Haloacetyl azides useful as AEP inhibitors

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 the treatment and / or prevention in a patient, particularly compounds that inhibit AEP activity.

[0002] The present invention relates to a compound of formula I [ka] (In the formula, R 1 is halo or alkyl; R 2 is H or halo; R 3 is H or halo; R 4 is H or halo; Or, R 3 and R 4 forms a 3- 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 -C(O)- or -S(O) 2 - and; m is 0 or 1; n is 0 or 1. The present invention provides novel compounds of the formula:

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

[0004] 2. Background of the Invention Alzheimer's Disease (AD) Alzheimer's disease occurs primarily in older adults and is the most common form of dementia, affecting over 5 million people in the United States alone. This number is expected to triple by 2050. The prevalence of AD is age-related, and patients often require institutionalization during the later stages of the disease. AD will continue to be a major public health problem in the coming years due to its severity, increasing prevalence, long duration, and high cost of treatment.

[0005] Currently approved treatments for AD modulate neurotransmission (e.g., Aricept, an acetylcholinesterase inhibitor). These drugs provide short-term relief from some symptoms, but do not alter the underlying pathology or course of the disease. There is a large unmet medical need for more effective treatments for AD at all stages, especially novel treatments that slow or delay disease progression.

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

[0007] Most advanced drugs in development for AD aim to modify disease progression by lowering Aβ. These drugs include Aβ-specific antibodies that promote Aβ clearance, as well as small molecule inhibitors and modulators of the proteolytic enzymes responsible for Aβ production, namely β- and γ-secretase. Strategies to mitigate tau aggregation are needed to complement Aβ-lowering approaches. Several companies are pursuing tau-directed monoclonal antibodies for AD. However, tau antibodies are still in early-stage clinical development and are unlikely to emerge as standard of care for AD in the next few years.

[0008] AD belongs to a larger group of neurodegenerative dementia diseases known as tauopathies. This group of diseases includes frontotemporal dementia (FTD-MAPT), progressive supranuclear palsy (PSP) and corticobasal degeneration (CBD). A common feature of tauopathies is the presence of fibrillar tau aggregates in cells. Therapeutic approaches targeting tau 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 intraneuronal neurofibrillary tangles is one of the defining pathologies of AD. Tau protein, the main component of tangles, is a highly soluble protein that normally associates with and stabilizes microtubules. In AD and other tauopathies, tau undergoes conformational changes that lead to its aggregation (Vaquer-Alicea et al. Acta Neuropath 2021). Tau aggregation is central to disease pathogenesis in tauopathies, as demonstrated by multiple lines of evidence: (1) Many MAPT mutations associated with frontotemporal dementia promote tau fibril assembly; (2) cognitive performance in AD patients is inversely correlated with tau aggregate burden; (3) multimodal imaging of AD patients shows that tau aggregate deposition correlates with regional brain atrophy and functional impairment; (4) Injection of fibrillar, but not monomeric, tau produces tangles in mouse brains; (5) Tau exogenously added to cells must aggregate in order to “seed” intracellular tau pathology.

[0010] Thus, tau aggregation is a toxic gain of function in disease, and thus prevention of tau aggregation is expected to protect against neurodegeneration in AD and primary tauopathies.

[0011] Tau cleavage by AEP The microtubule-binding region (MBTR) of tau is a critical part of the molecule that nucleates fibril assembly. Structural studies have shown that the centrally located MBTR is normally covered by the N- and C-terminal regions of the molecule, thereby preventing tau-tau interactions in solution. Post-translational modifications such as phosphorylation and proteolytic cleavage open the tau molecule, exposing the MBTR and promoting aggregation. PHF tau isolated from AD brains contains a large proportion of truncated tau species, suggesting that tau cleavage is essential for the tau aggregation process.

[0012] The lysosomal cysteine ​​proteinase AEP / legumain 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 the AEP-derived tau fragment 1-368 in neurons is strongly neurotoxic, whereas overexpression of full-length tau is not. In addition to cleaving tau, AEP may indirectly affect tau aggregation by promoting tau phosphorylation: AEP is known to (indirectly) inhibit protein phosphatase 2A, a key enzyme regulating 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 overactivated in AD (Wang et al. Mol Cell 2017; Basurto-Islas et al. J Biol Chem 2013). Whether due to upregulation or overactivation of AEP, AEP-cleaved tau N368 fragments are enriched in AD brain tissue (Zhang et al. Nat Med 2014). Tau N368 / total tau ratio was significantly decreased in CSF from AD patients and strongly negatively correlated with 18F-GTP1 tau PET signal (Blennow et al. Brain 2020).

