amounts of pyridine (24.8 mg, 25.2 μl, 314 μmol) and 2-chloro-2-fluoroacetyl chloride (41.9 mg, 27.9 μl, 314 μmol) were added, and stirring was continued at 0°C for 1 hour. Next, the mixture was concentrated under vacuum, and the residue was purified by silica gel flash chromatography (25 g, ethyl acetate / n-heptane, gradient 0:100~100:0, then methanol / ethyl acetate, gradient 0:100~10:90) to obtain the marked compound as a white solid (133 mg, yield 99%). MS(ESI)m / z:507.2 [MH] - .

[0147] Example 17 2-[(2-chloroacetyl)-[[(2S)-1-[4-(4-methoxyphenyl)phenyl]sulfonylpyrrolidine-2-carbonyl]amino]amino]acetamide [ka]

[0148] (S)-2-(2-(((4'-methoxy-[1,1'-biphenyl]-4-yl)sulfonyl)prolyl)hydrazineyl)acetamide (Int-18, 50 mg, 116 μmol) was suspended in dichloromethane (1.5 mL), and pyridine (10.1 mg, 10.2 μl, 127 μmol) was added. The solution was cooled to 0°C (ice bath), and a solution of 2-chloroacetyl chloride (14.5 mg, 10.2 μl, 127 μmol) in dichloromethane (100 μl) was added. The reaction mixture was stirred at 0°C for 30 minutes. Then, it was diluted with dichloromethane (5 mL), washed with water (5 mL), the organic layer was dried on sodium sulfate, and concentrated under vacuum. The residue was purified by silica gel flash chromatography (12 g, ethyl acetate / n-heptane, gradient 0:100~100:0, then methanol / ethyl acetate, gradient 0:100~20:80). After concentration of the product-containing fraction, the marked compound was obtained as a white solid (55 mg, yield 91%). MS(ESI)m / z:509.1[M+H] + .

[0149] Example 18 2-[(2-chloroacetyl)-[[(2S)-1-[1-[4-(trifluoromethoxy)phenyl]cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide [ka]

[0150] 2-(2-((1-(4-(trifluoromethoxy)phenyl)cyclopropane-1-carbonyl)-L-prolyl)hydrazinyl)acetamide (Int-12, 35 mg, 84 μmol) was dissolved in dichloromethane (1 mL), and pyridine (8.7 mg, 8.8 μl, 110 μmol) was added. The solution was cooled to 0°C (ice bath), and a solution of 2-chloroacetyl chloride (12.7 mg, 8.9 μl, 110 μmol) in dichloromethane (80 μL) was added. The reaction mixture was stirred at 0°C for 30 minutes. Thereafter, further amounts of pyridine (8.7 mg, 8.8 μl, 110 μmol) and 2-chloroacetyl chloride (12.7 mg, 8.9 μl, 110 μmol) in dichloromethane (80 μL) were added, and stirring was continued at 0°C for 30 minutes. Next, the mixture was concentrated under vacuum, and the residue was purified by silica gel flash chromatography (12 g, ethyl acetate / n-heptane, gradient 0:100~100:0, then methanol / ethyl acetate, gradient 0:100~10:90) to obtain the marked compound as a white solid (36 mg, yield 87%). MS(ESI)m / z:491.2[M+H] + .

[0151] Example 19 2-[(2,2-dichloroacetyl)-[[(2S)-1-[1-[4-(trifluoromethoxy)phenyl]cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide [ka]

[0152] 2-(2-((1-(4-(trifluoromethoxy)phenyl)cyclopropane-1-carbonyl)-L-prolyl)hydrazinyl)acetamide (Int-12, 25 mg, 60 μmol) was dissolved in dichloromethane (1.2 mL), and pyridine (7.2 mg, 7.3 μl, 90 μmol) was added. The solution was cooled to 0°C (ice bath), and 2,2-dichloroacetyl chloride (13.3 mg, 90 μmol) was added. The reaction mixture was stirred at 0°C for 3 hours. This was then concentrated under vacuum, and the residue was purified by preparative HPLC (YMC-Triart C18, 12 nm, 5 μm, 100 × 30 mm, acetonitrile / water + 0.1% triethylamine, gradient 20:80~98:2) to obtain the marked compound as a white solid (12 mg, yield 37%). MS(ESI)m / z:525.3[M+H] + .

