Tetrahydropyranoxazine derivatives with selective BACE1 inhibitory activity

Abstract

The present invention provides a compound having an effect of inhibiting the production of β-amyloid protein, especially an effect of inhibiting BACE1, and the compound can be used as a therapeutic agent or preventive agent for diseases induced by the production, secretion and / or deposition of β-amyloid protein. A compound of formula (I), wherein R 3 Each independently is an alkyl group optionally substituted by halogen, cyano, alkoxy, haloalkoxy or non-aromatic carbocyclic group, etc.; t is an integer of 0-3; R 5 is a hydrogen atom or a halogen; R 6 is selected from a hydrogen atom, a halogen and a substituted or unsubstituted alkyl group; Ring B is a substituted or unsubstituted pyrazine, etc.; or a pharmaceutically acceptable salt thereof.

Description

Technical Field

[0001] The present invention relates to a compound having an activity of inhibiting the production of β-amyloid protein (amyloid β) and useful as an agent for treating or preventing a disease induced by the production, secretion and / or deposition of β-amyloid protein. Background Art

[0002] A peptide consisting of about 40 amino acid residues called β-amyloid is widely observed in the brains of Alzheimer's patients, and accumulates outside nerve cells to form insoluble particles (senile specks). These senile specks are believed to kill nerve cells and cause Alzheimer's disease, and therefore, therapeutic agents for Alzheimer's disease, such as degraders of β-amyloid and amyloid vaccines, are being studied.

[0003] Secretase is an enzyme that cuts off a protein called β-amyloid precursor protein (APP) in cells and generates β-amyloid protein. The enzyme that controls the generation of the N-terminus of β-amyloid protein is called β-secretase (β-site APP-cleaving enzyme 1, BACE1). Inhibition of this enzyme is believed to lead to a reduction in the production of β-amyloid protein, and due to this inhibition, a therapeutic or preventive agent for Alzheimer's disease will be produced.

[0004] Patent documents 1 to 10 disclose compounds having structures similar to the compounds of the present invention. Each of these documents discloses compounds that can be used as therapeutic agents for Alzheimer's disease, Alzheimer's-related symptoms, diabetes, etc., but each substantially disclosed compound has a structure different from the compound of the present invention.

[0005] Citation List

[0006] Patent Literature

[0007] .

[0008] Brief description of the invention

[0009] Technical issues

[0010] The present invention provides compounds having an effect of reducing the production of β-amyloid protein, particularly selective BACE1 inhibitory activity, and useful as agents for treating diseases induced by the production, secretion and / or deposition of β-amyloid protein.

[0011] Advantageous Effects of the Invention

[0012] The compound of the present invention has selective BACE1 inhibitory activity and is useful as an agent for treating and / or preventing diseases induced by the production, secretion or deposition of β-amyloid protein, such as Alzheimer dementia.

[0013] Technical solutions to the problem

[0014] The present invention provides, for example, inventions described in the following items.

[0015] (1) A compound of formula (I) or a pharmaceutically acceptable salt thereof:

[0016] [Chemical formula 1]

[0017]

[0018] in

[0019] R 3 Each is independently an alkyl group optionally substituted by one or more groups selected from halogen, cyano, alkoxy, haloalkoxy and non-aromatic carbocyclic group; or a heterocyclic group optionally substituted by an alkyl group;

[0020] Two R's attached to the same carbon atom 3 Together with the carbon atoms to which they are attached, they may form a 3- to 5-membered non-aromatic carbocyclic ring, wherein the 3- to 5-membered non-aromatic carbocyclic ring is optionally substituted with one or more groups selected from halogen, alkyl and haloalkyl;

[0021] t is an integer from 0 to 3;

[0022] R 5 is a hydrogen atom or a halogen;

[0023] R 6 is a hydrogen atom, a halogen, or a substituted or unsubstituted alkyl group;

[0024] [Chemical formula 2]

[0025]

[0026] Where R 7a is halogen; cyano; alkoxy optionally substituted by one or more groups selected from cyano, halogen, hydroxyl, non-aromatic carbocyclic group and aromatic heterocyclic group; alkyl optionally substituted by one or more halogen; non-aromatic carbocyclic group optionally substituted by one or more groups selected from cyano and halogen; non-aromatic heterocyclic group optionally substituted by one or more groups selected from cyano and aromatic heterocyclic group; alkenyloxy optionally substituted by one or more groups selected from cyano, halogen and hydroxyl; alkynyloxy optionally substituted by one or more groups selected from cyano, halogen and hydroxyl; or aromatic heterocyclic group optionally substituted by one or more alkyl; and

[0027] R 7b It is a hydrogen atom, a halogen, an alkyl group, a halogenated alkyl group or an amino group.

[0028] (1) A compound of formula (I) or a pharmaceutically acceptable salt thereof:

[0029] [Chemical formula 3]

[0030]

[0031] in

[0032] R 3 are each independently selected from an alkyl group optionally substituted by halogen, cyano, alkoxy, haloalkoxy or a non-aromatic carbocyclic group; and an aromatic heterocyclic group optionally substituted by an alkyl group;

[0033] t is an integer from 0 to 3;

[0034] R 5 is a hydrogen atom or a halogen;

[0035] R 6 is a hydrogen atom, a halogen, or a substituted or unsubstituted alkyl group;

[0036] [Chemical formula 4]

[0037]

[0038] Where R 7a is halogen; cyano; alkoxy optionally substituted by cyano, halogen, hydroxyl or aromatic heterocyclic group; alkyl optionally substituted by halogen; non-aromatic carbocyclic group optionally substituted by cyano or halogen; non-aromatic heterocyclic group optionally substituted by cyano or aromatic heterocyclic group; aromatic heterocyclic group; and

[0039] R 7b It is a hydrogen atom, a halogen, an alkyl group, a halogenated alkyl group or an amino group.

[0040] (2) The compound according to item (1) or (1)' or a pharmaceutically acceptable salt thereof, wherein

[0041] [Chemical formula 5]

[0042] .

[0043] (2) The compound according to item (1) or (1) or a pharmaceutically acceptable salt thereof, wherein

[0044] [Chemical formula 6]

[0045] .

[0046] (3) A compound according to any one of the following items or a pharmaceutically acceptable salt thereof: (1), (2), (2)' and (1)', wherein R 6 It is fluorine or chlorine.

[0047] (3-2) A compound according to any one of the following items or a pharmaceutically acceptable salt thereof: (1), (2), (2)' and (1)', wherein R 6 It's fluorine.

[0048] (4) A compound according to any one of the following items or a pharmaceutically acceptable salt thereof: (1) to (3), (1)', (2)' and (3-2), wherein R 5 It is a hydrogen atom or a fluorine atom.

[0049] (4-2) The compound according to item (4) or a pharmaceutically acceptable salt thereof, wherein R 5 It's a hydrogen atom.

[0050] (5) A compound according to any one of the following items or a pharmaceutically acceptable salt thereof: (1) to (4), (1)', (2)', (3-2) and (4-2), wherein R 3 Each is independently alkyl optionally substituted with one or more halogens.

[0051] (5-2) The compound according to item (5) or a pharmaceutically acceptable salt thereof, wherein R 3 It's methyl.

[0052] (5-3) The compound according to item (5) or a pharmaceutically acceptable salt thereof, wherein R 3 It is a halomethyl group.

[0053] (6) A compound according to any one of the following items or a pharmaceutically acceptable salt thereof: (1) to (5), (1)', (2)', (3-2), (4-2), (5-2) and (5-3), wherein

[0054] [Chemical formula 7]

[0055] .

[0056] (7) A compound according to any one of the following items or a pharmaceutically acceptable salt thereof: (1) to (6), (1)', (2)', (3-2), (4-2), (5-2) and (5-3), wherein R 7a is cyano or alkoxy optionally substituted by one or more halogens and R 7b It is a hydrogen atom, a halogen or an amino group.

[0057] (7-2) The compound according to item (7) or a pharmaceutically acceptable salt thereof, wherein R 7a is methoxy optionally substituted by one or more halogens.

[0058] (7-3) The compound according to item (7) or a pharmaceutically acceptable salt thereof, wherein R 7a It is a C1-C2 alkoxy group.

[0059] (7-4) The compound according to item (7) or a pharmaceutically acceptable salt thereof, wherein R 7a It is a C1-C2 alkoxy group substituted by one or more halogens.

[0060] (8) A compound according to any one of the following items or a pharmaceutically acceptable salt thereof: (1) to (5), (1)', (2)', (3-2), (4-2), (5-2) and (5-3), wherein

[0061] [Chemical formula 8]

[0062] .

[0063] (8-2) A compound according to any one of the following items or a pharmaceutically acceptable salt thereof: (1) to (5), (1)', (2)', (3-2), (4-2), (5-2), (5-3) and (8), wherein R 7a is an alkyl group optionally substituted by one or more halogens and R 7b is a hydrogen atom or an alkyl group.

[0064] (9) A compound according to any one of the following items or a pharmaceutically acceptable salt thereof: (1) to (8), (1)', (2)', (3-2), (4-2), (5-2), (5-3), (7-2), (7-3), (7-4) and (8-2), wherein

[0065] [Chemical formula 9]

[0066]

[0067] Where R 3 is alkyl optionally substituted by one or more halogens,

[0068] R 5 is a hydrogen atom,

[0069] R 6 It is halogen.

[0070] [Chemical formula 10]

[0071]

[0072] and R 7a is alkoxy optionally substituted by one or more halogens.

[0073] (9-2) The compound according to any one of the following items: (1) to (9), (1)', (2)', (3-2), (4-2), (5-2), (5-3), (7-2), (7-3), (7-4) and (8-2), or a pharmaceutically acceptable salt thereof, wherein the compound is represented by formula (IA):

[0074] [Chemical formula 11]

[0075]

[0076] Each symbol is the same as defined in item (1) above.

[0077] (9-2)' A compound represented by formula (IB) or a pharmaceutically acceptable salt thereof:

[0078] [Chemical formula 12]

[0079]

[0080] wherein each symbol is the same as defined in any of the above items (1) to (9), (1)', (2)', (3-2), (4-2), (5-2), (5-3), (7-2), (7-3), (7-4) and (8-2).

[0081] (9-3) The compound according to any one of the following items: (1) to (9), (1)', (2)', (3-2), (4-2), (5-2), (5-3), (7-2), (7-3), (7-4), (9-2)' and (8-2), or a pharmaceutically acceptable salt thereof, wherein the compound is represented by formula (IA):

[0082] [Chemical formula 13]

[0083]

[0084] Where R 7a is a C1-C3 alkoxy group optionally substituted by one or more halogens; R 3 is C1-C3 alkyl optionally substituted by one or more halogens; and t is 0 or 1.

[0085] (9-4) A compound according to any one of the following items or a pharmaceutically acceptable salt thereof: (9), (9-2), (9-2)′ and (9-3), wherein R 7a It is a C1-C2 alkoxy group.

[0086] (9-5) A compound according to any one of the following items or a pharmaceutically acceptable salt thereof: (9), (9-2), (9-2)′ and (9-3), wherein R 7a It is a C1-C2 alkoxy group substituted by one or more halogens.

[0087] (10) A compound according to any one of the following items or a pharmaceutically acceptable salt thereof: (1) to (9), (9-2)', (1)', (2)', (3-2), (4-2), (5-2), (5-3), (7-2), (7-3), (7-4), (8-2), (9-2), (9-3), (9-4) and (9-5), which is selected from:

[0088] [Chemical formula 14]

[0089] .

[0090] (10) The compound according to the above (9-2) or a pharmaceutically acceptable salt thereof, which is selected from:

[0091] [Chemical formula 15]

[0092] .

[0093] (10)''A compound according to any one of the following items or a pharmaceutically acceptable salt thereof: (1) to (9), (1)', (2)', (3-2), (4-2), (5-2), (5-3), (7-2), (7-3), (7-4), (8-2), (9-2), (9-2)', (9-3), (9-4) and (9-5), which is selected from: I-007, I-008, I-009, I-010, I-047, I-059, I-067 and I-076.

[0094] (10)''' The compound according to the above item (10)' or a pharmaceutically acceptable salt thereof, which is selected from the optical isomers of any one of the compounds described in the above "Chemical Formula 15".

[0095] (11) A pharmaceutical composition comprising a compound according to any one of the following items or a pharmaceutically acceptable salt thereof: (1) to (10), (1)', (2)', (10)', (10)'', (3-2), (4-2), (5-2), (5-3), (7-2), (7-3), (7-4), (8-2), (9-2), (9-2)', (9-3), (9-4) and (9-5).

[0096] (12) A pharmaceutical composition having BACE1 inhibitory activity, comprising the compound according to item (11) or a pharmaceutically acceptable salt thereof.

[0097] (13) A pharmaceutical composition according to item (11) or (12), for treating or preventing Alzheimer's disease, mild cognitive impairment or prodromal Alzheimer's disease, for preventing the progression of Alzheimer's disease, mild cognitive impairment or prodromal Alzheimer's disease, or for preventing the progression of Alzheimer's disease in an asymptomatic patient at risk of developing Alzheimer's disease.

[0098] (14) A compound or a pharmaceutically acceptable salt thereof according to any one of the following items: (1) to (10), (1)', (2)', (10)', (10)'', (3-2), (4-2), (5-2), (5-3), (7-2), (7-3), (7-4), (8-2), (9-2), (9-2)', (9-3), (9-4) and (9-5), for use in a method for inhibiting BACE1 activity.

[0099] (15) A compound according to any one of the following items or a pharmaceutically acceptable salt thereof: (1) to (10), (1)', (2)', (10)', (10)'', (10)''', (3-2), (4-2), (5-2), (5-3), (7-2), (7-3), (7-4), (8-2), (9-2), (9-2)', (9-3), (9-4) and (9-5), for use in treating or preventing Alzheimer's disease, mild cognitive impairment or prodromal Alzheimer's disease, for preventing the progression of Alzheimer's disease, mild cognitive impairment or prodromal Alzheimer's disease, or for preventing the progression of asymptomatic patients at risk of developing Alzheimer's disease.

[0100] (16) A method for inhibiting BACE1 activity, comprising administering a compound according to any one of the following items or a pharmaceutically acceptable salt thereof: (1) to (10), (1)', (2)', (10)', (10)'', (3-2), (4-2), (5-2), (5-3), (7-2), (7-3), (7-4), (8-2), (9-2), (9-2)', (9-3), (9-4) and (9-5).

[0101] (17) A method for treating or preventing Alzheimer's disease, mild cognitive impairment or prodromal Alzheimer's disease, for preventing the progression of Alzheimer's disease, mild cognitive impairment or prodromal Alzheimer's disease, or for preventing the progression of Alzheimer's disease in an asymptomatic patient at risk of developing Alzheimer's disease, comprising administering a compound according to any one of the following items or a pharmaceutically acceptable salt thereof: (1) to (10), (1)', (2)', (10)', (10)'', (3-2), (4-2), (5-2), (5-3), (7-2), (7-3), (7-4), (8-2), (9-2), (9-2)', (9-3), (9-4) and (9-5).

[0102] (18) A BACE1 inhibitor comprising a compound according to any one of the following items or a pharmaceutically acceptable salt thereof: (1) to (10), (1)', (2)', (10)', (10)'', (3-2), (4-2), (5-2), (5-3), (7-2), (7-3), (7-4), (8-2), (9-2), (9-2)', (9-3), (9-4) and (9-5).

[0103] (19) Use of a compound according to any one of the following items or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for inhibiting BACE1 activity: (1) to (10), (1)', (2)' (10)', (10)'', (3-2), (4-2), (5-2), (5-3), (7-2), (7-3), (7-4), (8-2), (9-2), (9-2)', (9-3), (9-4) and (9-5).

[0104] (20) The pharmaceutical composition according to item (11) or (12), which is used for treating or preventing a disease induced by the production, secretion or deposition of β-amyloid protein.

[0105] (21) A method for treating or preventing a disease induced by the production, secretion or deposition of β-amyloid protein, comprising administering a compound according to any one of the following items or a pharmaceutically acceptable salt thereof: (1) to (10), (1)', (2)', (10)', (10)'', (3-2), (4-2), (5-2), (5-3), (7-2), (7-3), (7-4), (8-2), (9-2), (9-2)', (9-3), (9-4) and (9-5).

[0106] (22) A compound according to any one of the following items or a pharmaceutically acceptable salt thereof: (1) to (10), (1)', (2)', (10)', (10)'', (3-2), (4-2), (5-2), (5-3), (7-2), (7-3), (7-4), (8-2), (9-2), (9-2)', (9-3), (9-4) and (9-5), for use in treating or preventing a disease induced by the production, secretion or deposition of β-amyloid protein.

[0107] (23) Use of a compound according to any one of the following items or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for treating or preventing a disease induced by the production, secretion or deposition of β-amyloid protein: (1) to (10), (1)', (2)', (10)', (10)'', (3-2), (4-2), (5-2), (5-3), (7-2), (7-3), (7-4), (8-2), (9-2), (9-2)', (9-3), (9-4) and (9-5).

[0108] (24) The pharmaceutical composition according to item (11) or (12), which is used for treating or preventing Alzheimer's disease.

[0109] (25) A method for treating or preventing Alzheimer's disease, comprising administering a compound according to any one of the following items or a pharmaceutically acceptable salt thereof: (1) to (10), (1)', (2)', (10)', (10)'', (3-2), (4-2), (5-2), (5-3), (7-2), (7-3), (7-4), (8-2), (9-2), (9-2)', (9-3), (9-4) and (9-5).

[0110] (26) A compound or a pharmaceutically acceptable salt thereof according to any one of the following items: (1) to (10), (1)', (2)', (10)', (10)'', (3-2), (4-2), (5-2), (5-3), (7-2), (7-3), (7-4), (8-2), (9-2), (9-2)', (9-3), (9-4) and (9-5), for use in treating or preventing Alzheimer's disease.

[0111] (27) Use of a compound according to any one of the following items or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for treating or preventing Alzheimer's disease: (1) to (10), (1)', (2)', (10)', (10)'', (3-2), (4-2), (5-2), (5-3), (7-2), (7-3), (7-4), (8-2), (9-2), (9-2)', (9-3), (9-4) and (9-5).

[0112] (28) A pharmaceutical composition comprising a compound of any one of the following items or a pharmaceutically acceptable salt thereof: (1) to (10), (1)', (2)', (10)', (10)'', (3-2), (4-2), (5-2), (5-3), (7-2), (7-3), (7-4), (8-2), (9-2), (9-2)', (9-3), (9-4) and (9-5), which is for oral administration.

[0113] (29) The pharmaceutical composition of (28), which is in the form of tablets, powders, granules, capsules, pills, films, suspensions, emulsions, elixirs, syrups, lemonades, spirits, aromatic waters, extracts, decoctions or tinctures.

[0114] (30) The pharmaceutical composition of (29), which is a sugar-coated tablet, a film-coated tablet, an enteric-coated tablet, a sustained-release tablet, a troche tablet, a sublingual tablet, a buccal tablet, a chewable tablet, an orally disintegrating tablet, a dry syrup, a soft capsule, a microcapsule or a sustained-release capsule.