[0014] AEP may contribute to AD pathogenesis beyond promoting tau aggregation: the AEP cleavage fragment tau N368 was recently shown to enhance BACE1 expression and Aβ production via binding to the BACE1 transcription factor STAT1 (Zhang et al. Mol Psych 2018). Summary of the Invention [Problem to be solved by the invention]

[0015] The present invention relates to a compound of formula I: [ka] (In the formula, R 1 is halo or alkyl; R 2 is H or halo; R 3 is H or halo; R 4 is H or halo; Or, R 3 and R 4 forms a 3- 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 -C(O)- or -S(O) 2 - and; m is 0 or 1; n is 0 or 1) The present invention provides novel compounds of the formula: DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

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

[0017] The term "alkoxy" refers to a group in which R' is C 1~6 represents a group of the formula -O-R', which is an alkyl group. 1~6 Examples of -alkoxy groups include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy and tert-butoxy. Particular 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 site of acetylenic unsaturation (i.e., having at least one moiety of the formula C≡C). In some embodiments, unless otherwise specified, an alkynyl contains from 2 to 6 carbon atoms, or from 2 to 4 carbon atoms. Examples of alkynyl groups include, but are not limited to, ethynyl (or acetylenyl), prop-1-ynyl, prop-2-ynyl (or propargyl), but-1-ynyl, but-2-ynyl, and but-3-ynyl. A particular example is ethynyl.

[0019] The term "alkynylalkoxy" means 1~6 At least one hydrogen atom of the alkoxy group is C 2~6 C replaced by an alkynyl group 1~6Represents an alkoxy group. Particular examples are ethynylmethoxy, ethynylethoxy, prop-2-ynoxy, propynylmethoxy, propynylethoxy. Particular example is prop-2-ynoxy.

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

[0021] The term "cycloalkyl" refers to a monocyclic or polycyclic saturated or partially unsaturated non-aromatic hydrocarbon. In some embodiments, unless otherwise stated, a cycloalkyl contains 3 to 8 carbon atoms, 3 to 6 carbon atoms, or 3 to 5 carbon atoms. In some embodiments, a cycloalkyl is a saturated monocyclic or polycyclic hydrocarbon. In other embodiments, a cycloalkyl contains one or more double bonds (e.g., a cycloalkyl fused to an aryl or heteroaryl ring, or a non-aromatic monocyclic hydrocarbon containing one or two double bonds). Polycyclic cycloalkyl groups may contain spiro, fused, or bridged polycyclic moieties, each ring being a saturated or partially unsaturated non-aromatic hydrocarbon. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, octahydropentalenyl, spiro[3.3]heptanyl, and the like. Bicyclic means a ring system consisting of two saturated carbocyclic rings with two shared carbon atoms. Examples of monocyclic cycloalkyl are cyclopropyl, cyclobutanyl, cyclopentyl, cyclohexyl or cycloheptyl. Particular 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 iodo. Particular halogens are fluoro and chloro.

[0023] The term "haloalkoxy" means 1~6- at least one hydrogen atom of an alkoxy group is replaced by the same or different halogen atom 1~6 -represents an alkoxy group. Examples of haloalkoxy are difluoromethoxy, trifluoromethoxy, difluoroethoxy and trifluoroethoxy. A particular example is trifluoromethoxy.

[0024] The term "pharmaceutically acceptable salt" refers to a salt that retains the biological effectiveness and properties of the free base or free acid, and is not biologically or otherwise undesirable. Salts are formed with 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 from the addition of inorganic or organic bases to the free acid. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, and magnesium salts. Salts derived from organic bases include, but are not limited to, salts of 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, polyamine resins. Compounds of formula I can also exist in zwitterionic form. Particularly preferred pharma- ceutically acceptable salts of compounds of formula I are salts formed with formic acid and with hydrochloric acid to give hydrochloride, dihydrochloride or trihydrochloride salts.

[0025] The abbreviation uM means micromolar and is equivalent to the symbol μM.

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

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

[0028] The compounds of formula I may contain several asymmetric centers and may exist in the form of optically pure enantiomers, mixtures of enantiomers, such as racemates, optically pure diastereoisomers, mixtures of diastereoisomers, diastereomeric racemates or mixtures of diastereomeric racemates.

[0029] According to the Cahn-Ingold-Prelog rules, the asymmetric carbon atom can be of the "R" or "S" configuration.

