# Patent

## Title
Chromane compounds

## Abstract

The present invention provides a hydrate of N-[(4S)-2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-6′-yl]-5-chloropyridine-2-carboxamide which is useful as an active ingredient of a pharmaceutical composition, in particular, a pharmaceutical composition for preventing or treating diseases or conditions associated with and/or mediated by β-secretase activity, hydrolysis of a β-secretase site of a β-amyloid precursor protein, and/or β-amyloid protein accumulation, including a pharmaceutical composition for preventing or treating diseases including, but not limited to, Glaucoma, MCI (Mild cognitive impairment) or Alzheimer's disease, especially, Alzheimer's disease.

## Background
CROSS REFERENCE TO RELATED APPLICATIONS
This application is a divisional of U.S. patent application Ser. No. 14/192,667 filed Feb. 27, 2014 (now U.S. Pat. No. 8,975,415), which is a continuation of International Application No. PCT/US2013/043016 having an international filing date of May 29, 2013, which claims priority benefit of U.S. Provisional Patent Application Nos. 61/653,321 filed May 30, 2012 and 61/782,038 filed Mar. 14, 2013, the disclosures of which are incorporated herein by reference in their entirety.


INCORPORATION BY REFERENCE
The content of the following submission on ASCII text file (ST.25 text format) is incorporated herein by reference in its entirety: a computer readable form (CRF) of the Sequence Listing (file name is “322732001040_Sequence_Listing.txt”; date recorded: May 9, 2013; and the size of the ASCII text file in bytes is 4,096 bytes).
TECHNICAL FIELD
The present invention relates to a chromane compound which is useful as an active ingredient of a pharmaceutical composition, in particular, a pharmaceutical composition for preventing or treating diseases or conditions associated with and/or mediated by β-secretase activity, hydrolysis of a β-secretase site of an amyloid precursor protein, and/or β-amyloid protein accumulation, including a pharmaceutical composition for preventing or treating diseases including, but not limited to, Glaucoma, MCI (Mild cognitive impairment) or Alzheimer's disease, especially, Alzheimer's disease.
BACKGROUND OF THE INVENTION
Alzheimer's disease is a progressive mental deterioration in a human resulting, inter alia, in loss of memory, confusion and disorientation. Alzheimer's disease accounts for the majority of senile dementia and is a leading cause of death in adults (Non-Patent Document 1). Histologically, the brain of persons afflicted with Alzheimer's disease is characterized by a distortion of the intracellular neurofibrils and the presence of senile plaques composed of granular or filamentous argentophilic masses with an amyloid protein core, largely due to the accumulation of β-amyloid protein (Aβ) in the brain. Aβ accumulation plays a role in the pathogenesis and progression of the disease (Non-Patent Document 2) and is a proteolytic fragment of amyloid precursor protein (APP). APP is cleaved initially by β-secretase followed by γ-secretase to generate Aβ (Non-Patent Document 3 and 4).
It is known that inhibition of BACE may have a therapeutic effect in the prevention of dementia after stroke recovery (Non-Patent Document 5). It is reported that inhibition of BACE1 (beta-secretase 1) may have a therapeutic effect in Down syndrome (Non-Patent Document 6). The relationship between BACE1 mRNA levels and Parkinson's disease (PD) and Dementia with Lewy bodies (DLB) is also reported (Non-Patent Document 7 and 8).
In Patent Document 1, it is described that compounds (A) which are BACE inhibitors and are useful as therapeutic agents in the treatment, prevention, and amelioration of a disease or disorder characterized by elevated β-amyloid deposits or β-amyloid levels in a patient.




(for the symbols in the formula, refer to the patent publication).

In Patent Document 2, it is described that compounds (B) which are useful for inhibition of β-secretase enzymatic activity and for therapy and/or prophylaxis of neurodegenerative diseases associated therewith, particularly Alzheimer's Disease.




(for the symbols in the formula, refer to the patent publication).

In Patent Document 3, it is described that compounds (C) which are BACE inhibitors and are useful as therapeutic agents in the treatment, prevention, and amelioration of a disease or disorder characterized by elevated β-amyloid deposits or β-amyloid levels in a patient.




(for the symbols in the formula, refer to the patent publication).

In Patent Document 4, it is described that compounds (D) which are BACE inhibitors and are useful as therapeutic agents in the treatment, prevention, and amelioration of a disease or disorder characterized by elevated β-amyloid deposits or β-amyloid levels in a patient.




(for the symbols in the formula, refer to the patent publication).

In Patent Document 5 and 6, it is described that compounds (E) have BACE1 inhibitory activity and are useful as prophylactic or therapeutic agent for a neurodegenerative disease caused by Aβ and typified by Alzheimer type dementia, and pharmaceutical use thereof.




(for the symbols in the formula, refer to the patent publication).

In Patent Document 7, it is described that compounds (F) are useful for the modulation of the beta-secretase activity and are useful for the treatment of Alzheimer's disease and beta-secretase and/or plaque mediated disorders.




(for the symbols in the formula, refer to the patent publication).

In Patent Document 8, it is described that compounds (G) are useful for the modulation of the beta-secretase activity and are useful for the treatment of Alzheimer's disease and beta-secretase and/or plaque mediated disorders.




(for the symbols in the formula, refer to the patent publication).

In Patent Document 9, it is described that compounds (H) are useful for the modulation of the beta-secretase activity and are useful for the treatment of Alzheimer's disease and beta-secretase and/or plaque mediated disorders.




(for the symbols in the formula, refer to the patent publication).

In Patent Document 10, it is described that compounds (I) are useful for inhibition of β-secretase enzyme activity and the therapy and/or prophylaxis of neurodegenerative diseases associated therewith, such as Alzheimer's disease.




(for the symbols in the formula, refer to the patent publication).

In Patent Document 11, it is described that compounds (J) are inhibitors of beta-secretase-2 (BACE2) and the compounds may therefore be useful in the treatment of type 2 diabetes and other metabolic disorders.




(for the symbols in the formula, refer to the patent publication).

In Patent Document 12, it is described that compounds (K) are useful for inhibition of β-secretase enzyme activity and the therapy and/or prophylaxis of neurodegenerative diseases associated therewith, such as Alzheimer's disease.




(for the symbols in the formula, refer to the patent publication).

In Patent Document 13, it is described that compounds (L) are inhibitors of β-secretase and hence inhibit the formation of amyloid β (Aβ) peptides and are useful for treatment and/or prevention of Aβ-related pathologies such as Alzheimer's disease, and so on.




(for the symbols in the formula, refer to the patent publication).

In any of these Patent Documents, there is no specific disclosure of the compound of the present invention.
REFERENCES


    Patent Document 1: Pamphlet of International Publication WO 2010/021680
    Patent Document 2: Pamphlet of International Publication WO 2011/072064
    Patent Document 3: Pamphlet of International Publication WO 2011/106414
    Patent Document 4: Pamphlet of International Publication WO 2010/105179
    Patent Document 5: Pamphlet of International Publication WO 2010/013302
    Patent Document 6: Pamphlet of International Publication WO 2010/013794
    Patent Document 7: Pamphlet of International Publication WO 2010/030954
    Patent Document 8: Pamphlet of International Publication WO 2011/115938
    Patent Document 9: Pamphlet of International Publication WO 2011/115928
    Patent Document 10: Pamphlet of International Publication WO2011/123674
    Patent Document 11: Pamphlet of International Publication WO2010/128058
    Patent Document 12: Pamphlet of International Publication WO2012/071458
    Patent Document 13: Pamphlet of International Publication WO2012/087237
    Non-Patent Document 1: Anderson, R. N., et al., Natl. Vital Stat. Rep. 49:1-87 (2001)
    Non-Patent Document 2: Selkoe, D. J., Nature 399: 23-31 (1999)
    Non-Patent Document 3: Lin, X., et al., Proc. Natl. Acad. Sci. USA 97:1456-1460 (2000)
    Non-Patent Document 4: De Stropper, B., et al., Nature 391:387-390 (1998)
    Non-Patent Document 5: Wen Y., et al., Brain Res. 1009 (1-2):1-8 (2004)
    Non-Patent Document 6: Miners J. S., et al., J. Alzheimer's Dis. 23 (1):101-108 (2011)
    Non-Patent Document 7: Coulson D T., et al., J. Alzheimer's Dis. 22 (4):1111-1122 (2010)
    Non-Patent Document 8: Halliday G M., et al., J. Neural Transm. 118 (5):713-719 (2011)





BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 Drawing shows VCD spectra of Ex. 228b compound.
FIG. 2 Drawing shows VCD spectra of Ex. 229b compound.
FIG. 3 Drawing shows VCD spectra of Reference Example 225a compound.



SUMMARY OF THE INVENTION
The present invention provides a compound which is useful as an active ingredient of a pharmaceutical composition, in particular, a pharmaceutical composition for preventing or treating diseases or conditions associated with and/or mediated by β-secretase activity, hydrolysis of a β-secretase site of a β-amyloid precursor protein, and/or β-amyloid protein accumulation, including a pharmaceutical composition for preventing or treating diseases including, but not limited to, Glaucoma, MCI (Mild cognitive impairment) or Alzheimer's disease, especially, Alzheimer's disease.
Means for Solving the Problems
The present inventors have extensively studied compounds having beta-secretase inhibitory activity, and as a result, they have found that chromane compounds which are the compounds of the present invention have excellent beta-secretase inhibitory activity, and are therefore useful as agents for preventing or treating diseases or conditions associated with and/or mediated by β-secretase activity, hydrolysis of a β-secretase site of a β-amyloid precursor protein, and/or β-amyloid protein accumulation, including a pharmaceutical composition for preventing or treating diseases including, but not limited to, Glaucoma, MCI (Mild cognitive impairment) or Alzheimer's disease, especially, Alzheimer's disease, thereby completing the present invention.
The present invention relates to compounds of the formula (I) or a salt thereof:




wherein

A1 is O, S, —C(RA11RA12)-T-, or -T-C(RA11RA12)—;
A2 is —C(RA21RA22)—;
T is a single bond, O, or S;
RA11, RA12, RA21 and RA22 are, independently, H or halogen; or
RA11, RA12, RA21 and RA22 are combined with each other to form an aryl group, which is unsubstituted or substituted;
B is a hetero ring group which is unsubstituted or substituted, or cycloalkyl which is unsubstituted or substituted;
X and Y are independently selected from the group consisting of H, lower alkyl, which is unsubstituted or substituted, and cycloalkyl, which is unsubstituted or substituted; or
X and Y are combined with each other to form a cycloalkyl group, which is unsubstituted or substituted; and
R1, R2, R3 and R4 are independently selected from the group consisting of H, halogen, lower alkyl, which is unsubstituted or substituted, lower alkenyl, which is unsubstituted or substituted, —N(H)-(hetero ring group), wherein said hetero ring group is unsubstituted or substituted, —N(H)—C(O)-(hetero ring group), wherein said hetero ring group is unsubstituted or substituted, cycloalkenyl, which is unsubstituted or substituted, aryl, which is unsubstituted or substituted, and a hetero ring group, which is unsubstituted or substituted.
Further, unless specifically described otherwise, in the case where the symbols in any of the formulae in the present specification are also used in other formulae, the same symbols denote the same meanings.
Furthermore, the present invention relates to pharmaceutical compositions, comprising compounds of formula (I) or a salt thereof, as described herein, and a pharmaceutically acceptable carrier. Moreover, the present invention relates to pharmaceutical compositions for preventing or treating diseases or conditions associated with and/or mediated by β-secretase activity, hydrolysis of a β-secretase site of a β-amyloid precursor protein, and/or β-amyloid protein accumulation, including compounds of formula (I) or a salt thereof, as described herein, that are agents for preventing or treating diseases or conditions associated with and/or mediated by β-secretase activity, hydrolysis of a β-secretase site of a β-amyloid precursor protein, and/or β-amyloid protein accumulation, including compounds of formula (I) or a salt thereof.
Furthermore, the present invention relates to use of compounds of formula (I) or a salt thereof, as described herein, for preparation of a pharmaceutical composition (e.g., medicament) for preventing or treating diseases or conditions associated with and/or mediated by β-secretase activity, hydrolysis of a β-secretase site of a β-amyloid precursor protein, and/or β-amyloid protein accumulation, use of compounds of formula (I) or a salt thereof for preventing or treating diseases or conditions associated with and/or mediated by β-secretase activity, hydrolysis of a β-secretase site of a β-amyloid precursor protein, and/or β-amyloid protein accumulation, and methods for preventing or treating diseases or conditions associated with and/or mediated by β-secretase activity, hydrolysis of a β-secretase site of a β-amyloid precursor protein, and/or β-amyloid protein accumulation, including administering to a subject in need thereof an effective amount of the compounds of formula (I) or a salt thereof.
The present invention also relates to compounds of formula (I) or a salt thereof, as described herein, for use in the prevention or treatment of diseases or conditions associated with and/or mediated by β-secretase activity, hydrolysis of a β-secretase site of a β-amyloid precursor protein, and/or β-amyloid protein accumulation, the compounds of formula (I) or a salt thereof for preventing or treating diseases or conditions associated with and/or mediated by β-secretase activity, hydrolysis of a β-secretase site of a β-amyloid precursor protein, and/or β-amyloid protein accumulation. The present invention also relates to a method for preventing or treating diseases or conditions associated with and/or mediated by β-secretase activity, hydrolysis of a β-secretase site of a β-amyloid precursor protein, and/or β-amyloid protein accumulation, including administering to a subject an effective amount of the compounds of formula (I) or a salt thereof.
Effects of the Invention
The compounds of formula (I) or a salt thereof have beta-secretase inhibitory activity, and therefore can be used as an agent for preventing or treating diseases or conditions associated with and/or mediated by β-secretase activity, hydrolysis of a β-secretase site of a β-amyloid precursor protein, and/or β-amyloid protein accumulation, including, but not limited to, diseases such as Glaucoma, MCI (Mild cognitive impairment) or Alzheimer's disease, especially, Alzheimer's disease, or the like. In some embodiments, the compounds of formula (I) or a salt thereof can be used as an agent for preventing or treating diseases or conditions including, but not limited to, stroke, cerebrovascular dementia, Down syndrome, Parkinson's disease (PD), and dementia with Lewy bodies (DLB).
DETAILED DESCRIPTION
The present invention will be explained in more detail herein below. Further, “the compounds of formula (I) or a salt thereof” may be denoted as “the compounds (I) of the present invention” or “the compounds (I)” below in some cases.
In the present specification, the term “lower alkyl” refers to a straight (linear) or branched chain alkyl having 1 to 6 carbon atoms (hereinafter simply referred to as C1-6), for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl, and the like. In another embodiment, it is C1-4 alkyl, and in a further embodiment, C1-3 alkyl.
The term “lower alkenyl” refers to a straight (linear) or branched chain C2-6 alkenyl, for example, vinyl, propenyl, butenyl, pentenyl, 1-methylvinyl, 1-methyl-2-propenyl, 1,3-butadienyl, 1,3-pentadienyl, or the like. In another embodiment, it is C2-4 alkenyl, and in a still another embodiment, C2-3 alkenyl.
The term “lower alkynyl” refers to a linear or branched chain C2-6 alkynyl, for example, ethynyl, propynyl, butynyl, pentynyl, 1-methyl-2-propynyl, 1,3-butadiynyl, 1,3-pentadiynyl, or the like. In another embodiment, it is C2-4 alkynyl.
The term “cycloalkyl” refers to a C3-10 saturated hydrocarbon ring group, which may have a bridge. It is, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, adamantyl, or the like, in another embodiment, C3-8 cycloalkyl, and in a further embodiment, C3-6 cycloalkyl.
The term “cycloalkenyl” refers to a C4-15 hydrocarbon ring group having at least one double bond in the ring (provided that an aromatic hydrocarbon ring group is excluded), which may have a bridge, and includes a ring group fused (e.g., condensed) with a benzene ring at a double bond site. It is, for example, cyclopentenyl, cyclopentadienyl, cyclohexenyl, cyclohexadienyl, 1-tetrahydronaphthyl, 1-indenyl, 9-fluorenyl, or the like. In another embodiment, it is C5-10 cycloalkenyl, in a further embodiment, C5-8 cycloalkenyl, and in a further embodiment, C5-7 cycloalkenyl.
The term “aryl” refers to a C6-14 monocyclic to tricyclic aromatic hydrocarbon ring group, and includes a ring group fused with C5-8 cycloalkene at its double bond site. It is, for example, phenyl, naphthyl, 5-tetrahydronaphthyl, 4-indenyl, 1-fluorenyl, or the like. And the term “aryl” does not encompass aryl rings containing hetero atoms (such as S, N, O).
The term “hetero ring” means a ring group containing i) a monocyclic 3- to 8-membered hetero ring containing 1 to 4 hetero atoms selected from oxygen, sulfur, and nitrogen, and in another embodiment, a 5- to 7-membered hetero ring containing 1 to 4 hetero atoms selected from oxygen, sulfur, and nitrogen, and ii) a bicyclic or tricyclic hetero ring (in which the bicyclic or tricyclic heterocyclic ring may include a spiro ring) containing 1 to 5 hetero atoms selected from oxygen, sulfur, and nitrogen, formed by condensation or ring-fusion of the monocyclic hetero ring with one or two rings selected from the group consisting of a monocyclic hetero ring, a benzene ring, C5-8 cycloalkane, and C5-8 cycloalkene. The ring atom, sulfur or nitrogen, may be oxidized to form an oxide or a dioxide.
Examples of the “hetero ring” group include the following embodiments:
(1) Monocyclic Saturated Hetero Ring Groups, which mean monocyclic 3- to 8-membered saturated rings containing 1 to 4 hetero atoms selected from oxygen, sulfur, and nitrogen, and in another embodiment, 5- to 7-membered hetero rings containing 1 to 4 hetero atoms selected from oxygen, sulfur, and nitrogen.
(a) those containing 1 to 4 nitrogen atoms, for example, azepanyl, diazepanyl, aziridinyl, azetidinyl, pyrrolidinyl, imidazolidinyl, piperidyl, piperazolidinyl, piperazinyl, azocanyl, hexamethyleneimino, homopiperazinyl, and the like;
(b) those containing 1 to 3 nitrogen atoms and 1 to 2 sulfur atoms and/or 1 to 2 oxygen atoms, for example, thiomorpholinyl, thiazolidinyl, isothiazolidinyl, oxazolidinyl, morpholinyl, and the like;
(c) those containing 1 to 2 sulfur atoms, for example, tetrahydrothiopyranyl and the like;
(d) those containing 1 to 2 sulfur atoms and 1 to 2 oxygen atoms, for example, oxathiolanyl and the like; and
(e) those containing 1 to 2 oxygen atoms, for example, oxiranyl, oxetanyl, dioxolanyl, tetrahydrofuranyl, tetrahydropyranyl, 1,4-dioxanyl, and the like;
(2) Monocyclic Unsaturated Hetero Ring Groups, which mean monocyclic 3- to 8-membered unsaturated rings containing 1 to 4 hetero atoms selected from oxygen, sulfur, and nitrogen, and in another embodiment, 5- to 7-membered hetero rings containing 1 to 4 hetero atoms selected from oxygen, sulfur, and nitrogen.
(a) those containing 1 to 4 nitrogen atoms, for example, pyrrolyl, 2-pyrrolinyl, imidazolyl, 2-imidazolinyl, pyrazolyl, 2-pyrazolinyl, pyridyl, dihydropyridyl, tetrahydropyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazolyl, tetrazolyl, triazinyl, dihydrotriazinyl, azepinyl, and the like;
(b) those containing 1 to 3 nitrogen atoms and 1 to 2 sulfur atoms and/or 1 to 2 oxygen atoms, for example, thiazolyl, isothiazolyl, thiadiazolyl, dihydrothiazinyl, oxazolyl, isoxazolyl, oxadiazolyl, oxazinyl, and the like;
(c) those containing 1 to 2 sulfur atoms, for example, thienyl, thiepinyl, dihydrodithiopyranyl, dihydrodithionyl, 2H-thiopyranyl, and the like;
(d) those containing 1 to 2 sulfur atoms and 1 to 2 oxygen atoms, for example, dihydroxythiopyranyl and the like; and
(e) those containing 1 to 2 oxygen atoms, for example, furyl, dihydrofuryl, pyranyl, 2H-pyranyl, oxepinyl, dioxolyl, and the like;
(3) Fused Polycyclic Saturated Hetero Ring Groups, which mean bicyclic or tricyclic saturated hetero rings (in which the bicyclic or tricyclic heterocyclic ring may include a spiro ring) containing 1 to 5 hetero atoms selected from oxygen, sulfur, and nitrogen, formed by condensation or ring-fusion of the monocyclic saturated hetero ring with one or two rings selected from the group consisting of a monocyclic saturated hetero ring, and C5-8 cycloalkane.
(a) those containing 1 to 5 nitrogen atoms, for example, quinuclidinyl, 7-azabicyclo[2.2.1]heptyl, 3-azabicyclo[3.2.2]nonanyl, 2,8-diazaspiro[4.5]decan-8-yl, 2,3,6,8-tetraazaspiro[4.5]decan-8-yl, and the like;
(b) those containing 1 to 4 nitrogen atoms and 1 to 3 sulfur atoms, and/or 1 to 3 oxygen atoms, for example, trithiadiazaindenyl, dioxoloimidazolidinyl, 6-oxa-2,8-diazaspiro[4.5]decan-8-yl, 6-thia-2,8-diazaspiro[4.5]decan-8-yl, and the like; and
(c) those containing 1 to 3 sulfur atoms and/or 1 to 3 oxygen atoms, for example, 2,6-dioxabicyclo[3.2.2]oct-7-yl, 2-oxa-6-thiaspiro[4.5]decan-8-yl, and the like;
(4) Fused Polycyclic Unsaturated Hetero Ring Groups, which mean bicyclic or tricyclic unsaturated hetero rings (in which the bicyclic or tricyclic heterocyclic ring may include a spiro ring) containing 1 to 5 hetero atoms selected from oxygen, sulfur, and nitrogen, formed by condensation or ring-fusion of the monocyclic hetero ring with one or two rings selected from the group consisting of a monocyclic hetero ring, a benzene ring, C5-8 cycloalkane, and C5-8 cycloalkene.
(a) those containing 1 to 5 nitrogen atoms, for example, indolyl, isoindolyl, indolinyl, indolizinyl, benzimidazolyl, dihydrobenzimidazolyl, tetrahydrobenzimidazolyl, quinolyl, tetrahydroquinolyl, isoquinolyl, tetrahydroisoquinolyl, indazolyl, imidazopyridyl, benzotriazolyl, tetrazolopyridazinyl, carbazolyl, acridinyl, quinoxalinyl, dihydroquinoxalinyl, tetrahydroquinoxalinyl, phthalazinyl, dihydroindazolyl, benzopyrimidinyl, naphthyridinyl, quinazolinyl, cinnolinyl, pyridopyrrolidinyl, triazolopiperidinyl, 9,10-dihydroacridinyl, 2,8-diazaspiro[4.5]deca-3-en-8-yl, 2,3,6,8-tetraazaspiro[4.5]deca-1-en-8-yl, and the like;
(b) those containing 1 to 4 nitrogen atoms, and 1 to 3 sulfur atoms and/or 1 to 3 oxygen atoms, for example, benzothiazolyl, dihydrobenzothiazolyl, benzothiadiazolyl, imidazothiazolyl, imidazothiadiazolyl, benzoxazolyl, dihydrobenzoxazolyl, dihydrobenzoxazinyl, benzoxadiazolyl, benzoisothiazolyl, benzoisoxazolyl, thiazolopiperidinyl, 10H-phenothiazinyl, 6-oxa-2,8-diazaspiro[4.5]deca-3-en-8-yl, 6-thia-2,8-diazaspiro[4.5]deca-3-en-8-yl, and the like;
(c) those containing 1 to 3 sulfur atoms, for example, benzothienyl, benzodithiopyranyl, dibenzo[b,d]thienyl, and the like;
(d) those containing 1 to 3 sulfur atoms and 1 to 3 oxygen atoms, for example, benzoxathiopyranyl, 2-oxa-6-thiaspiro[4.5]deca-3-en-8-yl, and the like; and
(e) those containing 1 to 3 oxygen atoms, for example, benzodioxolyl, benzofuranyl, dihydrobenzofuranyl, isobenzofuranyl, chromanyl, chromenyl, isochromenyl, dibenzo[b,d]furanyl, methylenedioxyphenyl, ethylenedioxyphenyl, xanthenyl, and the like; etc.
Further, the terms “aryl”, “cycloalkyl”, and “hetero ring” groups as described above are meant to be monovalent groups, but these may be divalent or higher groups in some cases. For example, when aryl in the R2 is substituted, this aryl is described by monovalent group, but this aryl means divalent or higher groups.
The term “nitrogen-containing hetero ring” group refers to one containing at least one nitrogen atom, including, but not limited to, such as groups in (1)(a), (1)(b), (2)(a), (2)(b), (3)(a), (3)(b), (4)(a), and (4)(b), among the “hetero ring” groups above.
The term “oxygen-containing monocyclic saturated hetero ring” group refers to one containing at least one oxygen atom, including, but not limited to, such as groups containing at least one oxygen atom in (1)(b) or such as groups in (1)(d), and (1)(e), among the “(1) Monocyclic Saturated Hetero Ring Groups” above.
The term “cyclic ether” group refers to one containing only at least one oxygen atom as hetero atom, including, but not limited to, such as groups in, (1)(e), among the “oxygen-containing monocyclic saturated hetero ring” group above.
The term “nitrogen-containing monocyclic hetero ring” group refers to one containing at least one nitrogen atom, including, but not limited to, such as groups in (1)(a), (1)(b), (2)(a), and (2)(b), among the “Monocyclic Saturated Hetero Ring Groups” and “Monocyclic Unsaturated Hetero Ring Groups” above.
The term “halogen” means F, Cl, Br, or I.
In the present specification, the term “substituted” represents being substituted with 1 to 5 substituents. In some embodiments, the term “substituted” represents being substituted with 1, 2, 3, 4 or 5 substituents. Further, if a plurality of substituents are included, the substituents may be the same as or different from one another.
In some embodiments, the term “substituted with one or more substituents” represents being substituted with 1 to 5 substituents. In some embodiments, the term “substituted with one or more substituents” represents being substituted with 1, 2, 3, 4 or 5 substituents.
In the present specification, both B and




represent a group that shares a carbon atom with the chromane ring to which it is attached, as shown in formula (I). For example, when B is oxetanyl, this means that





is





and when B is cyclopropyl, this means that





is





In some embodiments, the B group may be substituted at one or more available positions, as described herein.

“RA11, RA12, RA21 and RA22 are combined with each other to form an aryl group” indicates that RA11, RA12, RA21 and RA22 combined with each carbon atom to which they are bonded to form a C6-14 monocyclic to tricyclic aromatic hydrocarbon ring group, and includes a ring group fused with C5-8 cycloalkene at its double bond site. It is, for example, phenyl, naphthyl, 5-tetrahydronaphthyl, 4-indenyl, 1-fluorenyl, or the like.
For example, when RA11, RA12, RA21 and RA22 are combined with each other to form phenyl, a structure of the compound of formula (I) is as below.




“X and Y are combined with each other to form a cycloalkyl group” indicates that X and Y combined with a carbon atom to which they are bonded to form a C3-10 saturated hydrocarbon ring group, which may have a bridge. It is, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, adamantyl, or the like, in another embodiment, C3-8 cycloalkyl, and in a further embodiment, C3-6 cycloalkyl.
For example, when X and Y are combined with each other to form cyclobutyl or cyclopentyl structures of the compound of formula (I) are as below.