[0115] (31) A pharmaceutical composition comprising a compound of any one of the following items or a pharmaceutically acceptable salt thereof: (1) to (10), (1)', (2)', (10)', (10)'', (3-2), (4-2), (5-2), (5-3), (7-2), (7-3), (7-4), (8-2), (9-2), (9-2)', (9-3), (9-4) and (9-5), which is for parenteral administration.

[0116] (32) The pharmaceutical composition of (31) for cutaneous, subcutaneous, intravenous, intraarterial, intramuscular, intraperitoneal, transmucosal, inhalation, nasal, ocular, inner ear or vaginal administration.

[0117] (33) The pharmaceutical composition of (31) or (32), which is an injection, infusion, eye drops, nasal drops, ear drops, aerosol, inhalation, lotion, soak, liniment, mouthwash, enema, ointment, plaster, gel, cream, patch, paste, external powder or suppository.

[0118] (34) A pharmaceutical composition comprising a compound of any one of the following items or a pharmaceutically acceptable salt thereof: (1) to (10), (1)', (2)', (10)', (10)'', (10)''', (3-2), (4-2), (5-2), (5-3), (7-2), (7-3), (7-4), (8-2), (9-2), (9-2)', (9-3), (9-4) and (9-5), for use in pediatric or elderly patients.

[0119] (35) A pharmaceutical composition comprising a compound of any one of the following items or a pharmaceutically acceptable salt thereof in combination with an acetylcholinesterase inhibitor, an NMDA antagonist or other drugs for Alzheimer's disease: (1) to (10), (1)', (2)', (10)', (10)'', (3-2), (4-2), (5-2), (5-3), (7-2), (7-3), (7-4), (8-2), (9-2), (9-2)', (9-3), (9-4) and (9-5).

[0120] (36) A pharmaceutical composition comprising a compound of any one of the following items or a pharmaceutically acceptable salt thereof: (1) to (10), (1)', (2)', (10)', (10)'', (10)''', (3-2), (4-2), (5-2), (5-3), (7-2), (7-3), (7-4), (8-2), (9-2), (9-2)', (9-3), (9-4) and (9-5), for use in combination therapy with an acetylcholinesterase inhibitor, an NMDA antagonist or other drugs for Alzheimer's disease. DETAILED DESCRIPTION

[0121] Hereinafter, the present invention is described with reference to the embodiments. It should be understood that throughout this specification, unless otherwise stated, the expression of the singular includes the concept of its plural form. Therefore, it should be understood that, unless otherwise stated, the articles in the singular (e.g., "a," "an," "the," etc. in English) include the concept of its plural form. In addition, it should be understood that, unless otherwise stated, the terms used herein are used with the meanings commonly used in the art. Therefore, unless otherwise defined, all technical and scientific terms used herein have the same meanings as those commonly understood by those skilled in the art to which the present invention belongs. If there is a contradiction, this specification (including definitions) shall prevail.

[0122] Each meaning of the terms used herein is described below. Unless otherwise specified, each term is used with the same meaning when used alone and in combination.

[0123] In the present specification, the term "consisting of" means having only components.

[0124] In the present specification, the term "comprising" means not to be limited to components, and does not exclude undescribed factors.

[0125] In the present specification, "halogen" includes fluorine, chlorine, bromine and iodine. Fluorine and chlorine are preferred. Fluorine is more preferred.

[0126] In this specification, "alkyl" includes a straight or branched alkyl group having a carbon number of 1 to 15, such as a carbon number of 1 to 10, such as a carbon number of 1 to 6, and such as a carbon number of 1 to 4, preferably a carbon number of 1 to 3, and more preferably a carbon number of 1 or 2. Examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, hexyl, isohexyl, n-heptyl, isoheptyl, n-octyl, isooctyl, n-nonyl and n-decyl. Examples are methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl and n-pentyl. In one embodiment, "alkyl" is methyl, ethyl, n-propyl, isopropyl or tert-butyl. Methyl is preferred.

[0127] The term "haloalkyl" includes groups in which one or more hydrogen atoms bonded to one or more carbon atoms of the above-mentioned "alkyl" are replaced by one or more of the above-mentioned "halogen". Examples are

[0128] Monofluoromethyl, monofluoroethyl, monofluoropropyl,

[0129] Difluoromethyl, difluoroethyl, difluoropropyl,

[0130] trifluoromethyl, trifluoroethyl, trifluoropropyl, pentafluoropropyl,

[0131] Monochloromethyl, monochloroethyl, monochloropropyl,

[0132] Dichloromethyl, dichloroethyl, dichloropropyl,

[0133] Trichloromethyl, trichloroethyl, trichloropropyl, pentachloropropyl,

[0134] 1-fluoroethyl, 2-fluoroethyl, 1,1-difluoroethyl, 2,2-difluoroethyl, 2,2,2-trifluoroethyl,

[0135] 1-chloroethyl, 2-chloroethyl, 1,1-dichloroethyl, 2,2-dichloroethyl, 2,2,2-trichloroethyl,

[0136] 1,2-dibromoethyl, 1,1,1-trifluoropropan-2-yl, and 2,2,3,3,3-pentafluoropropyl;

[0137] Examples are monofluoromethyl, difluoromethyl, trifluoromethyl, 1-fluoroethyl, 1,1-difluoroethyl and 2,2-difluoroethyl. Examples are monofluoromethyl, difluoromethyl, 1-fluoroethyl, 1,1-difluoroethyl and 2,2-difluoroethyl. Preference is given to difluoromethyl, trifluoromethyl and 2,2-difluoroethyl.

[0138] The term "alkenyl" includes straight or branched alkenyls having one or more double bonds in any available position, ranging from 2 to 15 carbons, such as 2 to 10 carbons, such as 2 to 6 carbons, and such as 2 to 4 carbons. Examples include vinyl, allyl, propenyl, isopropenyl, butenyl, isobutenyl, isoprenyl, butadienyl, pentenyl, isopentenyl, pentadienyl, hexenyl, isohexenyl, hexadienyl, heptenyl, octenyl, nonenyl, decenyl, undecenyl, dodecenyl, tridecenyl, tetradecenyl, and pentadecenyl. Examples are vinyl, allyl, propenyl, isopropenyl, and butenyl.

[0139] The term "alkynyl" includes alkynyl groups having a carbon number of 2 to 15, such as a carbon number of 2 to 10, such as a carbon number of 2 to 8, such as a carbon number of 2 to 6, and such as a straight or branched alkynyl group having a carbon number of 2 to 4, with one or more triple bonds at an optional position. Specific examples are ethynyl, propynyl, butynyl, pentynyl, hexynyl, heptynyl, octynyl, nonynyl and decynyl. These may further have a double bond at any available position. Examples are ethynyl, propynyl, butynyl and pentynyl.

[0140] The term "alkylene" includes straight or branched divalent carbon chains having carbon numbers of 1 to 15, such as carbon numbers of 1 to 10, such as carbon numbers of 1 to 6, and such as carbon numbers of 1 to 3. Examples are methylene, dimethylene and trimethylene.

[0141] One or more hydrogen atoms of the alkylene group in the compounds of formula (I), (IA) or (IB) may be replaced by hydrogen isotopes. 2 H (deuterium) substitution.

[0142] The term "alkoxy" includes groups in which the oxygen atom is substituted by the above-mentioned "alkyl". Examples are methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, isobutoxy, sec-butoxy, pentyloxy, isopentyloxy and hexyloxy;

[0143] In one embodiment, "alkoxy" is methoxy, ethoxy, n-propoxy, isopropoxy or tert-butoxy.

[0144] The term "alkenyloxy" includes groups in which an oxygen atom is substituted by the above-mentioned "alkenyl". Examples are vinyloxy, allyloxy, 1-propenyloxy, 2-butenyloxy, 2-pentenyloxy, 2-hexenyloxy, 2-heptenyloxy and 2-octenyloxy.

[0145] The term "alkynyloxy" includes groups in which the oxygen atom is substituted by the above-mentioned "alkynyl". Examples are ethynyloxy, 1-propynyloxy, 2-propynyloxy, 2-butynyloxy, 2-pentynyloxy, 2-hexynyloxy, 2-heptynyloxy and 2-octynyloxy;

[0146] In one embodiment, "alkynyloxy" is ethynyloxy, 1-propynyloxy and 2-propynyloxy.

[0147] The term "carbocycle" includes non-aromatic carbocycles and aromatic carbocycles.

[0148] The term "non-aromatic carbocycle" includes a saturated carbocycle or an unsaturated non-aromatic carbocycle that is monocyclic or composed of two or more rings. Two or more rings of "non-aromatic carbocycle" include fused cyclic groups in which a non-aromatic monocyclic carbocycle or two or more rings of non-aromatic carbocycle are fused to the ring of the above-mentioned "aromatic carbocycle";

[0149] In addition, "non-aromatic carbocycle" also includes cyclic groups having a bridge or cyclic group to form a spiro ring, as shown below:

[0150] [Chemical formula 16]

[0151]

[0152] The term "non-aromatic monocyclic carbocycle" includes groups having 3 to 16 carbon atoms, such as 3 to 12 carbon atoms, such as 3 to 8 carbon atoms, and such as 3 to 5 carbon atoms. Examples are cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, cyclooctane, cyclononane, cyclodecane, cyclopropenane, cyclobutenane, cyclopentenane, cyclohexenane, cycloheptenane and cyclohexadienane.

[0153] Examples of the non-aromatic carbocyclic ring composed of two or more rings include groups having 6 to 14 carbon atoms, and examples are indane, indenane, acenaphthene, tetralin, and fluorene.

[0154] The term "aromatic carbocyclic ring" includes a monocyclic ring or an aromatic hydrocarbon ring composed of two or more rings. Examples are aromatic hydrocarbon groups having a carbon number of 6 to 14, and specific examples are benzene, naphthalene, anthracene, and phenanthrene;

[0155] In one embodiment, the "aromatic carbocycle" is benzene;

[0156] In one embodiment, "carbocycle" is cyclopropane, cyclobutane and cyclopentane.

[0157] The term "heterocycle" includes non-aromatic heterocycles and aromatic heterocycles.

[0158] The term "non-aromatic heterocycle" includes a single ring or a non-aromatic group consisting of two or more rings containing one or more heteroatoms independently selected from oxygen, sulfur and nitrogen atoms.

[0159] The “non-aromatic heterocycle” of two or more rings includes a fused cyclic group in which a non-aromatic monocyclic heterocycle or a non-aromatic heterocycle of two or more rings is fused to the ring of the above-mentioned “aromatic carbocycle”, “non-aromatic carbocycle” and / or “aromatic heterocycle”.

[0160] In addition, "non-aromatic heterocycle" also includes cyclic rings having a bridge or cyclic group to form a spirocycle, as shown below:

[0161] [Chemical formula 17]

[0162]

[0163] The term "non-aromatic monocyclic heterocycle" includes 3 to 8 membered rings, and for example 4, 5 or 6 membered rings. Examples are dioxane, thiirane, oxirane, oxetane, oxathiolane, azetidine, thiazolidine, pyrrolidine, pyrroline, imidazolidine, imidazoline, pyrazolidine, pyrazoline, piperidine, piperazine, morpholinyl, morpholine, thiomorpholine, dihydropyridine, tetrahydropyridine, tetrahydrofuran, tetrahydropyran, dihydrothiazoline, tetrahydrothiazoline, tetrahydroisothiazoline, dihydrooxazine, hexahydroazepine, tetrahydroaziridine, tetrahydropyridazine, hexahydropyrimidine, dioxolane, dioxazine, aziridine, dioxoline, oxepane, thiolane, thiine and thiazine.

[0164] Examples of the non-aromatic heterocyclic ring of two or more rings include 9- to 14-membered groups, and examples are indoline, isoindoline, chroman, and isochroman.

[0165] The term "aromatic heterocycle" includes a single ring or an aromatic ring consisting of two or more rings containing one or more heteroatoms independently selected from oxygen, sulfur and nitrogen atoms.

[0166] The "aromatic heterocycle" of two or more rings includes a fused cyclic group in which an aromatic monocyclic heterocyclic group or a non-aromatic heterocycle composed of two or more rings is fused to the ring of the above-mentioned "aromatic carbocycle".

[0167] The term "aromatic monocyclic heterocycle" includes 5 to 8 membered groups, and for example 5 to 6 membered rings. Examples are pyrrole, imidazole, pyrazole, pyridine, pyridazine, pyrimidine, pyrazine, triazole, triazine, tetrazole, furan, thiophene, isoxazole, oxazole, oxadiazole, isothiazole, thiazole and thiadiazole.

[0168] Examples of aromatic bicyclic heterocycles include 9 to 10 membered rings and examples are indoline, isoindoline, indazoline, indolizine, quinoline, isoquinoline, cinnoline, phthalazine, quinazoline, naphthyridine, quinoxaline, purine, pteridine, benzimidazole, benzisoxazole, benzoxazole, benzoxadiazole, benzisothiazole, benzothiazole, benzothiadiazole, benzofuran, isobenzofuran, benzothiophene, benzotriazole, imidazopyridine, triazolopyridine, imidazothiazole, pyrazinopyridazine, oxazolopyridine and thiazolopyridine.

[0169] Examples of the aromatic heterocyclic ring of three or more rings include 13- to 14-membered groups, and examples are carbazole, acridine, xanthene, phenothiazine, phenoxathiine, phenoxazine, and dibenzofuran.

[0170] In one embodiment, "heterocycle" is 1,4-oxathiinane.

[0171] Examples of the substituent of the "substituted or unsubstituted alkyl group" are one or more groups selected from the following substituent group α.

[0172] Substituent group α is a group consisting of halogen, hydroxyl, cyano, and alkoxy.

[0173] The substituent of the "substituted or unsubstituted alkyl group" is, for example, halogen or the like.

[0174] R 2 The substituent of the "substituted or unsubstituted alkyl group" is, for example, halogen or the like.

[0175] R 3 The substituent of the "substituted or unsubstituted alkyl group" is, for example, halogen, alkoxy and the like.

[0176] Examples of the substituent of the "substituted or unsubstituted alkoxy group" are one or more groups selected from the following substituent group β.

[0177] Substituent group β is a group consisting of halogen, hydroxyl, cyano, optionally substituted alkyl (substituents: hydroxyl, non-aromatic carbocyclic group, cyano non-aromatic carbocyclic group, halogenated non-aromatic carbocyclic group, non-aromatic heterocyclic group, alkyl non-aromatic heterocyclic group, halogenated non-aromatic heterocyclic group, aromatic heterocyclic group).

[0178] R 2 The substituent of the "substituted or unsubstituted alkyl group" is, for example, halogen or the like.

[0179] R 3 The substituent of the "substituted or unsubstituted alkyl group" is, for example, halogen, alkoxy and the like.

[0180] R 7aThe substituent of the “substituted or unsubstituted alkoxy group” is, for example, an optionally substituted alkyl group (substituents: hydroxy, non-aromatic carbocyclic group, cyano non-aromatic carbocyclic group, halogenated non-aromatic carbocyclic group, non-aromatic heterocyclic group, alkyl non-aromatic heterocyclic group, halogenated non-aromatic heterocyclic group, aromatic heterocyclic group) and the like.

[0181] The "substituted or unsubstituted non-aromatic carbocyclic group" and the "substituted or unsubstituted non-aromatic heterocyclic group" may be substituted with "oxo". This includes groups in which two hydrogen atoms attached to the same carbon atom are substituted with oxo, as shown below:

[0182] [Chemical formula 18]

[0183] .

[0184] Examples of the substituent of the "substituted or unsubstituted carbocycle" or "substituted or unsubstituted heterocycle" include groups selected from Substituent Group α.

[0185] The compounds of formula (I), (IA) or (IB) are not limited to specific isomers and include all possible isomers, such as keto-enol isomers, imine-enamine isomers, diastereomers, optical isomers and rotational isomers, racemates and mixtures thereof. For example, the compounds of formula (I) include the following tautomers.

[0186] [Chemical formula 19]

[0187]

[0188] In this specification, the group is represented by the following formula:

[0189] [Chemical formula 20]

[0190]

[0191] It means a group represented by the following formula:

[0192] [Chemical formula 21]

[0193] .

[0194] One or more hydrogen, carbon and / or other atoms of the compounds of formula (I), (IA) or (IB) may be replaced by isotopes of hydrogen, carbon and / or other atoms, respectively. Examples of isotopes include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, iodine and chlorine, respectively, such as 2 H (D), 3 H. 11 C. 13 C. 14 C. 15 N. 18 O.17 O. 31 P. 32 P. 35 S. 18 F. 123 I and 36 Cl. Compounds of formula (I), (IA) or (IB) also include compounds substituted with such isotopes. Compounds substituted with such isotopes may also be used as pharmaceuticals, and include all radiolabeled compounds of compounds of formula (I), (IA) or (IB). The present invention includes "radiolabeling methods" for making "radiolabeled compounds", and the methods may be used as tools for research in metabolic pharmacokinetic studies, binding assays and / or diagnostics.

[0195] Radiolabeled compounds of compounds of formula (I), (IA) or (IB) may be prepared by methods known in the art. For example, tritiated compounds of formula (I), (IA) or (IB) may be prepared by introducing tritium into a particular compound of formula (I), (IA) or (IB), such as by catalyzing dehalogenation with tritium. The method may comprise reacting a suitable halogenated precursor of a compound of formula (I), (IA) or (IB) with tritium gas in the presence of a suitable catalyst such as Pd / C in the presence or absence of a base. Other suitable methods for preparing tritiated compounds may be found in Isotopes in the Physical and Biomedical Sciences, Volume 1, Labeled Compounds (Part A), Chapter 6 (1987). Tritiated compounds may be prepared by using a catalytic dehalogenation reaction having a catalytic dehalogenation reaction ... 14 C carbon starting materials to prepare 14 C-labeled compounds.

[0196] As pharmaceutically acceptable salts of the compounds of formula (I), (IA) or (IB), examples include salts with alkali metals (e.g. lithium, sodium and potassium), alkaline earth metals (e.g. calcium and barium), magnesium, transition metals (e.g. zinc and iron), ammonia, organic bases (e.g. trimethylamine, triethylamine, dicyclohexylamine, ethanolamine, diethanolamine, triethanolamine, meglumine, diethanolamine, ethylenediamine, pyridine, picoline, quinoline) and amino acids, and salts with inorganic acids (e.g. hydrochloric acid, sulfuric acid, nitric acid, carbonic acid, hydrobromic acid, phosphoric acid and hydroiodic acid) and organic acids (e.g. formic acid, acetic acid, propionic acid, trifluoroacetic acid, citric acid, lactic acid, tartaric acid, oxalic acid, maleic acid, fumaric acid, succinic acid, mandelic acid, glutaric acid, malic acid, benzoic acid, phthalic acid, ascorbic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid and ethanesulfonic acid). Specific examples are salts with hydrochloric acid, sulfuric acid, phosphoric acid, tartaric acid, succinic acid or methanesulfonic acid. These salts can be formed by conventional methods.