[0153] Example 20 2-[(2R)&(2S)-2-chloropropanoyl-[[(2S)-1-[1-[4-(trifluoromethoxy)phenyl]cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide [ka]

[0154] 2-(2-((1-(4-(trifluoromethoxy)phenyl)cyclopropane-1-carbonyl)-L-prolyl)hydrazinyl)acetamide (Int-12, 40 mg, 97 μmol) was dissolved in dichloromethane (1 mL), and pyridine (13.1 mg, 13.3 μl, 166 μmol) was added. The solution was cooled to 0°C (ice bath), and 2-chloropropionyl chloride (18.4 mg, 14.4 μl, 145 μmol) was added. The reaction mixture was stirred at 0°C for 1.5 hours. This was then concentrated under vacuum, and the residue was purified by preparative HPLC (YMC-Triart C18, 12 nm, 5 μm, 100 × 30 mm, acetonitrile / water + 0.1% triethylamine, gradient 20:80~98:2) to obtain the marked compound as a white solid (17 mg, yield 35%). MS(ESI)m / z:505.3[M+H] + .

[0155] AEP-activated enzyme assay procedure: This assay uses the fluorescence-generating substrate, Z-Ala-Ala-Asn-Rh110-(D-Pro) (Biosyntan, Berlin, Ord.Nr.80773), which fluoresces when cleaved by legmine and can be monitored when excited at 492 nm and emission at 530 nm. Prior to assay measurement, legmine (in-house, construct name: hLGMN(V18-Y433)_VD-8xHis-S2-r) is activated by diluting it from 1.1 mg / ml to 0.1 mg / ml (11-fold) in activation buffer (50 mM sodium acetate, 100 mM NaCl, pH 4.0) and incubating at 37°C for 2 hours. The activated enzyme solution stock is stored at -80°C. For measurement, the following solutions are then dispensed into a 384-well microplate (Corning 384 untreated, black / clear, catalog number: 3540). As a negative control, 8.5 μL of assay buffer (20 mM citrate, 60 mM Na2HPO4, 1 mM EDTA, 0.1% CHAPS, pH 5.8, 0.5 mM TCEP (freshly added)) or 8.5 μL of activated enzyme solution (0.5 nM regmine (final) in assay buffer (20 mM citrate, 60 mM Na2HPO4, 1 mM EDTA, 0.1% CHAPS, pH 5.8, 0.5 mM TCEP (freshly added))) and 1.5 μL of pre-diluted compound solution (100 μM to 0.1 nM, 1% DMSO in water (final)). After incubation at 25°C for 30 minutes, add 5 μL of substrate solution (0.5 μM Z-Ala-Ala-Asn-Rh110-(D-Pro) (final)) in assay buffer (20 mM citrate, 60 mM Na2HPO4, 1 mM EDTA, 0.1% CHAPS, pH 5.8, 0.5 mM TCEP (freshly added)), and measure the fluorescence using a plate reader with excitation at 490 nm and emission at 530 nm, for example, using a PHERAstar or Spectramax instrument in kinetic mode (analyze the first 30 minutes).

[0156] The results of the AEP active enzyme assay are provided in Table 1 for the compound of formula I. [Table 1]

[0157] Example A The compound of formula I can be used as an active ingredient in a manner known per se to produce tablets of the following composition: Per tablet Active ingredient 200 mg Microcrystalline cellulose 155 mg Corn starch 25 mg Talc 25 mg Hydroxypropylmethylcellulose 20 mg 425 mg

[0158] Example B The compound of formula I can be used as an active ingredient in a manner known per se to produce capsules of the following composition: Per capsule Active ingredient 100.0 mg Corn starch 20.0 mg Lactose 95.0 mg Talc 4.5 mg Magnesium stearate 0.5 mg 220.0 mg

Claims

1. Equation I 【Chemistry 1】 (In the formula, R １ is either halo or alkyl; R ２ is H or halo; R ３ is H or halo; R ４ Is it H or a halo? Or, R ３ and R ４ It forms a three-membered or four-membered cycloalkyl ring; R ５ is H, halo, haloalkoxy, alkynyl, alkynylalkoxy, or alkoxyphenyl; R ６ H is; R ７ is H; W is -C(O)- or -S(O) ２ - and; m is either 0 or 1; n is either 0 or 1. Compounds of and pharmaceutically acceptable salts thereof.