“Amyloid precursor protein,” or “APP,” as used herein, refers to an amyloid precursor polypeptide comprising a β-secretase cleavage site.
A “β-secretase cleavage site” is an amino acid sequence that is cleaved by an active memapsin 2 (also referred to as beta-secretase 1 or BACE-1, or active fragment thereof, such as described in U.S. Pat. No. 6,545,127). Specific β-secretase cleavage sites have also been previously set forth and discussed in detail in U.S. Application No. 20040121947, and International Application No. PCT/US02/34324 (Publication No. WO 03/039454), which are herein incorporated by reference for all purposes in their entirety, and include the Swedish mutation sequence, and the native amyloid precursor protein cleavage sequence. Thus, β-secretase inhibitors may be tested for their ability to decrease the hydrolysis of the β-secretase cleavage site of a substrate, such as the amyloid precursor protein, compounds of amyloid precursor protein, or fragments of amyloid precursor protein.
A “beta-secretase inhibitor” (i.e. β-secretase inhibitor) refers to a compound capable of reducing the proteolytic activity of memapsin-2 relative to the activity in the absence of inhibitor.
“Memapsin-2,” as used herein, refers to proteins identified by National Center for Biotechnology Information (“NCBI”) accession number NP—036236 (sometimes referred to as “β-site APP-cleaving enzyme 1” or “BACE1” or generically as “β-secretase” or “beta-secretase”), including homologs, isoforms and subdomains thereof that retain proteolytic activity. Sequence identities of active memapsin 2 proteins and protein fragments (and nucleic acid coding sequences thereof) have been previously disclosed and discussed in detail in U.S. Application No. 20040121947, and International Application No. PCT/US02/34324 (Publication No. WO 03/039454, International publication WO 01/00663, U.S. Pat. No. 6,545,127), which are herein incorporated by reference for all purposes in their entirety.
“Amyloid beta (Aβ or Abeta)” refers to a peptide of 36-43 amino acids. While best known as a component of amyloid plaques in association with Alzheimer's disease, as Aβ is the main component of certain deposits found in the brains of patients with Alzheimer's disease. The different Aβ isoforms (for example, Aβ40, Aβ42, and so on) refer to cleavage products of transmembranous APP via the β-secretase pathway. The cleavage by β-secretase (BACE1) liberates the Aβ N-terminus, together with sAPPβ and a C-terminal fragment C99. C99 is subsequently cleaved by γ-secretase to yield Aβ.
“Diseases or conditions associated with and/or mediated by β-secretase activity, hydrolysis of a β-secretase cleavage site of an amyloid precursor protein, and/or β-amyloid protein accumulation” as used herein, includes, but is not limited to, diseases such as Glaucoma, MCI (Mild cognitive impairment) or Alzheimer's disease. In another embodiment, the term includes, but is not limited to, MCI (Mild cognitive impairment) or Alzheimer's disease. In another embodiment, the term includes, but is not limited to, Alzheimer's disease. In another embodiment, the term includes, but is not limited to, MCI (Mild cognitive impairment). In some embodiments, the compounds of formula (I) or a salt thereof can be used as agent for preventing or treating diseases or conditions including, but not limited to, stroke, cerebrovascular dementia, Down syndrome, Parkinson's disease (PD), and dementia with Lewy bodies (DLB).
As used herein and in the appended claims, the singular forms “a,” “an,” and “the” include plural reference unless the context clearly indicates otherwise.
The term, “effective amount,” and cognates of this term, as used herein, refer to an amount that results in a desired pharmacological and/or physiological effect for a specified condition (e.g., disease, disorder, etc.) or one or more of its symptoms and/or to completely or partially prevent the occurrence of the condition or symptom thereof and/or may be therapeutic in terms of a partial or complete cure for the condition and/or adverse effect attributable to the condition. In reference to conditions mediated by memapsin 2 (β-secretase) or diseases or conditions associated with and/or mediated by β-secretase activity, hydrolysis of a β-secretase cleavage site of an amyloid precursor protein, and/or β-amyloid protein accumulation, a pharmaceutically or therapeutically effective amount comprises an amount sufficient to, among other things, cause antagonism or inhibition of memapsin 2 (β-secretase). In reference to glaucoma, a pharmaceutically or therapeutically effective amount comprises an amount sufficient to, among other things, decrease intraocular pressure; and/or halt, reverse, and/or diminish the loss of retinal ganglion cells (RGCs). In certain embodiments, the pharmaceutically effective amount is sufficient to prevent the condition, as in being administered to an individual prophylactically.
The “effective amount” will vary depending on the composition being administered, the condition being treated/prevented, the severity of the condition being treated or prevented, the age and relative health of the individual, the route and form of administration, the judgment of the attending medical or veterinary practitioner, and other factors appreciated by the skilled artisan in view of the teaching provided herein.
The “subject” means the animal which needs its prevention or treatment and the human who needs its prevention or treatment, in some embodiments it means the human who needs its prevention or treatment.
When used with respect to methods of treatment/prevention and the use of the compounds and compositions thereof described herein, a subject “in need thereof” may be an individual who has been diagnosed with or previously treated for the condition to be treated. With respect to prevention, the subject in need thereof may also be an individual who is at risk for a condition (e.g., a family history of the condition, life-style factors indicative of risk for the condition, etc.).
In some variations, the subject has been identified as having one or more of the conditions described herein. In some embodiments, the subject has been identified as susceptible to one or more of the conditions as described herein. The susceptibility of a subject may be based on any one or more of a number of risk factors and/or diagnostic approaches appreciated by the skilled artisan, including, but not limited to, genetic profiling, family history, medical history (e.g., appearance of related conditions), lifestyle or habits.
Examples of the embodiment of the substituent acceptable in the “RA11, RA12, RA21 and RA22 are combined with each other to form an aryl group, which is substituted” include, but are not limited to, e.g., halogen.
Examples of the embodiment of the substituent acceptable in the “a hetero ring group, which is substituted” in B include, but are not limited to, e.g., halogen.
Examples of the embodiment of the substituent acceptable in the “cycloalkyl, which is substituted” in B include, but are not limited to, e.g., halogen.
Examples of the embodiment of the substituent acceptable in the “lower alkyl, which is substituted” in X and Y include, but are not limited to, the groups shown in i) to iii) below.
i) halogen,
ii) cycloalkyl, or
iii) aryl.
Examples of the embodiment of the substituent acceptable in the “cycloalkyl, which is substituted” in X and Y include, but are not limited to, the groups shown in i) to iii) below.
i) halogen,
ii) cycloalkyl, or
iii) aryl.
Examples of the embodiment of the substituent acceptable in the “X and Y are combined with each other to form a cycloalkyl group, which is substituted” include, but are not limited to, the groups shown in i) to iii) below.
i) halogen,
ii) cycloalkyl, or
iii) aryl.
Examples of the embodiment of the substituent acceptable in the “a hetero ring group, which is substituted” in R1, R2, R3 and R4 include, but are not limited to, the groups shown in i) to vi) below.
i) halogen,
ii) lower alkyl, which is unsubstituted or substituted with one or more substituents selected from the group consisting of halogen and —O-(lower alkyl),
iii) lower alkynyl, which is unsubstituted or substituted with one or more substituents selected from the group consisting of —O-(lower alkyl) and cycloalkyl,
iv) —O-(lower alkyl), wherein said lower alkyl is unsubstituted or substituted with halogen,
v) cycloalkyl, or
vi) —CN.
Examples of the embodiment of the substituent acceptable in the “lower alkyl, which is substituted” in R1, R2, R3 and R4 include, but are not limited to, —O-(lower alkyl), or aryl, wherein said aryl is unsubstituted or substituted with lower alkyl.
Examples of the embodiment of the substituent acceptable in the “lower alkenyl, which is substituted” in R1, R2, R3 and R4 include, but are not limited to, —O-(lower alkyl).
Examples of the embodiment of the substituent acceptable in the “—N(H)-(hetero ring group), wherein said hetero ring group is substituted” in R1, R2, R3 and R4 include, but are not limited to, halogen or —O-(lower alkyl).
Examples of embodiments of substituents acceptable in the “—N(H)—C(O)-(hetero ring group), wherein said hetero ring group is substituted” in R1, R2, R3 and R4 include, but are not limited to, the groups shown in i) to vii) below.
i) halogen,
ii) lower alkyl, which is unsubstituted or substituted with one or more substituents selected from the group consisting of halogen, —O-(lower alkyl), and a hetero ring group,
iii) —CN,
iv) —O-(lower alkyl), wherein said lower alkyl is unsubstituted or substituted with halogen,
v) cycloalkyl,
vi) aryl, or
vii) a hetero ring group.
Examples of other embodiments of substituents acceptable in the “—N(H)—C(O)-(hetero ring group), wherein said hetero ring group is substituted” in R1, R2, R3 and R4 include, but are not limited to, the groups shown in i) to vii) below.
i) halogen,
ii) lower alkyl, which is unsubstituted or substituted with one or more substituents selected from the group consisting of halogen, —O-(lower alkyl), and a nitrogen-containing monocyclic hetero ring group,
iii) —CN,
iv) —O-(lower alkyl), wherein said lower alkyl is unsubstituted or substituted with halogen,
v) cycloalkyl,
vi) aryl, or
vii) a nitrogen-containing monocyclic hetero ring group.
Examples of other embodiments of the substituent acceptable in the “—N(H)—C(O)-(hetero ring group), wherein said hetero ring group is substituted” in R1, R2, R3 and R4 include, but are not limited to, the groups shown in i) to iv) below.
i) halogen,
ii) lower alkyl,
iii) —CN, or
iv) —O-(lower alkyl).
Examples of embodiments of substituents acceptable in the “cycloalkenyl, which is substituted” in R1, R2, R3 and R4 include, but are not limited to, lower alkyl.
Examples of the embodiment of the substituent acceptable in the “aryl, which is substituted” in R1, R2, R3 and R4 include, but are not limited to, the groups shown in i) to ix) below.
i) —OH,
ii) halogen,
iii) lower alkyl, which is unsubstituted or substituted with halogen,
iv) —O-(lower alkyl), wherein said lower alkyl is unsubstituted or substituted with halogen,
v) —S-(lower alkyl),
vi) cycloalkyl,
vii) —CN,
viii) lower alkenyl, which is unsubstituted or substituted with —CN, or
ix) —C(O)—N(H)-(lower alkyl).
Examples of the embodiment of groups of compounds of formula (I) of the present invention are shown below.
(1)
(1-1)
A1 is O or S;
A2 is —C(RA21RA22)—; and
RA21 and RA22 are H.
(1-2)
(1-2-1)
A1 is O;
A2 is —C(RA21RA22)—; and
RA21 and RA22 are H.
(2)
(2-1)
(2-1-1)
B is a hetero ring group, wherein said hetero ring group is unsubstituted or substituted with halogen, or cycloalkyl, wherein said cycloalkyl is unsubstituted or substituted with halogen.
(2-1-2)
B is a hetero ring group or cycloalkyl, wherein said cycloalkyl is unsubstituted or substituted with halogen.
(2-1-3)
B is an oxygen-containing monocyclic saturated hetero ring group or cycloalkyl, wherein said cycloalkyl is unsubstituted or substituted with halogen.
(2-1-4)
B is an oxygen-containing monocyclic saturated hetero ring group or C3-6 cycloalkyl, wherein said C3-6 cycloalkyl is unsubstituted or substituted with halogen.
(2-1-5)
B is an oxygen-containing monocyclic saturated hetero ring group or C3-6 cycloalkyl, wherein said C3-6 cycloalkyl is unsubstituted or substituted with F.
(2-1-6)
B is a cyclic ether group or cycloalkyl, wherein said cycloalkyl is unsubstituted or substituted with halogen.
(2-1-7)
B is a cyclic ether group or C3-6 cycloalkyl, wherein said C3-6 cycloalkyl is unsubstituted or substituted with halogen.
(2-1-8)
B is a cyclic ether group or C3-6 cycloalkyl, wherein said C3-6 cycloalkyl is unsubstituted or substituted with F.
(2-1-9)
B is oxetanyl, tetrahydropyranyl, cyclopropyl, cyclobutyl, or 3,3-difluorocyclobutan-1-yl.
(2-1-10)
B is oxetanyl, tetrahydrofuranyl, cyclopropyl, cyclobutyl, or 3,3-difluorocyclobutan-1-yl.
(2-2)
(2-2-1)
B is a hetero ring group.
(2-2-2)
B is an oxygen-containing monocyclic saturated hetero ring group.
(2-2-3)
B is a cyclic ether group.
(2-2-4)
B is oxetanyl or tetrahydropyranyl.
(2-2-5)
B is oxetanyl.
(2-2-6)
B is oxetanyl or tetrahydrofuranyl.
(2-3)
(2-3-1)
B is cycloalkyl, wherein said cycloalkyl is unsubstituted or substituted with halogen.
(2-3-2)
B is C3-6 cycloalkyl, wherein said C3-6 cycloalkyl is unsubstituted or substituted with halogen.
(2-3-3)
B is C3-6 cycloalkyl, wherein said C3-6 cycloalkyl is unsubstituted or substituted with F.
(2-3-4)
B is cyclopropyl, cyclobutyl, or 3,3-difluorocyclobutan-1-yl.
(2-3-5)
B is cyclopropyl.
(3)
(3-1)
X is lower alkyl; and
Y is lower alkyl.
(3-2)
X is methyl; and
Y is methyl.
(4)
(4-1)
(4-1-1)
R1, R2, R3 and R4 are independently selected from the group consisting of
H,
halogen, and
—N(H)—C(O)-(hetero ring group), wherein said hetero ring group is unsubstituted or substituted with one or more substituents selected from the group consisting of halogen,
lower alkyl which is unsubstituted or substituted with halogen,
—CN, and
—O-(lower alkyl).
(4-1-2)
R1 and R4 are H;
R3 is H or halogen; and
R2 is —N(H)—C(O)-(hetero ring group), wherein said hetero ring group is unsubstituted or substituted with one or more substituents selected from the group consisting of
halogen,
lower alkyl which is unsubstituted or substituted with halogen,
—CN, and
—O-(lower alkyl).
(4-1-3)
R1 and R4 are H;
R3 is H or halogen; and
R2 is —N(H)—C(O)-(nitrogen-containing monocyclic hetero ring group), wherein said nitrogen-containing monocyclic hetero ring group is unsubstituted or substituted with one or more substituents selected from the group consisting of
halogen,
lower alkyl which is unsubstituted or substituted with halogen,
—CN, and
—O-(lower alkyl).
(4-1-4)
R1 and R4 are H;
R3 is H or halogen; and
R2 is selected from the group consisting of
—N(H)—C(O)-(pyridyl), wherein said pyridyl is unsubstituted or substituted with one or more substituents selected from the group consisting of
halogen,
lower alkyl which is unsubstituted or substituted with halogen,
—CN, and
—O-(lower alkyl),
—N(H)—C(O)-(pyrazinyl), wherein said pyrazinyl is unsubstituted or substituted with one or more substituents selected from the group consisting of
halogen,
lower alkyl which is unsubstituted or substituted with halogen,
—CN, and
—O-(lower alkyl), and
—N(H)—C(O)-(pyrimidinyl), wherein said pyrimidinyl is unsubstituted or substituted with one or more substituents selected from the group consisting of
halogen,
lower alkyl which is unsubstituted or substituted with halogen,
—CN, and
—O-(lower alkyl).
(4-1-5)
R1 and R4 are H;
R3 is H or halogen; and
R2 is selected from the group consisting of
—N(H)—C(O)-(pyridyl), wherein said pyridyl is unsubstituted or substituted with one or more substituents selected from the group consisting of
halogen,
lower alkyl, and
—CN,
—N(H)—C(O)-(pyrazinyl), wherein said pyrazinyl is unsubstituted or substituted with —O-(lower alkyl) or
lower alkyl which is unsubstituted or substituted with halogen, and
—N(H)—C(O)-(pyrimidinyl), wherein said pyrimidinyl is unsubstituted or substituted with halogen.
(4-1-6)
R1 and R4 are H;
R3 is H or halogen; and
R2 is —N(H)—C(O)-(pyridyl), wherein said pyridyl is unsubstituted or substituted with one or more substituents selected from the group consisting of
halogen,
lower alkyl, and
—CN.
(4-1-6-1)
R1 and R4 are H;
R3 is H or halogen; and
R2 is —N(H)—C(O)-(pyridyl), wherein said pyridyl is unsubstituted or substituted with halogen.
(4-1-7)
R1 and R4 are H;
R3 is H or halogen; and
R2 is —N(H)—C(O)-(pyrazinyl), wherein said pyrazinyl is unsubstituted or substituted with —O-(lower alkyl) or
lower alkyl which is unsubstituted or substituted with halogen.
(4-1-8)
R1 and R4 are H;
R3 is H or halogen; and
R2 is —N(H)—C(O)-(pyrimidinyl), wherein said pyrimidinyl is unsubstituted or substituted with halogen.
(4-2)
The groups of any one of (4-1),
wherein R3 is H.
Furthermore, still other embodiments of the compounds of formula (I) of the present invention include the compounds including a combination of two or more of the groups described in (1) to (4) above, specifically, the following compounds.
(5) The compound of formula (I), wherein B is as described in (2).
(6) The compound as described in (5), wherein X and Y are as described in (3).
(7) The compound as described in (5) or (6), wherein R1, R2, R3 and R4 are as described in (4).
(8) The compound as described in (5), (6) or (7), wherein A1 and A2 are as described in (1).
Furthermore, still other embodiments of the compounds of formula (I) of the present invention include the compounds including a combination of two or more of the groups described in (1) to (4) above, specifically, the following compounds.
(9) The compound of formula (I), wherein A1 and A2 are as described in (1-2-1), B is as described in (2-1-3), X and Y are as described in (3-1) and R1, R2, R3 and R4 are as described in (4-1-2).
(10) The compound of formula (I), wherein A1 and A2 are as described in (1-2-1), B is as described in (2-1-3), X and Y are as described in (3-1) and R1, R2, R3 and R4 are as described in (4-1-5).
(11) The compound of formula (I), wherein A1 and A2 are as described in (1-2-1), B is as described in (2-2-2), X and Y are as described in (3-1) and R1, R2, R3 and R4 are as described in (4-1-5).
(12) The compound of formula (I), wherein A1 and A2 are as described in (1-2-1), B is as described in (2-3-1), X and Y are as described in (3-1) and R1, R2, R3 and R4 are as described in (4-1-5).
(13) The compound of formula (I), wherein A1 and A2 are as described in (1-2-1), B is as described in (2-2-5), X and Y are as described in (3-1) and R1, R2, R3 and R4 are as described in (4-1-5).
(14) The compound of formula (I), wherein A1 and A2 are as described in (1-2-1), B is as described in (2-2-5), X and Y are as described in (3-2) and R1, R2, R3 and R4 are as described in (4-1-5).
(15) The compound of formula (I), wherein A1 and A2 are as described in (1-2-1), B is as described in (2-2-5), X and Y are as described in (3-2) and R1, R2, R3 and R4 are as described in (4-1-6).
(16) The compound of formula (I), wherein A1 and A2 are as described in (1-2-1), B is as described in (2-2-5), X and Y are as described in (3-2) and R1, R2, R3 and R4 are as described in (4-1-6-1).
(17) The compound of formula (I), wherein A1 and A2 are as described in (1-2-1), B is as described in (2-2-5), X and Y are as described in (3-2) and R1, R2, R3 and R4 are as described in (4-1-7).
(18) The compound of formula (I), wherein A1 and A2 are as described in (1-2-1), B is as described in (2-2-5), X and Y are as described in (3-2) and R1, R2, R3 and R4 are as described in (4-1-8).
(19) The compound of formula (I), wherein A1 and A2 are as described in (1-2-1), B is as described in (2-2-5), X and Y are as described in (3-2) and R1, R2, R3 and R4 are as described in (4-1-6), wherein R3 is H.
(20) The compound of formula (I), wherein A1 and A2 are as described in (1-2-1), B is as described in (2-2-5), X and Y are as described in (3-2) and R1, R2, R3 and R4 are as described in (4-1-6-1), wherein R3 is H.
(21) The compound of formula (I), wherein A1 and A2 are as described in (1-2-1), B is as described in (2-2-5), X and Y are as described in (3-2) and R1, R2, R3 and R4 are as described in (4-1-7), wherein R3 is H.
(22) The compound of formula (I), wherein A1 and A2 are as described in (1-2-1), B is as described in (2-2-5), X and Y are as described in (3-2) and R1, R2, R3 and R4 are as described in (4-1-8), wherein R3 is H.
(23) The compound of formula (I), wherein A1 and A2 are as described in (1-2-1), B is as described in (2-3-5), X and Y are as described in (3-1) and R1, R2, R3 and R4 are as described in (4-1-5).
(24) The compound of formula (I), wherein A1 and A2 are as described in (1-2-1), B is as described in (2-3-5), X and Y are as described in (3-2) and R1, R2, R3 and R4 are as described in (4-1-5).
(25) The compound of formula (I), wherein A1 and A2 are as described in (1-2-1), B is as described in (2-3-5), X and Y are as described in (3-2) and R1, R2, R3 and R4 are as described in (4-1-6).
(26) The compound of formula (I), wherein A1 and A2 are as described in (1-2-1), B is as described in (2-3-5), X and Y are as described in (3-2) and R1, R2, R3 and R4 are as described in (4-1-6-1).
(27) The compound of formula (I), wherein A1 and A2 are as described in (1-2-1), B is as described in (2-3-5), X and Y are as described in (3-2) and R1, R2, R3 and R4 are as described in (4-1-7).
(28) The compound of formula (I), wherein A1 and A2 are as described in (1-2-1), B is as described in (2-3-5), X and Y are as described in (3-2) and R1, R2, R3 and R4 are as described in (4-1-8).
(29) The compound of formula (I), wherein A1 and A2 are as described in (1-2-1), B is as described in (2-3-5), X and Y are as described in (3-2) and R1, R2, R3 and R4 are as described in (4-1-6), wherein R3 is H.
(30) The compound of formula (I), wherein A1 and A2 are as described in (1-2-1), B is as described in (2-3-5), X and Y are as described in (3-2) and R1, R2, R3 and R4 are as described in (4-1-6-1), wherein R3 is H.
(31) The compound of formula (I), wherein A1 and A2 are as described in (1-2-1), B is as described in (2-3-5), X and Y are as described in (3-2) and R1, R2, R3 and R4 are as described in (4-1-7), wherein R3 is H.
(32) The compound of formula (I), wherein A1 and A2 are as described in (1-2-1), B is as described in (2-3-5), X and Y are as described in (3-2) and R1, R2, R3 and R4 are as described in (4-1-8), wherein R3 is H.
(33) The compound of formula (I), wherein A1 and A2 are as described in (1-2-1), B is as described in (2-1-3), X and Y are as described in (3-1) and R1, R2, R3 and R4 are as described in (4-1-3).
(34) The compound of formula (I), wherein A1 and A2 are as described in (1-2-1), B is as described in (2-1-3), X and Y are as described in (3-1) and R1, R2, R3 and R4 are as described in (4-1-4).
(35) The compound as described in (5) to (34), wherein the compound of formula (I) is the compound of formula (II) as below.




Examples of the specific compounds encompassed by the present invention include the following compounds. Nomenclature of some compounds described herein may be identified using IUPAC or other naming conventions including ACD/Name ver. 12.02, available from Advanced Chemistry Development, Inc., Toronto, Ontario, Canada.

    N-(2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-6′-yl)-5-chloropyridine-2-carboxamide,
    N-(2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-6′-yl)-5-fluoropyridine-2-carboxamide,
    N-(2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-6′-yl)-5-chloro-3-fluoropyridine-2-carboxamide,
    N-(2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-6′-yl)-5-bromopyrimidine-2-carboxamide,
    N-(2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-6′-yl)-5-chloro-3-methyl pyridine-2-carboxamide,
    N-(2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-6′-yl)-5-cyanopyridine-2-carboxamide,
    N-(2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-6′-yl)-5-methoxypyrazine-2-carboxamide,
    N-[(4S)-2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-6′-yl]-5-chloropyridine-2-carboxamide,
    N-[(4′R)-2′′-amino-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-6′-yl]-5-methoxypyrazine-2-carboxamide,
    N-[(4′R)-2′′-amino-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-6′-yl]-5-(difluoromethyl) pyrazine-2-carboxamide,
    N-[(4S)-2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-6′-yl]-5-methoxypyrazine-2-carboxamide,
    N-[(4S)-2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-6′-yl]-5-bromopyridine-2-carboxamide,
    N-[(4′R)-2′′-amino-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-6′-yl]-5-chloropyridine-2-carboxamide, and
    N-[(4′R)-2′′-amino-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-6′-yl]-5-fluoropyridine-2-carboxamide.


Other examples of the specific compounds encompassed by the present invention include the following compounds. Nomenclature of some compounds described herein may be identified using IUPAC or other naming conventions including ACD/Name ver. 12.02, available from Advanced Chemistry Development, Inc., Toronto, Ontario, Canada.

    N-[(4S)-2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-6′-yl]-5-chloropyridine-2-carboxamide,
    N-[(4′R)-2′′-amino-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-6′-yl]-5-methoxypyrazine-2-carboxamide, and
    N-[(4′R)-2′′-amino-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-6′-yl]-5-(difluoromethyl) pyrazine-2-carboxamide.


The present invention relates to a hydrate of the compound or a salt, wherein said compound is

    N-[(4S)-2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-6′-yl]-5-chloropyridine-2-carboxamide.


In some embodiments,
A1 is O;
A2 is CH2;
B is




or cyclopropyl;

X and Y are both methyl, or X and Y are both H;
R1, R3, and R4 are H; and
R2 is —N(H)—C(O)-(hetero ring group), wherein the hetero ring group of R2 is selected from the group consisting of each of




each of which is substituted with 1 or 2 substituents independently selected from the group consisting of halogen, cyano, unsubstituted —O-lower alkyl, unsubstituted lower alkyl, lower alkyl substituted with one or more halogen, —O-lower alkyl substituted with one or more halogen, lower alkyl substituted with —OCH3, unsubstituted lower alkynyl, and unsubstituted cycloalkyl. In some embodiments, B is





and X and Y are both methyl. In some embodiments, B is cyclopropyl, and X and Y are both H. In some embodiments, R2 is selected from the group consisting of





and wherein R5 and R6 are independently selected from the group consisting of hydrogen, fluoro, chloro, bromo, cyano, —OCH3, methyl, —CHF2, —OCHF2, —OCH2CHF2, —CH2OCH3, —C≡C—CH3, and cyclopropyl. In some embodiments, R5 is chloro or —OCH3, and R6 is hydrogen or fluoro. In some embodiments, R5 is —CHF2 or —OCHF2, and R6 is hydrogen.

In some embodiments,
A1 is O;
A2 is CH2; and
R2 is —N(H)—C(O)-(hetero ring group), wherein the hetero ring group is




In some embodiments, the compound is selected from the group consisting of




or a salt thereof.

In some embodiments, the compound is




or a salt thereof.

In some embodiments, the compound is




or a salt thereof.

In some embodiments, the compound is




or a salt thereof.

In some embodiments, the compound is selected from the group consisting of




or a salt thereof.

In some embodiments, the compound is




or a salt thereof.

In some embodiments, the compound is




or a salt thereof.

In some embodiments, the compound is




or a salt thereof.

In some embodiments, the compound is selected from the group consisting of
















or a salt thereof.

The compound of the formula (I) may exist in the form of tautomers or geometrical isomers depending on the kind of substituents. In the present specification, the compound of the formula (I) shall be described in only one form of isomer, yet the present invention includes such an isomer, isolated forms of the isomers, or a mixture thereof.
In addition, the compound of the formula (I) may have asymmetric carbon atoms or axial asymmetry in some cases, and correspondingly, it may exist in the form of optical isomers. The present invention includes both an isolated form of the optical isomers of the compound of the formula (I) or a mixture thereof.
Moreover, the present invention also includes a pharmaceutically acceptable prodrug of the compound of the formula (I). The pharmaceutically acceptable prodrug is a compound having a group that can be converted into an amino group, a hydroxyl group, a carboxyl group, or the like through solvolysis or under physiological conditions. Examples of the group forming the prodrug include the groups described in Prog. Med., 5, 2157-2161 (1985) and Pharmaceutical Research and Development, Drug Design, Hirokawa Publishing Company (1990), Vol. 7, 163-189, which is incorporated by reference herein in its entirety.
Furthermore, the salt of the compound of the formula (I) is a pharmaceutically acceptable salt of the compound of the formula (I) and may form an acid addition salt or a salt with a base depending on the kind of substituents. Specific examples thereof include acid addition salts with inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, phosphoric acid, and the like, and with organic acids such as formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, mandelic acid, tartaric acid, dibenzoyltartaric acid, ditoluoyltartaric acid, citric acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, aspartic acid, glutamic acid, and the like, and salts with inorganic bases such as sodium, potassium, magnesium, calcium, aluminum, and the like or organic bases such as methylamine, ethylamine, ethanolamine, lysine, ornithine, and the like, salts with various amino acids or amino acid derivatives such as acetylleucine and the like, ammonium salts, etc.
In addition, the present invention also includes various hydrates or solvates, and polymorphic crystal substances of the compound of the formula (I) and a salt thereof. In addition, the present invention also includes compounds labeled with various radioactive or non-radioactive isotopes.
(Preparation Methods)
The compound of the formula (I) and a salt thereof can be prepared using the characteristics based on the basic structure or the type of substituents thereof and by applying various known synthesis methods. During the preparation, replacing the relevant functional group with a suitable protective group (a group that can be easily converted into the functional group) at the stage from starting material to an intermediate may be effective depending on the type of the functional group in the production technology in some cases. The protective group for such a functional group may include for example, the protective groups described in “Greene's Protective Groups in Organic Synthesis (4th Ed., 2006)”, P. G. M. Wuts and T. W. Greene, and one of these may be selected and used as necessary depending on the reaction conditions. In this kind of method, a desired compound can be obtained by introducing the protective group, by carrying out the reaction and by eliminating the protective group as necessary.
In addition, the prodrug of the compound of the formula (I) can be produced by introducing a specific group or by carrying out the reaction using the obtained compound of the formula (I) at the stage from a starting material to an intermediate, just as in the case of the above-mentioned protective group. The reaction can be carried out using methods known to those skilled in the art, such as ordinary esterification, amidation, dehydration, and the like.
Hereinbelow, the representative preparation methods for the compound of the formula (I) will be described. Each of the production processes may also be carried out with reference to the References appended in the present description. Unless otherwise indicated, in any of the foregoing schemes, A1, A2, R1, R2, R3, R4, X, Y, and B are as described for formula (I) or any applicable variation thereof. Further, the preparation methods of the compound of the formula (I) are not limited to the examples as shown below.
(Production Process 1)




A compound (I) can be obtained by subjecting a compound (15) to addition and cyclization reactions.
In the addition reaction, the compound (15) and an equivalent amount or an excess amount of iodine and silver cyanate or silver thiocyanate are used, and a mixture thereof is stirred under any temperature condition from cooling to heating and refluxing, preferably at 0° C. to 200° C., and still more preferably at 20° C. to 120° C., usually for 0.1 hours to 5 days, in a solvent which is inert to the reaction or without a solvent. Examples of the solvent as used herein are not particularly limited, but include alcohols such as methanol, ethanol, tert-butanol, and the like, aromatic hydrocarbons such as benzene, toluene, xylene, and the like, ethers such as diethyl ether, tetrahydrofuran, dioxane, dimethoxyethane, and the like, halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane, chloroform, and the like, N,N-dimethylformamide, dimethylsulfoxide, ethyl acetate, acetonitrile, and a mixture thereof.
In the cyclization reaction, the crude mixture after the addition reaction and an equivalent amount or an excess amount of NH3 dissolved in solvent such as H2O or ethanol (EtOH), and so on, are used, and a mixture thereof is stirred under any temperature condition from cooling to heating to refluxing, preferably at 0° C. to 200° C., and still more preferably at 20° C. to 120° C., usually for 0.1 hours to 5 days, in a solvent which is inert to the reaction or without a solvent. Examples of the solvent as used herein are not particularly limited, but include alcohols such as methanol, ethanol, tert-butanol, and the like, aromatic hydrocarbons such as benzene, toluene, xylene, and the like, ethers such as diethyl ether, tetrahydrofuran, dioxane, dimethoxyethane, and the like, halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane, chloroform, and the like, N,N-dimethylformamide, dimethylsulfoxide, ethyl acetate, acetonitrile, water and a mixture thereof. It may be advantageous in some cases for the smooth progress of the reaction to carry out the reaction in the presence of an organic base such as triethylamine, N,N-diisopropylethylamine, N-methylmorpholine, and the like, or an inorganic base such as sodium tert-butoxide, potassium carbonate, sodium bis(trimethylsilyl)amide, sodium carbonate, potassium hydroxide, and the like.
In some embodiments of Production Process 1, the compound (I) is a compound of formula (Ie):




wherein A11 represents O or S, and A21 represents CH2.

(Production Process 2)





A compound (I) can be obtained by subjecting a compound (16) and NH3 to a substitution reaction.
In this reaction, the compound (16) and an equivalent amount or an excess amount of NH3 dissolved in solvent such as H2O or EtOH, and so on, are used, and a mixture thereof is stirred under any temperature condition from cooling to heating and refluxing, preferably at 0° C. to 200° C., and still more preferably at 20° C. to 120° C., usually for 0.1 hours to 5 days, in a solvent which is inert to the reaction or without a solvent. It may be advantageous in some cases for the smooth progress of the reaction to carry out the reaction under microwave irradiation. Examples of the solvent as used herein are not particularly limited, but include alcohols such as methanol, ethanol, tert-butanol, and the like, aromatic hydrocarbons such as benzene, toluene, xylene, and the like, ethers such as diethyl ether, tetrahydrofuran, dioxane, dimethoxyethane, and the like, halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane, chloroform, and the like, N,N-dimethylformamide, dimethylsulfoxide, ethyl acetate, acetonitrile, and a mixture thereof. It may be advantageous in some cases for the smooth progress of the reaction to carry out the reaction in the presence of tert-butyl hydroperoxide, and the like.
(Other Production Processes)
Furthermore, several substituents in the formula (I) can also be easily converted into other functional groups by using the compound of the present invention (I) as a starting material by means of the reactions apparent to a person skilled in the art, or modified methods thereof. The reaction can be carried out by any combination of the processes that can be usually employed by a person skilled in the art, such as hydrolysis, alkylation, halogenation, hydrogenation, and the like. Several examples thereof are presented below.
(Production Process 3)




(wherein RB represents H or lower alkyl, or two RB's are combined with each other to form C2-7 alkylene, Lv represents a leaving group, and R2a represents a group in R2 with the exception that R2a cannot be H or halogen. In some embodiments, R2a represents aryl or a hetero ring group which has aromaticity in R2. Moreover, said aryl may be substituted with substituents acceptable in the “aryl” of R1, R2, R3 and R4, and said hetero ring group may be substituted with substituents acceptable in the “a hetero ring group” of R1, R2, R3 and R4).

First, the compound (17) can be obtained by subjecting the compound (Ia) to a cross-coupling reaction with a borylation reagent.
In this reaction, a mixture of the compound (Ia) and a borylation reagent in equivalent amounts, or with either thereof in an excess amount is stirred under any temperature condition from cooling to heating, and preferably −20° C. to 60° C., usually for 0.1 hours to 5 days, in a solvent which is inert to the reaction, in the presence of an organometallic compound. The solvent as used herein is not particularly limited, but examples thereof include aromatic hydrocarbons such as benzene, toluene or xylene, and the like, halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane, chloroform, and the like, ethers such as diethyl ether, tetrahydrofuran, dioxane, dimethoxyethane, and the like, DMF, DMSO, EtOAc, acetonitrile, water, and a mixture thereof. Examples of the borylation reagent include bis(pinacolato)diboron, and the like. It may be advantageous in some cases for the smooth progress of the reaction to carry out the reaction in the presence of inorganic base, such as potassium acetate or potassium phenolate, and the like. Examples of an organometallic compound include palladium catalysts, such as [1,1′-Bis(diphenylphosphino)ferrocene]palladium(II) dichloride. It may be advantageous in some cases for the smooth progress of the reaction to carry out the reaction after protecting —NH2 of a compound (Ia).
Moreover, the compound (Ib) can be obtained by subjecting the compound (17) and the R2a-Lv to a coupling reaction. Herein, examples of the leaving group Lv include halogen, a trifluoromethanesulfonyloxy group, and the like.
In this reaction, a mixture of the compound (17) and an equivalent amount or an excess amount of R2a-Lv is stirred under any temperature condition from cooling to heating and refluxing, and preferably at 0° C. to 80° C., usually for 0.1 hours to 5 days, in a solvent which is inert to the reaction or without a solvent by using a catalyst used for Suzuki-Miyaura cross-coupling reaction. The solvent as used herein is not particularly limited, but examples thereof include aromatic hydrocarbons such as benzene, toluene, xylene, and the like, ethers such as dimethyl ether, diethyl ether, tetrahydrofuran, dioxane, dimethoxyethane, and the like, halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane, chloroform, and the like, N,N-dimethylformamide, dimethylsulfoxide, ethyl acetate, acetonitrile, and a mixture thereof. The catalyst as used herein is not particularly limited, but tetrakis(triphenylphosphine)palladium(0), palladium(II) acetate, dichloro[1,1′-bis(diphenylphosphino)ferrocene]palladium(II), bis(triphenylphosphine)palladium(II) chloride, tris(dibenzylideneacetone)dipalladium(0)-2-dicyclohexylphosphino-2′,6′-dimethoxybiphenyl or the like can be used. In addition, metal palladium(0) can also be used to carry out the coupling reaction. It may be advantageous in some cases for the smooth progress of the reaction to carry out the reaction after protecting —NH2 of a compound (17).
(Production Process 4)




A compound (Ib) can be obtained by subjecting a compound (Ia) and R2—B(ORB)2 to a coupling reaction. This reaction can be conducted by the same condition of the said reaction of (Production Process 3). It may be advantageous in some cases for the smooth progress of the reaction to carry out the reaction after protecting —NH2 of a compound (Ia).
(Production Process 5)




(wherein R′ represents a hetero ring group which may be substituted with substituents acceptable in the “—N(H)-(hetero ring group)” of R1, R2, R3 and R4.

A compound (Ic) among the compounds (I) of the present invention can be obtained by subjecting a compound (Ia) and NR22H2 to a substitution reaction.
In this reaction, the compound (Ia) and an equivalent amount or an excess amount of NR22H2 are used, and a mixture thereof is stirred under any temperature condition from cooling to heating and refluxing, preferably at 0° C. to 200° C., and still more preferably at 20° C. to 120° C., usually for 0.1 hours to 5 days, in a solvent which is inert to the reaction or without a solvent. It may be advantageous in some cases for the smooth progress of the reaction to carry out the reaction under microwave irradiation. Examples of the solvent as used herein are not particularly limited, but include alcohols such as methanol, ethanol, tert-butanol, and the like, aromatic hydrocarbons such as benzene, toluene, xylene, and the like, ethers such as diethyl ether, tetrahydrofuran, dioxane, dimethoxyethane, and the like, halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane, chloroform, and the like, N,N-dimethylformamide, dimethylsulfoxide, ethyl acetate, acetonitrile, and a mixture thereof. It may be advantageous in some cases for the smooth progress of the reaction to carry out the reaction in the presence of an organic base such as triethylamine, N,N-diisopropylethylamine, N-methylmorpholine, and the like, or an inorganic base such as sodium tert-butoxide, potassium carbonate, caesium carbonate, sodium bis(trimethylsilyl)amide, sodium carbonate, potassium hydroxide, and the like. It may be advantageous in some cases for the smooth progress of the reaction to carry out the reaction after protecting —NH2 of a compound (Ia).
Moreover, the reaction may be carried out using a catalyst which is not particularly limited, but includes catalysts used for Ullmann reaction, Buchwald-Hartwig reaction, or the like. The catalyst for Buchwald-Hartwig reaction as used herein is not particularly limited, but a suitable combination of tris(dibenzylideneacetone)palladium (0), tetrakis(triphenylphosphine)palladium (0), or the like with 4,5-bis(diphenylphosphino)-9,9′-dimethylxanthene (Xantphos), 2-dicyclohexylphosphino-2′,6′-dimethoxybiphenyl (SPhos), 2-dicyclohexylphosphino-2′,4′,6′-triisopropylbiphenyl (XPhos), and the like can be used. The catalyst for Ullmann reaction as used herein is not particularly limited, but a suitable combination of copper(I) iodide, or the like with (1R*, 2R*)-N,N′-dimethylcyclohexane-1,2-diamine, 1,10-phenanthroline and the like can be used.
(Production Process 6)




(wherein R21 represents a hetero ring group which may be substituted with substituents acceptable in the “—N(H)—C(O)-(hetero ring group)” of R1, R2, R3 and R4, Boc represents a tert-butoxylcarbonyl group.)