[0197] The compounds of the present invention represented by formula (I), (IA) or (IB) or their pharmaceutically acceptable salts may form solvates (e.g., hydrates, etc.) and / or crystalline polymorphs. The present invention includes those various solvates and crystalline polymorphs. "Solvates" may be those in which any number of solvent molecules (e.g., water molecules, etc.) are coordinated with the compounds represented by formula (I), (IA) or (IB). When the compound represented by formula (I) or a pharmaceutically acceptable salt is placed in the atmosphere, the compound may absorb water, causing the adsorbed water to attach or form a hydrate. Recrystallization of the compound represented by formula (I) or a pharmaceutically acceptable salt may produce crystalline polymorphs.

[0198] The compounds of the present invention represented by formula (I), (IA) or (IB) or their pharmaceutically acceptable salts can form prodrugs. The present invention also includes various prodrugs. Prodrugs are derivatives of the compounds of the present invention having chemically or metabolically degradable groups, and are compounds that are converted into pharmaceutically active compounds of the present invention by solvolysis or under physiological conditions in vivo. Prodrugs include compounds that are converted into compounds represented by formula (I), (IA) or (IB) by enzymatic oxidation, reduction, hydrolysis, etc. under physiological conditions in vivo and compounds that are converted into compounds represented by formula (I), (IA) or (IB) by hydrolysis by gastric acid, etc. Methods for selecting and preparing suitable prodrug derivatives are described in, for example, Design of Prodrugs, Elsevier, Amsterdam 1985. The prodrug itself can be an active compound.

[0199] When the compound of formula (I), (IA) or (IB) or a pharmaceutically acceptable salt thereof has a hydroxyl group, the prodrug includes an acyloxy derivative and a sulfonyloxy derivative, which can be prepared by reacting the compound having a hydroxyl group with a suitable acyl halide, a suitable acid anhydride, a suitable sulfonyl chloride, a suitable sulfonyl anhydride and a mixed acid anhydride or with a condensing agent. Examples are CH 3 COO-, C 2 H 5 COO-, t-BuCOO-, C 15 H 31 COO-, PhCOO-, (m-NaOOCPh)COO-, NaOOCCH 2 CH 2 COO-、CH 3 CH(NH 2 )COO-、CH 2 N(CH 3 ) 2 COO-、CH 3 SO 3 -、CH 3 CH 2 SO3 -、CF 3 SO 3 -、CH 2 FSO 3 -、CF 3 CH 2 SO 3 -、p-CH 3 -O-PhSO 3 -、PhSO 3 - and p-CH 3 PhSO 3 -.

[0200] Compounds of formula (I), (IA) or (IB) can be prepared by the methods described below together with synthetic methods known to those skilled in the art.

[0201] The starting materials are commercially available or can be prepared according to known methods.

[0202] During any of the following synthesis, it may be necessary or preferred to protect sensitive or reactive groups on any molecule. In such cases, these protections may be achieved by conventional protecting groups, such as those described in Greene's Protective Group in Organic Synthesis, John Wily & Sons, 2007.

[0203] Those skilled in the art will appreciate that the compounds described below will be produced as mixtures of diastereomers and / or enantiomers which may be separated at the relevant stages of the following procedures using conventional techniques such as crystallization, silica gel chromatography, chiral or achiral high performance liquid chromatography (HPLC) and chiral supercritical fluid (SFC) chromatography to provide the single enantiomers of the invention.

[0204] During all subsequent steps, the order of the steps to be performed can be appropriately changed. In each step, an intermediate can be isolated and then used in the next step. All reaction times, reaction temperatures, solvents, reagents and protecting groups etc. are only exemplary and are not limited as long as they do not adversely affect the reaction.

[0205] General Procedure A

[0206] [Chemical formula 22]

[0207]

[0208] wherein P is a protecting group such as benzoyl or benzyl, and the other symbols are the same as defined in (1) above.

[0209] General Procedure A is a method for preparing Compound A-9 from Compound A-1 through multiple steps from Step 1 to Step 8. One skilled in the art will appreciate that the protecting group P can be selected according to the reaction conditions used in the subsequent steps.

[0210] Step 1

[0211] Compound A-2 can be prepared by means of a 1,3-dipolar cycloaddition reaction. This type of reaction can be carried out using similar conditions described in J. Am. Chem. Soc., 1960, 82, 5339-5342 or J. Org. Chem. 1998, 63, 5272-5274. These 1,3-dipolar cycloaddition reactions can be carried out using a suitable dehydrating agent (such as, for example, phenyl isocyanate, phenyl diisocyanate or (Boc) 2 O) and a suitable base (such as, for example, triethylamine, dipropylethylamine or N-methylmorpholine) from cyclic compound A-1 and the corresponding nitrile oxide compound (generated in situ from the corresponding nitroalkane). Alternatively, the nitrile oxide compound can be generated in situ from the corresponding chlorinated oxime with a suitable base (such as, for example, triethylamine, dipropylethylamine or N-methylmorpholine). The solvent used in this step is not particularly limited as long as it does not interfere with the reaction. Examples of the solvent include tetrahydrofuran, 1,4-dioxane, 1,2-dimethoxyethane, diethyl ether, toluene, benzene and mixed solvents thereof. The reaction temperature is preferably room temperature to 120°C. The reaction time is not particularly limited and is generally 5 minutes to 24 hours, preferably 30 minutes to 24 hours.

[0212] Step 2

[0213] Compound A-3 can be prepared by nucleophilic addition of a suitable aryl lithium reagent or Grignard reagent to compound A-2. This type of reaction can be carried out using similar conditions described in J.Am.Chem.Soc., 2005, 127, 5376-5384. Preferably, the aryl lithium reagent or Grignard reagent can be prepared from the corresponding aromatic halide using a suitable base (such as, for example, n-, sec- or tert-butyl lithium, isopropyl magnesium bromide or metallic magnesium), which can then be reacted with a Lewis acid (such as, for example, BF 3 -OEt 2 ) is reacted with compound A-2 to obtain compound A-3. The solvent used in this step is not particularly limited as long as it does not interfere with the reaction. Examples of the solvent include tetrahydrofuran, 1,4-dioxane, 1,2-dimethoxyethane, diethyl ether, toluene, benzene, and mixed solvents thereof. The reaction temperature is preferably from -78°C to room temperature. The reaction time is not particularly limited and is generally from 5 minutes to 24 hours, preferably from 30 minutes to 24 hours.

[0214] Step 3

[0215] Compound A-4 can be prepared by a reduction cleavage reaction of the N-O bond of compound A-3. The reduction cleavage can be carried out using zinc and a suitable acid such as acetic acid, formic acid or hydrochloric acid. The solvent used in this step is not particularly limited as long as it does not interfere with the reaction. Examples of the solvent include methanol, ethanol, tetrahydrofuran, water and a mixed solvent thereof. The reaction temperature is preferably -20 ° C to the solvent reflux temperature. The reaction time is not particularly limited, and is generally 5 minutes to 24 hours, preferably 30 minutes to 24 hours.

[0216] Alternatively, the reaction can be carried out under hydrogen using a metal catalyst such as platinum oxide. The solvent used in this step is not particularly limited as long as it does not interfere with the reaction. Examples of the solvent include methanol, ethanol, water and mixed solvents thereof. The reaction temperature is preferably from room temperature to 50°C. The reaction time is not particularly limited and is generally from 5 minutes to 24 hours, preferably from 30 minutes to 24 hours.

[0217] In addition, this type of reaction can also be carried out using lithium aluminum hydride. The solvent used in this step is not particularly limited as long as it does not interfere with the reaction. Examples of the solvent include tetrahydrofuran, 1,4-dioxane, 1,2-dimethoxyethane, diethyl ether and a mixed solvent thereof. The reaction temperature is preferably -20°C to room temperature. The reaction time is not particularly limited and is generally 5 minutes to 24 hours, preferably 30 minutes to 24 hours.

[0218] Step 4

[0219] Compound A-5 can be prepared by forming the corresponding thiourea in situ from compound A-4, followed by a cyclization reaction. Such reactions are known to those skilled in the art and can be carried out under the conditions described in WO2014 / 065434. The thiourea can be obtained in situ from compound A-4 using a suitable isothiocyanate (such as, for example, benzoyl isothiocyanate or benzyl isothiocyanate), and then cyclized by adding a reagent such as, for example, m-CPBA, hydrogen peroxide, or a carbodiimide reagent (such as DCC, DIC, or EDC). Alternatively, the cyclization reaction can be carried out using an alkylating agent such as iodomethane and a suitable base such as sodium hydride, sodium bicarbonate, and potassium carbonate. The solvent used in this step is not particularly limited as long as it does not interfere with the reaction. Examples of the solvent include chloroform, dichloromethane, dichloroethane, tetrahydrofuran, and mixed solvents thereof. The reaction temperature is generally 0°C to 60°C. The reaction time is not particularly limited and is generally 5 minutes to 24 hours, preferably 30 minutes to 24 hours.

[0220] Step 5

[0221] Compound A-6 can be prepared by deprotection of compound A-5. The deprotection reaction is known to those skilled in the art and can be carried out under the conditions described in Green's Protective Groups in Organic Synthesis, Fourth Edition. When the protecting group is a benzoyl group, the deprotection reaction can be carried out under acidic conditions (such as sulfuric acid or hydrochloric acid), or under alkaline conditions (such as hydrazine, DBU or sodium hydroxide). The solvent used in this step is not particularly limited as long as it does not interfere with the reaction. Examples of the solvent include dichloromethane, tetrahydrofuran, 1,4-dioxane, methanol, toluene, benzene and mixed solvents thereof. The reaction temperature is preferably room temperature to 100°C. The reaction time is not particularly limited, and is generally 5 minutes to 24 hours, preferably 30 minutes to 24 hours.

[0222] Step 6

[0223] Compound A-7 can be prepared by nitration of compound A-6. A typical method comprises treating compound A6 dissolved in sulfuric acid and trifluoroacetic acid with a nitronium ion source such as, for example, potassium nitrate or nitric acid. The reaction temperature is preferably -20°C to 0°C. The reaction time is not particularly limited and is generally 5 minutes to 5 hours, preferably 30 minutes to 2 hours.

[0224] Step 7

[0225] Compound A-8 can be prepared by reducing compound A-7. The reduction can be carried out by a suitable catalyst (such as, for example, palladium on carbon) under a hydrogen atmosphere, or using a reducing agent (such as, for example, iron, zinc or tin (II) chloride). The solvent used in this step is not particularly limited as long as it does not interfere with the reaction. Examples of the solvent include tetrahydrofuran, methanol, ethanol, water and mixed solvents thereof. The reaction temperature is generally room temperature to 80°C, preferably room temperature to 60°C. The reaction time is not particularly limited, and is generally 5 minutes to 24 hours, preferably 30 minutes to 24 hours.

[0226] Step 8

[0227] Compound A-9 can be prepared by the amide coupling reaction of a suitable acid salt of compound A-8 with the corresponding carboxylic acid. The acid salt of compound A can be prepared with an equivalent of a suitable acid such as HCl before the condensation reaction. The reaction can be carried out by methods known to those skilled in the art, and suitable coupling conditions can be found in Chem. Rev. 2011, 111, 6557-6602, including: a) reaction using a condensation reagent; b) reaction using an acyl chloride or acyl fluoride. The reaction must be carried out without adding any base for chemically selective amide coupling at the aniline position.

[0228] Reaction a) can be carried out using a condensation reagent such as dicyclohexylcarbodiimide (DCC), diisopropylcarbodiimide (DIC), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC hydrochloride), O-(7-aza-1H-benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) and 1H-benzotriazol-1-yloxy-tris(pyrrolidino)phosphonium hexafluorophosphate (PyBOP). When uronium or phosphonium salts such as HATU and PyBOP are used, the reaction can be carried out in the presence of a base such as triethylamine and diisopropylethylamine. The reaction can be accelerated by using a catalyst such as 1-hydroxy-benzotriazole (HOBt) and 1-hydroxy-7-aza-benzotriazole (HOAt). The solvent used in the reaction is not particularly limited as long as it does not interfere with the reaction. Examples of the solvent include dichloromethane, N,N-dimethylformamide (DMF), N-methylpyrrolidone (NMP) and tetrahydrofuran. The reaction temperature is usually 0° C. to 50° C. and preferably room temperature.

[0229] Reaction b) can be carried out in the presence of a base such as triethylamine, diisopropylethylamine, pyridine and N,N-dimethyl-4-aminopyridine in a solvent such as dichloromethane, tetrahydrofuran and ethyl acetate by using a commercially available acid chloride or an acid chloride synthesized by a method known to those skilled in the art. The reaction temperature is usually 0°C to 60°C, and preferably 0°C to room temperature. The reaction time is not particularly limited, and is usually 5 minutes to 24 hours, preferably 20 minutes to 6 hours.

[0230] General Procedure B

[0231] [Chemical formula 23]

[0232]

[0233] Where R 3 ' and R 3 '' are each independently selected from an alkyl group optionally substituted by halogen, cyano, alkoxy, haloalkoxy or non-aromatic carbocyclic group and a heterocyclic group optionally substituted by an alkyl group, and other symbols are the same as defined above.

[0234] General Procedure B is a method for preparing compound B-5 from compound B-1 through multiple steps. Compound B-5 can be prepared according to the method described in General Procedure A using compound B-4.

[0235] Step 1

[0236] Compound B-2 can be prepared by 1,3-dipolar cycloaddition reaction. This type of reaction can be carried out using similar conditions described in J.Am.Chem.Soc. 1960, 82, 5339-5342 or J.Org.Chem.1998, 63, 5272-5274. These 1,3-dipolar cycloaddition reactions can be carried out using a suitable dehydrating agent (such as, for example, phenyl isocyanate, phenyl diisocyanate or (Boc) 2 O) and a suitable base (such as, for example, triethylamine, diisopropylethylamine or N-methylmorpholine) are carried out from the cyclic compound B-1 and the corresponding nitrile oxide compound generated in situ from the corresponding nitroalkane. Alternatively, the nitrile oxide compound can be generated in situ from the corresponding chlorinated oxime with a suitable base (such as, for example, triethylamine, diisopropylethylamine or N-methylmorpholine). The solvent used in this step is not particularly limited as long as it does not interfere with the reaction. Examples of the solvent include tetrahydrofuran, 1,4-dioxane, 1,2-dimethoxyethane, ether, toluene, benzene and mixed solvents thereof. The reaction temperature is preferably room temperature to 120°C. The reaction time is not particularly limited and is generally 5 minutes to 24 hours, preferably 30 minutes to 24 hours.

[0237] Step 2

[0238] When R3' is a hydrogen atom, compound B-3 can be prepared by carbonyl reduction of compound B-2. This type of reaction can use a suitable metal hydride, such as, for example, DIBAL-H, lithium tri-tert-butoxyaluminum hydride or bis(2-methoxyethoxy) Sodium aluminum hydride, by addition of nucleophilic hydride to compound B-2. The solvent used in this step is not particularly limited as long as it does not interfere with the reaction. Examples of the solvent include dichloromethane, tetrahydrofuran, 1,4-dioxane, 1,2-dimethoxyethane, ether, toluene, benzene and a mixed solvent thereof. The reaction temperature is preferably -78 ° C to room temperature. The reaction time is not particularly limited, and is generally 5 minutes to 24 hours, preferably 30 minutes to 24 hours.

[0239] When R 3 ' is not a hydrogen atom, compound B-3 can be prepared by nucleophilic addition to compound B-2. This type of reaction can be performed using a suitable nucleophilic reagent (such as, for example, an organolithium, magnesium, zinc or silyl reagent) in the presence or absence of a Lewis acid (such as, for example, BF 3 -OEt 2 、AlCl 3 or TiCl 4). The solvent used in this step is not particularly limited as long as it does not interfere with the reaction. Examples of the solvent include dichloromethane, tetrahydrofuran, 1,4-dioxane, 1,2-dimethoxyethane, ether, toluene, benzene, and mixed solvents thereof. The reaction temperature is preferably -78°C to room temperature. The reaction time is not particularly limited and is generally 5 minutes to 24 hours, preferably 30 minutes to 24 hours.

[0240] Step 3

[0241] When R 3 When '' is a hydrogen atom, compound B-4 can be prepared by reduction of compound B-3. This type of reaction can be prepared using a suitable reducing agent such as triethylsilane, sodium borohydride, in the presence or absence of a Lewis acid such as BF 3 -OEt 2 The solvent used in this step is not particularly limited as long as it does not interfere with the reaction. Examples of the solvent include dichloromethane, acetonitrile, tetrahydrofuran, 1,4-dioxane, 1,2-dimethoxyethane, ether, toluene, benzene, and a mixed solvent thereof. The reaction temperature is preferably -20°C to room temperature. The reaction time is not particularly limited and is generally 5 minutes to 24 hours, preferably 30 minutes to 24 hours.

[0242] When R 3 When '' is not a hydrogen atom, compound B-4 can be prepared by nucleophilic addition to compound B-2. This type of reaction can be carried out using a suitable nucleophilic reagent (such as, for example, an organolithium, magnesium, zinc or silyl reagent) in the presence or absence of a Lewis acid (such as, for example, BF 3 -OEt 2 、AlCl 3 or TiCl 4 ). The solvent used in this step is not particularly limited as long as it does not interfere with the reaction. Examples of the solvent include dichloromethane, tetrahydrofuran, 1,4-dioxane, 1,2-dimethoxyethane, ether, toluene, benzene, and mixed solvents thereof. The reaction temperature is preferably -78°C to room temperature. The reaction time is not particularly limited and is generally 5 minutes to 24 hours, preferably 30 minutes to 24 hours.

[0243] General Program C

[0244] [Chemical formula 24]

[0245]

[0246] Where P is a protecting group, such as benzoyl or benzyl, and R 3 ''' is ethyl or cyclopropyl, and the other symbols are the same as defined in the above (1).

[0247] General Procedure C is a method for preparing compound C-5 from compound B-1 through multiple steps. Compound C-3 and compound C-5 can be prepared according to the method described in General Procedure A from compounds C-2 and C-5.

[0248] Step 1

[0249] Compound C-1 can be prepared by nucleophilic addition of an allyl group to the carbonyl group of compound B-2. This type of reaction can be performed using suitable commercially available or in situ generated allyl reagents (such as, for example, allylsilane, lithium, magnesium, zinc reagents) in the presence or absence of a Lewis acid (such as, for example, BF 3 -OEt 2 、AlCl 3 or TiCl 4 ). The solvent used in this step is not particularly limited as long as it does not interfere with the reaction. Examples of the solvent include dichloromethane, tetrahydrofuran, 1,4-dioxane, 1,2-dimethoxyethane, ether, toluene, benzene, and mixed solvents thereof. The reaction temperature is preferably -78°C to room temperature. The reaction time is not particularly limited and is generally 5 minutes to 24 hours, preferably 30 minutes to 24 hours.

[0250] Step 2

[0251] Compound C-2 can be prepared by reduction of compound C-1. This type of reaction can be prepared using a suitable reducing agent such as triethylsilane or sodium borohydride in the presence or absence of a Lewis acid such as BF 3 -OEt 2 The solvent used in this step is not particularly limited as long as it does not interfere with the reaction. Examples of the solvent include dichloromethane, acetonitrile, tetrahydrofuran, 1,4-dioxane, 1,2-dimethoxyethane, ether, toluene, benzene, and a mixed solvent thereof. The reaction temperature is preferably -20°C to room temperature. The reaction time is not particularly limited and is generally 5 minutes to 24 hours, preferably 30 minutes to 24 hours.