2. R １ The compound according to claim 1, wherein the compound is a halo.

3. R ５ The compound according to claim 1, wherein the compound is H, halo, or haloalkoxy.

4. The compound according to claim 1, wherein W is -C(O)-.

5. The compound according to claim 1, wherein m is 1 and n is 0.

6. R ３ and R ４ The compound according to claim 1, wherein it forms a three-membered or four-membered cycloalkyl ring.

7. R １ But it's a halo; R ２ However, it is H or halo; R ３ But it's a halo; R ４ But is it a halo? Or, R ３ and R ４ However, it forms a three-membered or four-membered cycloalkyl ring; R ５ However, it is H, halo, or haloalkoxyl; W is -C(O)-; m is 1; n is 0, The compound according to claim 1, and a pharmaceutically acceptable salt.

8. R １ But it's a halo; R ２ However, it is H or halo; R ３ and R ４ However, it forms a three-membered or four-membered cycloalkyl ring; R ５ However, it is a halo or haloalkoxyl; W is -C(O)-; m is 1; n is 0, The compound according to claim 1, and a pharmaceutically acceptable salt.

9. R １ However, it is either F or Cl; R ２ However, it is either H or F; R ３ and R ４ However, it forms a three-membered cycloalkyl ring; R ５ However, it is chloro or trifluoromethoxy; W is -C(O)-; m is 1; n is 0, The compound according to claim 1, and a pharmaceutically acceptable salt.

10. 2-[(2-fluoroacetyl)-[[(2S)-1-(2,2-difluoro-2-phenyl-acetyl)pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-fluoroacetyl)-[[(2S)-1-(3-phenylpropanoyl)pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-fluoroacetyl)-[[(2S)-1-(1-phenylcyclobutanecarbonyl)pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-fluoroacetyl)-[[(2S)-1-[1-(4-chlorophenyl)cyclobutanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-fluoroacetyl)-[[(2S)-1-(2-phenylacetyl)pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-fluoroacetyl)-[[(2S)-1-[1-(4-chlorophenyl)cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-fluoroacetyl)-[[(2S)-1-(benzenesulfonyl)pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-fluoroacetyl)-[[(2S)-1-(1-phenylcyclopropanecarbonyl)pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-fluoroacetyl)-[[(2S)-1-benzylsulfonylpyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-fluoroacetyl)-[[(2S)-1-(2-phenylethylsulfonyl)pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-fluoroacetyl)-[[(2S)-1-[1-(4-ethynylphenyl)cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-fluoroacetyl)-[[(2S)-1-[1-[4-(trifluoromethoxy)phenyl]cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-fluoroacetyl)-[[(2S)-1-[4-(4-methoxyphenyl)phenyl]sulfonylpyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-fluoroacetyl)-[[(2S)-1-[1-(4-prop-2-inoxyphenyl)cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[((2R)-2-chloro-2-fluoro-acetyl)-[[(2S)-1-[4-(4-methoxyphenyl)phenyl]sulfonylpyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[((2S)-2-chloro-2-fluoro-acetyl)-[[(2S)-1-[4-(4-methoxyphenyl)phenyl]sulfonylpyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[((2R)-2-chloro-2-fluoro-acetyl)-[[(2S)-1-[1-[4-(trifluoromethoxy)phenyl]cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[((2S)-2-chloro-2-fluoro-acetyl)-[[(2S)-1-[1-[4-(trifluoromethoxy)phenyl]cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-chloroacetyl)-[[(2S)-1-[4-(4-methoxyphenyl)phenyl]sulfonylpyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-chloroacetyl)-[[(2S)-1-[1-[4-(trifluoromethoxy)phenyl]cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2,2-dichloroacetyl)-[[(2S)-1-[1-[4-(trifluoromethoxy)phenyl]cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2R)-2-chloropropanoyl-[[(2S)-1-[1-[4-(trifluoromethoxy)phenyl]cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2S)-2-chloropropanoyl-[[(2S)-1-[1-[4-(trifluoromethoxy)phenyl]cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; A compound according to claim 1, selected from, and a pharmaceutically acceptable salt thereof.

11. 2-[(2-fluoroacetyl)-[[(2S)-1-(2,2-difluoro-2-phenyl-acetyl)pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-fluoroacetyl)-[[(2S)-1-[1-(4-chlorophenyl)cyclobutanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-fluoroacetyl)-[[(2S)-1-[1-(4-chlorophenyl)cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-fluoroacetyl)-[[(2S)-1-[1-[4-(trifluoromethoxy)phenyl]cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[((2R)-2-chloro-2-fluoro-acetyl)-[[(2S)-1-[1-[4-(trifluoromethoxy)phenyl]cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[((2S)-2-chloro-2-fluoro-acetyl)-[[(2S)-1-[1-[4-(trifluoromethoxy)phenyl]cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-chloroacetyl)-[[(2S)-1-[1-[4-(trifluoromethoxy)phenyl]cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; A compound according to claim 1, selected from, and a pharmaceutically acceptable salt thereof.