A compound (19) can be obtained by subjecting a compound (18) which is obtained by the protection reaction of (Ia) and lithium bis(trimethylsilyl)amide to an amination reaction.
In this reaction, the compound (18) and an equivalent amount or an excess amount of lithium bis(trimethylsilyl)amide are used, and a mixture thereof is stirred under any temperature condition from cooling to heating and refluxing, preferably at 0° C. to 200° C., and still more preferably at 20° C. to 120° C., usually for 0.1 hours to 5 days, in a solvent which is inert to the reaction or without a solvent in the presence of a palladium catalyst. The palladium catalyst can be prepared in situ from bis(dibenzylideneacetone)palladium (0) and tri-tert-butylphosphonium tetrafluoroborate. Examples of the solvent as used herein are not particularly limited, but include aromatic hydrocarbons such as benzene, toluene, xylene, and the like, ethers such as diethyl ether, tetrahydrofuran, dioxane, dimethoxyethane, and the like, halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane, chloroform, and the like, N,N-dimethylformamide, dimethylsulfoxide, and a mixture thereof.
A compound (20) can be obtained by subjecting a compound (19) to an amination reaction. For the reaction, the compound (19) and an equivalent amount or an excess amount of R21—C(═O)—OH are used, and a mixture thereof is stirred in a range of from cooling to heating, preferably at a temperature from −20° C. to 60° C., usually for about 0.1 hours to 5 days, in a solvent which is inert to the reaction, in the presence of a condensing agent. The solvent as used herein is not particularly limited, but examples thereof include aromatic hydrocarbons such as benzene, toluene, xylene, and the like, halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane, chloroform, and the like, ethers such as diethyl ether, tetrahydrofuran, dioxane, dimethoxyethane, and the like, DMF, DMSO, EtOAc, acetonitrile, or water, and a mixture thereof. Examples of the condensing agent include, but are not limited to, CDI, diphenylphosphoryl azide, phosphorus oxychloride, WSC (Water-Soluble Carbodiimide, trademark, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, and the like), and DCC (dicyclohexylcarbodiimide). It may be in some cases preferable for the reaction to use an additive for example, 1-hydroxybenzotriazole. It is in some cases advantageous for smooth progress of the reaction to carry out the reaction in the presence of organic bases such as triethylamine, N,N-diisopropylethylamine, N-methylmorpholine, DBU, DMAP, and the like, or inorganic bases such as potassium carbonate, sodium carbonate, potassium hydroxide, and the like.
Furthermore, it is also possible to use a method in which a reactive derivative of R21—C(═O)—OH is used, and reacted with the compound (19). Examples of the reactive derivative of the carboxylic acid include acid halides that can be obtained by the reaction with a halogenating agent such as phosphorus oxychloride, thionyl chloride, and the like, mixed acid anhydrides that can be obtained by the reaction with isobutyl chloroformate or the like, activated esters that can be obtained by condensation with 1-hydroxybenzotriazole or the like, etc. The reaction of the reactive derivative with the compound (19) can be carried out in a range of from cooling to heating, and preferably from −20° C. to 60° C., in a solvent which is inert to the reaction, such as halogenated hydrocarbons, aromatic hydrocarbons, ethers, and the like.
A compound (Id) can be obtained by subjecting a compound (20) to a deprotection reaction. The deprotection reaction can be carried out with reference to, for example, “Greene's Protective Groups in Organic Synthesis (4th Ed., 2006)”, P. G. M. Wuts and T. W. Greene.
Moreover, each reaction in (Production Process 6) can be conducted with using a compound protected by protective group except a Boc group, and each reaction may be conducted with using a non-protective compound.
(Starting Material Synthesis 1)




A compound (2) can be obtained by subjecting a compound (1) and X—(C═O)—Y to a cyclization reaction.
In this reaction, the compound (1) and an equivalent amount or an excess amount of X—(C═O)—Y are used, and a mixture thereof is stirred under any temperature condition from cooling to heating and refluxing, preferably at 0° C. to 200° C., and still more preferably at 20° C. to 120° C., usually for 0.1 hours to 5 days, in a solvent which is inert to the reaction or without a solvent. Examples of the solvent as used herein are not particularly limited, but include alcohols such as methanol, ethanol, tert-butanol, and the like, aromatic hydrocarbons such as benzene, toluene, xylene, and the like, ethers such as diethyl ether, tetrahydrofuran, dioxane, dimethoxyethane, and the like, halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane, chloroform, and the like, N,N-dimethylformamide, dimethylsulfoxide, ethyl acetate, acetonitrile, and a mixture thereof. It may be advantageous in some cases for the smooth progress of the reaction to carry out the reaction in the presence of an organic acid or organic base. Examples of the organic acid as used herein are not particularly limited, but include acetic acid, trifluoroacetic acid and the like, and examples of the organic base as used herein are not particularly limited, but include pyrrolidine, triethylamine, N,N-diisopropylethylamine, N-methylmorpholine, and the like, or an inorganic base such as sodium tert-butoxide, potassium carbonate, sodium bis(trimethylsilyl)amide, sodium carbonate, potassium hydroxide, caesium carbonate, and the like.
A compound (3) can be obtained by subjecting a compound (2) and HCHO or Lv-CH2—OH to a substitution reaction in the presence of a base. Herein, examples of the leaving group Lv include a benzotriazolyl group, and the like.
In this reaction, the compound (2) and an equivalent amount or an excess amount of HCHO or Lv-CH2—OH are used, and a mixture thereof is stirred under any temperature condition from cooling to heating and refluxing, preferably at 0° C. to 200° C., and still more preferably at 20° C. to 120° C., usually for 0.1 hours to 5 days, in a solvent which is inert to the reaction or without a solvent. Examples of the solvent as used herein are not particularly limited, but include alcohols such as methanol, ethanol, tert-butanol, and the like, aromatic hydrocarbons such as benzene, toluene, xylene, and the like, ethers such as diethyl ether, tetrahydrofuran, dioxane, dimethoxyethane, and the like, halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane, chloroform, and the like, N,N-dimethylformamide, dimethylsulfoxide, ethyl acetate, acetonitrile, water, and a mixture thereof. Examples of the organic base as used herein are not particularly limited, but include pyrrolidine, triethylamine, N,N-diisopropylethylamine, N-methylmorpholine, and the like, or an inorganic base such as sodium tert-butoxide, potassium carbonate, sodium bis(trimethylsilyl)amide, calcium hydroxide, sodium carbonate, potassium hydroxide, caesium carbonate, and the like.
A compound (4) can be obtained by subjecting a compound (3) to a modified Mitsunobu reaction as described in Warren et al. J. Chem. Soc., Perkin Trans. 1, 2001, 2983.
In this reaction, a compound (3) is treated under any temperature condition from cooling to heating, and preferably −20° C. to 80° C., usually for 0.1 hours to 3 days, in a solvent which is inert to the reaction, in the presence of zinc bis(dimethyldithiocarbamate), an azo compound and a phosphorous compound. Examples of the solvent as used herein are not particularly limited, but include ethers such as diethyl ether, tetrahydrofuran, dioxane, dimethoxyethane, and the like, halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane, chloroform, and the like, aromatic hydrocarbons such as benzene, toluene, xylene, and the like, N,N-dimethylformamide, dimethylsulfoxide, and a mixture thereof. As the azo compound, diesters of azodicarboxylic acid, such as, diethyl azodicarboxylate, or diisopropyl azodicarboxylate can be used, and as the phosphorous compound, for example, triphenylphosphine is suitably used.
(Starting Material Synthesis 2)




A compound (6) can be obtained by subjecting the compound (5) to Wittig reaction.
In this reaction, the compound (5) is treated under any temperature condition from cooling to heating, and preferably −20° C. to 80° C., usually for 0.1 hours to 3 days, in a solvent which is inert to the reaction, in the presence of an equivalent amount or an excess amount of methyltriphenylphosphonium halide such as methyltriphenylphosphonium bromide in the presence of a base. Examples of the solvent as used herein are not particularly limited, but include ethers such as diethyl ether, tetrahydrofuran, dioxane, dimethoxyethane, and the like, aromatic hydrocarbons such as benzene, toluene, xylene, and the like, N,N-dimethylformamide, dimethylsulfoxide, and a mixture thereof. Examples of the base as used herein are not particularly limited, but include sodium bis(trimethylsilyl)amide, n-butyllithium, potassium tert-butoxide, sodium ethoxide, sodium methoxide, sodium hydride, and the like.
(Starting Material Synthesis 3)




(wherein RProc represents a protective group, B1 represents a cycloalkyl group which may be substituted).

A compound (9) can be obtained by subjecting a compound (7) and a compound (8) to a substitution reaction. Herein, the reaction is conducted after converting —OH group of a compound (7) to a leaving group, such as mesyloxy group (methanesulfonyloxy group).
In this reaction, a mesylate derivative of the compound (7) and an equivalent amount or an excess amount of compound (8) are used, and a mixture thereof is stirred under any temperature condition from cooling to heating and refluxing, preferably at 0° C. to 200° C., and still more preferably at 20° C. to 120° C., usually for 0.1 hours to 5 days, in a solvent which is inert to the reaction or without a solvent. Examples of the solvent as used herein are not particularly limited, but include aromatic hydrocarbons such as benzene, toluene, xylene, and the like, ethers such as diethyl ether, tetrahydrofuran, dioxane, dimethoxyethane, and the like, halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane, chloroform, and the like, N,N-dimethylformamide, dimethylsulfoxide, ethyl acetate, acetonitrile, and a mixture thereof. It may be advantageous in some cases for the smooth progress of the reaction to carry out the reaction in the presence of an acid or a base. Examples of the organic acid as used herein are not particularly limited, but include, acetic acid, trifluoroacetic acid, p-toluenesulfonic acid, methanesulfonic acid and the like, examples of the inorganic acid as used herein are not particularly limited, but include, hydrochloric acid, sulfuric acid, potassium hydrogen sulfate and the like, and examples of the organic base as used herein are not particularly limited, but include, pyridine, 2,6-lutidine (2,6-dimethylpyridine), triethylamine, diisopropylethylamine, 1,8-diazabicyclo[5.4.0]undec-7-ene and the like, examples of the inorganic base as used herein are not particularly limited, but include, sodium carbonate, potassium carbonate, sodium bicarbonate, potassium phosphate, caesium carbonate and the like.
A compound (10) can be obtained by converting an ester group of a compound (9) to a carboxylic acid group by a hydrolysis reaction, and then performing a cyclization reaction.
First, the hydrolysis reaction can be carried out with reference to, for example, “Greene's Protective Groups in Organic Synthesis (4th Ed., 2006)”, P. G. M. Wuts and T. W. Greene.
Next, the hydrolysis product obtained from the compound (9) is stirred under any temperature condition from cooling to heating and refluxing, and preferably at 0° C. to 80° C., usually for 0.1 hours to 5 days, in a solvent which is inert to the reaction or without a solvent. Examples of the solvent as used herein are not particularly limited, but include aromatic hydrocarbons such as benzene, toluene, xylene, and the like, ethers such as diethyl ether, tetrahydrofuran, dioxane, dimethoxyethane, and the like, halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane, chloroform, and the like, N,N-dimethylformamide, dimethylsulfoxide, ethyl acetate, acetonitrile, and a mixture thereof. It is in some cases advantageous in advancing the reaction smoothly to carry out the reaction under an acidic condition. Examples of the acid as used herein are not particularly limited, but include organic acids such as p-toluenesulfonic acid, acetic acid, and the like, and inorganic acids such as hydrochloric acid, sulfuric acid, and the like.
(Starting Material Synthesis 4)




A compound (12) can be obtained by hydrolysis reaction.
In this reaction, the compound (11) is stirred under any temperature condition from cooling to heating and refluxing, and preferably at 0° C. to 80° C., usually for 0.1 hours to 5 days, in a solvent which is inert to the reaction or without a solvent in the presence of a base such as lithium hydroxide. Examples of the solvent as used herein are not particularly limited, but include aromatic hydrocarbons such as benzene, toluene, xylene, and the like, ethers such as diethyl ether, tetrahydrofuran, dioxane, dimethoxyethane, and the like, alcohols such as methanol, ethanol, tert-butanol, and the like, N,N-dimethylformamide, dimethylsulfoxide, acetonitrile, water and a mixture thereof.
A compound (13) can be obtained by subjecting a compound (12) and Lv-C(═O)—CH2-Lv to a substitution reaction and cyclization reaction.
In the substitution reaction, the compound (12) and an equivalent amount or an excess amount of Lv-C(═O)—CH2-Lv are used, and a mixture thereof is stirred under any temperature condition from cooling to heating and refluxing, preferably at 0° C. to 200° C., and still more preferably at 20° C. to 120° C., usually for 0.1 hours to 5 days, in a solvent which is inert to the reaction or without a solvent. Examples of the solvent as used herein are not particularly limited, but include aromatic hydrocarbons such as benzene, toluene, xylene, and the like, ethers such as diethyl ether, tetrahydrofuran, dioxane, dimethoxyethane, and the like, halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane, chloroform, and the like, N,N-dimethylformamide, dimethylsulfoxide, ethyl acetate, acetonitrile, and a mixture thereof. It may be advantageous in some cases for the smooth progress of the reaction to carry out the reaction in the presence of an acid or base. Examples of the organic acid as used herein are not particularly limited, but include, acetic acid, trifluoroacetic acid, p-toluenesulfonic acid, methanesulfonic acid and the like, examples of the inorganic acid as used herein are not particularly limited, but include, hydrochloric acid, sulfuric acid, potassium hydrogen sulfate and the like, and examples of the organic base as used herein are not particularly limited, but include, pyridine, 2,6-lutidine (2,6-dimethylpyridine), triethylamine, diisopropylethylamine, 1,8-diazabicyclo[5.4.0]undec-7-ene and the like, examples of the inorganic base as used herein are not particularly limited, but include, sodium carbonate, potassium carbonate, sodium bicarbonate, potassium phosphate, caesium carbonate and the like.
In the cyclization reaction, the compound after substitution reaction of a compound (12) is stirred under any temperature condition from cooling to heating and refluxing, preferably at 0° C. to 200° C., and still more preferably at 20° C. to 120° C., usually for 0.1 hours to 5 days, in a solvent which is inert to the reaction or without a solvent. Examples of the solvent as used herein are not particularly limited, but include alcohols such as methanol, ethanol, tert-butanol, 2-methylbutan-2-ol, and the like, aromatic hydrocarbons such as benzene, toluene, xylene, and the like, ethers such as diethyl ether, tetrahydrofuran, dioxane, dimethoxyethane, and the like, halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane, chloroform, and the like, N,N-dimethylformamide, dimethylsulfoxide, ethyl acetate, acetonitrile, and a mixture thereof. It may be advantageous in some cases for the smooth progress of the reaction to carry out the reaction in the presence of an acid or base. Examples of the organic acid as used herein are not particularly limited, but include, acetic acid, trifluoroacetic acid, p-toluenesulfonic acid, methanesulfonic acid and the like, examples of the inorganic acid as used herein are not particularly limited, but include, hydrochloric acid, sulfuric acid, potassium hydrogen sulfate and the like, and examples of the organic base as used herein are not particularly limited, but include, pyridine, 2,6-lutidine (2,6-dimethylpyridine), triethylamine, diisopropylethylamine, potassium tert-butoxide, 1,8-diazabicyclo[5.4.0]undec-7-ene and the like, examples of the inorganic base as used herein are not particularly limited, but include, sodium carbonate, potassium carbonate, sodium bicarbonate, potassium phosphate, caesium carbonate and the like.
A compound (14) can be obtained by a reaction of a compound (13) and Lawesson's reagent.
In this reaction, the compound (13) is stirred under any temperature condition from cooling to heating and refluxing, and preferably at 0° C. to 80° C., usually for 0.1 hours to 5 days, in a solvent which is inert to the reaction or without a solvent in the presence of Lawesson's reagent (2,4-bis(4-methoxyphenyl)-1,3,2,4-dithiadiphosphetane-2,4-disulfide). Examples of the solvent as used herein are not particularly limited, but include aromatic hydrocarbons such as benzene, toluene, xylene, and the like, ethers such as diethyl ether, tetrahydrofuran, dioxane, dimethoxyethane, and the like, halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane, chloroform, and the like, dimethylsulfoxide, acetonitrile, and a mixture thereof.
(Starting Material Synthesis 5)




A compound (21) can be obtained by Mannich reaction and elimination reaction of compound (2).
In this reaction, a mixture of the compound (2), N,N,N′,N′-tetramethylmethanediamine, and acetic acid is stirred under any temperature condition from cooling to heating and refluxing, and preferably at 0° C. to 80° C., usually for 0.1 hours to 5 days, in a solvent which is inert to the reaction or without a solvent. Examples of the solvent as used herein are not particularly limited, but include aromatic hydrocarbons such as benzene, toluene, xylene, and the like, ethers such as diethyl ether, tetrahydrofuran, dioxane, dimethoxyethane, and the like, halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane, chloroform, and the like, N,N-dimethylformamide, dimethylsulfoxide, ethyl acetate, acetonitrile, and a mixture thereof. Subsequently, acetic anhydride is added to the mixture and the mixture is stirred under any temperature condition from cooling to heating and refluxing, and preferably at 0° C. to 80° C., usually for 0.1 hours to 5 days, in a solvent which is inert to the reaction or without a solvent. Examples of the solvent as used herein are described above.
A compound (22) can be obtained by subjecting a compound (21) to a Corey-Chaykovsky type reaction.
In this reaction, the compound (21) and an equivalent amount or an excess amount of trimethylsulfoxonium iodide are used, and a mixture thereof is stirred under any temperature condition from cooling to heating and refluxing, preferably at 0° C. to 200° C., and still more preferably at 20° C. to 120° C., usually for 0.1 hours to 5 days, in a solvent which is inert to the reaction or without a solvent in the presence of a base. Examples of the solvent as used herein are not particularly limited, but include alcohols such as methanol, ethanol, tert-butanol, and the like, aromatic hydrocarbons such as benzene, toluene, xylene, and the like, ethers such as diethyl ether, tetrahydrofuran, dioxane, dimethoxyethane, and the like, halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane, chloroform, and the like, N,N-dimethylformamide, dimethylsulfoxide, ethyl acetate, acetonitrile, and a mixture thereof. It may be advantageous in some cases for the smooth progress of the reaction to use a pre-formed mixture of a base and trimethylsulfoxonium iodide and add the mixture to the compound (21). Examples of the inorganic base as used herein are not particularly limited, but include, sodium hydride, potassium hydroxide, sodium carbonate, potassium carbonate, sodium bicarbonate, potassium phosphate, caesium carbonate, and the like.
(Starting Material Synthesis 6)




A compound (11) can be obtained by subjecting a compound (6) to reaction with silver cyanate. This reaction can be carried out using similar conditions as for the reaction of a compound (15) with silver cyanate in (Production Process 1), except for use of excess tert-butanol in the presence of triethylamine, instead of excess NH3, in the cyclization step.
(Starting Material Synthesis 7)




A compound (23) can be obtained by subjecting a compound (3) to reaction with nucleophilic methylation reagent and dehydration reaction.
In this reaction, a mixture of the compound (3) and nucleophilic methylation reagent is stirred under any temperature condition from cooling to heating and refluxing, and preferably at −78° C. to 80° C., usually for 0.1 hours to 5 days, in a solvent which is inert to the reaction or without a solvent. Examples of the solvent as used herein are not particularly limited, but include aromatic hydrocarbons such as benzene, toluene, xylene, and the like, ethers such as diethyl ether, tetrahydrofuran, dioxane, dimethoxyethane, and the like and a mixture thereof. Examples of the nucleophilic methylation reagent as used herein are not particularly limited, but include methylmagnesium bromide, methylmagnesium chloride, methylmagnesium iodide, methyllithium, and the like.
After the methylation step, an acid is added to the reaction mixture. And, the reaction mixture is stirred under any temperature condition from cooling to heating and refluxing, and preferably at 0° C. to 80° C., usually for 0.1 hours to 5 days. Examples of the organic acid as used herein are not particularly limited, but include, acetic acid, trifluoroacetic acid, p-toluenesulfonic acid, methanesulfonic acid and the like, examples of the inorganic acid as used herein are not particularly limited, but include, hydrochloric acid, sulfuric acid, potassium hydrogen sulfate and the like.
A compound (24) can be obtained by subjecting a compound (23) to a cyclization reaction.
First, a mixture of the compound (23) and a sulfonyl halide is stirred under any temperature condition from cooling to heating and refluxing, and preferably at −18° C. to 50° C., usually for 0.1 hours to 5 days, in a solvent which is inert to the reaction or without a solvent in the presence of a base. Examples of the sulfonyl halide as used herein are not particularly limited, but include methanesulfonyl chloride, tosyl chloride, and the like. Examples of the solvent as used herein are not particularly limited, but include aromatic hydrocarbons such as benzene, toluene, xylene, and the like, ethers such as diethyl ether, tetrahydrofuran, dioxane, dimethoxyethane, and the like, halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane, chloroform, and the like, ethyl acetate, acetonitrile, and a mixture thereof. Examples of the organic base as used herein are not particularly limited, but include pyridine, triethylamine, N,N-diisopropylethylamine, N-methylmorpholine, methyllithium, n-butyllithium and the like, or an inorganic base such as potassium carbonate, sodium bis(trimethylsilyl)amide, sodium carbonate, potassium hydroxide, caesium carbonate, sodium hydroxide, sodium hydride and the like. In some cases, compound (23) can be pretreated with the base for the smooth progress of the reaction before addition of the sulfonyl halide.
After the sulfonylation step, a base is added to the reaction mixture. And, the reaction mixture is stirred under any temperature condition from cooling to heating and refluxing, and preferably at 0° C. to 80° C., usually for 0.1 hours to 5 days. Examples of an organic base as used herein are not particularly limited, but include pyridine, triethylamine, N,N-diisopropylethylamine, N-methylmorpholine, methyllithium, n-butyllithium and the like, or an inorganic base such as potassium carbonate, potassium bis(trimethylsilyl)amide, sodium carbonate, potassium hydroxide, caesium carbonate, sodium hydroxide, sodium hydride and the like.
The compounds of the formula (I) can be isolated and purified as their free compounds, salts, hydrates, solvates, or polymorphic crystal substances thereof. The salts of the compound of the formula (I) can be prepared by carrying out a conventional salt forming reaction.
Isolation and purification are carried out by employing ordinary chemical operations such as extraction, fractional crystallization, various types of fractional chromatography, and the like.
Various isomers can be prepared by selecting an appropriate starting compound or separated by using the difference in the physicochemical properties among the isomers. For example, the optical isomers can be obtained by means of a general method for designing optical resolution of racemic products (for example, fractional crystallization for inducing diastereomer salts with optically active bases or acids, chromatography using a chiral column or the like, and others), and further, the isomers can also be prepared from an appropriate optically active starting compound.
The pharmacological activity of the compounds of the formula (I) was confirmed by the tests shown below.
TEST EXAMPLE
Inhibition of Beta-Secretase Activity
Test Example 1
Measurement of BACE1 Inhibition by Fluorescence Resonance Energy Transfer (FRET)
Potency of test compounds were determined by measurement of their inhibition of BACE1 activity toward a fluorescent substrate. Experiments were performed by reference to the procedure as described in Ermolieff, et al. (Biochemistry 39:12450-12456 (2000), the teachings of which are incorporated hereby in their entirety). Briefly, the recombinant protease unit of BACE1 was prepared from E. coli expression as inclusion bodies, refolded, and purified as described in Lin, et al., (Proc. Nat. Acad. Sci. 97:1456-1460 (2000)). Fluorogenic substrate, MCA-SEVNLDAEFK(DNP)-NH2 (SEQ ID NO:1) was purchased. (M-2485, Bachem Americas, Torrance, Calif.). The substrate was derived from 10 amino acids of the human amyloid precursor protein (APP), with the Swedish variant amino acids at the beta-secretase cleavage site. The terminal amino acid was modified from arginine to lysine to facilitate derivatization with a functional group for detection by autofluorescence. The amino acid sequence of the “core” peptide of the substrate is SEVNLDAEFK (SEQ ID NO:2). The amino terminus was derivatized with (7-methoxycoumarin-4-yl)acetyl (MCA), and the epsilon amine of the lysine side chain of the terminal residue (K in sequence SEVNLDAEFK (SEQ ID NO:2)) was derivatized with 2,4-dinitrophenyl (DNP). Assays were performed in a buffer of 0.1 M sodium acetate, pH 4.4, 0.08% 3-[(3-Cholamidopropyl)dimethylammonio]propanesulfonate (CHAPS), 0.005% Tween80. BACE1 enzyme (final concentration 65 nM) was pre-incubated with test compounds for 15 minutes at room temperature. Fluorescence intensities were measured 60 minutes after addition of the substrate (final concentration 3 μM) by Tecan Safire2TM. An excitation wavelength of 328 nm and an emission wavelength of 393 nm were used. For the calculation of % inhibition, fluorescence intensity without compounds was defined as the value for 0% inhibition and fluorescence intensity without the enzyme was defined as the value for 100% inhibition. The values of IC50 were calculated by GraphPad Prism version 5.
Moreover, the inhibition constants, Ki, were determined as described in Ermolieff, et al. (Biochemistry 39:12450-12456 (2000)). Briefly, the hydrolysis of the fluorogenic substrate, for a series of mixtures with constant enzyme, but increasing inhibitor concentration was carried out in the same manner as described in the method for Test Example 1. Quantification of enzymes was achieved by active-site titration using a tight-binding inhibitor. The inhibition constant, Ki, was determined from plot of activity vs. inhibitor concentration based on the equation described in Ermolieff, et al. (Biochemistry 39:12450-12456 (2000)).
The results of the representative compounds are shown in [Table. 1] below.
The inhibition constants Ki of Example 89 and Example 98 compounds described in Pamphlet of International Publication WO2011/123674 were determined. In the result, the Ki value of Example 89 compound was 0.241 μM, and the Ki value of Example 98 compound was 4.087 μM.
Herein, the structure of Example 89 compound described in Pamphlet of International Publication WO2011/123674 is




and this compound is a racemic mixture.

The structure of Example 98 compound described in Pamphlet of International Publication WO2011/123674 is




and this compound is a racemic mixture.

The results of the representative compounds are shown in [Table. 1] below.
Test Example 2
Measurement of BACE1 Inhibition by Time Resolved Fluorescence Resonance Energy Transfer (TR-FRET)
Potency of compounds were also measured using another fluorogenic substrate, TruPoint BACE1 Substrate Eu-CEVNLDAEFK-QSY 7 (SEQ ID NO:3) (AD0258, PerkinElmer, Boston Mass.). This substrate also has Swedish variant amino acids at the β-secretase cleavage site, with a fluorescent europium (Eu) chelate coupled to one end and a quencher of europium fluorescence (QSY7) coupled to the other end via lysine. If the sample contains BACE1 activity, the Eu chelate and the quencher will be separated as the substrate is cleaved. The Eu-signal increases and it can be measured by time-resolved fluorometry, EnVisionTM, 30 minutes after the substrate (final concentration 300 nM) was added.
The experiment was basically conducted in a way similar to Test Example 1 above.
The results of the representative compounds are shown in [Table. 1] below.
Test Example 3
Measurement of Aβ Production Inhibition in Cell
The potency of compounds against BACE1 activity was determined in a cellular assay of Aβ production. Human SK-N-BE(2) neuroblastoma cells (ATCC No. CRL-2271) were plated at 96,000 cells/well/100 μL in 96-well plates in RPMI1640 medium/10% fetal bovine serum (FBS)/penicillin-streptomycin and cultured for 24 hours at 37° C., 5% CO2. Test compounds were dissolved in dimethyl sulfoxide and diluted with dimethyl sulfoxide and put into RPMI1640/5% FBS/penicillin-streptomycin media (final dimethyl sulfoxide concentration is 0.5%). The culture media in 96-well plates were replaced by 125 μL/well of the media containing test compounds. After incubation for 6 hours at 37° C., 5% CO2, 30 μL of the media were transferred into a fresh 96-well plate and used for Aβ40 assay by an enzyme-linked immunosorbent assay (ELISA) kit (#27718, Immuno-Biological Laboratories, Japan) according to the manufacturer's protocol. Cell viability was measured by CellTiter-GloTM Luminescent Cell Viability Assay (#7571, Promega) after removal of 30 μL of the media for the Aβ assay. CellTiter-Glo Substrate was dissolved into CellTiter-Glo Buffer and added to the plates in 95 μL/well. After shaking the plates for 2 minutes, the whole sample was transferred into a white 96-well plate and luminescence was measured for ATP quantification as the cell viability. Aβ concentration measured by ELISA was normalized by the viability of the corresponding cells. The values of IC50 were calculated by GraphPad Prism version 5.
The results of the representative compounds are shown in [Table. 1] below.
Test Example 4
Brain Aβ Reduction in Rats
Effects on brain Aβ reduction in rats were determined with reference to the method described in the WO2012/054510. It was confirmed that some of the compounds of the formula (I) exhibit brain Aβ reduction in rats. Concretely, the test was conducted with the method as below.
Formulation
Test compounds were prepared in a vehicle of 35% HPβCD in H2O. The test compound was formulated the same day as oral dosing. Doses (see [Table. 1]) were based on the free base equivalent. Sonication was used where required to facilitate the formulation.
Test Species
Male Sprague-Dawley rats (150-300 grams) were obtained from Charles River Japan (Atsugi, Japan) and were given approximately 4 days of acclimation. Food and water was made available ad libitum throughout the study. Animals were visually inspected for health before being included into the study group, and were randomly assigned to the treatment and control groups to achieve similar group mean body weights. The dosing solution (dose volume 5 mL/kg) was administered directly into the stomach using a rodent gavage needle. Control animals received oral administration of equivalent volume of the vehicle.
Sampling Methods
At a time post-dose (e.g., 3 hours; see [Table. 1]), animals were euthanized with isoflurane. Blood was collected from the inferior vena cava using syringe flushed with EDTA 2K and placed on ice. Plasma was separated using centrifugation at 15,000 rpm (20,400×g) for 5 minutes at 4° C. and subsequently stored at −80° C. After blood sampling, CSF (cerebrospinal fluid) was carefully withdrawn from the cisterna magna using a 29 gauge needle after a quick dissection to expose the atlantooccipital membrane. The CSF samples were centrifuged at 15,000 rpm (20,400×g) for 5 minutes to confirm free of blood contamination and stored at −80° C. Immediately after decapitation, the hippocampus were isolated on ice, and quickly frozen in liquid nitrogen and stored at −80° C.
Extraction of Brain Aβ42
Fragments of the hippocampus were weighed while frozen. A 10-fold volume (w:v) of TBS (Tris-buffered saline) supplemented with a Complete Mini protease inhibitor tablet (catalog number: 11 836 153 001, Roche Diagnostics, In, USA) was added. The hippocampus were homogenized using sonication on ice in microcentrifuge tube. Resulting homogenates were centrifuged at 100,000×g for 1 hour in a refrigerated centrifuge at 4° C. Supernatants were collected as soluble fraction.
Determination of Aβ42
Concentration of Aβ42 in the extract of hippocampus, plasma, and CSF were analyzed using ELISA (Human/Rat Aβ42 ELISA, catalog number 292-64501, Wako Pure Chemical Industries, Ltd. Japan). Each concentration of Aβ42 was divided by the mean concentration of Aβ42 of vehicle-treated group, and these ratios were converted to percentages.
The results of the representative compounds are shown in [Table. 1] below.
Test Example 5
hERG (Human Ether-a-go-go Related Gene) Analysis hERG Inhibition
The hERG potassium current was measured in a hERG-stably-expressing Chinese hamster ovary K1 (CHO) cells. The experiments were performed using an automated planar patch-clamp system QPatch HTX (Sophion Bioscience A/S). The application of pressure for forming gigaseals and whole-cell patch clamp configuration were established using the QPatch assay software. Patch-clamp experiments were performed in voltage-clamp mode and whole-cell currents were recorded. The following stimulation protocol was applied to investigate the effects of compounds on hERG potassium channel.
The membrane potential was held at −80 mV and repetitively (every 15 seconds) depolarized to +20 mV for 4800 milliseconds after the pulse to −50 mV for 20 milliseconds served to define the baseline, followed by repolarizing step to −50 mV for 5000 milliseconds to evaluate of the tail current amplitude. Experiments were conducted at room temperature.
Effects of compounds were determined from cumulative applications of increasing 6 concentrations and calculated as percent of blocked current. The data points were fitted with Hill equation to calculate half-maximal inhibition concentrations (IC50). The maximum compound concentration tested in the assay was 10 μM for some compounds. If less than 50% inhibition was achieved at the 10 μM compound concentration, the IC50 is reported as >10 μM.
The test solution includes:
Extracellular solution: 2 mM of CaCl2, 1 mM of MgCl2, 10 mM of HEPES, 4 mM of KCl, 145 mM of NaCl, and 10 mM of glucose; and pH adjusted to 7.4 with NaOH,
Intracellular solution: 5.374 mM of CaCl2, 1.75 mM of MgCl2, 10 mM of HEPES, 10 mM of EGTA, 120 mM of KCl, and 4 mM of ATP, and pH adjusted to 7.2 with KOH.
hERG selectivity
Selectivity of BACE1 inhibition over hERG inhibition was calculated by dividing hERG IC50 by BACE1 Ki. The results of the representative compounds are shown in [Table. 1] below. As mentioned above, some results from the hERG assay are necessarily reported as >10 μM. Using these values in the calculation of selectivity will necessarily cause the selectivity values to be characterized as “>” or “greater than” the calculated ratio.
The hERG IC50 values of Example 89 and Example 98 compounds described in Pamphlet of International Publication WO2011/123674 were determined. In the result, the IC50 value of Example 89 compound was 0.44 μM, and the IC50 value of Example 98 compound was 9.46 μM. Moreover, selectivity values of BACE1 inhibition over hERG inhibition of these compounds were calculated. In the result, the selectivity value of Example 89 compound was 1.8, and the selectivity value of Example 98 compound was 2.3.
It is considered to be desirable that compounds show their primary pharmacological effect with selectivity over hERG inhibition (Jamieson et al. J. Med. Chem. 2006, 49, 5029). Compounds with lower hERG selectivity are considered to have higher risk to cause QTc prolongation, which may eventually lead to drug-induced arrhythmia and sudden deaths. For example, hERG selectivity around or less than 10-fold over hERG inhibition are recognized to have a concern for high risk of QTc prolongation (Kongsamut et al. Eur. J. Pharmacol. 2002, 450, 37. and Minotti, Cardiotoxicity of Non-Cardiovascular Drugs, Wiley, 2010. p. 65), whereas compounds with selectivity around or larger than 100-fold are recognized to be more favorable (Pajouhesh et al. Bioorg. Med. Chem. Lett. 2012, 22, 4153; Micheli et al. J. Med. Chem. 2010, 53, 374).
The results of the representative compounds are shown in [Table. 1] below.
In [Table. 1], Ex means Example Number, and “Test Example X” refers to the protocol described above used to obtain the data. Moreover, RP prefixed before the numeral shows the compound of Reference Example.