[0252] Step 3

[0253] When R 3 When ''' is ethyl, compound C-5 can be obtained by hydrogenation of compound C-4. The hydrogenation can be carried out under a hydrogen atmosphere using a suitable catalyst (such as, for example, palladium on carbon). The solvent used in this step is not particularly limited as long as it does not interfere with the reaction. Examples of the solvent include tetrahydrofuran, methanol, ethanol, water, and mixed solvents thereof. The reaction temperature is generally room temperature to 80°C, and preferably room temperature to 60°C. The reaction time is not particularly limited, and is generally 5 minutes to 24 hours, preferably 30 minutes to 24 hours.

[0254] When R3 When ''' is a cyclopropyl group, compound C-5 can be obtained by cyclopropanation of compound C-4. This type of reaction can be carried out using a suitable reagent such as diazomethane with or without a suitable catalyst, or using Simmons-Smith reaction conditions (such as, for example, diiodomethane and diethylzinc). The solvent used in this step is not particularly limited as long as it does not interfere with the reaction. Examples of the solvent include dichloromethane, ether, toluene, benzene, or a mixed solvent thereof. The reaction temperature is generally -30°C to room temperature. The reaction time is not particularly limited, and is generally 5 minutes to 24 hours, preferably 30 minutes to 24 hours.

[0255] General Program D

[0256] [Chemical formula 25]

[0257]

[0258] Where P is a protecting group, such as benzoyl or benzyl, and R 3 '''' is an alkyl group or alkoxy group substituted by fluorine, and the other symbols are the same as defined in the above (1).

[0259] General Procedure D is a method for preparing compound D-3 from compound C-3 through multiple steps. Compound D-3 can be prepared according to the method described in General Procedure A from compound D-2.

[0260] Step 1

[0261] Compound D-1 can be prepared by ozonolysis of compound C-3, followed by reduction of the resulting aldehyde. The reaction can be carried out by methods known to those skilled in the art. Ozonolysis can be carried out under an ozone atmosphere, in a suitable solvent (such as dichloromethane, methanol and mixtures thereof), with a suitable reagent (such as triphenylphosphine, pyridine, dimethyl sulfide and trimethylamine), under a nitrogen atmosphere for reduction post-treatment. The temperature for generating ozonide is preferably -78°C, which can then be warmed to room temperature for reduction post-treatment. The reaction time is not particularly limited and is generally 30 minutes to 5 hours, preferably 30 minutes to 2 hours. The reduction of the generated aldehyde can be carried out in a one-pot process using a suitable reducing agent such as sodium borohydride or lithium aluminum hydride. The reaction temperature is preferably 0°C to room temperature. The reaction time is not particularly limited and is generally 30 minutes to 5 hours, preferably 30 minutes to 2 hours.

[0262] Step 2

[0263] When R 3 '''' is CF 3 , CHF 2 or CH 2F, compound D-2 can be obtained in two steps; oxidizing compound D-1 to an aldehyde or carboxylic acid and then fluorinating, or directly fluorinating compound D-1. The reaction can be carried out by methods known to those skilled in the art. For example, compound D-1 can be oxidized to the corresponding aldehyde under suitable oxidation conditions, such as, for example, TEMPO, Dess-Martin or Swern oxidation. The corresponding carboxylic acid can be obtained by oxidizing the resulting aldehyde, or directly oxidizing compound D-1 using suitable conditions such as, for example, Pinnick, TEMPO or Jones oxidation (oxadation). The solvent used in this step is not particularly limited as long as it does not interfere with the reaction. The reaction temperature is generally -78°C to room temperature. The reaction time is not particularly limited and is generally 5 minutes to 24 hours, preferably 30 minutes to 24 hours. The fluorination reaction can be carried out using suitable reagents such as, for example, DAST, Deoxofluor or sulfur tetrafluoride. The solvent used in this step is not particularly limited as long as it does not interfere with the reaction. Examples of the solvent include dichloromethane, tetrahydrofuran, 1,4-dioxane, 1,2-dimethoxyethane, diethyl ether, toluene, benzene, and mixed solvents thereof. The reaction temperature is preferably -78°C to 50°C. The reaction time is not particularly limited and is generally 5 minutes to 24 hours, preferably 30 minutes to 24 hours.

[0264] When R 3 When '''' is an alkoxy group, compound D-2 can be obtained by alkylation of the terminal alcohol of compound D-1. The reaction can be carried out using a suitable base such as sodium hydride and a corresponding electrophilic reagent such as an alkyl halide, a mesylate or a triflate. The solvent used in this step is not particularly limited as long as it does not interfere with the reaction. Examples of the solvent include acetone, acetonitrile, tetrahydrofuran, DMF, DMA, DMSO, toluene, and a mixed solvent thereof. The reaction temperature is preferably 0°C to 100°C. The reaction time is not particularly limited and is generally 5 minutes to 24 hours, preferably 30 minutes to 24 hours.

[0265] General Program E

[0266] [Chemical formula 26]

[0267]

[0268] Where P is a protecting group, such as benzoyl or benzyl, R 3 ''''' is ethyl or cyclopropyl, X is a leaving group such as halogen, mesylate or trifluoromethanesulfonate, and the other symbols are the same as those defined in (1) above.

[0269] General Procedure E is a method for preparing compound E-4 from compound D-1 through multiple steps. Compound E-4 can be prepared according to the method described in General Procedure A from compound E-2.

[0270] Step 1

[0271] Compound E-1 can be prepared by converting the terminal alcohol of compound D-1 into a leaving group. The reaction can be carried out by methods known to those skilled in the art. Compound E-1 can be obtained under suitable halogenation conditions, such as, for example, using SOX 2 、POX 3 (X = Cl or Br), or Appel reaction conditions such as triphenylphosphine with CX 4 (X = Cl or Br) or iodine. The solvent used in this step is not particularly limited as long as it does not interfere with the reaction. Examples of the solvent include dichloromethane, tetrahydrofuran, toluene and mixed solvents thereof. The reaction temperature is preferably 0°C to 100°C. The reaction time is not particularly limited and is generally 5 minutes to 24 hours, preferably 30 minutes to 24 hours.

[0272] Step 2

[0273] Compound E-2 can be prepared by converting the terminal alcohol of compound D-1 into a leaving group. The reaction can be carried out by methods known to those skilled in the art. Compound E-1 can be obtained under suitable halogenation conditions, such as, for example, using SOX 2 、POX 3 (X = Cl or Br), or Appel reaction conditions such as triphenylphosphine with CX 4 (X = Cl or Br) or iodine. The solvent used in this step is not particularly limited as long as it does not interfere with the reaction. Examples of the solvent include dichloromethane, tetrahydrofuran, toluene and mixed solvents thereof. The reaction temperature is preferably 0°C to 100°C. The reaction time is not particularly limited and is generally 5 minutes to 24 hours, preferably 30 minutes to 24 hours.

[0274] Step 3

[0275] Compound E-2 can be prepared by means of an elimination reaction of compound E-1. The reaction can be carried out by methods known to those skilled in the art. Compound E-2 can be obtained using a suitable base such as, for example, sodium tert-butoxide or potassium tert-butoxide, triethylamine, diisopropylethylamine, DBU or pyridine. The solvent used in this step is not particularly limited as long as it does not interfere with the reaction. Examples of the solvent include dichloromethane, tetrahydrofuran, toluene and mixed solvents thereof. The reaction temperature is preferably 0°C to 60°C. The reaction time is not particularly limited and is generally 5 minutes to 24 hours, preferably 30 minutes to 24 hours.

[0276] Step 4

[0277] When R 3 When ''''' is ethyl, compound E-3 can be obtained by hydrogenation of compound E-2. The hydrogenation can be carried out under a hydrogen atmosphere using a suitable catalyst (such as, for example, palladium on carbon). The solvent used in this step is not particularly limited as long as it does not interfere with the reaction. Examples of the solvent include tetrahydrofuran, methanol, ethanol, water, and a mixed solvent thereof. The reaction temperature is generally room temperature to 80°C, and preferably room temperature to 60°C. The reaction time is not particularly limited, and is generally 5 minutes to 24 hours, preferably 30 minutes to 24 hours.

[0278] When R 3 When ''''' is a cyclopropyl group, compound E-3 can be obtained by cyclopropanation of compound C-4. This type of reaction can be carried out using a suitable reagent such as diazomethane with or without a suitable catalyst, or using Simmons-Smith reaction conditions (such as, for example, diiodomethane with diethylzinc). The solvent used in this step is not particularly limited as long as it does not interfere with the reaction. Examples of the solvent include dichloromethane, ether, toluene, benzene or a mixed solvent thereof. The reaction temperature is generally -30°C to room temperature. The reaction time is not particularly limited and is generally 5 minutes to 24 hours, preferably 30 minutes to 24 hours.

[0279] General Program F

[0280] [Chemical formula 27]

[0281]

[0282] Where P is a protecting group, such as benzoyl or benzyl, R 3 '''''' is an alkyl or alkoxy group substituted by fluorine, and other symbols are the same as defined above.

[0283] General Procedure F is a method for preparing compound F-3 from compound E2 through multiple steps. Compound F-3 can be prepared according to the method described in General Procedure A from compound F-2.

[0284] Step 1

[0285] Compound F-1 can be prepared by ozonolysis of compound F-3, followed by reduction of the resulting aldehyde. The reaction can be carried out by methods known to those skilled in the art. Ozonolysis can be carried out under an ozone atmosphere, in a suitable solvent (such as dichloromethane, methanol and mixtures thereof), with a suitable reagent (such as triphenylphosphine, pyridine, dimethyl sulfide and trimethylamine) under a nitrogen atmosphere for reduction post-treatment. The temperature for generating ozonide is preferably -78°C, which can then be warmed to room temperature for reduction post-treatment. The reaction time is not particularly limited and is generally 30 minutes to 5 hours, preferably 30 minutes to 2 hours. The reduction of the generated aldehyde can be carried out in a one-pot process using a suitable reducing agent such as sodium borohydride or lithium aluminum hydride. The reaction temperature is preferably 0°C to room temperature. The reaction time is not particularly limited and is generally 30 minutes to 5 hours, preferably 30 minutes to 2 hours.

[0286] Step 2

[0287] When R 3 '''''' is CF 3 , CHF 2 or CH 2 F, compound F-2 can be obtained in two steps; oxidizing compound F-1 to an aldehyde or carboxylic acid and then fluorinating, or directly fluorinating compound F-1. The reaction can be carried out by methods known to those skilled in the art. For example, compound F-1 can be oxidized to the corresponding aldehyde under suitable oxidation conditions, such as, for example, TEMPO, Dess-Martin or Swern oxidation. The corresponding carboxylic acid can be obtained by oxidizing the resulting aldehyde, or directly oxidizing compound F-1 using suitable conditions such as, for example, Pinnick, TEMPO or Jones oxidation (oxadation). The solvent used in this step is not particularly limited as long as it does not interfere with the reaction. The reaction temperature is generally -78°C to room temperature. The reaction time is not particularly limited and is generally 5 minutes to 24 hours, preferably 30 minutes to 24 hours. The fluorination reaction can be carried out using suitable reagents such as, for example, DAST, Deoxofluor or sulfur tetrafluoride. The solvent used in this step is not particularly limited as long as it does not interfere with the reaction. Examples of the solvent include dichloromethane, tetrahydrofuran, 1,4-dioxane, 1,2-dimethoxyethane, diethyl ether, toluene, benzene, and mixed solvents thereof. The reaction temperature is preferably -78°C to 50°C. The reaction time is not particularly limited and is generally 5 minutes to 24 hours, preferably 30 minutes to 24 hours.

[0288] When R 3When '''''' is an alkoxy group, compound F-2 can be obtained by alkylation of the terminal alcohol of compound F-1. The reaction can be carried out using a suitable base (such as sodium hydride) and a corresponding electrophilic reagent (such as an alkyl halide, a mesylate or a triflate). The solvent used in this step is not particularly limited as long as it does not interfere with the reaction. Examples of the solvent include acetone, acetonitrile, tetrahydrofuran, DMF, DMA, DMSO, toluene, and a mixed solvent thereof. The reaction temperature is preferably 0°C to 100°C. The reaction time is not particularly limited and is generally 5 minutes to 24 hours, preferably 30 minutes to 24 hours.

[0289] The compounds of the present invention have BACE1 inhibitory activity and can effectively treat and / or prevent the following diseases induced by the production, secretion or deposition of β-amyloid protein, improve the symptoms of the diseases and prevent the progression of the diseases, such as Alzheimer's disease, precocious dementia, Alzheimer's type senile dementia, mild cognitive impairment (MCI), prodromal Alzheimer's disease (for example, MCI caused by Alzheimer's disease), Down syndrome, memory impairment, prion disease (Creutzfeldt-Jakob disease), Dutch hereditary cerebral hemorrhage with amyloidosis, amyloid cerebrovascular disease, other types of degenerative dementia, mixed dementia such as Alzheimer's disease coexisting with vascular dementia, dementia with Parkinson's disease, dementia with progressive supranuclear palsy, dementia with cortical basal degeneration, Alzheimer's disease with diffuse Lewy body disease, age-related macular degeneration, Parkinson's disease, amyloid angiopathy, etc.

[0290] Furthermore, the compounds of the invention are effective in preventing the progression of asymptomatic patients at risk of developing Alzheimer's disease (preclinical Alzheimer's disease).

[0291] "Asymptomatic patients at risk for Alzheimer's disease" include subjects who are cognitively and functionally normal but have underlying very early signs of Alzheimer's disease or typical age-related changes (e.g., mild white matter hyperintensity on MRI), and / or have low cerebrospinal fluid Aβ 1-42 For example, "asymptomatic patients at risk for Alzheimer's disease" include subjects whose Clinical Dementia Rating (CDR) or Clinical Dementia Rating-Japanese (CDR-J) score is 0 and / or whose Functional Assessment Staging System (FAST) stage is stage 1 or stage 2.

[0292] The compounds of the present invention not only have BACE1 inhibitory activity, but also have benefits as drugs. The compounds preferably have any one or more of the following excellent properties.

[0293] a) The compounds have weak inhibitory activity against CYP enzymes such as CYP1A2, CYP2C9, CYP2C19, CYP2D6, and CYP3A4.

[0294] b) The compound exhibits excellent pharmacokinetic properties, such as high bioavailability or low clearance rate.

[0295] c) The compounds have high metabolic stability.

[0296] d) The compound does not show irreversible inhibition of CYP enzymes (such as CYP3A4) within the concentration range of the measurement conditions described in this specification.

[0297] e) The compounds are not shown to be mutagenic.

[0298] f) The risk of the compounds to the cardiovascular system is low.

[0299] g) The compounds show high solubility.

[0300] h) The compounds show high brain distribution.

[0301] i) The compounds have high oral absorption.

[0302] j) The compound has a long half-life.

[0303] k) The compound has a high protein unbinding ratio.

[0304] 1) The compound is negative in the Ames test.

[0305] m) The compounds have high selectivity for BACE1 over BACE2.

[0306] n) The compound has a weak mechanism-based inhibition on the CYP enzyme. For example, the reactive metabolite of the compound has a weak inhibition on the CYP enzyme.

[0307] o) The compound produces few reactive metabolites.

[0308] p) The compound is a weak P-gp substrate.

[0309] Since the compound of the present invention has high inhibitory activity against BACE1 and / or high selectivity against other enzymes (e.g., BACE2), it can be a drug with reduced side effects. In addition, since the compound has a high effect of reducing the production of β-amyloid protein in a cell system, particularly a high effect of reducing the production of β-amyloid protein in the brain, it can be an excellent drug. In addition, by converting the compound into an optically active compound having appropriate stereochemistry, the compound can be a drug with a wider safety margin in terms of side effects.

[0310] When the pharmaceutical composition of the present invention is used, it can be administered orally or parenterally. Compositions for oral administration can be administered in conventional dosage forms, such as oral solid preparations (e.g., tablets, powders, granules, capsules, pills, films, etc.), oral liquid preparations (e.g., suspensions, emulsions, elixirs, syrups, lemonades, alcohols, aromatic waters, extractants, decoctions, tinctures, etc.), etc., which can be prepared and administered according to conventional methods. The tablets can be sugar-coated tablets, film-coated tablets, enteric-coated tablets, sustained-release tablets, lozenges, sublingual tablets, buccal tablets, chewable tablets, or orally disintegrating tablets. Powders and granules can be dry syrups. Capsules can be soft capsules, microcapsules, or sustained-release capsules.

[0311] The composition for parenteral administration can be administered in a common parenteral dosage form such as dermal, subcutaneous, intravenous, intraarterial, intramuscular, intraperitoneal, transmucosal, inhalation, nasal, ocular, inner ear or vaginal administration, etc. In the case of parenteral administration, any form commonly used, such as injection, drops, external preparations (e.g., eye drops, nasal drops, ear drops, aerosols, inhalants, lotions, infusions, liniments, mouthwashes, enemas, ointments, plasters, gels, creams, patches, pastes, external powders, suppositories, etc.), etc. can be preferably administered. The injection can be an emulsion of the type O / W, W / O, O / W / O, W / O / W, etc.

[0312] Due to their high oral absorbability, the compounds of the present invention may be preferably administered in oral dosage forms.

[0313] If necessary, the pharmaceutical composition can be prepared by mixing various pharmaceutical additives suitable for preparations with an effective amount of the compound of the present invention, such as excipients, adhesives, disintegrants and lubricants. In addition, by appropriately changing the effective amount, preparation and / or various pharmaceutical additives of the compound of the present invention, the pharmaceutical composition can be used for pediatric patients, elderly patients, critical cases or surgery. Pediatric pharmaceutical compositions are preferably applied to patients under 12 or 15 years old. In addition, pediatric pharmaceutical compositions can be applied to patients under 27 days of age after birth, 28 days of age to 23 months of age, 2 to 11 years old, 12 to 16 years old or 18 years old after birth. Elderly pharmaceutical compositions are preferably applied to patients 65 years old or older.

[0314] The dosage of the pharmaceutical composition of the present invention should be determined in consideration of the patient's age and weight, the type and extent of the disease, the route of administration, etc. The usual oral dosage for adults is 0.05-100 mg / kg / day, and preferably 0.1-10 mg / kg / day. For parenteral administration, the dosage varies greatly depending on the route of administration, and the usual dosage is 0.005-10 mg / kg / day, and preferably 0.01-1 mg / kg / day. The dosage can be administered once or more per day.

[0315] The compound of the present invention can be used in combination with other drugs for treating Alzheimer's disease, senile dementia, etc., such as acetylcholinesterase inhibitors (hereinafter referred to as concomitant drugs) to enhance the activity of the compound or reduce the dosage of the compound, etc. In this case, the administration time of the compound of the present invention and the concomitant drug is not limited, and they can be administered to the subject simultaneously or at regular intervals. In addition, the compound of the present invention and the concomitant drug can be administered as two different compositions containing each active ingredient or as a single composition containing two active ingredients.