[0030] Certain embodiments of the present invention also provide compounds of formula I as described herein and pharma- ceutically acceptable salts or esters thereof, particularly compounds of formula I as described herein and pharma- ceutically acceptable salts thereof, more particularly compounds of formula I as described herein.

[0031] An embodiment of the present invention comprises: R 1 is halo or alkyl; R 2 is H or halo; R 3 is H or halo; R 4 is H or halo; Or, R 3 and R 4 form a 3- 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 -C(O)- or -S(O) 2 - and; m is 0 or 1; Provided herein are compounds of formula I as described herein, and pharma- ceutically acceptable salts, wherein n is 0 or 1.

[0032] One embodiment of the present invention is 1 is halo.

[0033] One embodiment of the present invention is 5 is H, halo, or haloalkoxy.

[0034] Certain embodiments of the present invention provide compounds of formula I as described herein, wherein W is -C(O)-.

[0035] Certain embodiments of the present invention provide compounds of formula I as described herein, wherein m is 1 and n is 0.

[0036] One embodiment of the present invention is 3 and R 4 forms a 3- or 4-membered cycloalkyl ring.

[0037] An embodiment of the present invention comprises: R 1 is the halo; R 2 is H or halo; R 3 is the halo; R 4 is a halo; Or, R 3 and R 4 form a 3- or 4-membered cycloalkyl ring; R 5 is H, halo, or haloalkoxyl; W is -C(O)-; m is 1; Compounds of formula I as described herein, and pharma- ceutically acceptable salts, are provided wherein n is 0.

[0038] An embodiment of the present invention comprises: R 1 is the halo; R 2 is H or halo; R 3 and R 4 form a 3- or 4-membered cycloalkyl ring; R 5 is halo or haloalkoxyl; W is -C(O)-; m is 1; Compounds of formula I as described herein, and pharma- ceutically acceptable salts, are provided wherein n is 0.

[0039] An embodiment of the present invention comprises: R 1 is F or Cl; R 2 is H or F; R 3 and R 4 forms a three-membered cycloalkyl ring; R 5 is chloro or trifluoromethoxy; W is -C(O)-; m is 1; Compounds of formula I as described herein, and pharma- ceutically acceptable salts, are provided wherein 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-ynoxyphenyl)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; and pharma- ceutically 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-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; and pharma- ceutically acceptable salts thereof.

[0042] The most specific examples of formula I described herein are 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; and pharma- ceutically acceptable salts thereof.

[0043] The processes for preparing the compounds of formula I described herein are an object 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 R, W, n, and m are as defined above, can be obtained as shown in Scheme 1. 1 and R 2 The compound of formula II, wherein R 3 , R 4 , R 5 , R 6 , R 7, W, n, and m are as above, and depending on the nature of W, X is a suitable functional group selected from F, Cl, Br, OH, ON-succinimidyl (OSu). 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 for example in the art. When X=F, Cl, Br or OSu, the reaction can be carried out in the presence of a stoichiometric amount of a suitable chemically inert base, for example a tertiary amine such as triethylamine or diisopropylethylamine, in a suitable polar aprotic or non-polar solvent, for example dimethylformamide or dichloromethane. W=-S(O) 2 In the case of -, X may be selected from F, Cl, preferably Cl. The reaction is carried out in the presence of a stoichiometric amount of a suitable chemically inert base, for example a tertiary amine such as triethylamine or diisopropylethylamine, in a suitable polar aprotic or non-polar solvent, for example dimethylformamide or dichloromethane. [ka] Scheme 1

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

[0046] R 3 , R 4 , R 5 , R 6 , R 7 Intermediate compounds of formula III and IV, where W, n, and m are as defined above, are available by methods known in the art.

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

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

[0049] Another embodiment of the present invention provides pharmaceutical compositions or medicaments containing the compounds of the present invention and therapeutically inert carriers, diluents or excipients, as well as methods of using the compounds of the present invention to prepare such compositions and medicaments. In one example, the compounds of formula I can be formulated into galenic dosage forms by mixing at ambient temperature, at an appropriate pH, and to the desired degree of purity with a physiologically acceptable carrier, i.e., a carrier that is non-toxic to the recipient at the dosage and concentration used. The pH of the formulation will depend primarily on the particular application and the concentration of the compound, but is preferably anywhere in the range of about 3 to about 8. In one example, the compounds of formula I are formulated in acetate buffer at pH 5. In another embodiment, the compounds of formula I are sterile. The compounds can be stored, for example, as solid or amorphous compositions, as lyophilized formulations, or as aqueous solutions.