TABLE 1







Test


Test
Test




Test
Example 1
Test
Test
Example 4
Example 5 
Test



Example 1
BACE1
Example 2 
Example 3
Aβ42
hERG
Example 5



IC50 
MCA Ki
IC50 
IC50 
reduction 
IC50
hERG


Ex. 
(μM)
(μM)
(μM)
(μM)
(%)
(μM) 
selectivity






















RP
73.3








1a









RP
138








1b









RP
59.6








2









RP
33.8








3









RP
12.6


7.2

>10



4









RP
1.56


0.59

>10



5









RP
11.7


3.9

>10



6









RP
39.0








7a









RP
39.7








7b









RP
17.3




>10



9a









RP
24.4




>10



9b









RP
61.3








8









RP
7.21




>10



10









RP
15.6








11a









RP
17.4








11b









RP
38.7








12a









RP
81.3








12b









RP
11.1








13a









RP
19.3








13b









RP
19.0








14a









RP
44.2








14b









RP
1.72


1.1

>10



15a









RP
1.01


0.44

7.93



15b









RP
9.54




>10



16a









RP
4.57


4.8

>10



16b









RP
30.2




>10



17









RP
41.3








18









RP
2.31


0.43





19









RP


2.71
0.58





20









RP


1.35
0.41





21









RP


2.60
2.9





22









RP
0.519


0.90





23









RP
37.9

32.9






24









RP
36.9








25









RP
3.75


0.069





26









27
0.0822
0.0382

0.0094

>10
>262


28
0.154
0.117

0.019

>10
>86


RP
0.276
0.240

0.022

>10
>42


29









30
0.0975
0.0515

0.0023

>10
>194


RP
1.06
1.03

0.061

>10
>10


31









RP
0.418
0.378

0.022

2.17
6


32









RP
1.52


0.95





33









RP
1.24


0.26

>10



34









RP
26.0








35









RP
2.90


0.81





36









RP


55.4






37









RP
17.3








38









RP
46.9








40









RP
24.8


>30





41









RP
16.1


5.9

>10



42









RP
1.96


1.4

>10



43









RP
0.602


0.098





44









RP
1.04


0.057

>10



45









RP
8.94


1.4

>10



46









RP
2.33


1.4





47









RP
1.13


0.69





48









RP
48.6




>10



51a









RP
0.172
0.137

0.072

>10



51b









RP
0.161


0.075

4.83



52a









RP
0.157
0.123

0.099

>10



53b









RP
0.406


0.079

>10



54









RP



0.74

>10



56









RP
3.69


0.61

3.48



57









RP
1.48


0.18

5.81



58









RP
1.11


0.076

6.9



59









RP
1.48


0.16

2.78



60









RP



0.35

9.22



61









RP


1.33
1.0

4.43



62









RP
7.90


3.3

4.48



63









RP
6.12


2.3

1.67



64









RP


0.388
0.049

9.77



65









RP


135


>10



66









RP


96.5


>10



67









RP
0.345


0.50

6.05



68









RP
0.397


0.040

>10



69









RP
1.75


0.96

>10



70









RP
1.08


0.11

>10



71









RP
0.817


0.068

8.22



72









RP
0.827


0.070





73









RP
35.4








74









RP
3.67


1.1

>10



75









RP
13.4




>10



76









RP
1.43


0.92

>10



77









RP
2.05


1.3

>10



78









RP
1.69


0.45

>10



79









RP
1.91


0.71

>10



80









RP
1.89


0.51

>10



81









RP
0.348


0.036

>10



82









RP
0.285
0.255

0.047

>10



83









RP
0.735


0.41

>10



84









RP
0.319


0.077

>10



85









RP
0.143
0.107

0.058

7.94



86









RP
1.55


0.43





87









RP
2.51


1.4

>10



88









RP
4.67




>10



89









RP
0.901


0.50

>10



90









RP
0.656


0.34

>10



91









RP
0.124
0.0879

0.037

5.34



92









RP
0.629


0.37

>10



93









RP
0.862


0.38

>10



94









RP
0.740


0.41

>10



95









RP
0.126
0.868

0.022

>1



96









RP
20.2


2.6

>10



97









RP
64.1




>10



98









RP
2.97


1.9

>10



99









RP
2.62


0.25

>10



100









RP
7.65




5.28



101









RP
1.61


0.24

>10



102









RP
2.19


0.45

>10



103









RP


1.13
0.30

3.25



104









RP
2.94


0.34

7.9



105









RP
1.83


0.16

5.75



106









RP
1.06


0.46

2.91



107









RP
0.383
0.354
0.316
0.096

9.4



108









RP
0.152
0.120

0.048

>1



109









RP
0.137
0.0904

0.0092

>1



110









RP
0.287
0.252

0.14

1.98



111









RP
0.400


0.66

4.32



112









RP
1.11


0.47





113









RP
68.3








114









RP
3.31


1.1

>10



115









RP
1.27


0.70

>10



116









RP
4.40


3.0

>10



117









RP
1.81


0.99

>10



118









RP
3.56








119









RP
1.89


2.3





120









RP
3.37








121









RP
2.74


1.1





122









RP
3.39








123









RP
3.59








124









RP
2.31


0.58





125









RP
0.696


1.2





126









RP
2.27


1.8





127









RP
1.68


1.5





128









RP
2.21


7.3





129









RP
1.87


1.7





130









RP
2.02


3.2





131









RP
2.45


1.2





132









RP
1.67


2.8





133









RP
3.63








134









RP
3.27


1.8





135









RP
0.635


0.66





136









RP
0.773


0.83





137









RP
1.67


1.5





138









RP
0.252
0.215

0.14





139









RP
0.389


0.41





140









RP
0.903


1.30





141









RP
0.401


0.15

>1



142









RP
1.26








143









RP
0.879


0.46





144









RP
0.933








145









RP
0.226
0.188

0.21





146









RP
0.621


0.17





147









RP
0.614


0.48





148









RP


1.593
1.1





149









RP


0.759
0.45





150









RP
1.36


0.28





151









RP
0.688


0.11





152









RP
1.13


0.13





153









RP


0.313
0.032

>1



154









RP


0.786
0.63





155









RP
2.61


0.18





156









RP
2.80


0.66





157









RP
0.930


0.37





158









RP
2.12








159









RP
0.981








160









RP
1.41








161









RP
1.04








162









RP


1.064






163









RP


2.314






164









RP
0.760








165









RP
3.63








166









RP
0.869








167









RP


0.760
0.31





168









RP
1.41

0.859
0.90





169









RP
1.00

0.588
0.40





170









RP
2.24

3.04
3.1





171









RP
2.24

2.16






172









RP
0.401

0.239
0.073





173









RP
3.35

2.18
3.4





174









RP
3.23

1.55






175









RP


0.994
0.85





176









RP


2.12
1.5





177









RP
1.15

0.747
0.81





178









RP
3.24

2.06
1.4





179









RP
2.03

1.77
0.49





180









RP
2.59

2.13
0.82





181









RP


1.45
2.5





182









RP
1.08

0.880






183









RP
2.82

2.72






184









RP
2.05

1.27
0.76





185









RP


2.42






186









RP
3.16

1.92






187









RP
50.6




>10



188









RP
0.661
0.623

0.043

>10
>16


189









190
0.125
0.0835

0.0041

>10
>120


RP
0.248
0.215

0.027

>10
>47


191









RP
0.318
0.280

0.030

>10
>36


192









RP
1.04


0.081

>10
>120


193









RP
0.349
0.313

0.028

4.91
16


194









195
0.109
0.0645

0.0032

>10
>155


RP
0.380
0.342

0.053

3.65
11


196









197
0.118
0.0720

0.0057

>10
>139


RP
14.9




>1



198









RP
0.736


0.24

4.25



199









RP
0.128
0.0921

0.026

>10



200









RP
0.113
0.0711

0.0087





201









RP
0.145
0.113

0.026

7.73



202









RP
0.109
0.0676

0.0049





203









RP
0.388


0.24

>10



204









RP
0.126
0.0825

0.022

5.22



205









RP
4.91


>30

>10



206









RP
0.460


0.16

>10



207









RP
7.76


6.1

>10



208









RP
2.00


0.83

>10



209









RP


4.95


3.7



210









RP


19.4






211









RP
17.6




>10



212









RP


39.6
6.3

>10



213









RP
40.3








214









RP
1.21


0.24

>10



215









RP
55.8








216









218
0.0447
0.0103

0.00078
24%
20.67
2007







(3 mg/kg, 









2.5 hours)









31%









(10 mg/kg, 









2.5 hours)




219
0.0444
0.0102
0.0418
0.0022

>10
>980


220
0.0765
0.0429

0.0031
21%
>10
>233







(3 mg/kg, 









1.5 hours)









27%









(10 mg/kg, 









1.5 hours)




221
0.0818
0.0378

0.00016
24%
3.25
86







(1 mg/kg, 









3 hours)









42%









(3 mg/kg, 









3 hours)




222
0.138
0.0915

0.00062

>10
>109


223
0.0991
0.0612

0.0031
19%
20.77
339







(1 mg/kg, 









3 hours)









34%









(3 mg/kg, 









3 hours)




224
0.0914
0.0527

0.00063
21%
15.1
287







(1 mg/kg, 









3 hours)









35%









(3 mg/kg, 









3 hours)