[0316] The dosage of the concomitant drug can be appropriately selected based on the dosage used clinically. In addition, the mixing ratio of the compound of the present invention and the concomitant drug can be appropriately selected in consideration of the administration object, administration route, target disease, symptoms, combination, etc. For example, when the administration object is a human, the concomitant drug can be used in the range of 0.01 to 100 parts by weight relative to 1 part by weight of the compound of the present invention.

[0317] Examples of concomitant drugs are Donepezil hydrochloride, Tacrine, Galanthamine, Rivastigmine, Zanapezil, Memantine and Vinpocetine.

[0318] Example

[0319] The following Examples and Test Examples illustrate the present invention in more detail, but the present invention is not limited to these Examples.

[0320] In the examples, the meaning of each abbreviation is as follows:

[0321] Ac: Acetyl

[0322] Et: Ethyl

[0323] Bz: Benzoyl

[0324] DCC: Dicyclohexylcarbodiimide

[0325] DIC: Diisopropylcarbodiimide

[0326] iPr: isopropyl

[0327] Me: methyl

[0328] Ph: Phenyl

[0329] t-Bu: tert-butyl

[0330] TBS: tert-Butyldimethylsilyl

[0331] AIBN: Azobisisobutyronitrile

[0332] ADDP: 1,1'-(azodicarbonyl)dipiperidin

[0333] AIBN: 2,2'-azobis(isobutyronitrile)

[0334] (Boc) 2 O: Di-tert-butyl dicarbonate

[0335] BOMCl: benzyl chloromethyl ether oxymethyl chloride

[0336] DAST: N,N-Diethylaminosulfur trifluoride

[0337] DBU: 1,8-diazabicyclo[5.4.0]-7-undecene

[0338] DCM: Dichloromethane

[0339] DEAD: Diethyl azodicarboxylate

[0340] DIAD: diisopropyl azodicarboxylate

[0341] DIBAL: Diisobutylaluminum hydride

[0342] DIPEA: N,N-diisopropylethylamine

[0343] DMA: N,N-dimethylacetamide

[0344] DMAP: 4-dimethylaminopyridine

[0345] DMF: N,N-dimethylformamide

[0346] DMSO: dimethyl sulfoxide

[0347] EDC: 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide

[0348] HATU: O-(7-aza-1H-benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate

[0349] HOAt: 1-Hydroxy-7-aza-benzotriazole

[0350] HOBt: 1-Hydroxy-benzotriazole

[0351] LDA: Lithium diisopropylamide

[0352] LHMDS: Lithium bis(trimethylsilyl)amide

[0353] mCPBA: meta-chloroperbenzoic acid

[0354] NCS: N-chlorosuccinimide

[0355] NMP: N-Methylpyrrolidone

[0356] PPTS: Pyridinium p-toluenesulfonate

[0357] PyBOP: 1H-Benzotriazol-1-yloxy-tris(pyrrolidino)phosphonium hexafluorophosphate

[0358] TEMPO: 2,2,6,6-Tetramethylpiperidinyl 1-oxyl radical

[0359] TFA: trifluoroacetic acid

[0360] THF: Tetrahydrofuran

[0361] THP: 2-tetrahydropyranyl.

[0362] 1 H NMR spectra were recorded on a Bruker Advance 400 MHz spectrometer and the chemical shifts relative to the tetramethylsilane or residual solvent peaks are reported (CDCl 3 = 7.26 ppm, DMSO-d 6 = 2.50 ppm).

[0363] Analytical LC / MS (ESI positive or negative, retention time (RT)) data were recorded on a Shimadzu UFLC or Waters UPLC system under the following conditions:

[0364] Method A

[0365] Column: XBridge (registered trademark) C18 (5 μm, inner diameter (id) 4.6 x 50 mm) (Waters)

[0366] Flow rate: 3 mL / min

[0367] UV detection wavelength: 254 nm

[0368] Mobile phase: [A] 0.1% formic acid solution, and [B] 0.1% formic acid in acetonitrile solvent.

[0369] Gradient: A linear gradient from 10% to 100% solvent [B] was performed over 3 min and 100% solvent [B] was held for 1 min.

[0370] Method B

[0371] Column: Shim-pack XR-ODS (2.2 μm, id 50 x 3.0 mm) (Shimadzu)

[0372] Flow rate: 1.6 mL / min

[0373] Column temperature: 50℃

[0374] UV detection wavelength: 254 nm

[0375] Mobile phase: [A] 0.1% formic acid in water; [B] 0.1% formic acid in acetonitrile

[0376] Gradient: linear gradient from 10% to 100% solvent [B] over 3 min, and 100% solvent [B] for 1 min.

[0377] Method C

[0378] Column: BEH C18 (1.7 μm, 2.1 x 50 mm) (Waters)

[0379] Flow rate: 0.8 mL / min

[0380] UV detection wavelength: 254 nm

[0381] Mobile phase: [A] 95% H 2 O + 5% CH 3 10 mM CH in CN 3 COONH 4 , and [B] is acetonitrile.

[0382] Gradient: A linear gradient from 5% to 95% solvent [B] was performed over 1.3 min and 95% solvent [B] was held for 0.7 min.

[0383] Method D

[0384] Column: HSS T3 (1.8μm, 2.1 x 100mm) (Waters)

[0385] Flow rate: 0.7 mL / min

[0386] UV detection wavelength: 254 nm

[0387] Mobile phase: [A] 95% H 2 O + 5% CH 3 10 mM CH in CN 3 COONH 4 , and [B] is acetonitrile.

[0388] Gradient: A linear gradient from 0% to 95% solvent [B] was performed over 2.1 min and 95% solvent [B] was held for 0.5 min.

[0389] Example 1

[0390] Synthesis of Compound I-007

[0391] [Chemical formula 28]

[0392]

[0393] Step 1: Synthesis of compound 7-2

[0394] At 100 °C, 3,6-dihydro-2H-pyran 7-1 (6.20 g, 73.7 mmol) and Et 3 To a solution of N (10.2 ml, 73.7 mmol) in toluene (60 ml) was added ( Z )-ethyl 2-chloro-2-(hydroxyimino)acetate (22.3 g, 147 mmol). After stirring at reflux temperature for 4 hours, Et 3 N (10.2 ml, 73.7 mmol). After stirring at reflux temperature for 6 hours, the reaction mixture was added with ( Z )-ethyl 2-chloro-2-(hydroxyimino)acetate (11.2 g, 73.7 mmol) and Et 3 N (10.2 ml, 73.7 mmol). After stirring at reflux temperature for 5 hours, the reaction mixture was added with ( Z )-ethyl 2-chloro-2-(hydroxyimino)acetate (5.58 g, 36.9 mmol) and Et 3 N (5.1 ml, 36.9 mmol). After stirring at reflux temperature for 1 hour, the reaction mixture was cooled to room temperature. H 2 O, and the aqueous layer was extracted with EtOAc. The organic layer was washed with water and 2 SO4 Dry and filter. Concentrate the filtrate in vacuo. Add the crude product to a silica gel column and elute with 0% to 40% hexane / EtOAc. Evaporate the collected fractions to give compound 7-2 (5.10 g, 25.6 mmol, 35%) as an orange oil.

[0395]

[0396] Step 2: Synthesis of compound 7-3

[0397] At 0 °C, add NaBH 4 To a solution of (3.26 g, 86.0 mmol) in EtOH (140 ml) was added a solution of compound 7-2 (14.3 g, 71.9 mmol) in EtOH (140 mL). The reaction mixture was stirred at 40 ° C for 3 hours and treated with AcOH at 0 ° C. The reaction mixture was concentrated in vacuo. The crude product was added to a silica gel column and eluted with 50% to 70% hexane / EtOAc. The collected fractions were evaporated to give compound 7-3 (7.24 g, 46.1 mmol, 64%) as a colorless oil.

[0398]

[0399] Step 3: Synthesis of compound 7-4

[0400] At -78 °C, compound 7-3 (7.24 g, 46.1 mmol) was added to CH 2 Cl 2 90% DAST (13.5 ml, 92.0 mmol) was added to a solution of 4% 4-(4-hydroxy-2-nitropropene) (109 ml). The reaction mixture was stirred at room temperature for 2.5 hours and treated with aqueous potassium carbonate at 0°C. The mixture was washed with CHCl 3 Extraction and Na 2 SO 4 The crude product was added to a silica gel column and eluted with 20% to 40% hexane / EtOAc. The collected fractions were evaporated to give compound 7-4 (4.39 g, 27.6 mmol, 60%) as a yellow oil.

[0401]

[0402] Step 4: Synthesis of compound 7-5

[0403] To a solution of 1-bromo-2-fluorobenzene (4.39 g, 27.6 mmol) in toluene (176 mL) and THF (44 mL) was added at -78 °C. n-BuLi (1.64 M in n-hexane, 50.5 mL, 83.0 mmol), and the reaction mixture was stirred at the same temperature for 5 min. BF 3 -OEt 2 (4.2 ml, 33.1 mmol) and a solution of compound 7-4 (4.39 g, 27.6 mmol) in toluene (97 mL) were added, and the reaction mixture was stirred at the same temperature for 10 minutes. NH 4 Cl aqueous solution, and the water layer was extracted with EtOAc. The organic layer was washed with water and concentrated in vacuo. The crude product was added to a silica gel column and eluted with 10% to 20% hexane / EtOAc. The collected fractions were evaporated to obtain compound 7-5 (4.91 g, 19.2 mmol, 70%) as a yellow oil.

[0404]

[0405] Step 5: Synthesis of compound 7-6

[0406] To a solution of compound 7-5 (4.91 g, 19.2 mmol) in AcOH (49 ml) was added Zn (12.6 g, 192 mmol) at room temperature. After stirring at 60 ° C for 1 hour, the reaction mixture was cooled to room temperature and filtered through a Celite (registered trademark) pad. Aqueous potassium carbonate solution was added to the filtrate. The mixture was filtered through a Celite (registered trademark) pad, and the filtrate was extracted with EtOAc. The organic layer was washed with water and concentrated in vacuo. The crude product was added to a silica gel column and eluted with hexane / EtOAc 50%. The collected fractions were evaporated to give compound 7-6 (3.80 g, 14.8 mmol, 77%) as a colorless oil.

[0407]

[0408] Step 6: Synthesis of compound 7-7

[0409] At 0 °C, compound 7-6 (3.80 g, 14.8 mmol) was added to CH 2 Cl 2To a solution in 4% ethyl acetate (38 ml) was added benzoyl isothiocyanate (2.18 ml, 16.2 mmol). After stirring at room temperature for 19 hours, EDC-HCl (5.66 g, 29.5 mmol) was added to the reaction mixture at the same temperature. After stirring at 40 ° C for 3 hours, the reaction mixture was concentrated in vacuo. The crude product was added to a silica gel column and eluted with 10% to 40% hexane / EtOAc. The collected fractions were evaporated to give compound 7-7 (4.22 g, 10.9 mmol, 74%) as a white solid.

[0410]

[0411] Step 7: Synthesis of Compound 7-8

[0412] To a solution of compound 7-7 (4.22 g, 10.9 mmol) in MeOH (42 ml) was added DBU (1.81 ml, 12.0 mmol) at room temperature. After stirring at 60 °C for 7 hours, 2 mol / L HCl and Et 2 O. The organic layer was washed with H 2 O stripping. 2 CO 3 The aqueous layer was basified (pH = 8) and extracted with AcOEt. The organic layer was washed with water and concentrated in vacuo. The crude product was mixed with CHCl 3 Trituration together gave compound 7-8 (2.39 g, 8.47 mmol, 78%) as a yellow solid.

[0413]

[0414] Step 8: Synthesis of Compounds 7-9

[0415] To a solution of compound 7-8 (3.00 g, 10.6 mmol) in TFA (17.2 ml) was added sulfuric acid (4.25 ml, 80 mmol) at -15 °C. After stirring at the same temperature for 10 min, HNO 3 (0.712 ml, 15.9 mmol). After stirring at the same temperature for 15 minutes, the reaction mixture was washed with K 2 CO 3 The aqueous layer was extracted with AcOEt. The organic layer was washed with brine and 2 SO 4 Dry and filter. The filtrate was concentrated in vacuo to give compound 7-9 as a light yellow solid, which was used in the next step without purification.

[0416] Step 9: Synthesis of Compounds 7-10

[0417] At room temperature, in H 2 A solution of compound 7-9 and 10% Pd-C (674 mg, 3.00 mmol) in MeOH (101 ml) was stirred under an atmosphere. After stirring at the same temperature for 2 hours, the mixture was filtered through a Celite (registered trademark) pad. The filtrate was concentrated under vacuum. The crude product was purified by supercritical fluid chromatography (SFC) (Chiralpak (registered trademark) IC; 40% ethanol with 0.1% diethylamine) SFC to give compound 7-10 (1.35 g, 4.56 mmol, 44%) as a yellow solid.

[0418]

[0419] Step 10: Synthesis of Compound I-007

[0420] To a solution of compound 7-10 (31.6 mg, 0.106 mmol) in MeOH (2 ml) at 0°C, 5-(fluoromethoxy)pyrazine-2-carboxylic acid (18.3 mg, 0.106 mmol) and 2 mol / L HCl (0.053 ml, 0.106 mmol) were added. EDC-HCl (9.86 mg, 0.0370 mmol) was added to the reaction mixture at the same temperature. After stirring at room temperature for 30 minutes, the reaction mixture was washed with NaHCO 3 The aqueous layer was extracted with AcOEt. The organic layer was washed with H 2 O and brine, and washed with Na 2 SO 4 Dry and filter. The filtrate was concentrated in vacuo. The crude product was triturated with AcOEt / hexane to give compound I-007 (32.1 mg, 0.071 mmol, 67%) as a yellow solid.

[0421]

[0422] Example 2

[0423] Synthesis of Compound I-008

[0424] [Chemical formula 29]

[0425]

[0426] Step 1: Synthesis of compound 8-2

[0427] To a solution of compound 8-1 (4.10 g, 36.6 mmol) prepared according to a known method and phenyl isocyanate (12.0 ml, 110 mmol) in toluene (80 ml) at 110°C were added 2-(2-nitroethoxy)tetrahydro-2H-pyran (9.62 g, 54.9 mmol) and DIPEA (0.320 ml, 1.83 mmol) in toluene (30 ml). After stirring at reflux temperature for 2 hours, DIPEA (0.639 ml, 3.66 mmol) and phenyl isocyanate (12.0 ml, 110 mmol) were added to the reaction mixture. After stirring at reflux temperature for 4 hours, the reaction mixture was cooled to room temperature. The mixture was filtered and the filtrate was concentrated in vacuo. The crude product was added to a silica gel column and eluted with 10% to 30% hexane / EtOAc. The collected fractions were evaporated to give compound 8-2 (8.05 g, 29.9 mmol, 82 %) as an orange oil.

[0428]

[0429] Step 2: Synthesis of compound 8-3

[0430] To a solution of compound 8-2 (8.05 g, 29.9 mmol) in EtOH (81 ml) was added PPTS (1.50 g, 5.98 mmol) at room temperature. After stirring at 60 ° C for 2 hours, the reaction mixture was concentrated in vacuo. The crude product was added to a silica gel column and eluted with 10% to 80% hexane / EtOAc. The collected fractions were evaporated to give compound 8-3 (4.46 g, 29.9 mmol, 81%) as a brown oil.

[0431]

[0432] Step 3: Synthesis of compound 8-4

[0433] At -78 °C, compound 8-3 (4.93 g, 26.6 mmol) was added to CH 2 Cl 2 90% DAST (5.86 ml, 39.9 mmol) was added to the solution in 4% ethanol (49 ml). The reaction mixture was stirred at room temperature for 2 h and treated with aqueous potassium carbonate solution. The mixture was washed with CH 2 Cl 2 The organic layer was extracted with Na 2 SO 4The crude product was added to a silica gel column and eluted with 10% to 30% hexane / EtOAc. The collected fractions were evaporated to give compound 8-4 (4.50 g, 24.1 mmol, 90%) as a yellow oil.

[0434]

[0435] Step 4: Synthesis of compound 8-5

[0436] At -78 °C, compound 8-4 (4.50 g, 24.1 mmol) was added to CH 2 Cl 2 DIBAL (1.03 mol / L in hexane, 24.5 ml, 10.9 mmol) was added to the solution in 45 ml of 1% ethanol (45 ml). After stirring at the same temperature for 20 minutes, Rochelle salt was added to the reaction mixture. After stirring at room temperature for 3 hours, 2 mol / L HCl (pH = 4) was added to the mixture. NaCl was added to the mixture, and then CH 2 Cl 2 The combined organic layers were purified by Na 2 SO 4 Dried and filtered. The filtrate was concentrated under vacuum. The crude product was added to a silica gel column and eluted with 30% to 50% hexane / EtOAc. The collected fractions were evaporated to give compound 8-5 (2.65 g, 14.0 mmol, 58%) as a white solid, a diastereoisomer mixture.

[0437]

[0438] Step 5: Synthesis of compound 8-6

[0439] To a solution of compound 8-5 (2.55 g, 13.5 mmol) and triethylsilane (10.8 ml, 67.5 mmol) in DCM (41 ml) and MeCN (41 ml) was added BF 3 -OEt 2 (8.56 ml, 67.5 mmol). After stirring at the same temperature for 40 min, the reaction mixture was treated with aqueous sodium carbonate solution. The aqueous layer was washed with CH 2 Cl 2 The organic layer was extracted with Na 2 SO 4 Dry and filter. The filtrate was concentrated under vacuum to give compound 8-6 as a yellow oil, which was used in the next step without purification.

[0440] Step 6: Synthesis of compound 8-7

[0441] To a solution of 1-bromo-2-fluorobenzene (6.11 g, 34.9 mmol) in toluene (128 mL) and THF (16 mL) was added n -BuLi (1.64 M in n-hexane, 21.3 mL, 34.9 mmol), and the reaction mixture was stirred at the same temperature for 5 min. BF 3 -OEt 2 (1.77 ml, 14.0 mmol). After stirring at the same temperature for 10 minutes, a solution of crude compound 8-6 in THF (16 mL) and toluene (32 ml) was added to the mixture at -78°C, and the reaction mixture was stirred at the same temperature for 15 minutes. NH 4 Cl aqueous solution, and the water layer was extracted with EtOAc. The organic layer was washed with water and concentrated in vacuo. The crude product was added to a silica gel column and eluted with 0% to 20% hexane / EtOAc. The collected fractions were evaporated to give compound 8-7 (3.19 g, 11.8 mmol, 85%) as a yellow oil.

[0442]

[0443] Step 7: Synthesis of compound 8-8

[0444] To a solution of compound 8-7 (3.19 g, 11.9 mmol) in AcOH (31.9 ml) was added Zn (7.74 g, 118 mmol) at room temperature. After stirring at 60 °C for 2 hours, the reaction mixture was cooled to room temperature and filtered through a Celite (registered trademark) pad. The filtrate was treated with an aqueous potassium carbonate solution, and the mixture was extracted with EtOAc. The organic layer was washed with water and concentrated in vacuo to give compound 8-8 (3.07 g), which was used in the next reaction without further purification.