[0050] The compositions are formulated, dosed, and administered in a manner consistent with good medical practice. Factors to consider in this context include the particular disorder being treated, the particular mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the site of delivery of the agent, the method of administration, the administration schedule, and other factors known to the medical profession.

[0051] The compounds of the present invention may be administered by any suitable means, including oral, topical (including buccal and sublingual), rectal, vaginal, transdermal, parenteral, subcutaneous, intraperitoneal, intrapulmonary, intradermal, intrathecal and epidural and intranasal, and, if localized treatment is desired, intralesional administration. Parenteral administration includes intramuscular, intravenous, intraarterial, intraperitoneal or subcutaneous administration.

[0052] The compounds of the present invention may 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 of pharmaceutical preparations, such as diluents, carriers, pH adjusters, sweeteners, fillers, and additional active agents.

[0053] Typical formulations are prepared by mixing the compound of the present invention and carrier or excipient.Suitable carrier and excipient are well known to those skilled in the art and are described in detail in, 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, glidants, processing aids, colorants, sweeteners, fragrances, flavorings, diluents, and other known additives to provide for superior presentation of the drug (i.e., the compound of the present invention or a pharmaceutical composition thereof) or to aid in the manufacture of a pharmaceutical product (i.e., a medicament).

[0054] The compounds of formula I and their pharma- ceutically acceptable salts can be processed with pharma- ceutically inert, inorganic or organic adjuvants for the manufacture of tablets, coated tablets, sugar-coated tablets, hard gelatin capsules, injections or topical preparations.Lactose, corn starch or its derivatives, talc, stearic acid or its salts, etc., can be used as such adjuvants for tablets, sugar-coated tablets and hard gelatin capsules.

[0055] Suitable adjuvants for soft gelatine capsules are, for example, vegetable oils, waxes, fats, semisolid substances and liquid polyols etc.

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

[0057] Suitable adjuvants for injection solutions are, for example, water, alcohols, polyols, glycerol, vegetable oils, etc.

[0058] Suitable adjuvants for suppositories are, for example, natural or hardened oils, waxes, fats, semisolid or liquid polyols etc.

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

[0060] In addition, the pharmaceutical preparations may contain preservatives, solubilizers, viscosity-increasing substances, stabilizers, wetting agents, emulsifiers, sweeteners, colorants, flavorings, salts for varying the osmotic pressure, buffers, masking agents or antioxidants. They may also contain further therapeutically valuable substances.

[0061] The dosage can vary within a wide range and will of course be adapted to the individual requirements in each particular case. In general, for oral administration, a daily dosage of about 0.1 mg to 20 mg per kg of body weight, preferably about 0.5 mg to 4 mg per kg of body weight (e.g. about 300 mg per person), preferably divided into 1 to 3 individual administrations, which may, if appropriate, consist of, for example, equal amounts. For topical administration, the preparation may contain 0.001% to 15% by weight of medicament, and the required dose, which may be between 0.1 and 25 mg, may be administered either by a single administration per day or per week, or by multiple administrations (2 to 4 times) per day or by multiple administrations per week. However, it will be clear that the upper or lower limits given herein may be exceeded, where this is indicated.

[0062] The present invention also relates in particular to: A compound of formula I for use as a therapeutically active substance; A compound of formula I for use in the treatment of a disease modulated by AEP.

[0063] One embodiment of the invention is the use of a compound of formula I to treat or prevent 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 linked to chromosome 17 (FTDP-17), Ritiko-Bodig disease (Parkinson-Dementia Complex of Guam), gangliogliomas and gangliocytomas, meningioangiomatosis, postencephalitic parkinsonism, subacute sclerosing panencephalitis (SSPE), Pick's disease, or corticobasal degeneration.

[0064] One embodiment of the invention is the use of a compound of formula I to treat or prevent 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 invention is the use of a compound of formula I for treating or preventing Alzheimer's disease.

[0066] An embodiment of the invention is the use of a compound of formula I for the preparation of 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 linked to chromosome 17 (FTDP-17), Ritiko-Bodig disease (Parkinson-Dementia Complex of Guam), gangliogliomas and gangliocytomas, meningioangiomatosis, postencephalitic parkinsonism, subacute sclerosing panencephalitis (SSPE), Pick's disease, or corticobasal degeneration.

[0067] An embodiment of the invention is the use of a compound of formula I for the preparation of 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 invention is the use of a compound of formula I for the preparation of a medicament for treating or preventing Alzheimer's disease.