227
0.107
0.0792

0.0080

>10
>126


228a
30.3
30.2



>10
>0.3


229a 
47.6%









inhibition at









30 microM









As described above, it was confirmed that the representative compounds of the formula (I) have β-secretase inhibitory activities, Aβ production inhibitory activities, and Aβ reduction activities and can be therefore used for diseases or conditions associated with and/or mediated by β-secretase activity, hydrolysis of a β-secretase cleavage site of an amyloid precursor protein, and/or β-amyloid protein accumulation, such as Glaucoma, MCI (Mild cognitive impairment) or Alzheimer's disease, especially, Alzheimer's disease, or the like.
Determination of Absolute Stereochemistry by Vibrational Circular Dichroism (VCD) Spectrometry Measurements
The infrared and VCD spectra were recorded on a Bio tools ChiralIR-2XTM Vibrational Circular Dichroism (VCD) spectrometer.
The infrared and VCD spectra were measured in CDCl3 solution placed in a 100 μm path length cell with BaF2 windows at 4 cm−1 resolution and their data collection was performed for 5 hours.
Calculations
Conformational searches were executed by using CONFLEXTM ver.6 program.
The geometry optimizations and the calculations of theoretical infrared and VCD spectra were implemented using density functional theory with B3LYP functional and 6-31G(d) basis set on Gaussian 09.
By comparison of measured and calculated spectra, absolute stereochemistry of Ex. 228a and 228b, Ex. 229a and 229b, Ex. 225a and 225b compounds were assigned. Based on the absolute stereochemistry of Ex. 228b and Ex. 229b compound, the absolute stereochemistry of their precursor, Reference Example 226 compound was determined.
Powder X-Ray Diffraction
The powder X-ray diffraction was measured using RIGAKU RINT-TTRII diffractometer under the conditions of a tube: Cu, a tube current: 300 mA, a tube voltage, 50 kV, a sampling width: 0.02°, a scanning speed: 4°/minute, a wavelength: 1.54056 angstroms, and a measurement diffraction angle (2θ): 2.5 to 40°.
Furthermore, the term “about” in the characteristic peaks of powder X-ray diffraction shown at angles 2θ denotes ±0.2°, in another embodiment, ±0.1°. Each crystal can be characterized by a powder X-ray diffraction spectrum, but with the powder X-ray diffraction, crystal lattice intervals and overall patterns are important for identification of crystals in terms of the properties of the data, and since the relative intensity may vary slightly depending on the direction of crystal growth, the particle size, and the measurement conditions, it should not be strictly construed.
Formulations
The pharmaceutical composition containing one or two or more kinds of the compound represented by the formula (I) or salts thereof as an active ingredient can be prepared using excipients that are usually used in the art, that is, excipients for pharmaceutical preparation, carriers for pharmaceutical preparation, and the like.
A “pharmaceutically acceptable carrier” refers to an ingredient in a pharmaceutical composition, other than an active ingredient, which is nontoxic to a subject. A pharmaceutically acceptable carrier includes, but is not limited to, a buffer, excipient, stabilizer, or preservative.
Administration can be accomplished either by oral administration via tablets, pills, capsules, granules, powders, solutions, and the like, or parenteral administration, such as intraarticular, intravenous, or intramuscular injections, and the like, suppositories, ophthalmic solutions, eye ointments, transdermal liquid preparations, ointments, transdermal patches, transmucosal liquid preparations, transmucosal patches, inhalers, and the like.
The solid composition for use in the oral administration according to the present invention is used in the form of tablets, powders, granules, or the like. In such a solid composition, one or more active ingredient(s) are mixed with at least one inactive excipient. According to a conventional method, the composition may contain inactive additives, such as a lubricant such as magnesium stearate, a disintegrating agent such as sodium carboxymethyl starch and the like, a stabilizer, or a solubilization assisting agent. If necessary, tablets or pills may be coated with sugar or a film of a gastric or enteric coating substance.
The liquid composition for oral administration contains pharmaceutically acceptable emulsions, solutions, suspensions, syrups, elixirs, or the like, and also contains generally used inert diluents, for example, purified water or ethanol. In addition to the inert diluent, the liquid composition may also contain auxiliary agents, such as a solubilization assisting agent, a moistening agent, and a suspending agent, sweeteners, flavors, aromatics, and antiseptics.
The injections for parenteral administration include sterile aqueous or non-aqueous solution preparations, suspensions and emulsions. The aqueous solvent includes, for example, distilled water for injection and physiological saline. Examples of the non-aqueous solvent include alcohols such as ethanol. Such a composition may further contain a tonicity agent, an antiseptic, a moistening agent, an emulsifying agent, a dispersing agent, a stabilizing agent, or a solubilizing aid. These are sterilized, for example, by filtration through a bacteria retaining filter, blending of a bactericide, or irradiation. In addition, these can also be used by preparing a sterile solid composition, and dissolving or suspending it in sterile water or a sterile solvent for injection prior to its use.
The agent for external use includes ointments, plasters, creams, jellies, poultices, sprays, lotions, eye drops, eye ointments, and the like. The agents contain generally used ointment bases, lotion bases, aqueous or non-aqueous liquid preparations, suspensions, emulsions, and the like.
As the transmucosal agents such as an inhaler, a transnasal agent, and the like, those in the form of a solid, liquid, or semi-solid state are used, and can be prepared in accordance with a conventionally known method. For example, a known excipient, and also a pH adjusting agent, an antiseptic, a surfactant, a lubricant, a stabilizing agent, a thickening agent, or the like may be appropriately added thereto. For their administration, an appropriate device for inhalation or blowing can be used. For example, a compound may be administered alone or as a powder of formulated mixture, or as a solution or suspension in combination with a pharmaceutically acceptable carrier, using a conventionally known device or sprayer, such as a measured administration inhalation device, and the like. A dry powder inhaler or the like may be for single or multiple administration use, and a dry powder or a powder-containing capsule may be used. Alternatively, this may be in a form such as a pressurized aerosol spray which uses an appropriate ejection agent, for example, a suitable gas such as chlorofluoroalkane, hydrofluoroalkane, carbon dioxide, and the like, or other forms.
In oral administration, the daily dose is generally from about 0.001 to 100 mg/kg, preferably from 0.1 to 30 mg/kg, and more preferably 0.1 to 10 mg/kg, per body weight, administered in one portion or in 2 to 4 divided portions. In the case of intravenous administration, the daily dose is suitably administered from about 0.0001 to 10 mg/kg per body weight, once a day or two or more times a day. In addition, a transmucosal agent is administered at a dose from about 0.001 to 100 mg/kg per body weight, once a day or two or more times a day. The dose is appropriately decided in response to the individual case by taking the symptoms, the age, and the gender, and the like into consideration.
The compound of the formula (I) can be used in combination with various therapeutic or prophylactic agents for the diseases, in which the compound of the formula (I) is considered effective, as described above. The combined preparation may be administered simultaneously or separately and continuously, or at a desired time interval. The preparations to be co-administered may be a blend or prepared individually.
EXAMPLES
Hereinbelow, the preparation methods for the compound of the formula (I) will be described in more detail with reference to Examples. Further, the present invention is not limited to the preparation methods described in the specific Examples, Reference Examples and Preparation Examples as described below, but the compound of the formula (I) can be prepared by any combination of the preparation methods or the methods that are apparent to a skilled person in the art, particularly in view of the detailed teachings provided herein.
Furthermore, the following symbols are used in the Examples, Reference Examples, Preparation Examples, and Tables as described below.
Rf: Preparation Example Number,
RP: Reference Example Number,
Ex: Example Number,
No.: Compound No.,
Data: Physicochemical data,
ESI+: representing m/z values in ESI-MS (positive ions), and representing [M+H]+ peaks unless otherwise specified,
APCI/ESI+: m/z value in APCI/ESI-MS (positive ions), and representing [M+H]+ peaks unless otherwise specified,
EI: representing m/z values in EI-MS (positive ions), and representing [M]+ peaks unless otherwise specified,
CI+: representing m/z values in CI-MS (positive ions), and representing [M+H]+ peaks unless otherwise specified,
NMR-DMSO-d6: δ (ppm) in 1H-NMR in DMSO-d6,
NMR-CDCl3: δ (ppm) in 1H-NMR in CDCl3,
Structure: Structural formula (In case HCl is described in a structural formula, a compound represented by a structural formula forms a salt with HCl. Compounds having a double bond described by a cross line represents mixtures of a cis-compound and a trans-compound),
rel-: representing relative configuration,
Syn: Preparation method (in which E prefixed before the numeral shows that the compound is prepared by the similar preparation method as the compound having the Example Number, R prefixed before the numeral shows that the compound is prepared by the similar preparation method as the compound having the Preparation Example Number and RP prefixed before the numeral shows that the compound is prepared by the similar preparation method as the compound having the Reference Example Number),
Boc/BOC: tert-butoxycarbonyl,
CHCl3: chloroform,
CH2Cl2: dichloromethane,
CO2: carbon dioxide,
Cs2CO3: caesium carbonate,
CuBr: copper (I) bromide,
CuI: copper (I) iodide,
DAST: N,N-diethylaminosulfur trifluoride,
DBU: 1,8-diazabicyclo[5.4.0]undec-7-ene,
DIBAL-H: diisobutylaluminium hydride,
DMAP: N,N-dimethyl-4-aminopyridine,
DMF: N,N-dimethylformamide,
DMSO: dimethyl sulfoxide,
Et3N: triethylamine,
AcOEt/EtOAc: ethyl acetate,
EtOH: ethanol,
Et2O: diethyl ether,
HCOOH: formic acid,
HCl: hydrogen chloride,
H2O: hydrogen oxide,
HPLC: high performance liquid chromatography,
IPE, iPr2O: diisopropyl ether,
K2CO3: potassium carbonate,
K3PO4: potassium phosphate,
LiBH4: lithium borohydride,
MeCN: acetonitrile,
MsCl: methanesulfonyl chloride,
MeMgBr: methylmagnesium bromide,
MeOH: methanol,
MgSO4: anhydrous magnesium sulfate,
n-BuLi: n-butyllithium,
NMP: 1-methyl-2-pyrrolidone,
NaOH: sodium hydroxide,
NaHCO3: sodium hydrogen carbonate,
Na2CO3: sodium carbonate,
Na2S2O3: sodium thio sulfate,
Na2SO4: anhydrous sodium sulfate,
Na2SO4′ 10H2O: sodium sulfate decahydrate,
NH4Cl: ammonium chloride,
PdCl2(dppf): [1,1′-Bis(diphenylphosphino)ferrocene]palladium(II) dichloride,
Pd(OAc)2: palladium(II) acetate,
Pd(PPh3)4: tetrakis(triphenylphosphine)palladium(0),
PdCl2(PPh3)2: bis(triphenylphosphine)palladium (II) chloride,
PPh3: triphenylphosphine,
PtO2: platinum (IV) oxide,
SiO2: silicon dioxide,
THF: tetrahydrofuran,
TsOH.H2O: p-toluenesulfonic acid monohydrate,
TMSOTf: Trimethylsilyl trifluoromethanesulfonate.
Preparation Example 1
To a mixture of 6-bromo-4-methylene-4H-spiro[chromene-3,3′-oxetane] (351 mg, 1.31 mmol), silver cyanate (295 mg, 1.97 mmol), EtOAc (1.7 mL), and MeCN (3.5 mL) was added a mixture of iodine (500 mg, 1.97 mmol) and EtOAc (5.3 mL) in an ice-water bath. After stirring for 1.5 hours at the same temperature, the mixture was stirred for 30 minutes at ambient temperature. The mixture was filtered through celite pad (washed with EtOAc), and the filtrate was washed with saturated aqueous Na2S2O3 and brine, dried over MgSO4 and filtered. After concentration of the filtrate at reduced pressure, tert-butyl alcohol (4.4 mL) and triethylamine (0.183 mL, 1.31 mmol) were added to the residue, and the mixture was stirred overnight under reflux. The reaction mixture was cooled down to ambient temperature, concentrated at reduced pressure to give crude 6′-bromo-2H-dispiro[1,3-oxazolidine-4,4′-chromene-3′,3′′-oxetan]-2-one.
Preparation Example 2
A mixture of di-tert-butyl[6′-(3-methoxyprop-1-yn-1-yl)dispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-yl]imidodicarbonate (34.7 mg, 0.067 mmol) and 10% palladium on carbon (7 mg) in EtOH (1.4 mL) was stirred for 13 hours under a hydrogen atmosphere (4.5 kgf/cm2). The mixture was filtered off, and the filtrate was evaporated to give crude di-tert-butyl [6′-(3-methoxypropyl)dispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-yl]imidodicarbonate (34.7 mg).
Preparation Example 8
To a solution of 6′-bromo-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-amine (643 mg, 1.82 mmol) in THF (12.9 mL) were added 4-dimethylaminopyridine (11 mg, 0.091 mmol) and di-tert-butyl dicarbonate (1.19 g, 5.46 mmol). The mixture was stirred overnight at ambient temperature. The reaction mixture was evaporated off at reduced pressure. The residue was purified with column chromatography on silica gel (hexane-EtOAc, a linear gradient of EtOAc from 0 to 20%) to give di-tert-butyl (6′-bromo-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-yl)imidodicarbonate (890 mg).
Preparation Example 22
The mixture of 6′-bromodispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-2′′-amine (1.1 g, 3.4 mmol), di-tert-butyl dicarbonate (2.2 g, 10 mmol), and N,N-dimethylpyridin-4-amine (21 mg, 0.17 mmol) in THF (21 ml) was stirred for 3 hours at ambient temperature and for 5 hours at 50° C. The mixture was concentrated in vacuo. The residue was purified by silica gel chromatography (hexane:EtOAc=100:0-80:20) to give di-tert-butyl (6′-bromodispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-2′′-yl)imidodicarbonate (1.5 g).
Preparation Example 23
The mixture of 6′-bromodispiro[cyclobutane-1,3′-chromene-4′,4′′-[1,3]oxazol]-2′′-amine (185 mg, 0.57 mmol), di-tert-butyl dicarbonate (374 mg, 1.7 mmol), N,N-dimethylpyridin-4-amine (3.4 mg, 0.029 mmol), and N,N-diethylethanamine (173 mg, 1.7 mmol) in THF (20 mL) was stirred overnight at ambient temperature. The mixture was concentrated in vacuo, and the residue was purified by silica gel chromatography (hexane:EtOAc=100:0-80:20) to give tert-butyl (6′-bromodispiro[cyclobutane-1,3′-chromene-4′,4′′-[1,3]oxazol]-2′′-yl)carbamate (223 mg).
Preparation Example 24
To a solution of 6′-bromodispiro[oxetane-3,3′-chromene-4′,4′′-[1,3]thiazol]-2′′-amine (214 mg, 0.627 mmol) in THF (2.1 mL) were added di-tert-butyl dicarbonate (411 mg, 1.88 mmol) and 4-dimethylaminopyridine (3.8 mg, 0.031 mmol). The mixture was stirred overnight at ambient temperature, and di-tert-butyl dicarbonate (68.4 mg, 0.314 mmol) was added to the reaction mixture. After stirring for 2 hours at ambient temperature, the mixture was partitioned between EtOAc and 10 wt. % aqueous citric acid. The organic layer was washed with brine, dried over MgSO4 and silica gel and filtered. The filtrate was evaporated off, and purification of the residue with column chromatography on silica gel (Hexane-EtOAc, a linear gradient of EtOAc from 0 to 50%) afforded di-tert-butyl (6′-bromodispiro[oxetane-3,3′-chromene-4′,4′′-[1,3]thiazol]-2′′-yl)imidodicarbonate (279 mg).
Preparation Example 26
The mixture of 4-bromo-2-iodophenol (3.30 g, 11.04 mmol), 1-bromo-5-chloropentan-2-one (75% purity, 3.9 g, 14.66 mmol), and K2CO3 (2.3 g, 16.64 mmol) in acetone (66 mL) was stirred for 48 hours at ambient temperature. The insoluble material was removed by filtration and washed with EtOAc. The filtrate was evaporated in vacuo. The residue was purified by silicagel column chromatography (EtOAc-hexane, a linear gradient of EtOAc from 0 to 25%) afforded 1-(4-bromo-2-iodophenoxy)-5-chloropentan-2-one (2.12 g).
Preparation Example 27
A solution of potassium tert-butoxide (441 mg, 3.93 mmol) in THF (5 mL) was added to a suspension of 6-bromospiro[chromene-2,1′-cyclobutan]-4(3H)-one (500 mg, 1.87 mmol) and 1H-benzotriazole-1-methanol (586 mg, 3.93 mmol) in THF (5 mL) over 10 minutes in a dry ice-acetone bath under an argon atmosphere. The mixture was stirred for 0.5 hours in an ice bath, and then diluted with EtOAc (10 mL). After stirring for 0.5 hours, the mixture was filtered off. The filtrate was washed with 0.2 M aqueous NaOH (two times), water and brine, dried over MgSO4 and silicagel, filtered off. The filtrate was evaporated to give crude 6-bromo-3,3-bis(hydroxymethyl)spiro[chromene-2,1′-cyclobutan]-4(3H)-one (641 mg).
Preparation Example 29
A mixture of di-tert-butyl (6′-bromodispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-yl)imidodicarbonate (300 mg, 0.571 mmol), 3-methoxypyridin-2-amine (354 mg, 2.86 mmol), tris(dibenzylideneacetone)dipalladium(0) (105 mg, 0.114 mmol), (9,9-dimethyl-9H-xanthene-4,5-diyl)bis(diphenylphosphine) (198 mg, 0.343 mmol), Cs2CO3 (558 mg, 1.71 mmol) and dioxane (15 mL) was stirred for 48 hours at 100° C. The reaction mixture was cooled down to ambient temperature, and partitioned with CHCl3 and water. The organic layer was dried over Na2SO4, and filtered. The filtrate was concentrated at reduced pressure, to give crude di-tert-butyl {6′-[(3-methoxypyridin-2-yl)amino]dispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-yl}imidodicarbonate, which was used for the next reaction without further purification.
Preparation Example 30
To a mixture of (4-amino-6-bromo-4H-spiro[chromene-3,3′-oxetan]-4-yl)methanol (280 mg, 0.933 mmol), CH2Cl2 (10 mL) and saturated aqueous NaHCO3 (10 mL) was added a mixture of chloroacetyl chloride (0.083 mL, 1.02 mmol) and CH2Cl2 (1 mL) at ambient temperature. After stirring for 30 minutes at ambient temperature, chloroacetyl chloride (0.016 mL, 0.197 mmol) was added to the reaction mixture. The mixture was stirred for 10 minutes at ambient temperature, diluted with CH2Cl2 and separated. The organic layer was washed with water, dried over MgSO4 and filtered. Concentration of the filtrate at reduced pressure gave crude N-[6-bromo-4-(hydroxymethyl)-4H-spiro[chromene-3,3′-oxetan]-4-yl]-2-chloroacetamide, which was used for the next reaction without further purification.
Preparation Example 31
To a mixture of crude N-[6-bromo-4-(hydroxymethyl)-4H-spiro[chromene-3,3′-oxetan]-4-yl]-2-chloroacetamide (351 mg, 0.933 mmol) and 2-methylbutan-2-ol (6.3 mL) was added potassium tert-butoxide (356 mg, 3.17 mmol) at ambient temperature, and the mixture was stirred for 1 hour at the same temperature. MeOH (3.2 mL) was added to the reaction mixture, and the mixture was concentrated at reduced pressure. Purification of the residue with column chromatography on silica gel (Hexane-EtOAc, a linear gradient of EtOAc from 50 to 100%) afforded 6′-bromo-5H-dispiro[1,4-oxazinane-3,4′-chromene-3′,3′′-oxetan]-5-one (277 mg).
Preparation Example 36
To a suspension of methyl(triphenyl)phosphonium bromide (8.13 g, 22.3 mmol) in THF (44 mL) was added n-butyllithium (1.65 M in n-hexane, 13.5 mL, 22.3 mmol) in a dry ice-acetone bath under argon atmosphere. The mixture was stirred for 60 minutes in an ice bath. To the mixture was added a mixture of 6-bromo-2,2-dimethyl-4H-spiro[chromene-3,3′-oxetan]-4-one (2.21 g, 7.44 mmol) and THF (11 mL) in an ice bath. The mixture was stirred for 1 hour at ambient temperature. The reaction was quenched by adding water in an ice-water bath. The mixture was partitioned between EtOAc-hexane (1:2) and water. The organic layer was washed with brine, dried over MgSO4, filtered, and the filtrate was evaporated. Purification using silica gel column chromatography (EtOAc-hexane, a linear gradient of EtOAc from 0 to 20%) afforded 6-bromo-2,2-dimethyl-4-methylene-4H-spiro[chromene-3,3′-oxetane] (2.02 g).
Preparation Example 48
To a mixture of 6-bromo-2,2-dimethyl-2,3-dihydro-4H-chromen-4-one (1.00 g, 3.92 mmol) and dioxane (10 mL) were added formaldehyde (37 wt. % in water, 2.95 mL, 39.2 mmol) and Na2CO3 (831 mg, 7.84 mmol) at room temperature. After stirring overnight at the same temperature, the reaction mixture was filtered. The filtrate was diluted with CHCl3, washed with 1M aqueous HCl and water, dried over MgSO4 and filtered. The filtrate was concentrated at reduced pressure, and the residue was purified with column chromatography on silica gel (hexane-EtOAc, a linear gradient of EtOAc from 0 to 50%) to afford 6-bromo-3,3-bis(hydroxymethyl)-2,2-dimethyl-2,3-dihydro-4H-chromen-4-one (1.07 g).
Preparation Example 58
To a solution of 1-(4-bromo-2-iodophenoxy)-5-chloropentan-2-one (1.61 g, 3.86 mmol) in THF (35 mL) was added vinylmagnesium bromide (1M solution in THF, 4.3 mL) at −78° C. under an argon atmosphere. After stirring for 1 hour at −78° C., the mixture was gradually warmed up to −30° C. over 1 hour. The reaction was quenched by adding saturated aqueous NH4Cl solution and extracted with EtOAc. The combined organic layer were washed with brine, dried over Na2SO4, filtered, and the filtrate was evaporated. The residue was purified by silicagel column chromatography (EtOAc-hexane, a linear gradient of EtOAc from 0 to 20%) afforded 3-[(4-bromo-2-iodophenoxy)methyl]-6-chlorohex-1-en-3-ol (1.35 g).
Preparation Example 59
The mixture of 3-[(4-bromo-2-iodophenoxy)methyl]-6-chlorohex-1-en-3-ol (1.53 g, 3.43 mmol), Pd(OAc)2 (77 mg, 0.343 mmol), PPh3 (360 mg, 1.37 mmol), and K2CO3 (2.84 g, 20.55 mmol) in MeCN (45 mL) was heated at 85° C. for 18 hours under an argon atmosphere. After addition of Pd(OAc)2 (23 mg, 0.102 mmol) and PPh3 (108 mg, 0.412 mmol), the mixture was heated at 85° C. for 30 hours. The reaction mixture was cooled to room temperature, and the insoluble material was removed by filtration and washed with EtOAc. The filtrate was evaporated in vacuo. The residue was purified by silicagel column chromatography (EtOAc-hexane, a linear gradient of EtOAc from 0 to 15%) to afford a mixture of 6-bromo-4-methylene-4′,5′-dihydro-3′H,4H-spiro[chromene-3,2′-furan] and a reaction intermediate (598 mg). To the mixture dissolved in MeCN (30 mL) were added Pd(OAc)2 (23 mg, 0.102 mmol), PPh3 (108 mg, 0.412 mmol), and K2CO3 (947 mg, 6.85 mmol) and the reaction mixture was heated at 85° C. for 2 hours under an argon atmosphere. The reaction mixture was cooled to room temperature, and the insoluble material was removed by filtration and washed with AcOEt. The filtrate was evaporated in vacuo. The residue was purified by silicagel column chromatography (EtOAc-hexane, a linear gradient of EtOAc from 0 to 15%) to afford 6-bromo-4-methylene-4′,5′-dihydro-3′H,4H-spiro[chromene-3,2′-furan] (460 mg).
Preparation Example 60
To a mixture of 6′-bromo-5H-dispiro[1,4-oxazinane-3,4′-chromene-3′,3′′-oxetan]-5-one (275 mg, 0.808 mmol) and dioxane (11 mL) was added 2,4-bis(4-methoxyphenyl)-1,3,2,4-dithiadiphosphetane 2,4-disulfide (236 mg, 0.566 mmol) at ambient temperature. After stirring for 2 hours at 80° C., the reaction mixture was cooled down to ambient temperature and concentrated at reduced pressure. The residue was purified with column chromatography on silica gel (hexane-EtOAc, a linear gradient of EtOAc from 0 to 50%) to afford 6′-bromo-5H-dispiro[1,4-oxazinane-3,4′-chromene-3′,3′′-oxetane]-5-thione (259 mg).
Preparation Example 62
To a solution of ethyl 1-(hydroxymethyl)cyclobutanecarboxylate (1.0 g, 6.3 mmol) and N,N-diethylethanamine (1.5 g, 8.2 mmol) in CH2Cl2 (30 ml) was added methanesulfonyl chloride (869 mg, 7.6 mmol). The mixture was stirred for 6 hours at ambient temperature. After dilution with CHCl3 and H2O, the organic layer was washed with H2O, dried over MgSO4, and concentrated in vacuo to give ethyl 1-{[(methylsulfonyl)oxy]methyl}cyclobutanecarboxylate (1.2 g).
Preparation Example 63
To a mixture of 6′-bromo-2H-dispiro[1,3-oxazolidine-4,4′-chromene-3′,3′′-oxetan]-2-one (1.98 g, 6.07 mmol), EtOH (9.9 mL) and water (50 mL) was added lithium hydroxide monohydrate (2.68 g, 60.7 mmol), and the mixture was stirred overnight at 100° C. The reaction mixture was cooled down to ambient temperature and extracted with CH2Cl2. The organic layer was washed with water, dried over Na2SO4 and filtered. After concentration of the filtrate at reduced pressure, the residue was triturated with hexane, collected by filtration, washed with EtOAc-hexane (1:3) and dried at reduced pressure to afford (4-amino-6-bromo-4H-spiro[chromene-3,3′-oxetan]-4-yl)methanol (1.40 g).
Preparation Example 64
To a solution of di-tert-butyl (6′-bromodispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-yl)imidodicarbonate (300 mg, 0.571 mmol), 4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi-1,3,2-dioxaborolane (174 mg, 0.685 mmol), and PdCl2(dppf) (21 mg, 0.029 mmol) in dioxane (6 mL) was added potassium acetate (112 mg, 1.14 mmol). The mixture was stirred for 3 hours at 110° C. The resulting precipitate was removed by filtration and the filtrate was evaporated. Silicagel column chromatography (MeOH—CHCl3, a linear gradient of MeOH from 3 to 10%) afforded di-tert-butyl[6′-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)dispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-yl]imidodicarbonate (305 mg).
Preparation Example 66
To a mixture of di-tert-butyl (6′-bromodispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-2′′-yl)imidodicarbonate (200 mg, 0.393 mmol), bis(dibenzylideneacetone)palladium (0) (22.6 mg, 0.039 mmol), and tri-tert-butylphosphonium tetrafluoroborate (11.6 mg, 0.039 mmol) was added lithium bis(trimethylsilyl)amide (1 M in toluene, 1.96 mL, 1.96 mmol) at ambient temperature. After stirring for 1 hour at 100° C., the mixture was cooled down to ambient temperature. To the mixture were added 1M aqueous HCl (1.96 mL) and MeOH (1.96 mL) at ambient temperature, and the mixture was stirred for 30 minutes at the same temperature. The mixture was extracted with CHCl3, and the organic layer was dried over Na2SO4 and filtered. The filtrate was concentrated at reduced pressure and purified with column chromatography on silica gel (hexane-EtOAc, a linear gradient of EtOAc from 0 to 90%) to afford tert-butyl (6′-aminodispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-2′′-yl)carbamate (103 mg).
Preparation Example 67
Under argon atmosphere, to a mixture of di-tert-butyl (6′-bromo-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-yl)imidodicarbonate (1.00 g, 1.81 mmol), bis(dibenzylideneacetone)palladium (0) (104 mg, 0.181 mmol) and tri-tert-butylphosphonium tetrafluoroborate (53.5 mg, 0.181 mmol) was added lithium bis(trimethylsilyl)amide (1.0 M in toluene, 9.03 mL, 9.03 mmol) at ambient temperature. After stirring for 1 hour at 100° C., the reaction mixture was cooled down to ambient temperature. To the mixture were added 1.0 M hydrochloric acid (9.0 mL) and MeOH (9.0 mL) and the mixture was stirred for 30 minutes at ambient temperature. After extraction of the mixture with CHCl3, the organic layer was dried over Na2SO4 and filtered. The filtrate was concentrated at reduced pressure and the residue was purified by column chromatography on silica gel (hexane-EtOAc, a linear gradient of EtOAc from 0 to 90%) to give tert-butyl (6′-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-yl)carbamate (616 mg).
Preparation Example 69
Under argon atmosphere, a mixture of 4,6-dichloropyrimidine (500 mg, 3.36 mmol), 1-(trimethylsilyl)-1-propyne (0.497 mL, 3.36 mmol), tetrabutylammonium fluoride (1 M in THF, 3.36 mL, 3.36 mmol), triethylamine (1.54 mL, 11.1 mmol), tetrakis(triphenylphosphine)palladium(0) (194 mg, 0.168 mmol), copper(I) iodide (192 mg, 1.01 mmol) and toluene (20 mL) was stirred for 9 hours at 60° C. The mixture was cooled down to ambient temperature, and water was added to the mixture. The mixture was extracted with CHCl3, and the organic layer was dried over MgSO4 and filtered. The filtrate was concentrated at reduced pressure, and purification of the residue with column chromatography on silica gel (hexane-EtOAc, a linear gradient of EtOAc from 0 to 20%) afforded 4-chloro-6-(prop-1-yn-1-yl)pyrimidine (207 mg).
Preparation Example 70
A mixture of tert-butyl (6′-aminodispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-2′′-yl)carbamate (115 mg, 0.333 mmol), 5-fluoropyridine-2-carboxylic acid (62.3 mg, 0.433 mmol), N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (83.0 mg, 0.433 mmol), 1-hydroxybenzotriazole (58.5 mg, 0.433 mmol), N,N-diisopropylethylamine (0.074 mL, 0.433 mmol) and CH2Cl2 (1.2 mL) was stirred for 2.5 days at ambient temperature. The reaction mixture was purified with column chromatography on silica gel (hexane-EtOAc, a linear gradient of EtOAc from 10 to 90%) to afford tert-butyl (6′-{[(5-fluoropyridin-2-yl)carbonyl]amino}dispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-2′′-yl)carbamate (136 mg).
Preparation Example 73 
A mixture of tert-butyl (6′-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-yl)carbamate (878 mg, 2.25 mmol), 5-chloro-2-pyridinecarboxylic acid (476 mg, 2.93 mmol), N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (562 mg, 2.93 mmol), 1-hydroxybenzotriazole (396 mg, 2.93 mmol), N,N-diisopropylethylamine (0.502 mL, 2.93 mmol) and CH2Cl2 (8.78 mL) was stirred for 1.5 hours at ambient temperature. After concentration of the reaction mixture at reduced pressure, the residue was purified with column chromatography on silica gel (hexane-EtOAc, a linear gradient of EtOAc from 0 to 90%) to give tert-butyl (6′-{[(5-chloropyridin-2-yl)carbonyl]amino}-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-yl)carbamate (976 mg).
Preparation Example 76
A mixture of di-tert-butyl (6′-bromodispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-yl)imidodicarbonate (150 mg, 0.286 mmol) and 3-methoxyprop-1-yne (0.072 mL, 0.86 mmol) in Et3N (1.5 mL) was purged with argon. To the mixture was added Pd(PPh3)4 (13 mg, 0.011 mmol) and CuBr (4.9 mg, 0.034 mmol), and the mixture was refluxed for 3 hours under an argon atmosphere. The mixture was partitioned between CHCl3 and brine, and filtered through celite. The organic layer of the filtrate was dried over MgSO4, filtered off, and the filtrate was evaporated. Silicagel column chromatography (EtOAc-hexane, a linear gradient of EtOAc from 0 to 25%) afforded di-tert-butyl[6′-(3-methoxyprop-1-yn-1-yl)dispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-yl]imidodicarbonate (34.7 mg).
Preparation Example 78
The mixture of 6-bromo-4H-spiro[chromene-3,1′-cyclopropan]-4-one (8.0 g, 32 mmol), 2-methylpropane-2-sulfinamide (12 g, 99 mmol), and titanium(IV) tetraethanolate (22 g, 95 mmol) in THF (160 ml) was stirred for 48 hours at 80° C. To the mixture was added H2O (20 ml), filtered through Celite and washed by EtOAc (50 ml). The filtrate was concentrated in vacuo. The residue was purified by silica gel chromatography (hexane:EtOAc=100:0-0:100) to give N-(6-bromo-4H-spiro[chromene-3,1′-cyclopropan]-4-ylidene)-2-methylpropane-2-sulfinamide (6.5 g).
Preparation Example 83
A mixture of di-tert-butyl (6′-bromodispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-yl)imidodicarbonate (300 mg, 0.571 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydro-2H-pyran (360 mg, 1.71 mmol), bis(triphenylphosphine)palladium(II) dichloride (40.1 mg, 0.057 mmol) and Na2CO3 (182 mg, 1.71 mmol) in dioxane (3.6 mL) and water (0.9 mL) was stirred for 1 hour at 100° C. The mixture was cooled down to ambient temperature and partitioned between EtOAc and water. The organic layer was washed with brine, dried over MgSO4, diluted with hexane, and filtered through silica gel pad (eluted with 50% EtOAc in hexane). The filtrate was concentrated at reduced pressure to give crude di-tert-butyl[6′-(3,6-dihydro-2H-pyran-4-yl)dispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-yl]imidodicarbonate which was used for the next reaction without further purification.
Preparation Example 85
Under argon atmosphere, a mixture of di-tert-butyl[6′-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)dispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-yl]imidodicarbonate (213 mg, 0.372 mmol), 3-bromo-5-(3-methoxyprop-1-yn-1-yl)pyridine (252 mg, 1.12 mmol), Na2CO3 (158 mg, 1.49 mmol), tetrakis(triphenylphosphine)palladium(0) (21.5 mg, 0.019 mmol), dioxane (3.4 mL) and water (0.85 mL) was stirred for 3 hours at 110° C. The reaction mixture was cooled down to ambient temperature, and water was added to the mixture. The mixture was extracted with MeOH—CHCl3 (1:9), and the organic layer was dried over Na2SO4 prior to filtration. The filtrate was concentrated at reduced pressure to give crude di-tert-butyl {6′-[5-(3-methoxyprop-1-yn-1-yl)pyridin-3-yl]dispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-yl}imidodicarbonate, which was used for the next reaction without further purification.
Preparation Example 91
A mixture of di-tert-butyl[6′-(5-bromopyridin-3-yl)dispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-yl]imidodicarbonate (232 mg, 0.385 mmol), ethynyl(trimethyl)silane (0.160 mL, 1.16 mmol), bis(triphenylphosphine)palladium(II) dichloride (13.5 mg, 0.019 mmol), copper(I) iodide (7.3 mg, 0.039 mmol) and triethylamine (3.2 mL) was stirred overnight at ambient temperature and for 6 days at 50° C. Ethynyl(triisopropyl)silane (0.257 mL, 1.16 mmol) was added to the reaction mixture at ambient temperature, and the mixture was stirred overnight at 85° C. The mixture was cooled down to ambient temperature, diluted with EtOAc and washed with saturated aqueous NH4Cl. The organic layer was dried over MgSO4 and filtered. The filtrate was concentrated at reduced pressure.
To a mixture of the residue and THF (4.6 mL) was added tetrabutylammonium fluoride (1M in THF, 1.54 mL, 1.54 mmol), and the mixture was stirred overnight at ambient temperature. The mixture was partitioned with EtOAc and saturated aqueous NH4Cl, and the organic layer was washed with brine, dried over anhydrous MgSO4 and filtered. The filtrate was concentrated at reduced pressure, and purification of the residue with column chromatography on silica gel (hexane-EtOAc, a linear gradient of EtOAc from 0 to 50%) afforded di-tert-butyl[6′-(5-ethynylpyridin-3-yl)dispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-yl]imidodicarbonate (59.9 mg).
Preparation Example 92
To a solution of 6-bromo-4H-chromen-4-one (12 g, 53 mmol) in CH2Cl2 (24 mL) was added TMSOTf (12.5 mL, 69.18 mmol) at ambient temperature. After stirring for 1 hour, THF (210 mL) was added to the mixture at ambient temperature and cooled to −78° C. To the mixture was added n-propylmagnesium bromide (1.05M solution in THF, 66 mL, 69 mmol). After stirring for 1 hour at −78° C., 1M aqueous NH4Cl was added to the mixture. The mixture was warmed to ambient temperature and stirred overnight. The organic and the aqueous layers were separated, and the organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated in vacuo. The residue was purified by silicagel column chromatography (EtOAc-hexane, a linear gradient of EtOAc from 0 to 10%) to afford 6-bromo-2-propyl-2,3-dihydro-4H-chromen-4-one (11.42 g).
Preparation Example 94
A mixture of 4-(methoxymethyl)-1H-pyrazole (145 mg, 1.29 mmol), 6′-bromo-4′H-dispiro[cyclobutane-1,2′-chromene-3′,3′′-oxetan]-4′-one (200 mg, 0.647 mmol), trans-N,N′-dimethylcyclohexane-1,2-diamine (74 mg, 0.52 mmol) and K2CO3 (268 mg, 1.94 mmol) in NMP (2 mL) was purged with argon. To the mixture was added CuI (49 mg, 0.26 mmol), and the mixture was sealed and stirred for 1 hour at 150° C. and 0.5 hours at 170° C. under a microwave irradiation. The mixture was partitioned between EtOAc and saturated aqueous NH4Cl. The organic layer was washed with water (two times) and brine, dried over MgSO4, filtered, and the filtrate was evaporated. Silicagel column chromatography (EtOAc-hexane, a linear gradient of EtOAc from 0 to 45%) afforded 6′-[4-(methoxymethyl)-1H-pyrazol-1-yl]-4′H-dispiro[cyclobutane-1,2′-chromene-3′,3′′-oxetan]-4′-one (97 mg).
Preparation Example 96
To a mixture of 6-bromo-3,3-bis(hydroxymethyl)-2,2-dimethyl-2,3-dihydro-4H-chromen-4-one (6.76 g, 21.4 mmol), zinc bis(dimethyldithiocarbamate) (26.2 g, 85.8 mmol) and triphenylphosphine (8.44 g, 32.2 mmol) in THF (0.20 L) was added diisopropyl azodicarboxylate (1.9 M solution in toluene, 16.9 mL, 32.2 mmol) in an ice-water bath. The mixture was stirred overnight at ambient temperature. The mixture was diluted with toluene (0.20 L), and the mixture was filtered off. The filtrate was washed with 1 M aqueous NaOH (three times), water and brine, dried over MgSO4 and filtered. The filtrate was evaporated to give a crude product. The crude product was purified with column chromatography on silica gel (Hexane-EtOAc, a linear gradient of EtOAc from 0 to 20%) to afford 6-bromo-2,2-dimethyl-4H-spiro[chromene-3,3′-oxetan]-4-one (2.22 g).
Preparation Example 106
To a solution of ethyl 1-[(4-bromophenoxy)methyl]cyclobutanecarboxylate (1.1 g, 3.5 mmol) in EtOH (11 ml) was added 1M aqueous NaOH (11 ml, 11 mmol). The mixture was stirred for 7 hours at 60° C. The mixture was concentrated in vacuo, and to the solution was added 1M aqueous HCl.
The resulting precipitate was collected by filtration, washed with H2O and dried in vacuo to give crude 1-[(4-bromophenoxy)methyl]cyclobutanecarboxylic acid (0.91 g).
Preparation Example 108
To a mixture of 1-(5-bromo-2-hydroxyphenyl)ethanone (10.0 g, 46.5 mmol) and MeOH (0.20 L) were added 3-methylbutanal (7.54 mL, 69.8 mmol) and pyrrolidine (5.77 mL, 69.8 mmol) at ambient temperature, and the mixture was stirred for 3 days at the same temperature. After concentration of the reaction mixture at reduced pressure, the residue was diluted with EtOAc, acidified to pH 3-4 with 1M aqueous HCl, and extracted with EtOAc. The organic layer was washed with water and brine, dried over MgSO4, and filtered. After concentration of the filtrate at reduced pressure, purification of the residue with column chromatography on silica gel (EtOAc-hexane, a linear gradient of EtOAc from 0 to 10%) afforded 6-bromo-2-isobutyl-2,3-dihydro-4H-chromen-4-one (9.02 g).
Preparation Example 111
The mixture of 1-(5-bromo-2-hydroxyphenyl)ethanone (10 g, 46.50 mmol), propionaldehyde (6.7 mL, 93 mmol), pyrrolidine (3.9 mL, 47 mmol), and acetic acid (3.2 mL, 56 mmol) in toluene (20 mL) was heated to 60° C. for 18 hours. After cooling to room temperature, the mixture was concentrated in vacuo. The mixture was diluted with diethyl ether and 1M aqueous HCl. The phases were separated. The organic phase was washed with 1M aqueous NaOH, then brine. The organic phase was dried over Na2SO4, filtered, and concentrated in vacuo. Purification with silicagel column chromatography (EtOAc-hexane, a linear gradient of EtOAc from 1 to 10%) afforded 6-bromo-2-ethyl-2,3-dihydro-4H-chromen-4-one (3.92 g).
Preparation Example 112
The mixture of ethyl 1-{[(methylsulfonyl)oxy]methyl}cyclobutanecarboxylate (1.50 g, 6.3 mmol), 4-bromophenol (1.2 g, 7.0 mmol), and caesium carbonate (4.13 g, 7.0 mmol) in DMF (15 ml) was stirred for 6 hours at 135° C. After dilution with EtOAc and H2O, the organic layer was washed with H2O, dried over MgSO4, and concentrated in vacuo. The residue was purified by silica gel chromatography (hexane:EtOAc=100:0-70:30) to give ethyl 1-[(4-bromophenoxy)methyl]cyclobutanecarboxylate (919 mg).
Preparation Example 114
To sulfuric acid (5 mL) was added 1-[(4-bromophenoxy)methyl]cyclobutanecarboxylic acid (1.9 g, 6.6 mmol) at 0° C. in an ice bath. After stirring for 1 hour at room temperature, ice was added to the mixture portionwise. The mixture was diluted with water, and extracted with EtOAc. The organic layer was washed with brine, dried over MgSO4, and concentrated in vacuo. The residue was purified by silica gel chromatography (hexane:EtOAc=100:0-70:30) to give 6-bromo-4H-spiro[chromene-3,1′-cyclobutan]-4-one (649 mg).
Preparation Example 116
To a stirred solution of 3,5-dibromopyridine (251 mg, 1.061 mmol) and di-tert-butyl (6′-bromotrispiro[cyclobutane-1,2′-chromene-4′,4′′-[1,3]oxazole-3′,3′′′-oxetan]-2′′-yl)imidodicarbona to (200 mg, 0.354 mmol) in dioxane (1.6 ml) were added 4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi-1,3,2-dioxaborolane (93 mg, 0.364 mmol), potassium acetate (69 mg, 0.707 mmol) and PdCl2(PPh3)2 (50 mg, 0.071 mmol) at room temperature and the mixture was stirred at 100° C. for 8 hours before the starting molecule was completely consumed to give the corresponding boronate intermediate. To this mixture was added Na2CO3 (150 mg, 1.42 mmol) and H2O (400 μl) and the mixture was stirred at 100° C. for 6 hours before the boronate intermediate was completely consumed. The mixture was cooled to room temperature and evaporated to give a crude, which was purified with column chromatography (EtOAc in hexane=0 to 50%) to give di-tert-butyl[6′-(5-bromopyridin-3-yl)trispiro[cyclobutane-1,2′-chromene-4′,4′′-[1,3]oxazole-3′,3′′′-oxetan]-2′′-yl]imidodicarbonate (48 mg).
Preparation Example 117
The mixture of 4-bromo-1-[(3,3-dimethoxy-1-vinylcyclobutyl)methoxy]-2-iodobenzene (3.16 g, 6.98 mmol) and 1M aqueous HCl (14 mL) in THF (31 mL) was stirred for ambient temperature for 1 hour. Then the mixture was stirred for 4 hours at 50° C. The mixture was cooled to room temperature and added saturated aqueous NaHCO3 and extracted with EtOAc. The combined organic extracts were washed with brine, dried over Na2SO4, filtered, and evaporated under reduced pressure to give 3-[(4-bromo-2-iodophenoxy)methyl]-3-vinylcyclobutanone (2.91 g).
Preparation Example 119
To a solution of diisopropylamine (3.2 mL, 22.67 mmol) in THF (45 mL) was added n-BuLi (2.69 M in hexane, 7.7 mL, 20.71 mmol) at −78° C. under argon. The mixture was stirred for 10 minutes at 0° C., then cooled to −78° C. and added a solution of methyl 3,3-dimethoxycyclobutanecarboxylate (3.0 g, 17.22 mmol) in THF (10 mL). The mixture was stirred for 30 minutes at −78° C., then added a solution of acetaldehyde (1.9 mL, 33.86 mmol) in THF (10 mL). The mixture was stirred for 30 minutes at −78° C., and water was added. The aqueous layer was extracted with EtOAc. Combined organic layers were washed with brine, dried over Na2SO4, filtered, and evaporated under reduced pressure to give methyl 1-(1-hydroxyethyl)-3,3-dimethoxycyclobutanecarboxylate (3.33 g).
Preparation Example 128
Under ice cooling, to a solution of 2-(4-amino-6-bromo-4H-spiro[chromene-3,1′-cyclopropan]-4-yl)-2,2-difluoroethanol (2.24 g, 6.46 mmol) in acetone (45 mL) was added benzoyl isothiocyanate (1.16 g, 7.10 mmol), and the mixture was stirred for 2 hours at room temperature and stirred for 13 hours at 40° C. After concentration, the residue was purified by silica gel chromatography (EtOAc/hexane=1:99-30:70) followed by purification using silica gel chromatography (NH-silicagel, EtOH/CHCl3=0:100-10:90) to give N-{[6-bromo-4-(1,1-difluoro-2-hydroxyethyl)-4H-spiro[chromene-3,1′-cyclopropan]-4-yl]carbamothioyl}benzamide (684 mg).
Preparation Example 131
To an ice chilled solution of 6-bromo-4-methylene-3′H,4H-spiro[chromene-3,1′-cyclobutan]-3′-one (148 mg, 0.53 mmol) in CH2Cl2 (4.4 mL) was added DAST (0.20 mL, 1.53 mmol), and the mixture was stirred at room temperature for 4.5 hours. Another portion of DAST (0.10 mL, 0.76 mmol) was added to the reaction mixture and the mixture was stirred at room temperature for 19.5 hours. The mixture was cooled at 0° C. and added to saturated aqueous NaHCO3 and the resulting mixture was extracted with CHCl3. The combined organic extracts were dried over Na2SO4, filtered, and concentrated in vacuo. The residue was purified by silica gel chromatography (hexane/EtOAc=100:0-90:10) to give 6-bromo-3′,3′-difluoro-4-methylene-4H-spiro[chromene-3,1′-cyclobutane] (65 mg).
Preparation Example 132
To a solution of N-{[6-bromo-4-(1,1-difluoro-2-hydroxyethyl)-4H-spiro[chromene-3,1′-cyclopropan]-4-yl]carbamothioyl}benzamide (340 mg, 0.684 mmol) in MeOH (1.7 mL) was added methylamine (9.8M MeOH solution, 698 μL, 6.84 mmol). The mixture was stirred for 3 hours at ambient temperature. The mixture was concentrated azeotropically with toluene 3 times to give crude 1-[6-bromo-4-(1,1-difluoro-2-hydroxyethyl)-4H-spiro[chromene-3,1′-cyclopropan]-4-yl]thiourea (268 mg).
Preparation Example 133
Under ice cooling, to a solution of N-[6-bromo-4-(1,1-difluoro-2-hydroxyethyl)-4H-spiro[chromene-3,1′-cyclopropan]-4-yl]-2-methylpropane-2-sulfinamide (4.60 g, 10.5 mmol) in THF-EtOH (50% v/v, 46 mL) was added 4M HCl/dioxane (13.1 mL, 52.5 mmol), and the mixture was stirred for 3 hours at room temperature. Under ice cooling, saturated aqueous NaHCO3, H2O and brine were added to the mixture, and then the mixture was extracted with EtOAc. The organic layer was dried over MgSO4 and concentrated. The residue was purified by silica gel chromatography (EtOAc/hexane=50:50-100:0) to give 2-(4-amino-6-bromo-4H-spiro[chromene-3,1′-cyclopropan]-4-yl)-2,2-difluoroethanol (2.28 g).
Preparation Example 134
To a solution of methyl 3,3-dimethoxy-1-vinylcyclobutanecarboxylate (2.02 g, 10.09 mmol) in THF (20 mL) was added DIBAL-H (1.04M in toluene, 29 mL, 30.16 mmol) at 0° C. under argon, and the mixture was stirred for 30 minutes at 0° C. To the mixture was carefully added MeOH (29 mL) and Na2SO4.10H2O (29 g), and the mixture was stirred overnight. The mixture was filtered and evaporated under reduced pressure. And the residue was diluted with hexane/EtOAc=1:1 and filtrated through the pad of silica gel and concentrated in vacuo to give (3,3-dimethoxy-1-vinylcyclobutyl)methanol (1.42 g).
Preparation Example 137
Under ice cooling, to a solution of LiBH4 (458 mg, 21.0 mmol) in THF (30 mL) was added a solution of ethyl {6-bromo-4-[(tert-butylsulfinyl)amino]-4H-spiro[chromene-3,1′-cyclopropan]-4-yl}(difluoro)acetate (5.05 g, 10.5 mmol) in THF (20 mL), and the mixture was stirred for 15 minutes at same temperature and stirred for 2 hours at room temperature. After adding H2O and brine, the mixture was extracted with EtOAc. The organic layer was dried over MgSO4 and concentrated to obtain crude N-[6-bromo-4-(1,1-difluoro-2-hydroxyethyl)-4H-spiro[chromene-3,1′-cyclopropan]-4-yl]-2-methylpropane-2-sulfinamide. The desired compound (4.6 g) was applied to the next step without further purification.
Preparation Example 149
To a suspension of activated zinc (3.44 g, 52.5 mmol) in Et2O-THF (50% v/v, 80 ml) under reflux was slowly added a solution of ethyl bromodifluoroacetate (8.00 g, 39.4 mmol) and N-(6-bromo-4H-spiro[chromene-3,1′-cyclopropan]-4-ylidene)-2-methylpropane-2-sulfinamide (4.68 g, 13.1 mmol) in Et2O-THF (50% v/v, 80 ml) over 40 minutes, and the mixture was stirred for 4 hours at the same temperature. After cooling, the mixture was filtrated through celite pad and washed with EtOAc. To the filtrate were added saturated aqueous NH4Cl and EtOAc. After separation, the water layer was extracted with EtOAc. The combined organic layer was dried over MgSO4 and concentrated. The residue was purified by silica gel chromatography (EtOAc:hexane=20:80-100:0) to give ethyl {6-bromo-4-[(tert-butylsulfinyl)amino]-4H-spiro[chromene-3,1′-cyclopropan]-4-yl}(difluoro)acetate (6.08 g).
Preparation Example 151
To a solution of methyl 1-(1-hydroxyethyl)-3,3-dimethoxycyclobutanecarboxylate (3.28 g, 15.03 mmol) and pyridine (2.4 mL, 29.83 mmol) in CH2Cl2 (65 mL) was added trifluoromethanesulfonic anhydride (3.0 mL, 17.86 mmol) at −78° C. The mixture was stirred for 10 minutes and then warmed to 0° C. After stirring for 15 minutes at 0° C., DBU (9.0 mL, 60.18 mmol) was added to the reaction mixture and the resulting mixture was stirred for 1 hour at room temperature. The mixture was partially evaporated under reduced pressure and filtrated through the pad of silica gel and washed with CH2Cl2. The filtrate was washed with saturated aqueous NaHCO3, dried over Na2SO4, filtered, and evaporated. The residue was diluted with hexane/EtOAc=4:1 and added small amount of CHCl3 and filtrated through the pad of silica gel to give methyl 3,3-dimethoxy-1-vinylcyclobutanecarboxylate (2.21 g).
Preparation Example 152
Under ice cooling, to a solution of N-{[6-bromo-4-(1,1-difluoro-2-hydroxyethyl)-4H-spiro[chromene-3,1′-cyclopropan]-4-yl]carbamothioyl}benzamide (340 mg, 0.684 mmol) in CH2Cl2 (9 mL) was added 1-chloro-N,N,2-trimethylprop-1-en-1-amine (206 mg, 1.54 mmol), and the mixture was stirred for 17 hours at room temperature. Ice was added and the mixture was neutralized by 10% aqueous K2CO3 and extracted with CH2Cl2. The organic layer was washed with brine and dried over MgSO4 and concentrated. The residue was purified by silica gel chromatography (EtOAc/hexane=1:99-40:60) to afford N-(6′-bromo-5′′,5′′-difluoro-5′′,6′′-dihydrodispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]thiazin]-2′′-yl)benzamide (198 mg).
Preparation Example 154
To a solution of (4S)-6′-bromo-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-amine (4.41 g, 12.5 mmol) in THF (44 mL) were added di-tert-butyl dicarbonate (6.54 g, 30.0 mmol) and 4-dimethylaminopyridine (76 mg, 0.62 mmol). After stirring for 16 hours at ambient temperature, the mixture was concentrated at reduced pressure. The residue was purified by column chromatography on silica gel (hexane-EtOAc, a linear gradient of EtOAc from 10 to 30%) to afford di-tert-butyl[(4S)-6′-bromo-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-yl]imidodicarbonate (6.95 g).
Preparation Example 155
To a solution of (4′R)-6′-bromo-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-2′′-amine (1.54 g, 4.57 mmol) in tetrahydrofuran (21 ml) were added di-tert-butyl dicarbonate (2.49 g, 11.4 mmol) and N,N-dimethylpyridin-4-amine (28 mg, 0.23 mmol). The mixture was stirred at room temperature overnight and concentrated in vacuo. The residue was purified by silica gel column chromatography (hexane/ethyl acetate=3:1) to afford di-tert-butyl[(4′R)-6′-bromo-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-2′′-yl]imidodicarbonate (1.99 g).
Preparation Example 156
A 3-necked-flask was charged with dimethylsulfoxide (17 ml) and potassium hydroxide (0.84 g, 15 mmol). Then trimethylsulfoxonium iodide (3.3 g, 15 mmol) was added and the mixture was stirred at room temperature for 30 minutes. To this mixture, 6-bromo-2,2-dimethyl-3-methylene-2,3-dihydro-4H-chromen-4-one (2.0 g, 7.5 mmol) and dimethylsulfoxide (3 ml) were added. The mixture was stirred at room temperature for 15 hours and then water (30 ml) was added. The mixture was extracted with a mixture of hexane (70 ml) and ethyl acetate (70 ml). The organic layer was washed with water (50 ml) twice and then with brine (30 ml), dried over anhydrous magnesium sulfate, and concentrated in vacuo. The residue was purified by silica gel column chromatography (NH-silica gel, hexane/ethyl acetate=100:1-20:1) to afford 6-bromo-2,2-dimethyl-4H-spiro[chromene-3,1′-cyclopropan]-4-one (1.2 g).
Preparation Example 157
To a mixture of di-tert-butyl[(4S)-6′-bromo-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-yl]imidodicarbonate (3.40 g, 6.14 mmol), bis(dibenzylideneacetone)palladium(0) (353 mg, 0.614 mmol), and tri-tert-butylphosphonium tetrafluoroborate (179 mg, 0.617 mmol) was added lithium bis(trimethylsilyl)amide (1M solution in toluene, 31 mL, 31 mmol) at ambient temperature under argon atmosphere. After stirring for 1.5 hours at 60° C., the mixture was cooled in an ice-water bath and 1M aqueous HCl (31 mL) was added. After stirring for 10 minutes at ambient temperature, to the mixture was added CHCl3, and the mixture was filtered through a pad of celite. The filtrate was separated and the aqueous layer was extracted with CHCl3. The combined organic layer was dried over Na2SO4 and filtered. The filtrate was concentrated at reduced pressure. To the residue were added MeOH (34 mL) and silica gel (neutral; 17 g) at ambient temperature. After stirring for 1 hour at 40° C., the mixture was concentrated at reduced pressure. The residue was purified with column chromatography on silica gel (hexane-EtOAc, a linear gradient of EtOAc from 10 to 100%) to afford tert-butyl[(4S)-6′-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-yl]carbamate (2.38 g).
Preparation Example 158
To a mixture of di-tert-butyl[(4′R)-6′-bromo-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-2′′-yl]imidodicarbonate (1.98 g, 3.68 mmol), bis(dibenzylideneacetone)palladium(0) (212 mg, 0.369 mmol), and tri-tert-butylphosphonium tetrafluoroborate (108 mg, 0.371 mmol) was added lithium bis(trimethylsilyl)amide (1M solution in toluene, 18 ml, 18 mmol) at room temperature under argon atmosphere. The mixture was stirred at 60° C. for 2 hours and then quenched with saturated aqueous ammonium chloride. The mixture was extracted with chloroform. The organic layer was dried over anhydrous magnesium sulfate, and concentrated in vacuo. The residue was purified by silica gel column chromatography (hexane/ethyl acetate=100:0-50:50-0:100) to afford tert-butyl[(4′R)-6′-amino-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-2′′-yl]carbamate (1.37 g).
Preparation Example 159
N,N,N′,N′-tetramethylmethanediamine (4.8 g, 47 mmol) was added to a solution of 6-bromo-2,2-dimethyl-2,3-dihydro-4H-chromen-4-one (3.0 g, 12 mmol) and acetic acid (0.67 ml, 12 mmol) in tetrahydrofuran (43 ml), and the mixture was stirred at 70° C. for 24 hours. To the mixture was added acetic anhydride (4.4 ml, 47 mmol), and the mixture was stirred at 70° C. for 4 hours and concentrated in vacuo. The residue was directly purified by silica gel column chromatography (hexane/ethyl acetate=20:1) to afford 6-bromo-2,2-dimethyl-3-methylene-2,3-dihydro-4H-chromen-4-one (2.8 g).
Preparation Example 160
To a suspension of methyl(triphenyl)phosphonium bromide (20.8 g, 58.2 mmol) in tetrahydrofuran (168 ml) was added n-butyllithium (2.69 M solution in hexane, 21.6 ml, 58.2 mmol) under dry ice-acetone bath cooling and argon atmosphere. The mixture was stirred for 1 hour at 0° C. To the mixture was added 6-bromo-2,2-dimethyl-4H-spiro[chromene-3,1′-cyclopropan]-4-one (8.18 g, 29 mmol). The mixture was stirred for 1 hour at 0° C. The reaction was quenched by adding water and extracted with ethyl acetate. The organic layer was washed with brine, dried over anhydrous magnesium sulfate, and concentrated in vacuo. The residue was purified by silicagel column chromatography (hexane/ethyl acetate=20:1-10:1) to afford 6-bromo-2,2-dimethyl-4-methylene-4H-spiro[chromene-3,1′-cyclopropane] (7.69 g).
Preparation Example 161
To a mixture of tert-butyl[(4S)-6′-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-yl]carbamate (1.00 g, 2.57 mmol) and CH2Cl2 (10 mL) were added 5-chloro-2-pyridinecarboxylic acid (526 mg, 3.34 mmol), N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (640 mg, 3.34 mmol), 1-hydroxybenzotriazole (451 mg, 3.34 mmol) and N,N-diisopropylethylamine (0.571 mL, 3.34 mmol) at ambient temperature. After stirring overnight at the same temperature, the reaction mixture was purified with column chromatography on silica gel (hexane-EtOAc, a linear gradient of EtOAc from 0 to 90%) and then on NH-silica gel (hexane-EtOAc, a linear gradient of EtOAc from 0 to 90%) to give tert-butyl[(4S)-6′-{[(5-chloropyridin-2-yl)carbonyl]amino}-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-yl]carbamate (1.02 g).
Preparation Example 165
To a mixture of tert-butyl[(4′R)-6′-amino-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-2′′-yl]carbamate (100 mg, 0.268 mmol), 5-methoxypyrazine-2-carboxylic acid (45 mg, 0.30 mmol) and 1H-benzotriazol-1-ol (40 mg, 0.29 mmol) in dichloromethane (2 mL) was added N-[3-(dimethylamino)propyl]-N′-ethylcarbodiimide hydrochloride (57 mg, 0.30 mmol). The mixture was stirred at room temperature for 3 hours and directly purified by silica gel column chromatography (precolumn: NH-silica gel, main column: neutral silica gel, hexane/ethyl acetate=2:1-1:1-0:1) to afford tert-butyl[(4′R)-6′-{[(5-methoxypyrazin-2-yl)carbonyl]amino}-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-2′′-yl]carbamate (108 mg).
Preparation Example 167
To an ice-water cooled mixture of tert-butyl[(4′R)-6′-amino-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-2′′-yl]carbamate (430 mg, 1.15 mmol), 5-(difluoromethyl)pyrazine-2-carboxylic acid (221 mg, 1.27 mmol) and 1H-benzotriazol-1-ol (170 mg, 1.26 mmol) in chloroform (8.6 ml) was added N-[3-(dimethylamino)propyl]-N′-ethylcarbodiimide hydrochloride (244 mg, 1.28 mmol). The mixture was stirred at room temperature overnight and directly purified by silica gel column chromatography (precolumn: basic silica gel, main column: neutral silica gel, hexane/ethyl acetate=2:1-1:1-0:100) to afford tert-butyl[(4′R)-6′-({[5-(difluoromethyl)pyrazin-2-yl]carbonyl}amino)-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-2′′-yl]carbamate (373 mg).
Preparation Example 169
A mixture of 6-bromo-2,3-dihydro-4H-chromen-4-one (0.3 g, 1.3 mmol), paraformaldehyde (0.48 g), L-proline (61 mg, 0.53 mmol), and 0.2 M aqueous sodium hydroxide (6 mL) was stirred at room temperature for 16 hours. The mixture was extracted with CHCl3 and concentrated in vacuo. The residue was purified with silica-gel column chromatography (CHCl3/MeOH=100:0 to 90:10) to give 6-bromo-3,3-bis(hydroxymethyl)-2,3-dihydro-4H-chromen-4-one (0.33 g).
Preparation Example 171
To a solution of 6-bromo-4H-spiro[chromene-3,3′-oxetan]-4-one (143 mg, 0.531 mmol) in THF (2 mL) was added Tebbe reagent ((C5H5)2TiCH2ClAl(CH3)2, μ-Chloro[di(cyclopenta-2,4-dien-1-yl)]dimethyl(μ-methylene)titaniumaluminum, 0.5 M in toluene, 2 mL) under ice-water bath cooling. After the reaction mixture was stirred at the same temperature for 2 hours and then room temperature for 4 hours, 1M aqueous NaOH (1 mL) was added. After dilution with water and filtration with Celite, the insoluble material was washed with CHCl3. The aqueous phase was extracted with CHCl3 and combined organic layer was concentrated in vacuo. The residue was purified with silica-gel column chromatography to give 6-bromo-4-methylene-4H-spiro[chromene-3,3′-oxetane] (0.10 g).
Preparation Example 173
Under nitrogen, MeMgBr solution (2.9 M in 2-methyltetrahydrofuran solution, 3.06 L, 8.88 mol) was diluted by adding into THF (8 L) at 0-5° C. A solution of 6-bromo-3,3-bis(hydroxymethyl)-2,2-dimethyl-2,3-dihydro-4H-chromen-4-one (700 g, 2.22 mol) in THF (5 L) was added dropwise via a dropping funnel to the diluted MeMgBr solution maintaining the temperature below 5° C. After the addition was complete, the reaction mixture was allowed to warm to room temperature and stirred for 30 minutes. Then the reaction mixture was heated under reflux overnight. The mixture was cooled to room temperature followed by cooling with an ice-water bath and 6 M hydrochloric acid (3.7 L, 22.2 mol) was added dropwise via a dropping funnel over 30 minutes maintaining the temperature below 10° C. After the addition was complete, the mixture was allowed to warm to room temperature and stirred for 20 minutes. The mixture was extracted with toluene (5 L×2) and the combined extracts were washed with brine, dried over Na2SO4 and concentrated under reduced pressure. The residue was treated with a mixture of hexane/toluene (5:1, 2 L) and the resulting suspension was stirred for 30 minutes. The precipitate was collected by filtration, washed with hexane and dried under vacuum to give (6-bromo-2,2-dimethyl-4-methylene-3,4-dihydro-2H-chromene-3,3-diyl)dimethanol (500 g).
Preparation Example 174
Under nitrogen, to a solution of (6-bromo-2,2-dimethyl-4-methylene-3,4-dihydro-2H-chromene-3,3-diyl)dimethanol (800 g, 2.55 mol) and MsCl (877.9 g, 7.66 mol) in THF (4.0 L) was added Et3N (851 g, 8.41 mol) over 45 minutes maintaining the temperature below 0-10° C. The reaction mixture was allowed to warm to room temperature and stirred for 1 hour. EtOH (8 L) and NaOH (1021.8 g, 25.55 mol) were added and the reaction mixture was heated under reflux for 16 hours. Water (4 L) was added and a clear solution was obtained. Most of the solvent was removed under reduced pressure and the resulting residue was extracted with ethyl acetate (1.5 L×2). The combined extracts were washed with brine, dried over Na2SO4 and concentrated under reduced pressure. The residue was dissolved in MeOH (640 mL) at 60° C. and the resulting solution was allowed to cool to room temperature. The precipitation formed was filtered-off. The filtrate was concentrated under reduced pressure and the residue was re-dissolved in MeOH (500 mL) at 60° C. The resulting solution was allowed to cool to room temperature and then further to 0-5° C. The mixture was stirred at 0-5° C. overnight and the precipitated yellow solid was collected by filtration to give 6-bromo-2,2-dimethyl-4-methylene-4H-spiro[chromene-3,3′-oxetane] (197.2 g). The filtrate was concentrated under reduced pressure and the residue was purified by column chromatography on silica gel (petroleum ether/ethyl acetate=50:1) to give another batch of 6-bromo-2,2-dimethyl-4-methylene-4H-spiro[chromene-3,3′-oxetane] (143.6 g).
The compounds of Preparation Examples shown in Tables below were prepared using the respective corresponding starting materials in the same manner as the methods of Preparation Examples above. The structures and the preparation methods are shown in [Table. 2] below, and the physicochemical data for the compounds of Preparation Examples are shown in [Table. 3] below.