[0445] Step 8: Synthesis of Compound 8-9

[0446] At 0 °C, crude compound 8-8 (11.3 g) was added to CH 2 Cl 2 To the solution in 1% ethyl acetate (30.7 ml) was added benzoyl isothiocyanate (1.67 ml, 12.5 mmol). After stirring at room temperature for 14 hours, EDC-HCl (4.34 g, 22.7 mmol) was added to the reaction mixture. After stirring at 40 °C for 5 hours, the reaction mixture was concentrated in vacuo. The crude product was added to a silica gel column and washed with 20% CHCl 3The collected fractions were evaporated to give compound 8-9 (3.73 g, 9.32 mmol, 82%) as a yellow solid.

[0447]

[0448] Step 9: Synthesis of Compounds 8-10

[0449] To a solution of compound 8-9 (3.73 g, 9.32 mmol) in MeOH (37 ml) and THF (37 ml) was added hydrazine hydrate (4.53 ml, 93.0 mmol) at room temperature. After stirring at the same temperature for 14 hours, the reaction mixture was concentrated. The obtained residue was added to an amino silica gel column and eluted with 40% hexane / EtOAc. The collected fractions were evaporated to give compound 8-10 (2.30 g, 7.77 mmol, 83%) as a white solid.

[0450]

[0451] Step 10: Synthesis of Compounds 8-11

[0452] To a solution of compound 8-10 (2.30 g, 7.76 mmol) in TFA (12.6 ml) was added sulfuric acid (3.10 ml, 58.2 mmol) at -17 °C. After stirring at the same temperature for 10 min, HNO 3 (0.520 ml, 11.6 mmol). After stirring at the same temperature for 20 minutes, the reaction mixture was washed with K 2 CO 3 The aqueous solution was worked up and the aqueous layer was extracted with EtOAc. The organic layer was washed with water and concentrated in vacuo to give compound 8-11 (2.83 g), which was used in the next reaction without further purification.

[0453] Step 11: Synthesis of compounds 8-12

[0454] At room temperature, in H 2A solution of crude compound 8-11 (2.83 g, 7.76 mmol) and 10% Pd-C (566 mg) in MeOH (85 ml) was stirred under an atmosphere. After stirring at the same temperature for 2 hours, the mixture was filtered through a Celite (registered trademark) pad. The filtrate was concentrated under vacuum. The residue was triturated with AcOEt, and the resulting solid was collected, washed with AcOEt, and dried under reduced pressure to give compound 8-12 (1.50 g, 4.83 mmol, 62%) as a white solid. The filtrate was concentrated, and the residue was then purified by column chromatography (silica gel AcOEt / MeOH = 10 / 1) to give compound 8-12 (374 mg, 1.20 mmol, 16%) as a white solid.

[0455]

[0456] Step 12: Synthesis of Compound I-008

[0457] To a solution of compound 8-12 (50.1 mg, 0.161 mmol) in MeOH (3.0 ml) at 0°C, 5-(fluoromethoxy)pyrazine-2-carboxylic acid (27.7 mg, 0.161 mmol) and 2 mol / L HCl (0.080 ml, 0.177 mmol) were added. EDC-HCl (33.9 mg, 0.177 mmol) was added to the reaction mixture at the same temperature. After stirring at room temperature for 30 minutes, the reaction mixture was washed with NaHCO 3 The aqueous layer was extracted with AcOEt. The organic layer was washed with H 2 O and brine, and washed with Na 2 SO 4 Dry and concentrate in vacuo. The resulting residue was added to an amino silica gel column and eluted with 50% to 100% hexane / EtOAc. The collected fractions were evaporated. The residue was triturated with AcOEt / hexane to give compound 8-13 (45.5 mg, 0.098 mmol, 61%) as a white solid.

[0458]

[0459] Example 3

[0460] Synthesis of Compound I-009

[0461] [Chemical formula 30]

[0462]

[0463] Step 1: Synthesis of compound 9-5

[0464] Compound 9-5 was synthesized from compound 9-1 in a manner similar to the above scheme (Example 7, Step 1 to Step 4).

[0465] Step 2: Synthesis of compound 9-6

[0466] At 0 °C, compound 9-5 (8.85 g, 50.50 mmol) and allyltrimethylsilane (28.90 g, 253.0 mmol) were added to CH 2 Cl 2 To a solution in a mixture of 1% ethanol (177 ml) and MeOH (177 ml) was added boron trifluoride etherate complex (32.0 ml, 253.0 mmol). The mixture was stirred at room temperature for 1 hour and treated with saturated aqueous sodium bicarbonate solution. The mixture was extracted with EtOAc, and the organic layer was purified by MgSO 4 The mixture was dried and filtered. The filtrate was concentrated in vacuo. The crude product was added to a silica gel column and eluted with 30% hexane / EtOAc. The collected fractions were evaporated to give compound 9-6 (6.43 g, 32.28 mmol, 64%) as a colorless oil.

[0467]

[0468] Step 3: Synthesis of compound 9-7

[0469] Compound 9-7 was prepared in a manner similar to the above scheme (Example 1, Step 7) (yield; 63%).

[0470]

[0471] Step 4: Synthesis of compound 9-8

[0472] Compound 9-8 (yield; crude) was prepared in a manner similar to the above scheme (Example 1, Step 8).

[0473]

[0474] Step 5: Synthesis of compound 9-9

[0475] Compound 9-9 was prepared in a manner similar to the above scheme (Example 1, Step 9) (yield; 70%).

[0476]

[0477] Step 6: Synthesis of Compounds 9-10

[0478] At -78°C, 3 Compound 9-9 (6.40 g, 15.01 mmol) was blown into CH2 Cl 2 (224 ml) solution. After 1.5 hours, N 2 Bubble into the solution for 1.5 hours while gradually raising the reaction temperature to room temperature. Add PPh 3 (9.05 g, 34.50 mmol) and the resulting mixture was stirred at room temperature. After 1 hour, MeOH (64 ml) and NaBH were added at 0 °C. 4 (1.70 g, 45.0 mmol). After stirring at room temperature for 1 hour, the reaction mixture was washed with saturated NH 4 The mixture was extracted with EtOAc, and the organic layer was concentrated with MgSO 4 The mixture was dried and filtered. The filtrate was concentrated in vacuo. The crude product was added to a silica gel column and eluted with 70% hexane / EtOAc. The collected fractions were evaporated to give compound 9-10 (4.84 g, 11.24 mmol, 75%) as a white solid.

[0479]

[0480] Step 7: Synthesis of compounds 9-11

[0481] To a solution of compound 9-10 (4.41 g, 10.25 mmol) in THF (88 ml) was added PPh 3 (5.37 g, 20.49 mmol), imidazole (1.40 g, 20.49 mmol) and iodine (5.20 g, 20.49 mmol). The mixture was stirred at the same temperature for 1 hour and washed with 2% NaHSO 3 The mixture was extracted with EtOAc, and the organic layer was concentrated with MgSO 4 The crude product was added to a silica gel column and diluted with 20% CHCl 3 The collected fractions were evaporated to give compound 9-11 (5.22 g, 9.66 mmol, 94 %) as a white solid.

[0482]

[0483] Step 8: Synthesis of compounds 9-12

[0484] Compound 9-12 was prepared in a manner similar to the above scheme (Example 1, Step 11) (yield; 98%).

[0485]

[0486] Step 9: Synthesis of Compounds 9-13

[0487] Compounds 9-13 were prepared in a manner similar to the above scheme (Example 2, Step 1). The yield was not determined because the product was used in the next step without purification.

[0488]

[0489] Step 10: Synthesis of Compounds 9-14

[0490] Compound 9-14 was prepared in a manner similar to the above scheme (Example 1, Step 15). The yield was not determined because the product was used in the next step without purification.

[0491]

[0492] Step 11: Synthesis of Compounds 9-15

[0493] Compound 9-15 was prepared in a manner similar to the above scheme (Example 1, Step 16). The yield was not determined because the product was used in the next step without purification.

[0494]

[0495] Step 12: Synthesis of Compounds 9-16

[0496] Compound 16 was prepared in a manner similar to the above scheme (Example 1, Step 17). The crude product was purified by supercritical fluid chromatography (SFC) (Chiralpak (registered trademark) IC; 40% ethanol with 0.1% diethylamine) to give Compound 9-16. (Yield: 36%, 4 steps).

[0497]

[0498] Step 13: Synthesis of Compound I-009

[0499] Compound I-009 was prepared in a manner similar to the above scheme (Example 1, Step 18). (Yield: 77%).

[0500]

[0501] Example 4

[0502] Synthesis of Compound I-010

[0503] [Chemical formula 31]

[0504]

[0505] Step 1: Synthesis of compound 10-2

[0506] Compound 10-2 was prepared in a manner similar to the above scheme (Example 7, Step 5). (Yield: 77%).

[0507]

[0508] Step 2: Synthesis of compound 10-3

[0509] Compound 10-3 was prepared in a manner similar to the above scheme (Example 3, Step 1). (Yield: 32%).

[0510]

[0511] Step 3: Synthesis of compound 10-4

[0512] Compound 10-4 was prepared in a manner similar to the above scheme (Example 1, Step 15). The yield was not determined because the product was used in the next step without purification.

[0513]

[0514] Step 4: Synthesis of compound 10-5

[0515] Compound 10-5 was prepared in a manner similar to the above scheme (Example 1, Step 16). The yield was not determined because the product was used in the next step without purification.

[0516]

[0517] Step 5: Synthesis of compound 10-6

[0518] Compound 10-6 was prepared in a manner similar to the above scheme (Example 1, Step 17). The crude product was purified by supercritical fluid chromatography (SFC) (Chiralpak (registered trademark) IC; 40% isopropanol with 0.1% diethylamine) to give compound 10-6 (yield: 50%).

[0519]

[0520] Step 6: Synthesis of Compound I-010

[0521] Compound 10-7 was prepared in a manner similar to the above scheme (Example 1, Step 18). (Yield: 71%).

[0522]

[0523] Example 5

[0524] Synthesis of Compound I-011

[0525] [Chemical formula 32]

[0526]

[0527] Step 1: Synthesis of compound 11-2

[0528] Compound 11-2 was prepared in a manner similar to the above scheme (Example 1, Step 13). (Yield: 88%).

[0529]

[0530] Step 2: Synthesis of compound 11-3

[0531] Compound 11-3 was prepared in a manner similar to the above scheme (Example 1, Step 14). (Yield: 75%).

[0532]

[0533] Step 3: Synthesis of compound 11-4

[0534] Compound 11-4 was prepared in a manner similar to the above scheme (Example 1, Step 15). The yield was not determined because the product was used in the next step without purification.

[0535]

[0536] Step 4: Synthesis of compound 11-5

[0537] Compound 11-5 was prepared in a manner similar to the above scheme (Example 1, Step 16). The yield was not determined because the product was used in the next step without purification.

[0538]

[0539] Step 5: Synthesis of compound 11-6

[0540] Compound 11-6 was prepared in a manner similar to the above scheme (Example 1, Step 17). The crude product was purified by supercritical fluid chromatography (SFC) (Chiralpak (registered trademark) IC; 40% ethanol with 0.1% diethylamine) to give compound 11-6 (yield; 32%, 3 steps).

[0541]

[0542] Step 6: Synthesis of Compound I-011

[0543] Compound 11-8 was prepared in a manner similar to the above scheme (Example 1, Step 18). (Yield: 71%).

[0544]

[0545] The following compounds were prepared in a similar manner as described above. In the table, tR represents LC / MS retention time (minutes).

[0546]

[0547]

[0548]

[0549]

[0550]

[0551]

[0552]

[0553]

[0554]

[0555]

[0556]

[0557]

[0558]

[0559] Test Examples for the compounds of the present invention are described below.

[0560] Pharmacological Examples

[0561] The compounds provided in the present invention are inhibitors of β-site APP-cleaving enzyme 1 (BACE1). Inhibition of BACE1 (an aspartic protease) is believed to be relevant to the treatment of Alzheimer's disease (AD). It is believed that the production and accumulation of β-amyloid peptide (Aβ) derived from β-amyloid precursor protein (APP) play a key role in the onset and progression of AD. Aβ is produced by amyloid precursor protein (APP) by sequential cleavage at the N-terminus and C-terminus of the Aβ domain by BACE1 and γ-secretase, respectively.

[0562] It is expected that the compounds of the present invention exert their effects selectively on BACE1 relative to BACE2 due to their ability to selectively bind to BACE1 relative to BACE2 and inhibit the enzymatic activity of BACE1 relative to BACE2. The performance of such inhibitors is tested using a biochemical competitive radioligand binding assay, a biochemical fluorescence resonance energy transfer (FRET)-based assay, and the following cellular αLisa assay, which is suitable for identifying such compounds.

[0563] (Experimental Example 1: Biochemical competitive radioligand binding analysis of BACE1 and BACE2)

[0564] To probe the enzyme selectivity of BACE1 over BACE2, the binding affinity (Ki) to the respective purified enzyme was determined in a competitive radioligand binding assay, ie in competition with tritiated non-selective BACE1 / BACE 2 inhibitors.

[0565] In brief, in a test tube, the compound of interest is combined with a radioligand and a HEK 293-derived membrane containing BACE1 or BACE2. The competitive binding reaction is performed at pH 6.2 and incubated at room temperature until equilibrium is reached. The free radioligand is then separated from the bound radioligand by filtration using a Brandell 96 harvester. The filter is washed 4 times with wash buffer and the filter is punched into a scintillation tube. Ultima Gold scintillation cocktail is added and the sample is shaken. The next day, the vials are counted in a Tricarb scintillation counter to obtain the decays per minute (dpm) of the bound radioligand.

[0566] Calculate %CTL = (sample / HC) × 100, where HC is the high control, i.e., total binding of radioligand, allowing a curve to be fitted through the data points for different doses of the test compound. Calculate pIC 50 or IC 50 , and can be converted to K by the following formula i :

[0567] K i = IC 50 / (1+([RL] / K d )), where [RL] is the concentration of the radioligand used, and K d is the determined dissociation constant for the radioligand-membrane complex.

[0568] (Test Example 2)

[0569] (1) Analysis based on BACE1 biochemical FRET

[0570] The analysis is based on fluorescence resonance energy transfer analysis (FRET). The substrate used for the analysis is a 13-amino acid peptide derived from APP, which contains the "Swedish" Lys-Met / Asn-Leu mutation of the amyloid precursor protein (APP) β-secretase cleavage site. The substrate also contains two fluorophores: (7-methoxycoumarin-4-yl) acetic acid (Mca) is a fluorescence donor with an excitation wavelength of 320 nm and emission at 405 nm, and 2,4-dinitrophenol (Dnp) is a proprietary quenching receptor. The distance between the two groups is selected so that when light is excited, the donor fluorescence energy is significantly quenched by the receptor through resonance energy transfer. When cleaved by BACE1, the fluorophore Mca is separated from the quenching group Dnp, restoring the complete fluorescence yield of the donor. The increase in fluorescence is linearly related to the rate of protein decomposition.

[0571] Briefly, in a 384-well plate, in the absence or presence of compounds, a final concentration of 0.04 μg / mL of recombinant BACE1 protein was incubated with 20 μM substrate in an incubation buffer (final concentration: 33.3 mM citrate buffer pH 5.0, 0.033% PEG, 3% DMSO) at room temperature for 450 minutes. Then, the amount of protein degradation was directly measured by fluorescence measurement at T = 0'-120' and T = 450' (excitation at 320 nm and emission at 405 nm). The results are expressed in RFU (relative fluorescence units) as the difference between T450 and Tx (Tx is selected according to the reaction speed between 0 and 120 minutes).

[0572] The best fit curve was fitted to a plot of %Controlmin versus compound concentration using the minimum sum of squares method. This yielded the IC 50 value (inhibitory concentration causing 50% inhibition of activity).

[0573] LC = median low control value

[0574] = Low control: No enzyme reaction

[0575] HC = median high control value

[0576] = High Control: Reaction with enzyme

[0577] % effect = 100-[(sample-LC) / (HC-LC) *100]

[0578] %Control = (Sample / HC)*100

[0579] %Controlmin = (Sample-LC) / (HC-LC) *100

[0580] The compounds of the present invention are expected to have BACE1 inhibitory activity, and it is sufficient that the compound is able to inhibit the BACE1 receptor.

[0581] Specifically, according to the scheme shown above, IC50 is preferably 5000 nM or lower, more preferably 1000 nM or lower, further preferably 100 nM or lower.

[0582] (2) Analysis based on BACE2 biochemical FRET

[0583] The assay is a fluorescence resonance energy transfer (FRET) based assay. The substrate used for the assay contains the "Swedish" Lys-Met / Asn-Leu mutation of the amyloid precursor protein (APP) β-secretase cleavage site. The substrate also contains two fluorophores: (7-methoxycoumarin-4-yl)acetic acid (Mca) is a fluorescence donor with an excitation wavelength of 320 nm and emission at 405 nm, and

[0584] 2,4-Dinitrophenol (Dnp) is a proprietary quencher acceptor. The distance between the two groups is chosen so that upon photoexcitation, the donor fluorescence energy is significantly quenched by the acceptor via resonance energy transfer. Upon cleavage by β-secretase, the fluorophore Mca separates from the quencher group Dnp, restoring the full fluorescence yield of the donor. The increase in fluorescence is linearly related to the rate of protein degradation.

[0585] Briefly, in a 384-well plate, in the absence or presence of compounds, a final concentration of 0.04 μg / mL of recombinant BACE2 protein was incubated with 10 μM substrate in an incubation buffer (final concentration: 33.3 mM citrate buffer pH 5.0, 0.033% PEG, 2% DMSO) at room temperature for 450 minutes. Then, the amount of protein degradation was directly measured by fluorescence measurement at T = 0 and T = 450 (excitation at 320 nm and emission at 405 nm). The results are expressed in RFU (relative fluorescence units) as the difference between T450 and T0.

[0586] The best fit curve was fitted to a plot of % Controlmin versus compound concentration using the least squares method. This yielded the IC 50 value (inhibitory concentration causing 50% inhibition of activity).

[0587] LC = median low control value

[0588] = Low control: No enzyme reaction

[0589] HC = median high control value

[0590] = High Control: Reaction with enzyme

[0591] % effect = 100-[(sample-LC) / (HC-LC) *100]

[0592] %Control = (Sample / HC)*100

[0593] % Controlmin = (Sample-LC) / (HC-LC) *100.

[0594] The following exemplary compounds were tested essentially as described above and exhibited the following activities:

[0595]

[0596]

[0597] (Test Example 3-1: Effect of reducing β-amyloid protein in rat midbrain)

[0598] The compound of the present invention was suspended in 0.5% methylcellulose, the final concentration was adjusted to 2 mg / mL, and it was orally administered to male Crl: SD rats (7 to 9 weeks old) at 10 mg / kg. In the vehicle control group, only 0.5% methylcellulose was administered, and the administration test was performed with 3 to 8 animals per group. The brain was separated 3 hours after administration, the brain hemispheres were separated, their weights were measured, and the brain hemispheres were quickly frozen in liquid nitrogen and stored at -80°C until the extraction date. The frozen brain hemispheres were transferred to a homogenizer manufactured by Teflon (registered trademark) under ice cooling, and 4 volumes of extraction buffer (containing 1% CHAPS ({3- [(3-chloroamidopropyl) dimethylamino] -1-propanesulfonate}), 20 mmol / L Tris-HCl (pH 8.0), 150 mmol / L NaCl, complete (Roche) protease inhibitor) were added, and the up and down movements were repeated, and it was homogenized to dissolve for 2 minutes. The suspension was transferred to a centrifuge tube, allowed to stand on ice for 3 hours or longer, and then centrifuged at 100,000 x g for 20 minutes at 4°C. After centrifugation, the supernatant was transferred to an ELISA plate (Product No. 294-62501, WakoJunyaku Kogyo) for measuring beta-amyloid 40. ELISA measurement was performed according to the attached instructions. The reduction effect was calculated as a ratio compared to the brain beta-amyloid 40 level of the vehicle control group of each test.