[0069] An embodiment of the 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 linked to chromosome 17 (FTDP-17), Ritiko-Bodig disease (Parkinson-Dementia Complex of Guam), ganglioglioma and gangliocytoma, meningioangiomatosis, postencephalitic parkinsonism, subacute sclerosing panencephalitis (SSPE), Pick's disease, or corticobasal degeneration.

[0070] One embodiment of the 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] Certain embodiments of the invention are methods 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 linked to chromosome 17 (FTDP-17), Ritiko-Bodig disease (Parkinson-Dementia Complex of Guam), gangliogliomas and gangliocytomas, meningioangiomatosis, postencephalitic parkinsonism, subacute sclerosing panencephalitis (SSPE), Pick's disease, or corticobasal degeneration.

[0073] One embodiment of the 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 invention is a method for treating or preventing Alzheimer's disease.

[0075] An embodiment of the 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 linked to chromosome 17 (FTDP-17), Ritiko-Bodig disease (Parkinson-Dementia Complex of Guam), gangliogliomas and gangliocytomas, meningioangiomatosis, postencephalitic parkinsonism, subacute sclerosing panencephalitis (SSPE), Pick's disease, or corticobasal degeneration, comprising administering to a patient in need thereof an effective amount of a compound of formula I.

[0076] An embodiment of the 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 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] Certain embodiments of the present invention also provide compounds of formula I as described herein when prepared according to any one of the processes described.

[0079] Also an object of the present invention is a pharmaceutical composition comprising a compound of formula I as described herein and a therapeutically inert carrier.

[0080] The invention will now be illustrated by the following examples, which have no limiting character.

[0081] Where the preparations are obtained as mixtures of enantiomers, epimers and / or diastereoisomers, the pure enantiomers may be obtained by methods known to those skilled in the art, such as, for example, chiral chromatography or crystallization.

[0082] Abbreviation The following abbreviations were used in the experimental section: 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 = azabenzotriazole tetramethyluronium hexafluorophosphate.

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

[0084] Intermediates 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] tert-Butyl carbazate (49761 mg, 376 mmol) and N,N-diisopropylethylamine (65 mL, 376 mmol) 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 h. After cooling, the mixture was filtered and the filtrate was concentrated to give 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 in vacuo to give the title compound as a colorless oil (60.00 g, 214 mmol, 65% yield). 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 formed. The mixture was stirred at 25° C. for 5 h. The mixture was then filtered and the solid was washed with ethyl acetate (200 mL) and dried in vacuum to give the title compound as a white solid (42.0 g, 194 mmol, 94% yield). 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)prop-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 h. The reaction was then diluted with cold water (30 mL) and extracted with dichloromethane (2×15 mL). The combined organic layers were washed with brine (20 mL), dried over sodium sulfate and concentrated in vacuo to give a yellow oil. The crude product was purified by column chromatography (silica gel, 25 g, petroleum ether / ethyl acetate, gradient 10:1 to 1:2 (v / v)) to give the title compound as a yellow oil (1800 mg, 4.37 mmol, 86% yield). 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-oxo-ethyl)amino]carbamoyl]pyrrolidine-1-carboxylate (Int-4, 10.0 g, 24.3 mmol) in ammonia in methanol (7N, 10 mL, 700 mmol) was stirred at 25 °C for 14 h. The mixture was then concentrated to give a pale yellow oil. The pale yellow oil was dissolved in water (100 mL) and extracted with methyl tert-butyl ether (5 x 70 mL, the whole extraction process needed to stand for 20 min for emulsification, and the addition of a small amount of EtOH was beneficial for the separation). The aqueous solution was lyophilized to give the title 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). A solution of fluoroacetyl chloride (1645 mg, 17.0 mmol) in dichloromethane (15 mL) was then added dropwise at -78°C. The mixture was stirred at -78°C for 0.5 h. The mixture was warmed to 25°C and stirred for 1.5 h. It was then concentrated in vacuo to give the crude title 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 on charcoal (10%, 800 mg) was added. The resulting mixture was stirred at 25° C. for 4 h under a hydrogen atmosphere (hydrogen balloon). The mixture was then filtered through a celite pad, washed with methanol (50 mL) and the filtrate was concentrated in vacuo to give 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 [ka]