TABLE 2





Rf
Syn
Structure







 1
R1






 


 2
R2






 


 3
R2






 


 4
R22






 


 5
R22






 


 6
R22






 


 7
R22






 


 8
R8 or R22






 


 9
R22






 


 10
R22






 


 11
R22






 


 12
R22






 


 13
R22






 


 14
R22






 


 15
R22






 


 16
R22






 


 17
R22






 


 18
R22






 


 19
R22






 


 20
R22






 


 21
R22






 


 22
R22






 


 23
R23






 


 24
R24






 


 25
R24






 


 26
R26






 


 27
R27






 


 28
R29






 


 29
R29






 


 30
R30






 


 31
R31






 


 32
R36






 


 33
R36






 


 34
R36






 


 35
R36






 


 36
R36






 


 37
R36






 


 38
R36






 


 39
R36






 


 40
R36






 


 41
R36






 


 42
R36






 


 43
R36






 


 44
R36






 


 45
R36






 


 46
R36






 


 47
R36






 


 48
R48






 


 49
R48






 


 50
R48






 


 51
R48






 


 52
R48






 


 53
R48






 


 54
R48






 


 55
R48






 


 56
R48






 


 57
R48






 


 58
R58






 


 59
R59






 


 60
R60






 


 61
R62






 


 62
R62






 


 63
R63






 


 64
R64






 


 65
R64






 


 66
R66






 


 67
R66 or R67






 


 68a
R69






 


 68b
R69






 


 69
R69






 


 70
R70






 


 71
R70






 


 72
R70






 


 73
R70 or R73






 


 74
R70






 


 75
R70






 


 76
R76






 


 77
R76






 


 78
R78






 


 79
R83






 


 80
R83






 


 81
R83






 


 82
R83






 


 83
R83






 


 84
R83






 


 85
R85






 


 86
R85






 


 87
R85






 


 88
R85






 


 89
R85






 


 90
R83






 


 91
R91






 


 92
R92






 


 93
R92






 


 94
R94






 


 95
R96






 


 96
R96






 


 97
R96






 


 98
R96






 


 99
R96






 


100
R96






 


101
R96






 


102
R96






 


103
R96






 


104
R96






 


105
R96






 


106
R106






 


107
R106






 


108
R108






 


109
R108






 


110
R108






 


111
R111






 


112
R112






 


113
R112






 


114
R114






 


115
R114






 


116
R116






 


117
R117






 


118
R1






 


119
R119






 


120
R22






 


121
R22






 


122
R22






 


123
R22






 


124
R22






 


125
R22






 


126
R22






 


127
R22






 


128
R128






 


129
R30






 


130
R31






 


131
R131






 


132
R132






 


133
R133






 


134
R134






 


135
R59






 


136
R60






 


137
R137






 


138
R62






 


139
R63






 


140
R70






 


141
R70






 


142
R70






 


143
R70






 


144
R70






 


145
R70






 


146
R70






 


147
R70






 


148
R70






 


149
R149






 


150
R83






 


151
R151






 


152
R152






 


153
R112






 


154
R154






 


155
R155






 


156
R156






 


157
R157






 


158
R158






 


159
R159






 


160
R160






 


161
R161






 


162
R161






 


163
R161






 


164
R165






 


165
R165






 


166
R165






 


167
R167






 


168
R161






 


169
R169






 


170
R96






 


171
R171






 


172
R64






 


173
R173






 


174
R174






 


175
R64






 


176
R85





















TABLE 3





Rf
Data







 1
ESI+: 326, 328


 2
ESI+: 519


 3
ESI+: 565


 4
ESI+: 525, 527


 5
ESI+: 565, 567


 6
ESI+: 539, 541


 7
ESI+: 539, 541


 8
ESI+: 553, 555


 9
ESI+: 579, 581


 10
ESI+: 553, 555; a compound prepared from Reference



Example 9a


 11
ESI+: 553, 555; a compound prepared from Reference



Example 9b


 12
ESI+: 581, 583; a compound prepared from Reference



Example 13a


 13
ESI+: 581, 583; a compound prepared from Reference



Example 13b


 14
ESI+: 565, 567; a compound prepared from Reference



Example 12a


 15
ESI+: 565, 567; a compound prepared from Reference



Example 12b


 16
ESI+: 567, 569; a compound prepared from Reference



Example 11a


 17
ESI+: 567, 569; a compound prepared from Reference



Example 11b


 18
ESI+: 621, 623; a compound prepared from Reference



Example 14a


 19
ESI+: 539, 541; a compound prepared from Reference



Example 1a


 20
ESI+: 539, 541; a compound prepared from Reference



Example 1b


 21
ESI+: 527, 529


 22
ESI+: 509, 511


 23
ESI+: 423, 425


 24
ESI+: 541, 543


 25
ESI+: 569, 571


 26
ESI+: 439, 441 [M + Na]+


 27
APCI/ESI+: 327


 28
ESI+: 539


 29


 30
ESI+: 376, 378


 31
ESI+: 340, 342


 32
EI: 306, 308


 33
APCI/ESI+: 339


 34
APCI/ESI+: 307


 35
EI: 280, 282


 36
EI: 294, 296


 37
EI: 264, 266


 38
EI: 294, 296


 39
ESI+: 321, 323


 40
ESI+: 309, 311


 41
ESI+: 307, 309


 42
ESI+: 323, 325


 43
EI: 362, 364


 44
ESI+: 357, 359


 45
EI: 268, 270


 46
ESI+: 321, 323


 47
EI: 250, 252


 48
ESI+: 315, 317


 49
ESI+: 301, 303


 50
ESI+: 315, 317


 51
ESI+: 341, 343


 52
ESI+: 329, 331


 53
ESI+: 327, 329


 54
ESI+: 365, 367 [M + Na]+


 55
ESI+: 383, 385


 56
ESI+: 377, 379


 57
ESI+: 341, 343


 58
EI: 444, 446


 59
ESI+: 281, 283


 60
APCI/ESI+: 356, 358


 61
ESI+: 209


 62
ESI+: 237


 63
ESI+: 300, 302


 64


 65
ESI+: 601


 66
ESI+: 346


 67
ESI+: 390


 68a
ESI+: 196, 198


 68b
ESI+: 196, 198


 69
ESI+: 153, 155


 70
ESI+: 469


 71
ESI+: 485, 487


 72
ESI+: 513


 73
ESI+: 529, 531


 74
ESI+: 547, 549


 75
ESI+: 503, 505


 76
ESI+: 515


 77
ESI+: 561


 78
ESI+: 356, 358


 79
ESI+: 571


 80
ESI+: 576


 81
ESI+: 525


 82
ESI+: 538


 83


 84
APCI/ESI+: 309


 85


 86


 87


 88


 89
ESI+: 602, 604


 90


 91
ESI+: 548


 92
EI: 268, 270


 93
EI: 316, 318


 94
APCI/ESI+: 341


 95
ESI+: 331, 333 [M + Na]+


 96
EI: 296, 298


 97
ESI+: 297, 299


 98
ESI+: 345, 347 [M + Na]+


 99
ESI+: 311, 313


100
ESI+: 331, 333 [M + Na]+


101
ESI+: 347, 349 [M + Na]+


102
ESI+: 365, 367


103
ESI+: 359, 361


104
ESI+: 323, 325


105
EI: 282, 284


106
EI: 284, 286


107
EI: 288, 290


108
ESI+: 283, 285


109
EI: 322, 324


110
ESI+: 281, 283


111
EI: 254, 256


112
EI: 312, 314


113
EI: 302, 304


114
EI: 266, 268


115
EI: 270, 272


116


117
EI: 406, 408


118


119
ESI+: 241 [M + Na]+


120
ESI+: 615, 617; a compound prepared from Reference



Example 15a


121
ESI+: 615, 617; a compound prepared from Reference



Example 15b


122
ESI+: 473, 475


123
ESI+: 579, 581; a compound prepared from Reference



Example 16a


124
ESI+: 559, 561


125
ESI+: 579, 581; a compound prepared from Reference



Example 16b


126
ESI+: 559, 561


127
ESI+: 523, 525


128
ESI+: 497, 499


129
ESI+: 360, 362


130
ESI+: 324, 326


131
EI: 300, 302


132


133


134
ESI+: 195 [M + Na]+


135
EI: 278, 280


136
ESI+: 340, 342


137


138
ESI+: 273 [M + Na]+


139
ESI+: 284, 286


140
ESI+: 530, 532


141
ESI+: 574, 576


142
ESI+: 482


143
ESI+: 490


144
ESI+: 510, 512


145
ESI+: 499, 501


146
ESI+: 543, 545


147
ESI+: 476


148
ESI+: 520


149
ESI+: 480, 482


150
APCI/ESI+: 479


151
ESI+: 223 [M + Na]+


152
ESI+: 479, 481


153
EI: 452, 454


154
ESI+: 553, 555


155
ESI+: 559, 561 [M + Na]+


156
ESI+: 281, 283


157
ESI+: 390


158
ESI+: 374


159
CI+: 267, 269


160
ESI+: 279, 281


161
ESI+: 529, 531


162
ESI+: 573, 575


163
ESI+: 526


164
ESI+: 513


165
ESI+: 510


166
ESI+: 497


167
ESI+: 530


168
ESI+: 526


169
ESI+: 287, 289


170
EI: 268, 270


171


172
ESI+: 613


173
NMR-CDCl3: 1.33 (s, 6H), 2.32 (br s, 2H), 3.90 (d,



J = 11.1 Hz, 2H), 4.02 (d, J = 11.1 Hz, 2H), 5.51



(s, 1H), 5.86 (s 1H), 6.69 (d, J = 8.7 Hz, 1H), 7.31-7.27



(m, 1H), 7.68 (br s, 1 H)