12. 2-[(2-fluoroacetyl)-[[(2S)-1-[1-(4-chlorophenyl)cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[(2-fluoroacetyl)-[[(2S)-1-[1-[4-(trifluoromethoxy)phenyl]cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[((2R)-2-chloro-2-fluoro-acetyl)-[[(2S)-1-[1-[4-(trifluoromethoxy)phenyl]cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; 2-[((2S)-2-chloro-2-fluoro-acetyl)-[[(2S)-1-[1-[4-(trifluoromethoxy)phenyl]cyclopropanecarbonyl]pyrrolidine-2-carbonyl]amino]amino]acetamide; A compound according to claim 1, selected from, and a pharmaceutically acceptable salt thereof.

13. A method for preparing the compound according to any one of claims 1 to 12, comprising the reaction of a compound of formula II with a compound of formula III. 【Chemistry 2】 (In the formula, R １ , R ２ , R ３ , R ４ , R ５ , R ６ , R ７ (W, n, and m are as defined above, and X is a suitable functional group selected from F, Cl, Br, OH, and O-N-succinimidyl (OSu)). Methods that include...

14. A method for preparing the compound according to any one of claims 1 to 12, comprising the reaction of a compound of formula IV with a compound of formula V 【Transformation 3】 (In the formula, R １ , R ２ , R ３ , R ４ , R ５ , R ６ , R ７ (W, n, and m are as defined above, and Y is a suitable functional group selected from F, Cl, Br, OH, and O-N-succinimidyl (OSu)). Methods that include...

15. A compound according to any one of claims 1 to 12, for use as a therapeutically active substance.

16. A compound according to any one of claims 1 to 12, for use in the treatment of diseases regulated by AEP.

17. A pharmaceutical composition comprising a compound according to any one of claims 1 to 12 and a therapeutically inactive carrier.

18. A pharmaceutical composition comprising a compound according to any one of claims 1 to 12 for treating or preventing Alzheimer's disease, primary age-related tauopathy (PART) dementia, frontotemporal dementia (FTD-MAPT), chronic traumatic encephalopathy (CTE), progressive supranuclear palsy (PSP), corticobasal degeneration (CBD), frontotemporal dementia and parkinsonism associated with chromosome 17 (FTDP-17), Ritico-Bodig disease (Guam Parkinson's Dementia Complex), ganglioglioma and gangliocytoma, meningeal hemangioma, post-encephalitis parkinsonism, subacute sclerosing panencephalitis (SSPE), Pick's disease, or corticobasal degeneration.

19. A pharmaceutical composition comprising a compound according to any one of claims 1 to 12 for treating or preventing Alzheimer's disease.

20. Use of the compounds according to any one of claims 1 to 12 for preparing a medicament for treating or preventing Alzheimer's disease, primary age-related tauopathy (PART) dementia, frontotemporal dementia (FTD-MAPT), chronic traumatic encephalopathy (CTE), progressive supranuclear palsy (PSP), corticobasal degeneration (CBD), frontotemporal dementia and parkinsonism associated with chromosome 17 (FTDP-17), Ritico-Bodig disease (Guam Parkinson's Dementia Complex), ganglioglioma and gangliocytoma, meningeal hemangioma, post-encephalitis parkinsonism, subacute sclerosing panencephalitis (SSPE), Pick's disease, or corticobasal degeneration.

21. Use of the compound according to any one of claims 1 to 12 for preparing a medicament for treating or preventing Alzheimer's disease.

22. A compound according to any one of claims 1 to 12 for treating or preventing Alzheimer's disease, primary age-related tauopathy (PART) dementia, frontotemporal dementia (FTD-MAPT), chronic traumatic encephalopathy (CTE), progressive supranuclear palsy (PSP), corticobasal degeneration (CBD), frontotemporal dementia and parkinsonism associated with chromosome 17 (FTDP-17), Ritico-Bodig disease (Guam Parkinson's Dementia Complex), ganglioglioma and gangliocytoma, meningeal hemangioma, post-encephalitis parkinsonism, subacute sclerosing panencephalitis (SSPE), Pick's disease, or corticobasal degeneration.

23. A compound according to any one of claims 1 to 12 for treating or preventing Alzheimer's disease.

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