[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-t-butyl dicarbonate (4088 mg, 18.7 mmol) was added followed by triethylamine (4.73 g, 46.8 mmol). The mixture was stirred at 25° C. for 16 h. The reaction mixture was then 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 on charcoal (10% m / m, 100 mg) in tetrahydrofuran (60 mL) was stirred under a hydrogen atmosphere (balloon) at 25° C. for 48 h. The mixture was then filtered through a pad of celite, washed with tetrahydrofuran (50 mL) and the filtrate was concentrated in vacuo to give the title 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 h. The reaction mixture was then purified by reverse phase chromatography (Phenomenex luna C18 150*25 mm*10 um, eluent (water + 0.225% formic acid) / acetonitrile). After lyophilization, the title compound was obtained as a white solid (100 mg, 0.190 mmol, 46% yield). 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 h. The reaction mixture was then concentrated in vacuo to give the crude title 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-prolinate 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). Pyridine (3.1 g, 3.2 mL, 39.6 mmol) was then added, followed by 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.-23° C. for 3 h. The reaction mixture was then poured into ice-cold aqueous citric acid 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 in vacuo. The residue was purified by column chromatography (silica gel, 80 g, n-heptane / ethyl acetate, gradient 100:0 to 20:80 (v / v)) to give, after concentration of the product-containing fractions, the title compound as a white solid (2.55 g, 6.79 mmol, 86% yield). 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 h. The mixture was slowly acidified by addition of aqueous citric acid (5% m / m, to pH 3-4), extracted with ethyl acetate (2 x 20 mL) and the combined organic layers were dried over sodium sulfate and concentrated in vacuo to give the title 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 stirring was continued at 0° C. for 1 h. Then ethylaminoglycinate hydrochloride (85.6 mg, 553 μmol) and N,N-diisopropylethylamine (95 mg, 125 μl, 719 μmol) were added. The reaction mixture was allowed to warm to room temperature (23° C.) and stirred for 30 min. It was then poured into water (5 mL) and extracted with ethyl acetate (2×10 mL). The combined organic layers were washed with saturated aqueous sodium bicarbonate (20 mL), dried over sodium sulfate and concentrated in vacuo. The crude material was purified by column chromatography (silica gel, 12 g, n-heptane / ethyl acetate, gradient 100:0 to 0:100 (v / v)) to give the title 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)hydrazinyl)acetamide (Int-18)

[0114] A solution of ethyl (S)-(1-((4'-methoxy-[1,1'-biphenyl]-4-yl)sulfonyl)pyrrolidine-2-carboxamido)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 h and at room temperature (23° C.) for 15 h. The reaction mixture was then concentrated in vacuo, 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 in vacuo to give the title 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 h. It was then directly purified by preparative HPLC (Shim-pack C18 150*25*10 um, eluent (water + 0.225% formic acid) / acetonitrile, gradient 76:24 to 56:44 within 10 min). After lyophilization, the title compound was obtained as a white solid (30 mg, 0.07 mmol, 16% yield). 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 h. It was then directly purified by preparative HPLC (Phenomenex luna C18 150*25 mm*10 um, eluent (water + 0.225% formic acid) / acetonitrile, gradient 81:19 to 51:49 within 11 min). After lyophilization, the title compound was isolated as a white solid (13 mg, 0.03 mmol, 17% yield). 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 um, eluent (water + 0.225% formic acid) / acetonitrile). After lyophilization, the title compound was obtained as a white solid (22 mg, 0.05 mmol, 18% yield). 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. It was then directly purified by preparative HPLC (Phenomenex luna C18 150*25 mm*10 um, eluent (water + 0.225% formic acid) / acetonitrile). After lyophilization, the title compound was obtained as a white solid (23 mg, 0.05 mmol, 21% yield). 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 h. It was then directly purified by preparative HPLC (Phenomenex luna C18 150*25 mm*10 um, eluent (water + 0.225% formic acid) / acetonitrile, gradient 88:12 to 58:42). After lyophilization, the title compound was isolated as a white solid (22 mg, 0.06 mmol, 30% yield). 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 h. It was then directly purified by preparative HPLC (Phenomenex luna C18 150*25 mm*10 um, eluent (water + 0.225% formic acid) / acetonitrile, gradient 73:27 to 43:57 within 10 min). After lyophilization, the title compound was obtained as a white solid (18 mg, 0.04 mmol, 21% yield). 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 benzenesulfonyl 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 um, eluent (water + 0.225% formic acid) / acetonitrile). After lyophilization, the title compound was obtained as a white solid (29 mg, 0.08 mmol, 37% yield). 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 h. It was then directly purified by preparative HPLC (Phenomenex luna C18 150*25 mm*10 um, eluent (water + 0.225% formic acid) / acetonitrile). After lyophilization, the title compound was isolated as a white solid (23 mg, 0.06 mmol, 29% yield). 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 um, eluent (water + 0.225% formic acid) / acetonitrile). After lyophilization, the title compound was obtained as a white solid (32 mg, 0.08 mmol, 39% yield). 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 um, eluent (water + 0.225% formic acid) / acetonitrile). After lyophilization, the title compound was isolated as a white solid (40 mg, 0.10 mmol, 24% yield). 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. It was then directly purified by preparative HPLC (Phenomenex luna C18 150*25 mm*10 um, eluent (water + 0.225% formic acid) / acetonitrile). After lyophilization, the title compound was obtained as a white solid (100 mg, 0.24 mmol, 59% yield). 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 h. Then, additional portions 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 h. It was then concentrated in vacuo and the residue was purified by silica gel flash chromatography (12 g, ethyl acetate / n-heptane, gradient 0:100 to 100:0, then methanol / ethyl acetate, gradient 0:100 to 10:90) to give the title compound as a white solid (34 mg, 85% yield). 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)hydrazinyl)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 min. Then, a further portion of a solution 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) was added and stirring was continued at 0° C. for 30 min. It was then diluted with dichloromethane (5 mL) and washed with water (5 mL), the organic layer was dried over sodium sulfate and concentrated in vacuo. The residue was purified by silica gel flash chromatography (12 g, ethyl acetate / n-heptane, gradient 0:100 to 100:0, then methanol / ethyl acetate, gradient 0:100 to 10:90) to give, after concentration of the product-containing fractions, the title compound as a white solid (54 mg, 95% yield). MS (ESI) m / z: 493.2 [M+H] + .