174
NMR-CDCl3: 1.46 (s, 6H), 4.55 (d, J = 6.6 Hz, 2H), 4.71 (d,



J = 6.6 Hz, 2H), 5.33 (s, 1H), 5.73 (s, 1H), 6.70 (d,



J = 8.7 Hz, 1H), 7.28 (dd, J = 2.4, 8.7 Hz, 1H),



7.65 (d, J = 2.4 Hz, 1H),


175
ESI+: 557


176
ESI+: 554









Reference Example 1a,b
To an ice chilled solution of 6-bromo-4-methylene-4′,5′-dihydro-3′H,4H-spiro[chromene-3,2′-furan] (497 mg, 1.77 mmol) in EtOAc (2.5 ml) and MeCN (2.5 ml) was added silver cyanate (397 mg, 2.65 mmol) in an ice bath under an argon atmosphere. To the mixture was added iodine (673 mg, 2.65 mmol). After stirring for 30 minutes in the ice bath and 30 minutes at ambient temperature, the mixture was filtered through celite. The filtrate was washed with saturated aqueous Na2S2O3. The organic layer was washed with brine, dried over Na2SO4, filtered, and the filtrate was evaporated to give an oil. The oil was dissolved in THF (5 mL) and a 2M EtOH solution of NH3 (4.5 ml, 9.0 mmol) was added under ice bath cooling. The mixture was stirred overnight under ice bath cooling and for 1 hour at 70° C. After concentration in vacuo, the residue was diluted with saturated aqueous NaHCO3 and extracted with CHCl3. The organic layer was washed with brine, dried over Na2SO4, filtered, and the filtrate was evaporated off. The residue was purified by silica gel chromatography (CHCl3/EtOH=100:0-90:10) to give less polar diastereomer of 6′-bromo-4,5-dihydro-3H-dispiro[furan-2,3′-chromene-4′,4′′-[1,3]oxazol]-2′′-amine (178 mg) and polar diastereomer of 6′-bromo-4,5-dihydro-3H-dispiro[furan-2,3′-chromene-4′,4′′-[1,3]oxazol]-2′′-amine (367 mg).
Reference Example 6
Under an argon atmosphere, to a mixture of 6-bromo-2,2-dimethyl-4-methylene-4H-spiro[chromene-3,3′-oxetane] (504 mg, 1.71 mmol), silver cyanate (384 mg, 2.56 mmol) and EtOAc-MeCN (1:1, 5.0 mL) was added iodine (649 mg, 2.56 mmol) over 5 minutes in an ice-water bath. After stirring for 30 minutes at the same temperature, the mixture was filtered through celite pad. The filtrate was washed with saturated aqueous Na2S2O3 and brine, dried over MgSO4 and filtered. After concentration of the filtrate at reduced pressure, the residue was dissolved in THF (5.0 mL). The solution was added to 2 M EtOH solution of ammonia (10.7 mL, 21.4 mmol) in an ice-water bath. The mixture was stirred for 1 hour in the bath and 3 hours at 70° C. After cooling down to ambient temperature, NH-silica gel was added to the reaction mixture, and the mixture was concentrated at reduced pressure. The residue was purified with column chromatography on silica gel (CHCl3-EtOH, a linear gradient of EtOH from 0 to 20%) to afford 6′-bromo-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-amine (483 mg).
Reference Example 19
A mixture of di-tert-butyl {6′-[5-(3-methoxyprop-1-yn-1-yl)pyridin-3-yl]dispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-yl}imidodicarbonate (220 mg, 0.372 mmol), silica gel (neutral; 660 mg) and toluene (2.2 mL) was stirred for 3 hours at 100° C. The mixture was cooled down to ambient temperature, and concentrated at reduced pressure. Purification of the residue with column chromatography on silica gel (CHCl3-MeOH, a linear gradient of MeOH from 0 to 10%) afforded 6′-[5-(3-methoxyprop-1-yn-1-yl)pyridin-3-yl]dispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-amine (92.4 mg).
Example 27
A mixture of tert-butyl (6′-{[(5-chloropyridin-2-yl)carbonyl]amino}-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-yl)carbamate (217 mg, 0.410 mmol), silica gel (neutral; 651 mg), and toluene (4 mL) was stirred for 80 minutes at 120° C. The reaction mixture was cooled down to ambient temperature and concentrated at reduced pressure. Purification of the residue with column chromatography on silica gel (eluted with EtOH/CHCl3=0/100 to 20/80) afforded N-(2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-6′-yl)-5-chloropyridine-2-carboxamide (144 mg).
Reference Example 31
To a mixture of tert-butyl (6′-{[(5-fluoropyridin-2-yl)carbonyl]amino}dispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-2′′-yl)carbamate (135 mg, 0.288 mmol) and CH2Cl2 (5.4 mL) was added trifluoroacetic acid (1.30 mL, 17.0 mmol) at ambient temperature. After stirring for 2 hours at the same temperature, the mixture was concentrated at reduced pressure. The residue was partitioned between CHCl3 and saturated aqueous NaHCO3. The organic layer was washed with water, dried over Na2SO4 and filtered. The filtrate was concentrated at reduced pressure, and the residue was purified with column chromatography on silica gel (CHCl3-EtOH, a linear gradient of EtOH from 0 to 20%) to afford N-(2′′-aminodispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-6′-yl)-5-fluoropyridine-2-carboxamide (43.6 mg).
Reference Example 39
The mixture of di-tert-butyl[6′-(6-methoxypyridin-2-yl)dispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-yl]imidodicarbonate (160 mg, 0.289 mmol) and TsOH.H2O (275 mg, 1.45 mmol) in MeCN (3.2 mL) was stirred for 4 hours at 40° C. After dilution with CHCl3, the organic layer was washed with saturated aqueous NaHCO3 and brine, dried over Na2SO4, and concentrated in vacuo. The residue was purified by silica gel chromatography (CHCl3:MeOH=99:1-95:5) and then the residue was washed with EtOAc to afford 6′-(6-methoxypyridin-2-yl)dispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-amine (40 mg).
Reference Example 40
To a solution of 2-fluoro-6-iodobenzonitrile (1560 mg, 6.32 mmol) in THF (16 mL) at −78° C. was added n-butyllithium (2.64M solution in n-hexane, 2.39 ml, 6.32 mmol) dropwise. The mixture was stirred for 0.5 hour at −78° C. and to the solution was added a solution of N-(6-bromo-4H-spiro[chromene-3,1′-cyclopropan]-4-ylidene)-2-methylpropane-2-sulfinamide (1500 mg, 4.21 mmol) in THF (5 mL). The mixture was stirred for 1 hour at −78° C. and overnight at room temperature. To the mixture was added saturated aqueous NH4Cl. The mixture was extracted with EtOAc and the organic layer was concentrated in vacuo. The residue was purified by silica gel chromatography (hexane/AcOEt=100:0-0:100, then CHCl3:MeOH=85:15) to give 6′-bromo-4′′-fluorodispiro[cyclopropane-1,3′-chromene-4′,1′′-isoindol]-3′′-amine (469 mg).
Reference Example 41
6-Bromo-4-methylene-4H-spiro[chromene-3,3′-oxetane] (1.50 g, 5.62 mmol) was added to a mixture of silver thiocyanate (3.73 g, 22.5 mmol), iodine (2.85 g, 11.2 mmol) and toluene (15 mL) in an ice-water bath. After stirring overnight at ambient temperature, the mixture was filtered through celite pad (washed with EtOAc). The filtrate was washed with saturated aqueous Na2S2O3 and brine, dried over MgSO4 and filtered. The filtrate was concentrated at reduced pressure. A mixture of the residue and THF (15 mL) was added to ammonia (2.0 M in EtOH, 30 mL) in an ice-water bath. After stirring for 1 hour at the same temperature, the mixture was stirred for 2.5 days at ambient temperature. The mixture was partitioned with MeOH—CHCl3 (1:9) and water. The organic layer was dried over Na2SO4 and filtered. The filtrate was concentrated at reduced pressure, and purification of the residue with column chromatography on silica gel (CHCl3-EtOH, a linear gradient of EtOH from 0 to 5%) afforded 6′-bromodispiro[oxetane-3,3′-chromene-4′,4′′-[1,3]thiazol]-2′′-amine (1.26 g).
Reference Example 43
The mixture of di-tert-butyl (6′-bromodispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-yl)imidodicarbonate (150 mg, 0.286 mmol), 3-chloropyridin-2-amine (184 mg, 1.43 mmol), (1E,4E)-1,5-diphenylpenta-1,4-dien-3-one-palladium (3:2) (52 mg, 0.057 mmol), di-tert-butyl(2′,4′,6′-triisopropylbiphenyl-2-yl)phosphine (97 mg, 0.23 mmol), and Cs2CO3(279 mg, 0.857 mmol) in 1,4-dioxane (7.5 ml) was stirred for 48 hours at 100° C. The reaction mixture was cooled down to ambient temperature, and partitioned with CHCl3 and water. The organic layer was dried over Na2SO4, and filtered. The residue was dissolved in toluene, and was added silica gel (neutral), and stirred at 130° C. for 3 hours. The mixture was concentrated under reduced pressure, and purified by silica gel column chromatography (CHCl3/MeOH=100:0-85:15) to give an amorphous, which was washed with CHCl3/hexane to give 6′-(3-chloropyridin-2-yl)aminodispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetane]-2-amine (28 mg).
Reference Example 46
To a stirred mixture of palladium (II) acetate (7.9 mg, 0.035 mmol) and biphenyl-2-yl(di-tert-butyl)phosphine (5.3 mg, 0.018 mmol) in THF (5 mL) under argon atmosphere was added di-tert-butyl (6′-bromotrispiro[cyclobutane-1,2′-chromene-4′,4′′-[1,3]oxazole-3′,3′′′-oxetan]-2′′-yl)imidodicarbonate (100 mg, 0.177 mmol) followed by isobutylzinc bromide (0.5 M THF solution, 1.1 mL). The reaction mixture was stirred at room temperature for 17 hours, and then the reaction was quenched with H2O and brine. The resulting mixture was extracted with chloroform 3 times. The combined organic layer was dried over magnesium sulfate and concentrated under reduced pressure. The crude product was dissolved in toluene (6 mL). To the solution was added silica gel (neutral; 600 mg), and the resulting suspension was stirred at 100° C. for 1 hour. After cooling and concentration, the residue was purified by silica gel column chromatography (EtOH/CHCl3=0:100-20:80) followed by column chromatography (NH-silica gel, EtOAc/hexane=20:80-100:0) to afford 6′-isobutyltrispiro[cyclobutane-1,2′-chromene-4′,4′′-[1,3]oxazole-3′,3′′′-oxetan]-2′′-amine (20 mg).
Reference Example 48
To a stirred solution of di-tert-butyl[6′-(5-bromopyridin-3-yl)trispiro[cyclobutane-1,2′-chromene-4′,4′′-[1,3]oxazole-3′,3′′′-oxetan]-2′′-yl]imidodicarbonate (48 mg, 0.075 mmol) in Et3N (0.67 ml) were added ethynylcyclopropane (0.063 mL, 0.75 mmol), PdCl2(PPh3)2 (10 mg, 0.015 mmol) and CuI (5.7 mg, 0.030 mmol) at room temperature and the mixture was sealed and irradiated with microwave at 150° C. for 1 hour. To this mixture was added activated carbon (ca. 100 mg) and the mixture was stirred for 1 hour to remove palladium residues. The mixture was passed through a pad of Celite. The filtrate was evaporated to give a residue. To the solution of the residue in toluene (3 ml) was added silica gel (500 mg) and the mixture was refluxed for 2 hours. The mixture was passed through a pad of Celite and the filtrate was evaporated to give a crude product, which was purified with column chromatography (MeOH in CHCl3=0 to 10%) to give 6′-[5-(cyclopropylethynyl)pyridin-3-yl]trispiro[cyclobutane-1,2′chromene-4′,4′′-[1,3]oxazole-3′,3′′′-oxetan]-2′′-amine (5 mg).
Reference Example 49
A suspension of 6′-(2-methylimidazo[1,2-a]pyridin-6-yl)trispiro[cyclobutane-1,2′-chromene-4′,4′′-[1,3]oxazole-3′,3′′′-oxetan]-2′′-amine (66 mg, 0.16 mmol), acetic acid (27 μl, 0.06 mmol) and PtO2 (13 mg) in EtOH (3.3 ml) was stirred at room temperature under the hydrogen atmosphere at 3 atm for 8 hours. To the mixture was added acetic acid (63 μl), and the mixture was stirred at room temperature under hydrogen atmosphere at 3 atm for 38 hours. The mixture was filtrated through celite pad. And then the filtrate was concentrated under reduced pressure. The obtained residue was purified by silicagel column chromatography (28% NH4OH/EtOH/CHCl3=2:20:80) followed by trituration in iPr2O and filtration to give 6′-(2-methyl-5,6,7,8-tetrahydroimidazo[1,2-a]pyridin-6-yl)trispiro[cyclobutane-1,2′-chromene-4′,4′′-[1,3]oxazole-3′,3′′′-oxetan]-2′′-amine (56 mg).
Reference Example 51a,b
2′,2′-Dimethyl-6′-(pyrimidin-5-yl)dispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-amine (489 mg, 1.39 mmol) was subjected to chromatography using supercritical CO2/[MeOH with 0.1% diethylamine] (70:30) on Chiralcel OD-H column (Daicel, 10×250 mm) eluting at a flow rate 10 mL/minute (40° C. column temperature). The first peak (retention time=3.44 minutes) provided an enantiomer of 2′,2′-dimethyl-6′-(pyrimidin-5-yl)dispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-amine (220 mg), and the second peak (retention time=6.92 minutes) provided the other enantiomer of 2′,2′-dimethyl-6′-(pyrimidin-5-yl)dispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-amine (215 mg).
Reference Example 52a,b
6′-[5-(prop-1-yn-1-yl)pyridin-3-yl]dispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-2′′-amine (330 mg, 0.96 mmol) was subjected to chromatography using supercritical CO2/[EtOH with 0.1% diethylamine] (65/35) on Chiralcel OZ-H column (10×250 mm) eluting at a flow rate 10 mL/minute (40° C. column temperature). The first peak (retention time=3.62 minutes) was concentrated in vacuo, and dissolved in EtOAc (3 mL) and then hexane (20 mL) was added. The resulting precipitate was collected by filtration, washed with hexane and dried in vacuo to give an enantiomer of 6′-[5-(prop-1-yn-1-yl)pyridin-3-yl]dispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-2′′-amine (108 mg). The second peak (retention time=6.27 minutes) was concentrated in vacuo, and dissolved in EtOAc (3 mL) and hexane (20 mL) was added. The resulting precipitate was collected by filtration, washed with hexane and dried in vacuo to give the other enantiomer of 6′-[5-(prop-1-yn-1-yl)pyridin-3-yl]dispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-2′′-amine (80 mg).
Reference Example 53a,b
6′-[5-(prop-1-yn-1-yl)pyridin-3-yl]dispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-amine (400 mg, 1.39 mmol) was subjected to chromatography using supercritical CO2/[MeOH with 0.1% diethylamine) (70:30) on Chiralcel AD-H column (10×250 mm) eluting at a flow rate 10 mL/minute (40° C. column temperature). The first peak (retention time=5.81 minutes) provided an enantiomer of 6′-[5-(prop-1-yn-1-yl)pyridin-3-yl]dispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-amine (160 mg), and the second peak (retention time=9.25 minutes) provided the other enantiomer of 6′-[5-(prop-1-yn-1-yl)pyridin-3-yl]dispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-amine (170 mg)
Reference Example 54
A mixture of di-tert-butyl (6′-bromo-2′,2′-dimethyldispiro[oxetane-3,3′-chromene-4′,4′′-[1,3]thiazol]-2′′-yl)imidodicarbonate (268 mg, 0.471 mmol), pyrimidin-5-ylboronic acid (175 mg, 1.41 mmol), bis(triphenylphosphine)palladium(II) dichloride (33.0 mg, 0.047 mmol) and Na2CO3 (150 mg, 1.41 mmol) in dioxane-water (4:1, 5.4 mL) was stirred for 1.5 hours at 100° C. Tetrakis(triphenylphosphine)palladium(0) (272 mg, 0.235 mmol) was added to the mixture and the mixture was stirred for 1.5 hours at 100° C. To the mixture was added charcoal and the mixture was stirred for 30 minutes at 50° C. The mixture was filtered through celite pad (eluted with EtOAc) and the filtrate was washed with water and brine. The organic layer was dried over MgSO4 and filtered, and the filtrate was concentrated at reduced pressure. To the residue were added toluene (2.7 mL) and silica gel (neutral; 804 mg), and the mixture was stirred for 1 hour at 110° C. After concentration of the reaction mixture at reduced pressure, the residue was purified with column chromatography on silica gel (CHCl3-EtOH, a linear gradient of EtOH from 0 to 20%), then re-purified with column chromatography on amino silica gel (Hexane-EtOAc, a linear gradient of EtOAc from 0 to 90%). The purified product was dissolved in dioxane (5.0 mL) and HCl (4 M in dioxane, 0.049 mL, 0.198 mmol) was added. The mixture was stirred for 2 hours at ambient temperature. After concentration at reduced pressure, the residue was triturated with IPE, collected by filtration and washed with IPE. The solid was dried under reduced pressure at 30° C. to afford 2′,2′-dimethyl-6′-(pyrimidin-5-yl)dispiro[oxetane-3,3′-chromene-4′,4′′-[1,3]thiazol]-2′′-amine hydrochloride (62.6 mg).
Reference Example 55
Under argon atmosphere, to a mixture of di-tert-butyl (6′-bromodispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-yl)imidodicarbonate (300 mg, 0.571 mmol), dioxane (3.0 mL) and water (1.5 mL) were added 1H-indazol-4-ylboronic acid (185 mg, 1.14 mmol), K2CO3 (237 mg, 1.71 mmol) and bis(triphenylphosphine)palladium(II) dichloride (40 mg, 0.057 mmol), and the mixture was stirred for 3 hours at 100° C. The reaction mixture was cooled down to ambient temperature, partitioned between H2O and 10% MeOH in CHCl3. The organic layer was dried over Na2SO4 and filtered, and the filtrate was concentrated at reduced pressure. The residue was dissolved with dioxane (3.0 mL), and silica gel (neutral; 900 mg) was added to the mixture. After stirring for 2.5 hours at 110° C., the reaction mixture was cooled down to ambient temperature, and concentrated at reduced pressure. Purification of the residue with column chromatography on silica gel (CHCl3-EtOH, a linear gradient of EtOH from 0 to 20%) afforded 6′-(1H-indazol-4-yl)dispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-amine (142 mg).
Reference Example 61
The mixture of di-tert-butyl[6′-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)dispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-2′′-yl]imidodicarbonate (140 mg, 0.25 mmol), 5-bromo-1H-pyrrolo[2,3-b]pyridine (87 mg, 0.44 mmol), Pd(PPh3)4 (58 mg, 0.050 mmol), and Na2CO3 (80 mg, 0.76 mmol) in 1,4-dioxane-H2O (2.8 ml, 4:1) was stirred for 7 hours at 100° C. After dilution with EtOAc and H2O, the organic layer was washed with H2O, dried over MgSO4, and concentrated in vacuo. The residue was purified by silica gel chromatography (CHCl3:MeOH=100:0-85:15) to give white solid. The solid was washed by diisopropyl ether to give 6′-(1H-pyrrolo[2,3-b]pyridin-5-yl)dispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-2′′-amine (42 mg).
Reference Example 65
A mixture of di-tert-butyl[2′,2′-dimethyl-6′-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)dispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-yl]imidodicarbonate (282 mg, 0.470 mmol), 2-bromo-4-methoxypyridine (221 mg, 1.17 mmol), tetrakis(triphenylphosphine)palladium(0) (271 mg, 0.235 mmol) and Na2CO3 (149 mg, 1.41 mmol) in dioxane-water (4:1, 5.6 mL) was stirred for 8 hours at 100° C. The mixture was diluted with MeOH—CHCl3 (1:9) and washed with water, then the organic layer was dried over MgSO4 and filtered. The filtrate was concentrated at reduced pressure. To the residue were added toluene (2.8 mL) and silica gel (neutral; 846 mg), and the mixture was stirred for 3 hours at 120° C. After concentration of the reaction mixture at reduced pressure, the residue was purified with column chromatography on silica gel (CHCl3-EtOH, a linear gradient of EtOH from 0 to 20%) afforded 6′-(4-methoxypyridin-2-yl)-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-amine (146 mg).
Reference Example 75
A mixture of di-tert-butyl (6′-bromotrispiro[cyclobutane-1,2′-chromene-4′,4′′-[1,3]oxazole-3′,3′′′-oxetan]-2′′-yl)imidodicarbona to (300 mg, 0.531 mmol), 5-methylpyridine-3-boronic acid (145 mg, 1.06 mmol), bis(triphenylphosphine)palladium(II) dichloride (37 mg, 0.053 mmol) and Na2CO3 (169 mg, 1.59 mmol) in dioxane-H2O (4:1, 6.0 mL) was stirred for 1 hour at 100° C. To the mixture was added charcoal and the mixture was stirred for 10 minutes at 50° C. The mixture was filtered through celite pad (eluted with EtOAc) and the filtrate was washed with H2O and brine. The organic layer was dried over MgSO4 and filtered, and the filtrate was concentrated under reduced pressure. To the residue were added toluene (3.0 mL) and silica gel (neutral; 900 mg), and the mixture was stirred for 3 hours at 120° C. After concentration of the reaction mixture at reduced pressure, the residue was purified with column chromatography on silica gel (CHCl3-EtOH, a linear gradient of EtOH from 0 to 20%) to afford 6′-(5-methylpyridin-3-yl)trispiro[cyclobutane-1,2′-chromene-4′,4′′-[1,3]oxazole-3′,3′′′-oxetan]-2′′-amine (151 mg).
Reference Example 108
The mixture of 6′-bromodispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-amine (650 mg, 2.00 mmol), [5-(prop-1-yn-1-yl)pyridin-3-yl]boronic acid (644 mg, 4.00 mmol), (1E,4E)-1,5-diphenylpenta-1,4-dien-3-one-palladium (3:2) (92 mg, 0.10 mmol), dicyclohexyl(2′,6′-dimethoxybiphenyl-2-yl)phosphine (164 mg, 0.400 mmol), and K3PO4 (1.70 g, 8.00 mmol) in DMF (13 mL) was stirred for 16 hours at 110° C. under argon atmosphere. The precipitate formed was removed by filtration with celite and washed with CHCl3. The filtrate was concentrated in vacuo. The residue was purified by silica gel chromatography (CHCl3:MeOH=99:1-95:5) to give 6′-[5-(prop-1-yn-1-yl)pyridin-3-yl]dispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-amine (310 mg).
Reference Example 114
A mixture of 6′-bromo-5H-dispiro[1,4-oxazinane-3,4′-chromene-3′,3′′-oxetane]-5-thione (200 mg, 0.561 mmol) and ammonia (2 M in EtOH, 6.0 mL) was stirred for 1 week at ambient temperature. The reaction mixture was concentrated at reduced pressure, and the residue was purified with column chromatography on amino silica gel (CHCl3-EtOAc, a linear gradient of EtOAc from 0 to 50% then CHCl3-EtOH, a linear gradient of EtOH from 0 to 20%) to afford 6′-bromo-6H-dispiro[1,4-oxazine-3,4′-chromene-3′,3′′-oxetan]-5-amine (123 mg).
Reference Example 115
A mixture of di-tert-butyl (6′-bromotrispiro[cyclobutane-1,2′-chromene-4′,4′′-[1,3]oxazole-3′,3′′′-oxetan]-2′′-yl)imidodicarbonate (300 mg, 0.531 mmol), 4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi-1,3,2-dioxaborolane (150 mg, 0.584 mmol), potassium acetate (91.1 mg, 0.928 mmol) and bis(triphenylphosphine)palladium(II) dichloride (37 mg, 0.053 mmol) in dioxane (2.4 mL) was stirred for 7 hours at 100° C. To the mixture was added 3-bromo-2-cyanopyridine (243 mg, 1.33 mmol), Na2CO3 (225 mg, 2.12 mmol) and water (0.60 mL), and the mixture was stirred for 2 hours at 100° C. The mixture was treated with charcoal, and filtered off. The filtrate was partitioned between EtOAc and water. The organic layer was washed with brine, dried over MgSO4, filtered, and the filtrate was evaporated to give a pale brown oil. The oil was dissolved in toluene (3.0 mL) and to the mixture was added silica gel (neutral; 900 mg). The mixture was refluxed for 1 hour. The solvent was evaporated off. Silica gel column chromatography (CHCl3-EtOH, a linear gradient of EtOH from 0 to 10%) afforded a solid. The solid was triturated in Et2O, collected by filtration, washed with Et2O and dried at reduced pressure at 70° C. to give 3-(2′′-aminotrispiro[cyclobutane-1,2′-chromene-4′,4′′-[1,3]oxazole-3′,3′′′-oxetan]-6′-yl)pyridine-2-carbonitrile (141 mg).
Reference Example 116
A mixture of 3-bromo-2-fluoro-5-methylpyridine (202 mg, 1.06 mmol), 4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi-1,3,2-dioxaborolane (300 mg, 1.17 mmol), potassium acetate (208 mg, 2.12 mmol) and PdCl2(PPh3)2 (37 mg, 0.053 mmol) in dioxane (4 mL) was stirred for 3 hours at 100° C. To the mixture was added di-tert-butyl (6′-bromotrispiro[cyclobutane-1,2′-chromene-4′,4′′-[1,3]oxazole-3′,3′′′-oxetan]-2′′-yl)imidodicarbonate (300 mg, 0.531 mmol), sodium carbonate (225 mg, 2.12 mmol) and H2O (1 mL), and the mixture was stirred for 1.5 hours at 100° C. The mixture was treated with charcoal, and filtered off. The filtrate was partitioned between EtOAc and water. The organic layer was washed with brine, dried over MgSO4, filtered, and the filtrate was evaporated to give a pale brown oil. The oil was dissolved in toluene (5 mL) and to the mixture was added silicagel (neutral; 600 mg). The mixture was refluxed for 1 hour. The solvent was evaporated off. Silicagel column chromatography (CHCl3-EtOH, linear gradient of EtOH from 0 to 20%) afforded a solid. The solid was triturated in Et2O and collected by filtration, washed with Et2O and dried in vacuo at 70° C. to give 6′-(2-fluoro-5-methylpyridin-3-yl)trispiro[cyclobutane-1,2′-chromene-4′,4′′-[1,3]oxazole-3′,3′′′-oxetan]-2′′-amine (130 mg).
Reference Example 136
A mixture of di-tert-butyl (6′-bromotrispiro[cyclobutane-1,2′-chromene-4′,4′′-[1,3]oxazole-3′,3′′′-oxetan]-2′′-yl)imidodicarbonate (14.1 mg), 2-fluoro-5-methoxyphenylboronic acid (12.7 mg), PdCl2(PPh3)2 (5.3 mg) and 1M aqueous Na2CO3 (0.1 mL) in dioxane (0.4 mL) was stirred for 2 hours at 100° C. The mixture was filtered by using Chem Elut cartridges (Agilent Technologies) and washed with CHCl3. The filtrate was evaporated to give a brown oil. The oil was dissolved in toluene (0.5 mL) and to the mixture was added silicagel (neutral; 50 mg). The mixture was stirred for 1 hour at 100° C. The mixture was filtered and washed with CHCl3. The filtrate was evaporated. The residue was purified with HPLC (Column: Waters SunFireTM Prep C18 OBDTM 5 micrometer, 19×100 mm; MeOH/0.1% aqueous HCOOH 11/89 to 95/5(v/v)) and afforded 6′-(2-fluoro-5-methoxyphenyl)trispiro[cyclobutane-1,2′-chromene-4′,4′′-[1,3]oxazole-3′,3′′′-oxetan]-2′′-amine (1.5 mg).
Reference Example 146
A mixture of di-tert-butyl (6′-bromo-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-yl)imidodicarbonate (13.8 mg), 3-chloro-5-fluorophenylboronic acid (8.7 mg), Pd(PPh3)4 (2.9 mg) and 1M aqueous Na2CO3 (0.063 mL) in dioxane (0.25 mL) was stirred for 12 hours at 100° C. The mixture was filtered by using Chem Elut cartridges and washed with CHCl3. The filtrate was evaporated. The residue was purified with HPLC (Column: Waters SunFireTM Prep C18 OBDTM 5 micrometer, 19×100 mm; MeOH/0.1% aqueous HCOOH 11/89 to 95/5(v/v)) and afforded 6′-(3-chloro-5-fluorophenyl)-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-amine (4.6 mg).
Reference Example 174
A mixture of di-tert-butyl (6′-bromo-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-yl)imidodicarbonate (13.8 mg), indole-5-boronic acid (8.1 mg), PdCl2(dppf) (2.0 mg) and 1M aqueous K2CO3 (0.063 mL) in dioxane (0.25 mL) was stirred for 12 hours at 100° C. The mixture was filtered by using Chem Elut cartridges and washed with CHCl3. The filtrate was evaporated. The residue was purified with HPLC (Column: Waters SunFireTM Prep C18 OBDTM 5 micrometer, 19×100 mm; MeOH/0.1% aqueous HCOOH 11/89 to 95/5(v/v)) and afforded 6′-(1H-indol-5-yl)-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-amine (4.5 mg).
Reference Example 198
To a solution of N-[5′′,5′′-difluoro-6′-(pyrimidin-5-yl)-5′′,6′′-dihydrodispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]thiazin]-2′′-yl]benzamide (76 mg, 0.16 mmol) in ethanol (4 mL) were added N-hydroxymethanamine hydrochloride (133 mg, 1.59 mmol) and pyridine (126 mg, 1.59 mmol), and the mixture was stirred for 27 hours at 70° C. After cooling, H2O and brine were added and the mixture was extracted with chloroform. The organic layer was dried over MgSO4 and concentrated. The residue was purified by silica gel chromatography (EtOAc/hexane=50:50-100:0 and then EtOH/CHCl3=5:95) and a resultant solid was washed with iPr2O to give 5′′,5′′-difluoro-6′-(pyrimidin-5-yl)-5′′,6′′-dihydrodispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]thiazin]-2′′-amine (36 mg).
Reference Example 214
A suspension of 6′-bromo-5′′H-dispiro[cyclopropane-1,3′-chromene-4′,3′′-[1,4]oxazinane]-5′′-thione (391 mg, 1.15 mmol) in 2M solution of ammonia in ethanol (60 mL, 120 mmol) was treated with tert-butylhydroperoxide (5.0-6.0 M solution in decane, 4.60 mL). The mixture was stirred at room temperature for 1 hour, followed by the addition of methanol (10 mL). After stirring at room temperature for 5 hours, insoluble material was filtered off. The filtrate was poured into saturated aqueous sodium hydrogen carbonate and extracted with chloroform. The organic layer was washed three times with saturated aqueous sodium hydrogen carbonate, dried over sodium sulfate, and evaporated. The residue was purified by silica gel chromatography (NH-silica gel, hexane:EtOAc=100:0-0:100) to give 6′-bromo-6′′H-dispiro[cyclopropane-1,3′-chromene-4′,3′′-[1,4]oxazin]-5′′-amine (247 mg).
Reference Example 215
In the same manner as in the method of Preparation Example 83 and Reference Example 31, 6′-(pyrimidin-5-yl)-6′′H-dispiro[cyclopropane-1,3′-chromene-4′,3′′-[1,4]oxazin]-5′′-amine was prepared with using di-tert-butyl (6′-bromo-6′′H-dispiro[cyclopropane-1,3′-chromene-4′,3′′-[1,4]oxazin]-5′′-yl)imidodicarbonate as a starting material.
Reference Example 216
To a solution of 1-[6-bromo-4-(1,1-difluoro-2-hydroxyethyl)-4H-spiro[chromene-3,1′-cyclopropan]-4-yl]thiourea (268 mg, 0.682 mmol) in MeOH (2.7 mL) were added methyliodide (967 mg, 6.82 mmol) and 1M aqueous NaOH (0.68 mL, 0.68 mmol). The mixture was stirred for 3 hours at 60° C. After cooling, to the mixture were added H2O and brine. The mixture was extracted with CH2Cl2. The extract was dried over MgSO4 and concentrated.
Purification using silicagel column chromatography (EtOAc-hexane, a linear gradient of EtOAc from 1 to 100%) afforded a solid (177 mg). To a solution of the solid in MeOH (2.7 mL) was added 1M aqueous NaOH (0.68 mL, 0.68 mmol). The mixture was stirred for 5 hours at 60° C. After cooling, to the mixture were added H2O and brine. The mixture was extracted with EtOAc. The extract was dried over MgSO4 and concentrated. Purification using silicagel column chromatography (EtOAc-hexane, a linear gradient of EtOAc from 1 to 100%) afforded 6′-bromo-5′′,5′′-difluoro-5′′,6′′-dihydrodispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazin]-2′′-amine (145 mg).
Reference Example 217
To a solution of 6-bromo-2,2-dimethyl-4-methylene-4H-spiro[chromene-3,1′-cyclopropane] (6.72 g, 24 mmol) in ethyl acetate (67.2 ml) and acetonitrile (67.2 ml) was added silver cyanate (5.4 g, 36 mmol) under cooling with an ice bath under an argon atmosphere. To the mixture was added iodine (9.17 g, 36 mmol). The mixture was stirred at 0° C. for 2 hours and filtered. The cake was washed with ethyl acetate and the filtrate was partitioned between ethyl acetate and saturated aqueous sodium thiosulfate. The organic layer was washed with brine, dried over anhydrous magnesium sulfate, and concentrated in vacuo.
The residue was dissolved in tetrahydrofuran (69 mL) and the solution was added to 2 M ethanolic ammonia (151 mL, 302 mmol) under cooling with an ice bath. The mixture was stirred at room temperature overnight, and concentrated in vacuo.
The residue was dissolved in MeOH (30 ml) and saturated aqueous sodium bicarbonate was added. The mixture was stirred at room temperature for 1 hour and the resulting precipitate was collected and dried in vacuo. The residue was triturated with a mixture of ethyl acetate/diisopropyl ether and filtered to afford 6′-bromo-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-2′′-amine (8.12 g).
Example 218
A mixture of tert-butyl[(4S)-6′-{[(5-chloropyridin-2-yl)carbonyl]amino}-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-yl]carbamate (1.02 g, 1.93 mmol), silica gel (neutral; 3.06 g) and toluene (20.4 mL) was stirred for 3 hours at 120° C. The reaction mixture was cooled down to ambient temperature and concentrated at reduced pressure. The residue was purified with column chromatography on silica gel (CHCl3-EtOH, a linear gradient of EtOH from 0 to 15%) and then with NH-silica gel (Hexane-EtOAc, a linear gradient of EtOAc from 50 to 100%). The purified product was recrystallized from EtOH/H2O (1:1), and the solid was collected by filtration and dried at reduced pressure to give a hydrate of N-[(4S)-2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-6′-yl]-5-chloropyridine-2-carboxamide (547 mg) as a crystal.
Example 223
To a solution of tert-butyl[(4′R)-6′-{[(5-methoxypyrazin-2-yl)carbonyl]amino}-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-2′′-yl]carbamate (392 mg, 0.769 mmol) in chloroform (3 ml) was added trifluoroacetic acid (1.8 ml), and the mixture was stirred at room temperature for 2 hours. The mixture was concentrated in vacuo, and the residue was purified by silica gel column chromatography (precolumn: NH-silica gel, main column: neutral silica gel, chloroform/methanol=10:0-10:1). To the purified product was added a mixture of hexane/ethyl acetate (4:1) and the mixture was stirred at room temperature over night. The precipitate was collected, washed with mixture of hexane/ethyl acetate (4:1), and dried in vacuo to afford N-[(4′R)-2′′-amino-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-6′-yl]-5-me thoxypyrazine-2-carboxamide (246 mg).
Example 224
To a solution of tert-butyl[(4′R)-6′-({[5-(difluoromethyl)pyrazin-2-yl]carbonyl}amino)-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-2′′-yl]carbamate (373 mg, 0.704 mmol) in chloroform (6 ml) was added trifluoroacetic acid (2 ml). The mixture was stirred at room temperature for 2 hours and concentrated in vacuo. The residue was purified by silica gel column chromatography (precolumn: NH-silica gel, main column: neutral silica gel, chloroform/methanol=100:0-10:1). To the purified product was added a mixture of hexane and ethyl acetate (4:1), and the mixture was stirred at room temperature over night. The resulting precipitate was collected, washed with a mixture of hexane and ethyl acetate (4:1), and dried in vacuo to afford N-[(4′R)-2′′-amino-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-6′-yl]-5-(difluoromethyl)pyrazine-2-carboxamide (253 mg).
Reference Example 225a,b
To a solution of 6′-bromo-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-2′′-amine (3.7 g, 11 mmol) in methanol (50 ml) was added (+)-dibenzoyl-D-tartaric acid monohydrate (4.1 g, 11 mmol). The mixture was stirred at room temperature for 5 minutes and concentrated in vacuo. To the residue was added dioxane (25 ml). The mixture was heated under reflux for 5 minutes, cooled to room temperature and stirred at room temperature overnight. The resulting precipitate was collected, washed with dioxane, and dried in vacuo. The resulting powder was dissolved in saturated aqueous sodium hydrogen carbonate and chloroform. The mixture was extracted with chloroform, and the organic layer was dried over anhydrous magnesium sulfate, and concentrated in vacuo to afford (4′R)-6′-bromo-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-2′′-amine (1.5 g). The mother liquor was concentrated in vacuo and purified by NH-silica gel column chromatography (CHCl3/MeOH=20:1). The purified material was treated with (−)-dibenzoyl-L-tartaric acid monohydrate (2.0 g, 5.4 mmol) in the same manner as described above which lead to isolation of (4′S)-6′-bromo-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-2′′-amine (1.2 g).
Reference Example 226
A mixture of racemic 6′-bromo-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-amine (155 g, 0.44 mol) and L-camphorsulfonic acid (102 g, 0.44 mol) in ethanol (2.7 L) and water (340 mL) was heated at 50° C. till a clear solution was formed. The mixture was allowed to cool to room temperature and stood for 48 hours. The precipitate was collected by filtration, washed with ethanol and dried under reduced pressure to afford a salt with L-camphorsulfonic acid (65.0 g). The salt was dissolved in water (500 mL) and 10% aqueous Na2CO3 (400 mL) was added. The mixture was stirred for 1 hour and extracted with dichloromethane twice. The combined extracts were washed with brine (40 mL), dried over Na2SO4 and concentrated under reduced pressure to provide (4S)-6′-bromo-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-amine (38.0 g).
Example 228a,b
N-(2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-6′-yl)-5-chloro pyridine-2-carboxamide (352 mg, 0.821 mmol) was subjected to chromatography using supercritical CO2 (supercritical CO2/[EtOH with 0.1% diethylamine]=60:40) on Chiralcel OD-H column (10×250 mm) eluting at a flow rate 10 mL/minute (40° C. column temperature). After concentration of collected fractions of the first peak (retention time=5.23 minutes) at reduced pressure, recrystallization of the residue with EtOH/water (1:1) provided a hydrate of N-[(4R)-2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-6′-yl]-5-chloropyridine-2-carboxamide (153 mg, 44%) as a crystal. After concentration of collected fractions of the second peak (retention time=8.16 minutes) at reduced pressure, recrystallization of the residue with EtOH/water (1:1) provided a hydrate of N-[(4S)-2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-6′-yl]-5-chloropyridine-2-carboxamide (152 mg) as a crystal.
Example 229a,b
N-[2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-6′-yl]-5-methoxypyrazine-2-carboxamide (100 mg, 0.235 mmol) was chromatographed using supercritical CO2 (supercritical CO2/EtOH=60:40) on Chiralcel OD-H column (10×250 mm) eluting at a flow rate 10 mL/minute (40° C. column temperature). After concentration of collected fractions of the first peak (retention time=5.25 minutes) under reduced pressure, trituration of the residue with EtOAc/hexane provided N-[(4S)-2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-6′-yl]-5-methoxypyrazine-2-carboxamide (35 mg). After concentration of collected fractions of the second peak (retention time=8.08 minutes) under reduced pressure, trituration of the residue with EtOAc/hexane provided N-[(4R)-2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-6′-yl]-5-methoxypyrazine-2-carboxamide (35 mg).
Example 230
To a solution of tert-butyl[(4′R)-6′-{[(5-methoxypyrazin-2-yl)carbonyl]amino}-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-2′′-yl]carbamate (13.75 g, 26.98 mmol) in chloroform (140 ml) was added trifluoroacetic acid (68 ml) in an ice-water bath, and the mixture was stirred at room temperature for 3 hours. The mixture was concentrated in vacuo, and the residue was purified by silica gel column chromatography (pre-column: NH-silica gel; main column: neutral silica gel, chloroform/methanol=100:0-10:1). To the purified product was added saturated aqueous sodium hydrogen carbonate and extracted with chloroform. The organic layer were washed with brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated in vacuo. The residue was purified by silica gel column chromatography (pre-column: basic silica gel; main column: neutral silica gel, chloroform/methanol=100:0-10:1). The purified product was triturated with a mixture of hexane/ethyl acetate (4:1) (300 mL), and the mixture was stirred at 60° C. for 1 hour and room temperature for 4 days. The precipitate was collected, washed with mixture of hexane/ethyl acetate (4:1) (200 mL), and dried in vacuo at 50° C. to afford N-[(4′R)-2′′-amino-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-6′-yl]-5-me thoxypyrazine-2-carboxamide (8.38 g) as a crystal.
Example 231
To a solution of N-[(4′R)-2′′-amino-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-6′-yl]-5-(difluoromethyl)pyrazine-2-carboxamide (800 mg, 1.86 mmol) in MeOH (10 mL) was added 4M solution of hydrogen chloride in ethyl acetate (0.5 mL, 2 mmol) and the mixture was concentrated in vacuo. The residue was triturated with EtOH (10 mL), and the mixture was refluxed for 30 minutes and stirred at room temperature overnight. The precipitate was collected, washed with EtOH (2 mL), and dried under reduced pressure at 70° C. overnight to afford N-[(4′R)-2′′-amino-2′,2′-dimethyldispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-6′-yl]-5-(difluoromethyl)pyrazine-2-carboxamide hydrochloride (449 mg) as a crystal.
The compounds of Examples and Reference Examples shown in Tables below were prepared using the respective corresponding starting materials in the same manner as the methods of Examples or Reference Examples above. The structures and the preparation methods are shown in [Table. 4] below, and the physicochemical data for the compounds of Examples or Reference Examples are shown in [Table. 5] below.









TABLE 4





Ex
Syn
Structure







RP 1a
RP 1a,b






 


RP 1b
RP 1a,b






 


RP 2
RP6






 


RP 3
RP6






 


RP 4
RP6






 


RP 5
RP6






 


RP 6
RP6






 


RP 7a
RP 1a,b






 


RP 7b
RP 1a,b






 


RP 8
RP6






 


RP 9a
RP 1a,b






 


RP 9b
RP 1a,b






 


RP 10
RP6






 


RP 11a
RP 1a,b






 


RP 11b
RP 1a,b






 


RP 12a
RP 1a,b






 


RP 12b
RP 1a,b






 


RP 13a
RP 1a,b






 


RP 13b
RP 1a,b






 


RP 14a
RP 1a,b






 


RP 14b
RP 1a,b






 


RP 15a
RP 1a,b






 


RP 15b
RP 1a,b






 


RP 16a
RP 1a,b






 


RP 16b
RP 1a,b






 


RP 17
RP6






 


RP 18
RP6






 


RP 19
RP19






 


RP 20
RP19






 


RP 21
RP19






 


RP 22
RP19






 


RP 23
RP19






 


RP 24
RP19






 


RP 25
RP19






 


RP 26
RP19






 


27
E27






 


28
E27






 


RP 29
E27






 


30
E27






 


RP 31
RP31






 


RP 32
RP31






 


RP 33
RP39






 


RP 34
RP39






 


RP 35
RP39






 


RP 36
RP39






 


RP 37
RP39






 


RP 38
RP39






 


RP 39
RP39






 


RP 40
RP40






 


RP 41
RP41






 


RP 42
RP41






 


RP 43
RP43






 


RP 44
RP43






 


RP 45
RP43






 


RP 46
RP46






 


RP 47
RP46






 


RP 48
RP48






 


RP 49
RP49






 


RP 50
RP49






 


RP 51a
RP 51a,b






 


RP 51b
RP 51a,b






 


RP 52a
RP 52a,b






 


RP 52b
RP 52a,b






 


RP 53a
RP 53a,b






 


RP 53b
RP 53a,b






 


RP 54
RP 54






 


RP 55
RP 55






 


RP 56
RP 61






 


RP 57
RP 61






 


RP 58
RP 61






 


RP 59
RP 61






 


RP 60
RP 61






 


RP 61
RP 61






 


RP 62
RP 61






 


RP 63
RP 61






 


RP 64
RP 61






 


RP 65
RP 65






 


RP 66
RP 65






 


RP 67
RP 65






 


RP 68
RP 19






 


RP 69
RP75






 


RP 70
RP75






 


RP 71
RP75






 


RP 72
RP75






 


RP 73
RP75






 


RP 74
RP75






 


RP 75
RP75






 


RP 76
RP75






 


RP 77
RP75






 


RP 78
RP75






 


RP 79
RP75






 


RP 80
RP75






 


RP 81
RP75






 


RP 82
RP75






 


RP 83
RP75






 


RP 84
RP75






 


RP 85
RP75






 


RP 86
RP75






 


RP 87
RP75






 


RP 88
RP75






 


RP 89
RP75






 


RP 90
RP75






 


RP 91
RP75






 


RP 92
RP75






 


RP 93
RP75






 


RP 94
RP75






 


RP 95
RP75






 


RP 96
RP75






 


RP 97
RP75






 


RP 98
RP75






 


RP 99
RP75






 


RP 100
RP75






 


RP 101
RP75






 


RP 102
RP75






 


RP 103
RP75






 


RP 104
RP75






 


RP 105
RP75






 


RP 106
RP75






 


RP 107
RP75






 


RP 108
RP108






 


RP 109
RP108






 


RP 110
RP108






 


RP 111
RP108






 


RP 112
RP108






 


RP 113
RP108






 


RP 114
RP114






 


RP 115
RP115






 


RP 116
RP116






 


RP 117
RP116






 


RP 118
RP116






 


RP 119
RP136






 


RP 120
RP136






 


RP 121
RP136






 


RP 122
RP136






 


RP 123
RP136






 


RP 124
RP136






 


RP 125
RP136






 


RP 126
RP136






 


RP 127
RP136






 


RP 128
RP136






 


RP 129
RP136






 


RP 130
RP136






 


RP 131
RP136






 


RP 132
RP136






 


RP 133
RP136






 


RP 134
RP136






 


RP 135
RP136






 


RP 136
RP136






 


RP 137
RP136






 


RP 138
RP136






 


RP 139
RP146






 


RP 140
RP146






 


RP 141
RP146






 


RP 142
RP146






 


RP 143
RP146






 


RP 144
RP146






 


RP 145
RP146






 


RP 146
RP146






 


RP 147
RP146






 


RP 148
RP146






 


RP 149
RP146






 


RP 150
RP146






 


RP 151
RP146






 


RP 152
RP146






 


RP 153
RP146






 


RP 154
RP146






 


RP 155
RP146






 


RP 156
RP146






 


RP 157
RP146






 


RP 158
RP146






 


RP 159
RP146






 


RP 160
RP146






 


RP 161
RP146






 


RP 162
RP146






 


RP 163
RP146






 


RP 164
RP146






 


RP 165
RP146






 


RP 166
RP146






 


RP 167
RP146






 


RP 168
RP146






 


RP 169
RP174






 


RP 170
RP174






 


RP 171
RP174






 


RP 172
RP174






 


RP 173
RP174






 


RP 174
RP174






 


RP 175
RP174






 


RP 176
RP174






 


RP 177
RP174






 


RP 178
RP174






 


RP 179
RP174






 


RP 180
RP174






 


RP 181
RP174






 


RP 182
RP174






 


RP 183
RP174






 


RP 184
RP174






 


RP 185
RP174






 


RP 186
RP174






 


RP 187
RP174






 


RP 188
RP6






 


RP 189
E27






 


190
E27






 


RP 191
E27






 


RP 192
E27






 


RP 193
E27






 


RP 194
E27






 


195
E27






 


RP 196
E27






 