[0599] (Test Example 3-2: Effect of reducing β-amyloid protein in the brain of mice)

[0600] The compound of the present invention was dissolved in 20% hydroxy-β-cyclodextrin, the final concentration was adjusted to 2 mg / mL, and it was orally administered to male Crl:CD1 (ICR) mice (6 to 8 weeks old) at 1-10 mg / kg. In the vehicle control group, only 20% hydroxy-β-cyclodextrin was administered, and the administration test was performed with 3 to 6 animals per group. The brain was separated 1 to 6 hours after administration, the brain hemisphere was separated, its weight was measured, and the brain hemisphere was quickly frozen in liquid nitrogen and stored at -80°C until the extraction date.

[0601] The frozen hemispheres were transferred to a homogenization tube containing ceramic beads and incubated on ice for 20 minutes. The ceramic beads were in 8 volumes of extraction buffer (containing 0.4% DEA (diethylamine), 50 mmol / L NaCl, complete protease inhibitors (Roche)) by weight. Thereafter, MP BIO FastPrep (registered trademark) -24 was used to homogenize with lysis matrix D 1.4 mm ceramic beads (20 seconds, 6m / s). Then, the tube was spun down for 1 minute, the supernatant was transferred to a centrifuge tube, and centrifuged at 221,000xg, 4°C for 50 minutes. After centrifugation, the supernatant was transferred to a NuncMaxisorp (registered trademark) plate (Thermo Fisher Scientific) coated with an antibody against the N-terminus of β-amyloid protein for measuring total β-amyloid protein, and the plate was incubated overnight at 4°C. The plates were washed with TBS-T (Tris buffered saline containing 0.05% Triton X-100), and HRP-conjugated 4G8 dissolved in PBS (pH 7.4) containing 0.1% casein was added to the plates and incubated at 4°C for 1 hour. After washing with TBS-T, SuperSignal ELISA Pico chemiluminescent substrate (Thermo Scientific) was added to the plates. Chemiluminescent counts were then measured as quickly as possible by ARVO (registered trademark) MX 1420 multi-label analyzer (Perkin Elmer). The reduction effect was calculated as a ratio compared to the total brain beta amyloid protein level of the vehicle control group of each test.

[0602] (Test Example 4-1: CYP3A4 fluorescence MBI test)

[0603] The CYP3A4 fluorescent MBI test is a test that examines the enhancement of CYP3A4 inhibition of a compound caused by a metabolic reaction.

[0604] 7-Benzyloxytrifluoromethylcoumarin (7-BFC) is debenzylated by CYP3A4 enzyme (enzyme expressed in E. coli) and produces 7-hydroxytrifluoromethylcoumarin (7-HFC) as a fluorescent metabolite. The assay is performed using the 7-HFC generation reaction as the labeling reaction.

[0605] The reaction conditions are as follows: substrate, 5.6 μmol / L 7-BFC; prereaction time, 0 or 30 minutes; substrate reaction time, 15 minutes; reaction temperature, 25°C (room temperature); CYP3A4 content (expressed in Escherichia coli), 62.5 pmol / mL at the prereaction time and 6.25 pmol / mL at the reaction time (10-fold dilution); the concentrations of the compounds of the present invention are 0.625, 1.25, 2.5, 5, 10, and 20 μmol / L (6 points).

[0606] The enzyme and the compounds of the present invention solution in K-Pi buffer (pH 7.4) are added to 96 orifice plates as pre-reaction solution with pre-reaction composition. A portion of the pre-reaction solution is transferred to another 96 orifice plates, and is used in 10 times of dilutions of the substrate in K-Pi buffer. NADPH is added as a cofactor to initiate labeling reaction (without pre-incubation). After the scheduled time of labeling reaction, acetonitrile / 0.5 mol / L three (trishydroxyaminomethane) = 4 / 1 (v / v) solution is added to terminate the labeling reaction. On the other hand, NADPH is also added to the remaining pre-reaction solution to initiate pre-reaction (with pre-incubation). After the scheduled time of pre-reaction, a portion is transferred to another 96 orifice plates, and is used in 10 times of dilutions of the substrate in K-Pi buffer to initiate labeling reaction. After the scheduled time of labeling reaction, acetonitrile / 0.5 mol / L three (trishydroxyaminomethane) = 4 / 1 (v / v) solution is added to terminate the labeling reaction. The fluorescence value of 7-HFC as a metabolite in each index reaction plate was measured using a fluorescence plate reader (Ex = 420 nm, Em = 535 nm).

[0607] A sample in which DMSO was added to the reaction system instead of the solution of the compound of the present invention was used as a control (100%) because DMSO was used as a solvent for dissolving the compound of the present invention. The remaining activity (%) was calculated for each concentration of the compound of the present invention added as a solution, and IC 50 The values ​​were calculated by back extrapolation using a logistic model with concentration and inhibition rate. 50 The IC without preincubation was subtracted from the value 50 When the difference in values ​​was 5 μM or more, it was defined as positive (+), and when the difference was 3 μM or less, it was defined as negative (-).

[0608] (Test Example 4-2: CYP3A4 (MDZ) MBI test)

[0609] The CYP3A4 (MDZ) MBI assay is an assay to investigate the mechanistic inhibition (MBI) potential of compounds on CYP3A4 inhibition. CYP3A4 inhibition is evaluated using the 1-hydroxylation reaction of midazolam (MDZ) via pooled human liver microsomes as a labeled reaction.

[0610] The reaction conditions were as follows: substrate, 10 μmol / L MDZ; pre-reaction time, 0 or 30 min; substrate reaction time, 2 min; reaction temperature, 37°C; protein content of the combined human liver microsomes, 0.5 mg / mL at the pre-reaction time and 0.05 pmg / mL at the reaction time (diluted 10 times); concentration of the compound of the invention, 1, 5, 10, 20 μmol / L (4 points).

[0611] With pre-reaction composition, the human liver microsomes and the compound of the present invention solution merged in K-Pi buffer (pH 7.4) are added to 96-well plates as pre-reaction solution. A portion of the pre-reaction solution is transferred to another 96-well plate and diluted 10 times with the substrate in K-Pi buffer. NADPH is added as a cofactor to initiate the labeling reaction (without pre-incubation). After the scheduled time of the labeling reaction, methanol / acetonitrile=1 / 1 (v / v) solution is added to terminate the labeling reaction. On the other hand, NADPH is also added to the remaining pre-reaction solution to initiate the pre-reaction (with pre-incubation). After the scheduled time of the pre-reaction, a portion is transferred to another 96-well plate and diluted 10 times with the substrate in K-Pi buffer to initiate the labeling reaction. After the scheduled time of the labeling reaction, methanol / acetonitrile=1 / 1 (v / v) solution is added to terminate the labeling reaction. After centrifugation at 3000 rpm for 15 minutes, the 1-hydroxymidazolam in the supernatant is quantified by LC / MS / MS.

[0612] A sample in which DMSO was added to the reaction system instead of the solution of the compound of the present invention was used as a control (100%) because DMSO was used as a solvent for dissolving the compound of the present invention. The remaining activity (%) was calculated for each concentration of the compound of the present invention added as a solution, and IC 50 The value was calculated by back extrapolation using a logistic model with concentration and inhibition rate. The IC value of the shift was calculated as "IC value without preincubation (0 minutes) / IC value with preincubation (30 minutes)". When the IC value of the shift was 1.5 or more, it was defined as positive. When the IC value of the shift was less than 1.1, it was defined as negative.

[0613]

[0614] (Test Example 5: CYP inhibition test)

[0615] The CYP inhibition test is a test for evaluating the inhibitory effect of the compounds of the present invention on the typical substrate metabolism reactions of CYP enzymes in human liver microsomes. The labeling reaction of the five major human CYP enzymes (CYP1A2, 2C9, 2C19, 2D6 and 3A4) is used as follows: O-deethylation of 7-ethoxyresorufin (CYP1A2), methyl hydroxylation of tolbutamide (CYP2C9), 4'-hydroxylation of mephenytoin (CYP2C19), O-demethylation of dextromethorphan (CYP2D6) and hydroxylation of terfenadine (CYP3A4). Commercially available pooled human liver microsomes are used as enzyme sources.

[0616] The reaction conditions are as follows: substrate, 0.5 μmol / L ethoxyresorufin (CYP1A2), 100 μmol / L tolbutamide (CYP2C9), 50 μmol / L S-mephenytoin (CYP2C19), 5 μmol / L dextromethorphan (CYP2D6), 1 μmol / L terfenadine (CYP3A4); reaction time, 15 minutes; reaction temperature, 37°C; enzyme, pooled human liver microsomes 0.2 mg protein / mL; concentration of the compound of the present invention, 1, 5, 10, 20 μmol / L (4 points).

[0617] The five substrates, human liver microsomes and the compound of the present invention in a 50 mmol / L Hepes buffer solution were added to a 96-well plate as a reaction solution in the above composition. NADPH was added to the 96-well plate as a cofactor to initiate the labeling reaction. After incubation at 37°C for 15 minutes, methanol / acetonitrile = 1 / 1 (v / v) solution was added to terminate the labeling reaction. After centrifugation at 3000rpm for 15 minutes, resorufin (CYP1A2 metabolite) in the supernatant was quantified by a fluorescent plate reader or LC / MS / MS, and hydroxytoluamide (CYP2C9 metabolite), 4'-hydroxymephenytoin (CYP2C19 metabolite), dextromethorphan (CYP2D6 metabolite) and terfenadine alcohol metabolite (CYP3A4 metabolite) in the supernatant were quantified by LC / MS / MS.

[0618] A sample in which DMSO was added to the reaction system instead of the solution of the compound of the present invention was used as a control (100%) because DMSO was used as a solvent for dissolving the compound of the present invention. The residual activity (%) was calculated for each concentration of the compound of the present invention, and IC 50 Values ​​were calculated by back extrapolation using a logistic model with concentration and inhibition rate.

[0619] (Test Example 6: Ames Fluctuation Test)

[0620] 20 μL of each frozen stock of salmonella typhimurium (TA98 and TA100 strains) was inoculated into 10 mL of liquid nutrient medium (2.5% Oxoid nutrient broth, No. 2), and the culture was incubated at 37°C with shaking for 10 hours. 7.70 to 8.00 mL of TA98 culture was centrifuged (2000 × g, 10 minutes) to remove the medium, and the bacteria were suspended in 7.70 mL of Micro F buffer (K 2 HPO 4 :3.5g / L, KH 2 PO 4 :1g / L, (NH 4 ) 2 SO 4 :1g / L, trisodium citrate dihydrate: 0.25g / L, MgSO 4 7H 2O: 0.1 g / L) and add the suspension to 120 mL of exposure medium (Micro F buffer containing biotin: 8 μg / mL, histidine: 0.2 μg / mL, glucose: 8 mg / mL). 3.10-3.42 mL of TA100 culture was added to 130 mL of exposure medium to prepare the test bacterial solution. 588 μL of the test bacterial solution (or in the case of a metabolic activation system, a mixed solution of 498 μL of the test bacterial solution and 90 μL of the S9 mixture) was mixed with 12 μL of each of the following solutions: a DMSO solution of the compound of the present invention (diluted in several stages from the maximum dose of 50 mg / mL in a ratio of 2 to 3 times); DMSO as a negative control; a 50 μg / mL DMSO solution of 4-nitroquinoline-1-oxide as a positive control for TA98 without a metabolic activation system; a 0.25 μg / mL DMSO solution of 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide as a positive control for TA100 without a metabolic activation system; a 40 μg / mL DMSO solution of 2-aminoanthracene as a positive control for TA98 with a metabolic activation system; or a 20 μg / mL DMSO solution of 2-aminoanthracene as a positive control for TA100 with a metabolic activation system. The mixed solution was incubated at 37° C. with shaking for 90 minutes. 460 μL of the bacterial solution exposed to the compound of the present invention was mixed with 2300 μL of indicator medium (Micro F buffer containing biotin: 8 μg / mL, histidine: 0.2 μg / mL, glucose: 8 mg / mL, bromocresol purple: 37.5 μg / mL), and 50 μL of each was distributed to 48 wells / dose in a microplate, and cultured at 37°C for 3 days. The color of the wells containing bacteria that have acquired the ability to proliferate by mutation of the gene encoding amino acid (histidine) synthetase changes from purple to yellow due to pH changes. The number of yellow wells in a total of 48 wells per dose was counted, and mutagenicity was evaluated by comparison with the negative control group. (-) means that the mutagenicity is negative, and (+) means positive.

[0621] (Test Example 7: Solubility Test)

[0622] The solubility of each compound of the present invention is measured under the condition of adding 1% DMSO. A 10 mmol / L solution of the compound is prepared with DMSO, and 2 μL of the solution of the compound of the present invention is added to 198 μL of JP No. 1 fluid (water is added to 2.0 g of sodium chloride and 7.0 mL of hydrochloric acid to reach 1000 mL) and JP No. 2 fluid (1 volume of water is added to 1 volume of 3.40 g of potassium dihydrogen phosphate and 3.55 g of anhydrous disodium hydrogen phosphate dissolved in water to reach 1000 mL). The mixture is left to stand at 25° C. for 16 hours or shaken at room temperature for 1 hour, and the mixture is vacuum filtered. Use methanol / water=1 / 1 (v / v) or MeCN / MeOH / H 2 The filtrate was diluted ten- or one-hundred-fold by 0 (=1 / 1 / 2), and the compound concentration in the filtrate was measured by LC / MS or solid phase extraction (SPE) / MS by absolute calibration method.

[0623] (Test Example 8: Metabolic stability test)

[0624] Using commercially available pooled human liver microsomes, the compound of the present invention was reacted for a constant time, and the residual ratio was calculated by comparing the reacted sample with the unreacted sample, thereby evaluating the degree of metabolism in the liver.

[0625] The reaction (oxidation reaction) was carried out at 37°C in 0.2 mL buffer (50 mmol / L Tris-HCl pH 7.4, 150 mmol / L potassium chloride, 10 mmol / L magnesium chloride) containing 0.5 mg protein / mL human liver microsomes for 0 minutes or 30 minutes in the presence of 1 mmol / L NADPH. After the reaction, 50 μL of the reaction solution was added to 100 μL of methanol / acetonitrile = 1 / 1 (v / v), mixed and centrifuged at 3000 rpm for 15 minutes. The compound of the present invention in the supernatant was quantified by LC / MS / MS or solid phase extraction (SPE) / MS, and the remaining amount of the compound of the present invention after the reaction was calculated, and the amount of the compound at 0 minute reaction time was calculated as 100%.

[0626]

[0627] (Test Example 9: hERG test)

[0628] In order to evaluate the risk of prolonging the QT interval of the electrocardiogram, CHO cells expressing human ether-a-go-go related gene (hERG) channels were used to study the effects of the compounds of the present invention on delayed rectifier K+ current (I Kr ), the K+ current plays an important role in the ventricular repolarization process.

[0629] The cells were maintained at a membrane potential of -80 mV using the whole-cell patch clamp method using a fully automated patch clamp system (Qpatch; Sophion Bioscience A / S). After applying a leak potential of -50 mV, the I induced by a depolarizing pulse stimulation at +20 mV for 2 seconds and a repolarizing pulse stimulation at -50 mV for 2 seconds was recorded. Kr .

[0630] After the generated current stabilized, the extracellular solution containing the compound of the present invention dissolved at the target concentration (NaCl: 145 mmol / L, KCl: 4 mmol / L, CaCl 2 :2 mmol / L、MgCl 2 :1 mmol / L, HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid): 10 mmol / L, glucose: 10 mmol / L, pH=7.4) were applied to the cells at room temperature for 10 minutes. Kr The absolute value of the peak tail current was measured based on the current value at resting membrane potential using analysis software (Qpatch analysis software; Sophion Bioscience A / S). The inhibition percentage of the peak tail current before administration of the compound of the present invention was further calculated and compared with the vehicle administration group (0.1% dimethyl sulfoxide solution) to evaluate the effect of the compound of the present invention on I Kr impact.

[0631] The following data show the inhibitory effects of compounds of the invention at 3 μM.

[0632]

[0633] (Test Example 10: Powder Solubility Test)

[0634] An appropriate amount of the compounds of this invention is placed in an appropriate container. 200 μL of JP 1 fluid 1 (water is added to 2.0 g of sodium chloride and 7.0 mL of hydrochloric acid to reach 1000 mL), 200 μL of JP fluid 2 (1 volume of water is added to 1 volume of 3.40 g of potassium dihydrogen phosphate and 3.55 g of anhydrous disodium hydrogen phosphate dissolved in water to reach 1000 mL), 200 μL of fasting state simulated intestinal fluid (FaSSIF) and 200 μL of fed state simulated intestinal fluid (FeSSIF) are added to respective containers. When the total amount of the compounds of this invention is dissolved after adding the test fluid, the compound is appropriately added. The container is sealed and shaken at 37°C for 1 hour and / or 24 hours. The mixture is filtered and 100 μL of methanol is added to each filtrate (100 μL) to dilute the filtrate twice. If necessary, the dilution ratio can be changed. After confirming that there are no bubbles and precipitation in the diluted solution, the container is sealed and shaken. Quantification was performed by HPLC using absolute calibration.

[0635] (Test Example 11: Pharmacokinetic study)

[0636] Materials and methods used for oral absorption studies

[0637] (1) Animals: mice or rats

[0638] (2) Housing conditions: Mice or rats were allowed free access to tap water and solid food.

[0639] (3) Dosage and grouping: Oral or intravenous administration at a predetermined dose; grouping as follows (dose depends on the compound)

[0640] Oral administration: about 1 to 30 mg / kg (n = 2 to 3)

[0641] Intravenous administration: approximately 0.5 to 10 mg / kg (n = 2 to 3)

[0642] (4) Dosage preparation: For oral administration, in the form of solution or suspension; for intravenous administration, in the form of dissolved

[0643] (5) Administration method: In oral administration, the drug is administered by force using a syringe connected to a flexible feeding tube; in intravenous administration, the drug is administered from the tail vein using a syringe connected to a needle.

[0644] (6) Evaluation items: Blood was collected at a predetermined time, and the plasma concentration of the compound of the present invention was measured by LC / MS / MS

[0645] (7) Statistical analysis: Regarding the changes in plasma concentration of the compound of the present invention, the area under the plasma concentration-time curve (AUC) was calculated by the trapezoidal method, and the bioavailability (BA) of the compound of the present invention was calculated from the AUC of the oral administration group and the intravenous administration group.