[0141] Example 14 2-[(2-Fluoroacetyl)-[[(2S)-1-[1-(4-prop-2-ynoxyphenyl)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-ynoxyphenyl)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 h. It was then directly purified by preparative HPLC (Phenomenex luna C18 150*25 mm*10 um, eluent (water + 0.225% formic acid) / acetonitrile, gradient 83:17 to 50:50 within 11 min). After lyophilization, the title 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-fluoro-acetyl)-[[(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)hydrazinyl)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 min. Then, further portions 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 stirring was continued for 15 min at 0° C. It was then diluted with dichloromethane (5 mL), washed with water (5 mL) and the organic layer was dried over sodium sulfate and concentrated in vacuum. The residue was purified by silica gel flash chromatography (12 g, ethyl acetate / n-heptane, gradient 0:100 to 100:0, then methanol / ethyl acetate, gradient 0:100 to 10:90) to give, after concentration of the product-containing fractions, the title compound as a white solid (35 mg, 64% yield). MS (ESI) m / z: 527.1 [M+H] + .

[0145] Example 16 2-[((2R)&(2S)-2-chloro-2-fluoro-acetyl)-[[(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 h. Then, additional portions 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 h. It was then concentrated in vacuo and the residue was purified by silica gel flash chromatography (25 g, ethyl acetate / n-heptane, gradient 0:100 to 100:0, then methanol / ethyl acetate, gradient 0:100 to 10:90) to give the title compound as a white solid (133 mg, 99% yield). 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)hydrazinyl)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 min. It was then diluted with dichloromethane (5 mL), washed with water (5 mL) and the organic layer was dried over sodium sulfate and concentrated in vacuo. The residue was purified by silica gel flash chromatography (12 g, ethyl acetate / n-heptane, gradient 0:100 to 100:0, then methanol / ethyl acetate, gradient 0:100 to 20:80) to give, after concentration of the product-containing fractions, the title compound as a white solid (55 mg, 91% yield). 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 min. Then, another portion of a solution 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) was added and stirring was continued at 0° C. for 30 min. It was then concentrated in vacuo and the residue was purified by silica gel flash chromatography (12 g, ethyl acetate / n-heptane, gradient 0:100 to 100:0, then methanol / ethyl acetate, gradient 0:100 to 10:90) to give the title compound as a white solid (36 mg, 87% yield). 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 h. It was then concentrated in vacuo 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 to 98:2) to give the title compound as a white solid (12 mg, 37% yield). 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 h. It was then concentrated in vacuo 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 to 98:2) to give the title compound as a white solid (17 mg, 35% yield). MS(ESI)m / z:505.3[M+H] + .