[0646] (Test Example 12: Brain distribution study)

[0647] The compound of the present invention was intravenously administered to rats at a dose of about 0.5 mg / mL / kg. After 30 minutes, all blood was drawn from the abdominal aorta under isoflurane anesthesia to cause exsanguination.

[0648] The brain was removed and a 20-25% homogenate was prepared with distilled water.

[0649] The obtained blood was used as plasma after centrifugation. The control plasma was added to the brain sample at 1:1. The control brain homogenate was added to the plasma sample at 1:1. Each sample was measured using LC / MS / MS. The obtained area ratio (brain / plasma) was used for the brain Kp value.

[0650] (Test Example 13: Ames test)

[0651] The Ames test was conducted by using Salmonella (Salmonella typhimurium) TA 98, TA100, TA1535 and TA1537 and Escherichia coli WP2uvrA as test strains with or without metabolic activation in a pre-culture method to examine the presence or absence of gene mutagenicity of the compounds of the present invention.

[0652] (Test Example 14: P-gp substrate test)

[0653] 1. Cell lines:

[0654] a. MDR1 / LLC-PK1 (Becton Dickinson)

[0655] b. LLC-PK1 (Becton Dickinson).

[0656] 2. Reference substrate:

[0657] a. Digoxin (2 μM).

[0658] Methods and steps

[0659] 1. LLC-PK1 cells expressing MDR1 and their parental cells were routinely cultured in medium A (medium 199 (Invitrogen), supplemented with 10% FBS (Invitrogen), gentamicin (0.05 mg / mL, Invitrogen), and hygromycin B (100 μg / mL, Invitrogen)) at 37°C in 5% CO2 / 95% O2 atmosphere. For transport experiments, these cells were plated at 1.4 × 10 4 The density of cells / insert was seeded on Transwell (registered trademark) inserts (96 wells, pore size: 0.4 μm, Coaster), and medium B (medium 199 supplemented with 10% FBS and 0.05 mg / mL gentamicin) was added to the feeder tray. The cells were incubated in a CO2 incubator (5% CO2 / 95% O2 gas, 37°C), and the medium on the apical and basolateral sides was replaced every 48-72 hours after seeding. The cells were used between 4 and 6 days after seeding.

[0660] 2. Remove the medium in the culture inserts seeded with MDR1 expressing cells or parental cells by aspiration and wash with HBSS. The apical side (140 μL) or the basal side (175 μL) is replaced with a transport buffer containing a reference substrate and the present invention, and then an aliquot (50 μL) of the transport buffer in the donor side is collected to estimate the initial concentration of the reference substrate and the present invention. After incubation at 37°C for the planned time, an aliquot (50 μL) of the transport buffer in the donor and recipient sides is collected. Analyze in duplicate or triplicate.

[0661] 3. Quantification of the reference substrate and the present invention in an aliquot by LC / MS / MS.

[0662] calculate

[0663] The amount of permeability across monolayers of MDR1 expressing cells and parental cells was determined and the permeability coefficient (Pe) was calculated using Excel 2003 from the following equation:

[0664] Pe (cm / sec) = osmotic pressure (pmol) / cell membrane area (cm 2 ) / initial concentration (nM) / incubation time (sec)

[0665] Here, the permeation amount was calculated from the permeation concentration (nM, concentration on the receptor side) of the substance after a given incubation time (sec) multiplied by the volume (mL), and a cell membrane area of ​​0.1433 (cm2) was used.

[0666] Calculate the Efflux Ratio using the following equation:

[0667] Outflow ratio = base side to apical Pe / apical side to base side Pe

[0668] Calculate the net flux using the following equation:

[0669] Net flux = efflux ratio in MDR1-expressing cells / efflux ratio in parental cells.

[0670]

[0671] (Test Example 15: Inhibitory effect on P-gp transport)

[0672] Material

[0673] 1. Cell lines:

[0674] a. MDR1 / LLC-PK1 (Becton Dickinson)

[0675] b. LLC-PK1 (Becton Dickinson).

[0676] 2. Reference substrate:

[0677] a. [ 3 H]Digoxin (1 μM)

[0678] b. [ 14 C] Mannitol (1 μM).

[0679] 3. Reference inhibitor:

[0680] Verapamil (1 μM)

[0681] Methods and steps

[0682] 1. LLC-PK1 cells expressing MDR1 and their parental cells were cultured in medium A (medium 199 (Invitrogen) supplemented with 10% FBS (Invitrogen), gentamicin (0.05 mg / mL, Invitrogen), and hygromycin B (100 μg / mL, Invitrogen)) at 37°C in 5% CO. 2 / 95% O 2 For transport experiments, these cells were cultured at 1.4 × 10 4 The cells / insert were seeded on Transwell (registered trademark) inserts (96 wells, pore size: 0.4 μm, Coaster), and medium B (medium 199 supplemented with 10% FBS and 0.05 mg / mL gentamicin) was added to the feed tray. These cells were grown under CO2 Incubate in an incubator (5% CO 2 / 95% O 2 Gas, 37°C), and the apical and basolateral media were replaced every 48-72 hours after seeding. The cells were used between 6 and 9 days after seeding.

[0683] 2. Remove the medium in the culture inserts seeded with MDR1 expressing cells or parental cells by aspiration and wash with HBSS. The apical side (150 μL) or the basal side (200 μL) is replaced with transport buffer containing a reference substrate (with or without the compound of the present invention), and then an aliquot (50 μL) of the transport buffer in the donor side is collected to estimate the initial concentration of the reference substrate. After incubation at 37°C for the planned time, an aliquot (50 μL) of the transport buffer in the donor and receptor sides is collected. The analysis is performed in triplicate.

[0684] 3. Mix an aliquot (50 μL) of transport buffer with 5 mL of scintillation cocktail and measure the radioactivity using a liquid scintillation counter.

[0685] calculate

[0686] The permeability across monolayers of MDR1 expressing and parental cells was determined and the permeability coefficient (Pe) was calculated using Excel 2003 by the following equation:

[0687] Pe (cm / sec) = osmotic pressure (pmol) / cell membrane area (cm 2 ) / initial concentration (nM) / incubation time (sec)

[0688] The permeation amount was calculated by multiplying the permeation concentration (nM, concentration on the receptor side) of the substance after a given incubation time (sec) by the volume (mL), and a 0.33 (cm 2 ) of the cell membrane area.

[0689] Calculate the outflow ratio using the following equation:

[0690] Outflow ratio = base side to apex Pe / apex to base side Pe

[0691] Calculate the net flux using the following equation:

[0692] Net flux = efflux ratio in MDR1-expressing cells / efflux ratio in parental cells

[0693] The percentage of control is calculated as the ratio of the net efflux of the reference compound in the presence of the compound of the invention to the absence of the compound of the invention.

[0694] IC was calculated using the curve fitting program XLfit50 value.

[0695] (Test Example 16: P-gp substrate test using mdr1a / 1b (- / -) B6 mice)

[0696] Material

[0697] Animals: mdr1a / 1b (- / -) B6 mice (KO mice) or C57BL / 6J mice (wild-type mice).

[0698] Methods and steps

[0699] 1. The animals may be fed prior to administration of the compounds of the invention.

[0700] 2. The compound of the present invention is administered to three animals at each time point, and blood and brain samples are removed at selected time points (e.g., 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, or 24 hours) after administration. Blood (0.3-0.7 mL) is collected via trunk bleeding using a syringe containing an anticoagulant (EDTA and heparin). Blood and tissue (e.g., brain) samples are immediately placed on thawing ice.

[0701] 3. Centrifuge the blood samples (1780 × g, 10 minutes) to remove cells and obtain plasma. Then, transfer the plasma samples into clean tubes and store them in a -70°C freezer until analysis.

[0702] 4. Homogenize tissue (e.g., brain) samples at a ratio of 1:3 tissue weight to milliliters (ml) of distilled water, transfer to a clean tube, and store in a -70°C freezer until analysis.

[0703] 5. Prepare plasma and tissue (e.g., brain) samples using protein precipitation and analyze by LC / MS / MS. Calibrate the analytical method by including a standard curve constructed with blank plasma or brain samples and known amounts of the analyte. Include quality control samples to monitor the accuracy and precision of the method.

[0704] 6. Introduce the plasma and brain concentration values ​​(ng / mL and ng / g) into a suitable mathematical tool for calculating pharmacokinetic parameters. One public platform is the WinNonlin (registered trademark) pharmacokinetic software modeling program.

[0705] calculate

[0706] Kp; ratio of tissue to plasma concentration

[0707] Kp ratio = Kp in KO mice / Kp in wild-type mice

[0708] The ratio of AUC tissue / AUC plasma of KO / wild mice = {AUC tissue / AUC plasma (KO mice)} / {AUC tissue / AUC plasma (wild mice)}.

[0709] (Test Example 17: Cardiovascular study in anesthetized guinea pigs)

[0710] Animal species: Guinea pigs (Slc: Hartley, 4-5 weeks old, male), N = 4;

[0711] Study Design:

[0712] Dosage: 3, 10 and 30 mg / kg (in principle)

[0713] (The compound of the present invention is cumulatively administered)

[0714] preparation:

[0715] The composition of the vehicle; dimethylacetamide (DMA): polyethylene glycol 400 (PEG400): distilled water (DW) = 1:7:2 (in principle).

[0716] The compound of the invention was dissolved with DMA, and then PEG400 and DW were added. Finally, solutions of 1.5, 5 and 15 mg / mL were prepared.

[0717] Route and schedule of administration:

[0718] Intravenous infusion for 10 min (2 mL / kg);

[0719] 0 to 10 minutes: 3 mg / kg, 30 to 40 minutes: 10 mg / kg, 60 to 70 minutes: 30 mg / kg

[0720] Vehicle was administered on the same schedule as above.

[0721] Group composition:

[0722] Vehicle group and the compound of the present invention group (4 guinea pigs in each group).

[0723] Evaluation Methodology:

[0724] Assessment items:

[0725] Mean blood pressure [mmHg], heart rate (derived from blood pressure waveform [beats / min]), QTc (ms), and toxicokinetics.

[0726] Experimental steps:

[0727] Guinea pigs were anesthetized with urethane (1.4 g / kg, ip), and polyethylene tubes were inserted into the carotid artery (for blood pressure measurement and blood collection) and the jugular vein (for infusion of the test compound). Electrodes (lead 2) were connected subcutaneously. Blood pressure, heart rate, and electrocardiogram (ECG) were measured using a PowerLab (registered trademark) system (ADInstruments).

[0728] Toxicokinetics:

[0729] At each assessment point, approximately 0.3 mL of blood (approximately 120 μL of plasma) was drawn from the carotid artery with a syringe containing sodium heparin and immediately cooled on ice. Plasma samples were obtained by centrifugation (4°C, 10,000 rpm, 9,300 × g, 2 min). The plasma separation step was performed on ice or at 4°C. The obtained plasma (TK sample) was stored in a deep freezer (set temperature: -80°C).

[0730] Analysis method: The mean blood pressure and heart rate were calculated as the average of blood pressure and heart rate within 30 seconds at each evaluation time point. ECG parameters (QT interval [ms] and QTc) were obtained from the average waveform of 10 seconds of continuous beats at the evaluation time point. QTc [Fridericia formula; QTc = QT / (RR)1 / 3)] was calculated using the PowerLab (registered trademark) system. The incidence of arrhythmias was visually evaluated for all ECG records of all four animals (from 0.5 hours before drug administration to the end of the experiment).

[0731] Evaluation time point:

[0732] Before the first dose (pre-dose), and 10, 25, 40, 55, 70 and 85 min after the first dose.

[0733] QTc data analysis:

[0734] The percent change (%) of QTc from the pre-dose value was calculated (pre-dose value was taken as 100%). The relative QTc was compared with the vehicle value at the same assessment point.

[0735] (Test Example 18: Pharmacology in Beagle Dogs)

[0736] The test compound was tested to assess the effect on beta-amyloid distribution in the cerebrospinal fluid (CSF) of dogs following a single dose, combined with pharmacokinetic (PK) tracking and limited safety assessments.

[0737] In the case of the compounds shown below, 2 or 4 beagles (1 or 2 males, 1 or 2 females) were administered with vehicle (1 mL / kg of 20% cyclodextrin aqueous solution), and 4 beagles (2 males and 2 females) per administration group were administered with the test compound on an empty stomach at the doses shown in Table 20 in 20% cyclodextrin aqueous solution at concentrations in mg / mL, the same as the doses given in mg / kg.

[0738] CSF was taken directly from the lateral ventricle in awake animals via a cannula screwed into the skull and covered by subcutaneous tissue and skin before and after dosing. Eight hours after dosing, animals were allowed to eat their regular diet for 30 minutes. Blood was collected before and 8 hours and 25 hours after dosing for PK tracking (0.5, 1, 2, 4, 8, 25 and 49 hours), and serum samples were collected for biochemical analysis. CSF samples were used to measure Abeta 1-37, Abeta 1-38, Abeta 1-40 and Abeta 1-42. The results are summarized in the following table:

[0739] .

[0740] The % reduction is indicated at 8 h and at the last time point where a relevant reduction (>20% reduction) was observed.

[0741] (Test Example 19: Dansyl GSH capture test)

[0742] Dansylglutathione (GSH) capture is a test for studying reactive metabolites.

[0743] The reaction conditions were as follows: substrate, 50 μmol / L of the compound of the present invention; capture agent, 0.1 mmol / L dansyl GSH; protein content of the combined human liver microsomes, 1 mg / mL; pre-reaction time, 5 minutes; reaction time, 60 minutes; reaction temperature, 37°C.

[0744] A solution of the combined human liver microsomes and the compound of the present invention in K-Pi buffer (pH 7.4) was added to a 96-well plate as a pre-reaction solution in a pre-reaction composition. NADPH was added as a cofactor to initiate the reaction. After a predetermined reaction time, a portion was transferred to another 96-well plate, and an acetonitrile solution including 5 mmol / L dithiothreitol was added to terminate the reaction. After centrifugation at 3000 rpm for 15 minutes, the fluorescence peak area of ​​the metabolite captured by dansyl GSH was quantified by HPLC with fluorescence detection.

[0745] (Test Example 20: 14 C]-KCN capture assay)

[0746] [ 14 C]-potassium cyanide (KCN) capture is a test for studying reactive metabolites.

[0747] The reaction conditions are as follows: substrate, 10 or 50 μmol / L of the compound of the present invention; capture agent, 1 mmol / L [ 14 C]-KCN (11.7 μCi / tube); protein content of pooled human liver microsomes, 1 mg / mL; pre-reaction time, 5 min; reaction time, 60 min; reaction temperature, 37°C.

[0748] A solution of the combined human liver microsomes and the compound of the invention in K-Pi buffer (pH 7.4) was added as a pre-reaction solution to a 96-well plate in a pre-reaction composition. NADPH was added as a cofactor to initiate the reaction. After a predetermined time, the metabolic reaction was terminated and the [ 14 C]-KCN captured metabolites were extracted into 100 μL methanol solution. Quantification by radio-HPLC system [ 14 C]-KCN captured radioactive peak areas of metabolites.

[0749] Formulation Examples

[0750] The following formulation examples are merely illustrative and are not intended to limit the scope of the present invention.

[0751] Preparation Example 1: Tablet

[0752] Compound of the present invention 15 mg

[0753] Lactose 15 mg

[0754] Calcium stearate 3 mg

[0755] All the above ingredients (except calcium stearate) are mixed uniformly. The mixture is then crushed, granulated and dried to obtain particles of suitable size. Then, calcium stearate is added to the granules. Finally, tableting is performed under compression force.

[0756] Preparation Example 2: Capsules

[0757] Compound of the present invention 10 mg

[0758] Magnesium stearate 10 mg

[0759] Lactose 80 mg

[0760] The above ingredients are uniformly mixed to obtain powder or fine granules, and the obtained mixture is then filled into capsules.

[0761] Preparation Example 3: Granules

[0762] Compound of the present invention 30 g

[0763] Lactose 265 g

[0764] Magnesium stearate 5 g

[0765] After the above ingredients are uniformly mixed, the mixture is compressed. The compressed material is crushed, granulated and sieved to obtain particles of suitable size.

[0766] Preparation Example 4: Orally disintegrating tablet

[0767] The compound of the present invention and crystalline cellulose are mixed, granulated and tableted to obtain an orally disintegrating tablet.

[0768] Preparation Example 5: Dry syrup

[0769] The compound of the present invention and lactose are mixed, crushed, granulated and sieved to obtain a dry syrup of suitable size.

[0770] Preparation Example 6: Injection

[0771] The compound of the present invention is mixed with phosphate buffer to obtain an injection.

[0772] Preparation Example 7: Infusion

[0773] The compound of the present invention is mixed with phosphate buffer to obtain an injection.

[0774] Preparation Example 8: Inhalant

[0775] The compound of the present invention and lactose are mixed and finely crushed to obtain an inhalant.

[0776] Preparation Example 9: Ointment

[0777] The compound of the present invention is mixed with petrolatum to obtain an ointment.

[0778] Preparation Example 10: Patch

[0779] The compound of the present invention and a base such as an adhesive plaster or the like are mixed to obtain a patch.

[0780] [Industrial Applicability]

[0781] The compound of the present invention may be a drug useful as an agent for treating or preventing a disease caused by the production, secretion and / or deposition of β-amyloid protein.

Claims

1. A compound of formula (IA) or a pharmaceutically acceptable salt thereof: [Chemical formula 1] in R 3 is C1-C3 alkyl optionally substituted by one or more halogens; t is 0 or 1; and R 7a is C1-C3 alkoxy optionally substituted by one or more halogens.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein [Chemical formula 2] 3. Compound or its pharmaceutically acceptable salt: [Chemical formula 3] 4. Compound or its pharmaceutically acceptable salt: [Chemical formula 4] 5. Compound or its pharmaceutically acceptable salt: [Chemical formula 5] 6. Compound or its pharmaceutically acceptable salt: [Chemical formula 6] 7. A compound or a pharmaceutically acceptable salt thereof: [Chemical formula 7] 8. A compound or a pharmaceutically acceptable salt thereof: [Chemical formula 8] 9. A compound or a pharmaceutically acceptable salt thereof: [Chemical formula 9] 10. A compound or a pharmaceutically acceptable salt thereof: [Chemical formula 10] 11. A pharmaceutical composition comprising the compound according to any one of claims 1 to 10 or a pharmaceutically acceptable salt thereof.

12. A BACE1 inhibitor comprising the compound according to any one of claims 1 to 10 or a pharmaceutically acceptable salt thereof.

13. Use of the compound according to any one of claims 1 to 10 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for inhibiting BACE1 activity.

14. Use of a compound according to any one of claims 1 to 10 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating or preventing Alzheimer's disease, mild cognitive impairment or prodromal Alzheimer's disease, for preventing the progression of Alzheimer's disease, mild cognitive impairment or prodromal Alzheimer's disease, or for preventing the progression of asymptomatic patients at risk of Alzheimer's disease.

15. Use of the compound according to any one of claims 1 to 10 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating or preventing a disease induced by the production, secretion and / or deposition of β-amyloid protein.

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