[0155] AEP Activity Enzyme Assay Procedure: The assay uses a fluorogenic substrate, Z-Ala-Ala-Asn-Rh110-(D-Pro) (Biosyntan, Berlin, Ord.Nr.80773), which fluoresces when cleaved by legumain and can be monitored upon excitation at 492 nm, emission at 530 nm. Prior to the assay measurement, legumain (in-house, construct name: hLGMN(V18-Y433)_VD-8xHis-S2-r) is activated by diluting (11-fold) from 1.1 mg / ml to 0.1 mg / ml in activation buffer (50 mM sodium acetate, 100 mM NaCl, pH 4.0) and incubating for 2 hours at 37 °C. The activated enzyme solution stock is stored at -80 °C. For the measurement, the following solutions are then dispensed into a 384-well microplate (Corning384 untreated, black / clear, catalog number: 3540): Negative controls included 8.5 uL of assay buffer (20 mM citric acid, 60 mM Na2HPO4, 1 mM EDTA, 0.1% CHAPS, pH 5.8, 0.5 mM TCEP (freshly added)) or 8.5 uL of activated enzyme solution (0.5 nM legumain (final) in assay buffer (20 mM citric acid, 60 mM Na2HPO4, 1 mM EDTA, 0.1% CHAPS, pH 5.8, 0.5 mM TCEP (freshly added)) and 1.5 uL of pre-diluted compound solution (100 uM to 0.1 nM, 1% DMSO in water (final)). After 30 min incubation at 25° C., 5 uL of substrate solution (0.5 uM Z-Ala-Ala-Asn-Rh110-(D-Pro) (final)) in assay buffer (20 mM citric acid, 60 mM Na2HPO4, 1 mM EDTA, 0.1% CHAPS, pH 5.8, 0.5 mM TCEP (freshly added)) is added and fluorescence is measured in a plate reader (first 30 min is analyzed) using, for example, a PHERAstar or Spectramax instrument in kinetic mode, with excitation at 490 nm and emission at 530 nm.

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

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

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

Claims

1. Formula I 【Chemical 1】 (wherein R １ is halo or alkyl; R ２ is H or halo; R ３ is H or halo; R ４ is H or halo; Alternatively, R ３ and R ４ form a 3- 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 -C(O)- or -S(O)- ２ -; m is 0 or 1; n is 0 or 1) compounds, and pharmaceutically acceptable salts thereof.

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

3. R ５ The compound according to claim 1, wherein R 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 R and R form a 3- or 4-membered cycloalkyl ring.

7. R １ is a halo; R ２ is H or halo; R ３ is a halo; R ４ is a halo; Alternatively, R ３ and R ４ form a 3- or 4-membered cycloalkyl ring; R ５ is H, halo or haloalkoxyl; W is -C(O)-; m is 1; n is 0, The compound according to claim 1, and pharmaceutically acceptable salts thereof.

8. R １ is a halo; R ２ is H or halo; R ３ and R ４ form a 3- or 4-membered cycloalkyl ring; R ５ is a halo or haloalkoxyl; W is -C(O)-; m is 1; n is 0, The compound according to claim 1, and pharmaceutically acceptable salts thereof.

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

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-ynyloxyphenyl)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; The compound according to claim 1, selected from the group consisting of: 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; The compound according to claim 1, selected from the group consisting of: 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; The compound according to claim 1, selected from the group consisting of, and a pharmaceutically acceptable salt thereof.

13. A method for preparing a 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 【Chemical Formula 2】 (wherein 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, O-N-succinimidyl (OSu)) A method comprising.

14. A method for preparing a 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 【Chemical Formula 3】 (wherein 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, O-N-succinimidyl (OSu)) A method comprising.

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

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

17. A pharmaceutical composition comprising a compound according to any one of claims 1 to 12 and a therapeutically inert 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), Lytico-Bodig disease (Parkinson's dementia complex of Guam), ganglioglioma and gangliocytoma, meningovascular angiomatosis, postencephalitic parkinsonism, subacute sclerosing panencephalitis (SSPE), Pick's disease, or corticobasal degeneration.

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

20. Use of the compound 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 linked to chromosome 17 (FTDP-17), Lytico-Bodig disease (Parkinson-dementia complex of Guam), ganglioglioma and ganglionocytoma, meningeal angiomatosis, postencephalitic 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. The 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 linked to chromosome 17 (FTDP-17), Lytico-Bodig disease (Parkinson-dementia complex of Guam), ganglioglioma and ganglionocytoma, meningeal angiomatosis, postencephalitic parkinsonism, subacute sclerosing panencephalitis (SSPE), Pick's disease, or corticobasal degeneration.

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

Images