197
E27






 


RP 198
RP198






 


RP 199
RP55






 


RP 200
RP75






 


RP 201
RP75






 


RP 202
RP75






 


RP 203
RP75






 


RP 204
RP75






 


RP 205
RP75






 


RP 206
RP75






 


RP 207
RP75






 


RP 208
RP75






 


RP 209
RP75






 


RP 210
RP75






 


RP 211
RP75






 


RP 212
RP75






 


RP 213
RP75






 


RP 214
RP214






 


RP 215
RP215






 


RP 216
RP216






 


RP 217
RP217






 


218
E218






 


219
E218






 


220
E218






 


221
E223






 


222
E223






 


223
E223






 


224
E224






 


RP 225a
RP225 a,b






 


RP 225b
RP225 a,b






 


RP 226
RP226






 


227
E218






 


228a
E228a, b






 


228b
E228a, b






 


229a
E229a, b






 


229b
E229a, b






 


230
E230






 


231
E231





















TABLE 5





Ex
Data







RP
ESI+: 339, 341


1a
less polar diastereomer


RP
ESI+: 339, 341


1b
polar diastereomer


RP2
ESI+: 325, 327


RP3
ESI+: 365, 367


RP4
ESI+: 397


RP5
ESI+: 365


RP6
ESI+: 353, 355


RP
ESI+: 339, 341


7a


RP
ESI+: 339, 341


7b


RP8
ESI+: 323, 325


RP
ESI+: 353, 355


9a
a diastereomer with higher Rf value on TLC (CHCl3/EtOAc 9:1, NH silicagel)


RP
ESI+: 353, 355


9b
a diastereomer with lower Rf value on TLC (CHCl3/EtOAc 9:1, NH silicagel)


RP
ESI+: 379, 381


10


RP
ESI+: 367, 369


11a
a diastereomer with higher Rf value on TLC (CHCl3/EtOAc 9:1, NH silicagel)


RP
ESI+: 367, 369


11b
a diastereomer with lower Rf value on TLC (CHCl3/EtOAc 9:1, NH silicagel)


RP
ESI+: 365, 367


12a
a diastereomer with higher Rf value on TLC (CHCl3/EtOAc 9:1, NH silicagel)


RP
ESI+: 365, 367


12b
a diastereomer with lower Rf value on TLC (CHCl3/EtOAc 9:1, NH silicagel)


RP
ESI+: 381, 383


13a
a diastereomer with higher Rf value on TLC (CHCl3/EtOAc 9:1, NH silicagel)


RP
ESI+: 381, 383


13b
a diastereomer with lower Rf value on TLC (CHCl3/EtOAc 9:1, NH silicagel)


RP
ESI+: 421, 423


14a
a diastereomer with higher Rf value on TLC (CHCl3/EtOAc 9:1, NH silicagel)


RP
ESI+: 421, 423


14b
a diastereomer with lower Rf value on TLC (CHCl3/EtOAc 9:1, NH silicagel)


RP
ESI+: 415, 417


15a
a diastereomer with higher Rf value on TLC (CHCl3/EtOAc 9:1, NH silicagel)


RP
ESI+: 415, 417


15b
NMR-CDCl3: 3.43 (1H, dd, J = 9.0, 14.7 Hz), 3.53 (1H, dd, J = 3.1, 14.7 Hz), 3.99 (1H, d,



J = 8.5 Hz), 4.10-4.18 (2H, m), 4.47-4.55 (4H, m), 4.64 (1H, d, J = 6.7 Hz), 4.81 (1H, d, J =



6.3 Hz), 6.63 (1H, d, J = 8.7 Hz), 7.21 (1H, dd, J = 2.4, 8.7 Hz), 7.25-7.41 (6H, m)



a diastereomer with lower Rf value on TLC (CHCl3/EtOAc 9:1, NH silicagel)


RP
ESI+: 379, 381


16a
a diastereomer with higher Rf value on TLC (CHCl3/EtOAc 9:1, NH silicagel)


RP
ESI+: 379, 381


16b
a diastereomer with lower Rf value on TLC (CHCl3/EtOAc 9:1, NH silicagel)


RP
ESI+: 309, 311


17


RP
ESI+: 327, 329


18


RP
ESI+: 392


19
NMR-DMSO-d6: 3.37 (3H, s), 4.16-4.28 (5H, m), 4.37-4.39 (3H, m), 4.60 (1H, d, J = 5.4 Hz),



4.71 (1H, d, J = 11.4 Hz), 6.40 (2H, s), 6.88 (1H, d, J = 8.4 Hz), 7.48 (1H, d, J = 2.3 Hz),



7.54 (1H, dd, J = 8.5, 2.4 Hz), 8.02-8.03 (1H, m), 8.60 (1H, d, J = 1.9 Hz), 8.76 (1H,



d, J = 2.3 Hz)


RP
ESI+: 362


20
NMR-DMSO-d6: 2.10 (3H, s), 4.17 (2H, m), 4.23-4.28 (3H, m), 4.38 (1H, d, J = 6.4 Hz),



4.60 (1H, d, J = 5.3 Hz), 4.73 (1H, d, J = 11.4 Hz), 6.44 (2H, s), 6.88 (1H, d, J = 8.5 Hz),



7.51 (1H, dd, J = 5.3, 1.8 Hz), 7.55 (1H, d, J = 2.3 Hz), 7.58-7.61 (2H, m), 8.52 (1H, d, J =



5.2 Hz)


RP
ESI+: 363


21
NMR-DMSO-d6: 2.16 (3H, s), 4.10-4.17 (2H, m), 4.23-4.30 (3H, m), 4.38 (1H, d, J = 6.4 Hz),



4.59 (1H, d, J = 5.4 Hz), 4.75 (1H, d, J = 11.5 Hz), 6.46 (2H, s), 6.89 (1H, d, J = 8.6 Hz),



7.97-8.00 (2H, m), 8.17 (1H, d, J = 2.3 Hz), 9.09 (1H, d, J = 1.2 Hz)


RP
ESI+: 363


22


RP
ESI+: 348


23
NMR-DMSO-d6: 4.16-4.28 (5H, m), 4.38 (1H, d, J = 6.4 Hz), 4.49 (1H, s), 4.60 (1H, d, J =



5.4 Hz), 4.71 (1H, d, J = 11.4 Hz), 6.40 (2H, s), 6.88 (1H, d, J = 8.5 Hz), 7.48 (1H, d, J =



2.3 Hz), 7.53 (1H, dd, J = 8.4, 2.4 Hz), 8.04-8.05 (1H, m), 8.61 (1H, d, J = 1.9 Hz), 8.78



(1H, d, J = 2.3 Hz)


RP
ESI+: 329


24


RP
ESI+: 339


25


RP
ESI+: 369


26
NMR-DMSO-d6: 3.85 (3H, s), 4.07 (1H, d, J = 8.8 Hz), 4.13-4.16 (2H, m), 4.21-4.24 (2H,



m), 4.34 (1H, d, J = 6.3 Hz), 4.56-4.61 (2H, m), 6.31 (2H, s), 6.63 (1H, d, J = 8.8 Hz),



6.67 (1H, dd, J = 7.8, 5.0 Hz), 7.13 (1H, dd, J = 7.9, 1.4 Hz), 7.56 (1H, d, J = 2.6 Hz),



7.61-7.64 (2H, m), 7.97 (1H, s)


27
ESI+: 429, 431



NMR-DMSO-d6: 1.18 (3H, s), 1.73 (3H, s), 4.13-4.19 (2H, m), 4.30-4.32 (2H, m), 4.36



(1H, d, J = 6.8 Hz), 4.90 (1H, d, J = 5.6 Hz), 6.33 (2H, s), 6.66 (1H, d, J = 8.8 Hz), 7.57



(1H, dd, J = 8.8, 2.7 Hz), 7.70 (1H, d, J = 2.5 Hz), 8.13 (1H, dd, J = 8.5, 0.7 Hz), 8,18 (1H,



dd, J = 8.4, 2.4 Hz), 8.75 (1H, dd, J = 2.3, 0.8 Hz), 10.52 (1H, s)


28
ESI+: 413



NMR-DMSO-d6: 1.18 (3H, s), 1.73 (3H, s), 4.13-4.19 (2H, m), 4.30-4.32 (2H, m), 4.36



(1H, d, J = 6.8 Hz), 4.90 (1H, d, J = 5.6 Hz), 6.32 (2H, s), 6.66 (1H, d, J = 8.8 Hz), 7.55



(1H, dd, J = 8.8, 2.6 Hz), 7.70 (1H, d, J = 2.5 Hz), 7.96 (1H, td, J = 8.7, 2.9 Hz), 8.20 (1H,



dd, J = 8.7, 4.6 Hz), 8.70 (1H, d, J = 2.8 Hz), 10.46 (1H, s)


RP
ESI+: 403, 405


29
NMR-DMSO-d6: 0.37-0.43 (3H, m), 0.82-0.85 (1H, m), 3.58 (1H, d, J = 11.6 Hz), 4.09



(1H, d, J = 8.1 Hz), 4.31 (1H, d, J = 8.2 Hz), 4.34-4.38 (1H, m), 6.18 (2H, s), 6.73-6.75



(1H, m), 7.55-7.57 (2H, m), 8.29 (1H, dd, J = 10.2, 1.9 Hz), 8.62-8.63 (1H, m), 10.47 (1H,



s)


30
ESI+: 447, 449



NMR-DMSO-d6: 1.18 (3H, s), 1.73 (3H, s), 4.13-4.18 (2H, m), 4.31 (2H, d, J = 6.0 Hz),



4.36 (1H, d, J = 6.8 Hz), 4.89 (1H, d, J = 5.6 Hz), 6.34 (2H, s), 6.65-6.68 (1H, m),



7.52-7.55 (2H, m), 8.29 (1H, dd, J = 10.2, 2.0 Hz), 8.62-8.63 (1H, m), 10.51 (1H, s)


RP
ESI+: 369


31
NMR-DMSO-d6: 0.36-0.43 (3H, m), 0.82-0.88 (1H, m), 3.58 (1H, d, J = 11.6 Hz), 4.09



(1H, d, J = 8.1 Hz), 4.30-4.37 (2H, m), 6.17 (2H, s), 6.73 (1H, d, J = 8.8 Hz), 7.58 (1H, dd,



J = 8.8, 2.6 Hz), 7.71 (1H, d, J = 2.6 Hz), 7.96 (1H, td, J = 8.7, 2.9 Hz), 8.18-8.21 (1H, m),



8.70 (1H, d, J = 2.9 Hz), 10.41 (1H, s)


RP
ESI+: 385, 387


32
NMR-DMSO-d6: 0.36-0.44 (3H, m), 0.82-0.88 (1H, m), 3.58 (1H, d, J = 11.6 Hz), 4.09



(1H, d, J = 8.1 Hz), 4.31 (1H, d, J = 8.1 Hz), 4.35 (1H, dd, J = 11.5, 1.4 Hz), 6.17 (2H, s),



6.73 (1H, d, J = 8.8 Hz), 7.59 (1H, dd, J = 8.8, 2.6 Hz), 7.71 (1H, d, J = 2.6 Hz), 8.12 (1H,



dd, J = 8.4, 0.7 Hz), 8.18 (1H, dd, J = 8.5, 2.3 Hz), 8.75 (1H, dd, J = 2.3, 0.7 Hz), 10.47



(1H, s)


RP
ESI+: 371


33


RP
ESI+: 376


34


RP
ESI+: 319


35


RP
ESI+: 325


36


RP
ESI+: 338


37


RP
ESI+: 365


38


RP
ESI+: 354


39


RP
ESI+: 373, 375


40


RP
ESI+: 341, 343


41


RP
ESI+: 369, 371


42


RP
ESI+: 373, 375


43
NMR-DMSO-d6: 4.05-4.36 (6H, m), 4.57-4.65 (2H, m), 6.32 (2H, brs), 6.67-6.75 (2H, m),



7.39-7.42 (2H, m), 7.70-7.72 (1H, m), 7.97-7.99 (1H, m), 8.26 (1H, s)


RP
ESI+: 413, 415


44
NMR-DMSO-d6: 1.82-1.88 (2H, m), 2.13-2.24 (2H, m), 2.32-2.43 (1H, m), 2.92-2.98 (1H,



m), 3.95-4.06 (1H, m), 4.16-4.21 (2H, m), 4.44-4.51 (2H, m), 4.62-4.66 (1H, m), 6.29



(2H, brs), 6.71-6.74 (2H, m), 7.31-7.38 (2H, m), 7.70-7.71 (1H, m), 7.98-7.99 (1H, m),



8.25 (1H, s)


RP
ESI+: 409


45
NMR-DMSO-d6: 1.80-1.88 (2H, m), 2.12-2.40 (3H, m), 2.89-2.97 (1H, m), 3.76-3.94 (4H,



m), 4.17-4.24 (2H, m), 4.49-4.64 (3H, m), 6.29 (2H, brs), 6.67-6.72 (2H, m), 7.13 (1H, d,



J = 7.6 Hz), 7.57-7.70 (3H, m), 8.00 (1H, brs)


RP
ESI+: 343


46


RP
APCI/ESI+: 371


47


RP
ESI+: 428


48
NMR-DMSO-d6: 0.77-0.82 (2H, m), 0.90-0.96 (2H, m), 1.57-1.64 (1H, m), 1.80-1.95 (2H,



m), 2.12-2.34 (2H, m), 2.38-2.50 (1H, m), 2.93-3.04 (1H, m), 4.11-4.21 (3H, m),



4.49-4.53 (2H, m), 4.65 (1H, d, J = 5.5 Hz), 6.39 (2H, brs), 6.91 (1H, d, J = 8.5 Hz), 7.27



(1H, d, J = 2.2 Hz), 7.50 (1H, dd, J = 8.5, 2.4 Hz), 7.86-7.87 (1H, m), 8.49 (1H, d, J = 1.9 Hz),



8.65 (1H, d, J = 2.3 Hz)


RP
ESI+: 421


49


RP
APCI/ESI+: 421


50


RP
ESI+: 353


51a
retention time = 3.44 minutes


RP
ESI+: 353


51b
NMR-DMSO-d6: 1.22 (3H, s), 1.77 (3H, s), 4.20 (1H, d, J = 8.9 Hz), 4.29-4.34 (3H, m),



4.39 (1H, d, J = 6.9 Hz), 4.91 (1H, d, J = 5.6 Hz), 6.35 (2H, s), 6.85 (1H, d, J = 8.5 Hz),



7.35 (1H, d, J = 2.4 Hz), 7.54 (1H, dd, J = 8.4, 2.4 Hz), 8.97 (2H, s), 9.14 (1H, s)



retention time = 6.92 minutes


RP
ESI+: 346


52a
NMR-DMSO-d6: 0.38-0.46 (3H, m), 0.81-0.88 (1H, m), 2.11 (3H, s), 3.65 (1H, d, J = 11.7 Hz),



4.21 (1H, d, J = 8.1 Hz), 4.34 (1H, d, J = 8.1 Hz), 4.41-4.45 (1H, m), 6.21 (2H, brs),



6.87 (1H, d, J = 8.5 Hz), 7.37-7.41 (1H, m), 7.50-7.53 (1H, m), 7.91-7.92 (1H, t, J = 2.1 Hz),



8.51 (1H, d, J = 2.1 Hz), 8.70 (1H, d, J = 2.1 Hz);



retention time = 3.62 minutes


RP
ESI+: 346


52b
retention time = 6.27 minutes


RP
ESI+: 362


53a
retention time = 5.81 minutes


RP
ESI+: 362


53b
NMR-DMSO-d6: 2.11 (3H, s), 4.15-4.21 (2H, m), 4.24-4.27 (3H, m), 4.38 (1H, d, J = 6.4 Hz),



4.60 (1H, d, J = 5.3 Hz), 4.71 (1H, d, J = 11.4 Hz), 6.41 (2H, s), 6.87 (1H, d, J = 8.5 Hz),



7.46 (1H, d, J = 2.3 Hz), 7.51 (1H, dd, J = 2.3, 8.5 Hz), 7.92-7.94 (1H, m), 8.52 (1H,



d, J = 1.9 Hz), 8.70 (1H, d, J = 2.2 Hz);



retention time = 9.25 minutes


RP
ESI+: 369


54


RP
ESI+: 363


55


RP
ESI+: 362


56
NMR-DMSO-d6: 2.12 (3H, s), 4.10-4.17 (2H, m), 4.24-4.27 (3H, m), 4.38 (1H, d, J = 6.4 Hz),



4.59-4.61 (1H, m), 4.71 (1H, d, J = 11.4 Hz), 6.43 (2H, s), 6.83 (1H, d, J = 8.6 Hz),



7.24 (1H, dd, J = 5.0, 1.4 Hz), 7.756-7.762 (1H, m), 7.83 (1H, dd, J = 8.6, 2.4 Hz), 7.99



(1H, d, J = 2.3 Hz), 8.58 (1H, dd, J = 5.1, 0.8 Hz)


RP
ESI+: 348


57
NMR-DMSO-d6: 0.42 (3H, brs), 0.82 (3H, brs), 1.02-1.07 (2H, m), 1.98-2.02 (1H, m),



3.65 (1H, d, J = 11.3 Hz), 4.20-4.22 (1H, m), 4.32-4.36 (1H, m), 4.43 (1H, d, J = 11.3 Hz),



6.22 (2H, brs), 6.86 (1H, d, J = 8.4 Hz), 7.36 (1H, brs), 7.46-7.50 (2H, m), 8.32 (1H, s),



8.50 (1H, s)


RP
ESI+: 406


58
NMR-DMSO-d6: 0.41-0.46 (3H, m), 0.82-0.86 (1H, m), 3.65 (1H, d, J = 11.6 Hz), 4.21



(1H, d, J = 8.1 Hz), 4.34 (1H, d, J = 8.1 Hz), 4.44 (1H, d, J = 11.6 Hz), 4.92-4.99 (2H, m),



6.21 (2H, brs), 6.89 (1H, d, J = 8.5 Hz), 7.41-7.43 (1H, m), 7.51-7.54 (1H, m), 7.64-7.66



(1H, m), 8.34 (1H, d, J = 2.6 Hz), 8.43-8.46 (1H, m)


RP
ESI+: 388


59
NMR-DMSO-d6: 0.41-0.46 (3H, m), 0.82-0.88 (1H, m), 3.66 (1H, d, J = 11.7 Hz), 4.20



(1H, d, J = 8.16 Hz), 4.33 (1H, d, J = 8.16 Hz), 4.42-4.54 (3H, m), 6.20 (2H, brs),



6.29-6.58 (1H, m), 6.88 (1H, d, J = 8.5 Hz), 7.40-7.41 (1H, m), 7.51-7.53 (1H, m),



7.57-7.58 (1H, m), 8.29-8.30 (1H, m), 8.40-8.41 (1H, m)


RP
ESI+: 374


60
NMR-DMSO-d6: 0.40-0.45 (3H, m), 0.82-0.85 (1H, m), 3.66 (1H, d, J = 11.6 Hz), 4.20



(1H, d, J = 8.1 Hz), 4.34 (1H, d, J = 8.1 Hz), 4.44 (1H, d, J = 11.6 Hz), 6.21 (2H, brs), 6.90



(1H, d, J = 8.4 Hz), 7.21-7.58 (3H, m), 7.77-7.79 (1H, m), 8.42 (1H, d, J = 2.6 Hz), 8.66



(1H, d, J = 1.9 Hz)


RP
ESI+: 347


61
NMR-DMSO-d6: 0.41-0.46 (3H, m), 0.84-0.87 (1H, m), 3.62 (1H, d, J = 11.7 Hz), 4.21



(1H, d, J = 8.1 Hz), 4.34 (1H, d, J = 8.1 Hz), 4.42 (1H, d, J = 11.7 Hz), 6.20 (2H, brs),



6.48-6.50 (1H, m), 6.85 (1H, d, J = 8.4 Hz), 7.36-7.37 (1H, m), 7.43-7.46 (1H, m),



7.48-7.49 (1H, m), 8.03 (1H, d, J = 2.1 Hz), 8.37 (1H, d, J = 2.1 Hz), 11.6 (1H, brs)


RP
ESI+: 381, 383


62


RP
ESI+: 336


63


RP
ESI+: 438


64


RP
ESI+: 382


65


RP
ESI+: 417


66


RP
ESI+: 417


67


RP
ESI+: 378


68
NMR-DMSO-d6: 2.11 (3H, s), 3.34-3.40 (2H, m), 4.23-4.28 (2H, m), 4.43 (1H, d, J = 11.3 Hz),



4.54 (1H, d, J = 6.4 Hz), 4.63 (1H, d, J = 5.4 Hz), 4.74 (1H, d, J = 11.4 Hz), 6.87 (2H,



s), 6.90 (1H, d, J = 8.5 Hz), 7.42 (1H, d, J = 2.3 Hz), 7.53 (1H, dd, J = 8.6, 2.3 Hz), 7.87



(1H, t, J = 2.1 Hz), 8.52 (1H, d, J = 1.9 Hz), 8.66 (1H, d, J = 2.2 Hz)


RP
ESI+: 353


69
NMR-DMSO-d6: 1.22 (3H, s), 1.77 (3H, s), 4.20 (1H, d, J = 8.9 Hz), 4.29-4.35 (3H, m),



4.39 (1H, d, J = 6.9 Hz), 4.91 (1H, d, J = 5.6 Hz), 6.35 (2H, s), 6.85 (1H, d, J = 8.5 Hz),



7.35 (1H, d, J = 2.3 Hz), 7.54 (1H, dd, J = 8.6, 2.3 Hz), 8.97 (2H, s), 9.14 (1H, s)


RP
ESI+: 382


70


RP
ESI+: 382


71
NMR-DMSO-d6: 1.82-1.94 (2H, m), 2.14-2.33 (2H, m), 2.39-2.51 (1H, m), 2.95-3.03 (1H,



m), 4.06 (1H, d, J = 8.7 Hz), 4.17-4.20 (2H, m), 4.49-4.53 (2H, m), 4.65 (1H, d, J = 5.4 Hz),



6.38 (2H, s), 6.93 (1H, d, J = 8.5 Hz), 7.25-7.26 (1H, m), 7.37-7.40 (1H, m),



7.42-7.46 (1H, m), 7.96-8.01 (1H, m), 8.18-8.20 (1H, m)


RP
ESI+: 394


72
NMR-DMSO-d6: 1.81-1.94 (2H, m), 2.13-2.33 (2H, m), 2.38-2.51 (1H, m), 2.95-3.02 (1H,



m), 3.89 (3H, s), 4.10 (1H, d, J = 8.6 Hz), 4.17-4.20 (2H, m), 4.50-4.54 (2H, m), 4.65 (1H,



d, J = 5.4 Hz), 6.39 (2H, s), 6.92 (1H, d, J = 8.4 Hz), 7.28 (1H, d, J = 2.3 Hz), 7.43 (1H,



dd, J = 2.6, 2.0 Hz), 7.50 (1H, dd, J = 8.4, 2.3 Hz), 8.25 (1H, d, J = 2.7 Hz), 8.32 (1H, d, J =



1.8 Hz)


RP
ESI+: 416


73


RP
ESI+: 378


74


RP
ESI+: 378


75


RP
ESI+: 379


76


RP
ESI+: 356


77


RP
ESI+: 339


78


RP
ESI+: 356


79


RP
ESI+: 339


80


RP
ESI+: 368


81


RP
ESI+: 370


82
NMR-DMSO-d6: 1.22 (3H, s), 1.77 (3H, s), 4.18 (1H, d, J = 8.9 Hz), 4.24 (1H, d, J = 8.9 Hz),



4.32-4.33 (2H, m), 4.39 (1H, d, J = 6.9 Hz), 4.91 (1H, d, J = 5.6 Hz), 6.35 (2H, s),



6.82 (1H, d, J = 8.5 Hz), 7.27-7.28 (1H, m), 7.34-7.38 (1H, m), 7.44 (1H, ddd, J = 7.4, 4.9,



2.0 Hz), 7.99 (1H, ddd, J = 10.4, 7.5, 2.0 Hz), 8.18-8.20 (1H, m)


RP
ESI+: 382


83
NMR-DMSO-d6: 1.21 (3H, s), 1.77 (3H, s), 3.89 (3H, s), 4.18 (1H, d, J = 8.8 Hz),



4.28-4.33 (3H, m), 4.40 (1H, d, J = 6.9 Hz), 4.91 (1H, d, J = 5.6 Hz), 6.36 (2H, s), 6.81



(1H, d, J = 8.4 Hz), 7.30 (1H, d, J = 2.3 Hz), 7.44 (1H, dd, J = 2.7, 1.9 Hz), 7.47 (1H, dd, J =



8.5, 2.3 Hz), 8.25 (1H, d, J = 2.7 Hz), 8.32 (1H, d, J = 1.8 Hz)


RP
ESI+: 353


84
NMR-DMSO-d6: 1.12 (3H, t, J = 7.2 Hz), 1.59-1.74 (1H, m), 1.86-1.99 (1H, m), 4.20 (1H,



d, J = 6.9 Hz), 4.38 (1H, d, J = 6.3 Hz), 4.42 (1H, d, J = 6.9 Hz), 4.45-4.52 (1H, m),



4.54-4.68 (3H, m), 6.22 (2H, bs), 6.91 (1H, d, J = 8.4 Hz), 7.48 (1H, d, J = 2.3 Hz), 7.56



(1H, dd, J = 2.3, 8.4 Hz), 9.02 (2H, s), 9.13 (1H, s)



a compound prepared from Ex. 9a


RP
ESI+: 379


85
NMR-DMSO-d6: 1.51-1.55 (1H, m), 1.60-1.77 (4H, m), 1.87-1.96 (1H, m), 2.13-2.21 (1H,



m), 2.49-2.56 (1H, m), 4.19-4.24 (2H, m), 4.27-4.29 (2H, m), 4.43 (1H, d, J = 6.8 Hz),



4.83 (1H, d, J = 5.3 Hz), 6.36 (2H, s), 6.85 (1H, d, J = 8.5 Hz), 7.34 (1H, d, J = 2.3 Hz),



7.53 (1H, dd, J = 8.5, 2.4 Hz), 8.97 (2H, s), 9.14 (1H, s)


RP
ESI+: 390


86
NMR-DMSO-d6: 1.11 (3H, t, J = 7.1 Hz), 1.57-1.72 (1H, m), 1.84-1.97 (1H, m), 2.11 (3H,



s), 4.20 (1H, d, J = 6.9 Hz), 4.37 (1H, d, J = 6.3 Hz), 4.41 (1H, d, J = 6.9 Hz), 4.44-4.51



(1H, m), 4.52-4.66 (3H, m), 6.23 (2H, bs), 6.87 (1H, d, J = 8.5 Hz), 7.41 (1H, d, J = 2.4 Hz),



7.50 (1H, dd, J = 2.4, 8.5 Hz), 7.93-7.97 (1H, m), 8.52 (1H, d, J = 1.9 Hz), 8.71 (1H,



d, J = 2.3 Hz)



a compound prepared from Reference Example 9a


RP
ESI+: 353


87
NMR-DMSO-d6: 1.21 (3H, t, J = 7.3 Hz), 2.02-2.30 (2H, m), 3.95-4.13 (3H, m), 4.20-4.30



(1H, m), 4.33-4.45 (2H, m), 4.55 (1H, d, J = 5.5 Hz), 6.44 (2H, bs), 6.9 (1H, d, J = 8.5 Hz),



7.46 (1H, bs), 7.51-7.61 (1H, m), 8.99 (2H, bs), 9.13 (1H, s)



a compound prepared from Reference Example 9b


RP
ESI+: 365


88
NMR-DMSO-d6: 0.39-0.51 (2H, m), 0.67-0.81 (2H, m), 1.36-1.46 (1H, m), 3.80 (1H, d, J =



9.5 Hz), 4.15 (1H, d, J = 7.0 Hz), 4.50 (1H, d, J = 6.4 Hz), 4.64-4.66 (2H, m), 4.81 (2H,



s), 6.21 (2H, s), 6.92 (1H, d, J = 8.4 Hz), 7.53-7.57 (2H, m), 9.04 (2H, s), 9.13 (1H, s)



a compound prepared from Reference Example 12a


RP
ESI+: 365


89
a compound prepared from Reference Example 12b


RP
ESI+: 381


90
NMR-DMSO-d6: 1.01 (3H, d, J = 6.5 Hz), 1.07 (3H, d, J = 6.5 Hz), 1.88-1.95 (1H, m),



2.02-2.15 (2H, m), 4.02 (1H, d, J = 8.9 Hz), 4.11-4.14 (2H, m), 4.25 (1H, d, J = 5.5 Hz),



4.32 (1H, d, J = 6.5 Hz), 4.41 (1H, d, J = 6.6 Hz), 4.58 (1H, d, J = 5.5 Hz), 6.44 (2H, s),



6.88 (1H, d, J = 8.5 Hz), 7.45 (1H, d, J = 2.4 Hz), 7.54 (1H, dd, J = 8.5, 2.4 Hz), 8.98 (2H,



s),9.13(1H, s)



a compound prepared from Reference Example 13b


RP
ESI+: 367


91
NMR-DMSO-d6: 0.98 (3H, t, J = 7.2 Hz), 1.42-1.58 (1H, m), 1.62-1.77 (2H, m), 1.78-1.90



(1H, m), 4.19 (1H, d, J = 6.8 Hz), 4.37 (1H, d, J = 6.4 Hz), 4.42 (1H, d, J = 6.8 Hz),



4.53-4.71 (4H, m), 6.23 (2H, bs), 6.90 (1H, d, J = 8.4 Hz), 7.49 (1H, d, J = 2.3 Hz), 7.55



(1H, dd, J = 2.3, 8.4 Hz), 9.02 (2H, s), 9.13 (1H, s)



a compound prepared from Reference Example 11a


RP
ESI+: 404


92
NMR-DMSO-d6: 0.98 (3H, t, J = 7.2 Hz), 1.43-1.58 (1H, m), 1.61-1.75 (2H, m), 1.76-1.89



(1H, m), 2.11 (3H, s), 4.20 (1H, d, J = 7.0 Hz), 4.32-4.47 (2H, m), 4.51-4.78 (4H, m), 6.24



(2H, bs), 6.86 (1H, d, J = 8.5 Hz), 7.31-7.60 (2H, m), 7.97 (1H, bs), 8.52 (1H, d, J = 1.9 Hz),



8.72 (1H, bs)



a compound prepared from Reference Example 11a


RP
ESI+: 381


93
NMR-DMSO-d6: 0.98 (3H, d, J = 6.6 Hz), 1.01 (3H, d, J = 6.7 Hz), 1.62-1.68 (1H, m),



1.74-1.81 (1H, m), 1.93-2.03 (1H, m), 4.17 (1H, d, J = 7.0 Hz), 4.37 (1H, d, J = 6.5 Hz),



4.41 (1H, d, J = 7.0 Hz), 4.54 (1H, d, J = 6.5 Hz), 4.63-4.72 (3H, m), 6.23 (2H, s), 6.89



(1H, d, J = 8.5 Hz), 7.49 (1H, d, J = 2.3 Hz), 7.55 (1H, dd, J = 8.5, 2.3 Hz), 9.02 (2H, s),



9.14 (1H, s)



a compound prepared from Reference Example 13a


RP
ESI+: 367


94
NMR-DMSO-d6: 1.05 (3H, t, J = 7.4 Hz), 1.45-1.63 (1H, m), 1.73-1.89 (1H, m), 2.00-2.21



(2H, m), 3.94-4.15 (3H, m), 4.23 (1H, d, J = 5.5 Hz), 4.37 (1H, d, J = 6.5 Hz), 4.41 (1H, d,



J = 6.5 Hz), 4.56 (1H, d, J = 5.5 Hz), 6.44 (2H, bs), 6.89 (1H, d, J = 8.5 Hz), 7.46 (1H, bs),



7.55 (1H, dd, J = 2.2, 8.5 Hz), 8.99 (2H, bs), 9.13 (1H, s)



a compound prepared from Reference Example 11b


RP
ESI+: 421


95
NMR-DMSO-d6: 1.90-2.07 (1H, m), 2.16-2.20 (1H, m), 2.52-2.69 (2H, m), 4.18 (1H, d, J =



7.2 Hz), 4.40 (1H, d, J = 6.8 Hz), 4.47 (1H, d, J = 7.2 Hz), 4.55 (1H, d, J = 6.8 Hz),



4.58-4.68 (1H, m), 4.73 (1H, d, J = 9.5 Hz), 4.75 (1H, d, J = 9.5 Hz), 6.21 (2H, bs), 6.96



(1H, d, J = 8.5 Hz), 7.53 (1H, d, J = 2.2 Hz), 7.58 (1H, dd, J = 2.2, 8.5 Hz), 9.04 (2H, s),



9.14 (1H, s)



a compound prepared from Reference Example 14a


RP
ESI+: 458


96
NMR-DMSO-d6: 1.89-2.05 (1H, m), 2.11 (3H, s), 2.14-2.29 (1H, m), 2.50-2.69 (2H, m),



4.18 (1H, d, J = 7.2 Hz), 4.39 (1H, d, J = 6.8 Hz), 4.46 (1H, d, J = 7.2 Hz), 4.56 (1H, d, J =



6.8 Hz), 4.58-4.65 (1H, m), 4.69 (1H, d, J = 9.2 Hz), 4.74 (1H, d, J = 9.2 Hz), 6.22 (2H,



bs), 6.91 (1H, d, J = 8.5 Hz), 7.45 (1H, d, J = 2.3 Hz), 7.52 (1H, dd, J = 2.3, 8.5 Hz),



7.95-8.00 (1H, m), 8.52 (1H, d, J = 1.9 Hz), 8.72 (1H, d, J = 2.3 Hz)



a compound prepared from Reference Example 14a


RP
ESI+: 339


97
a compound prepared from Reference Example 1a


RP
ESI+: 339


98
a compound prepared from Reference Example 1b


RP
ESI+: 432


99


RP
ESI+: 309


100
NMR-DMSO-d6: 0.42-0.44 (3H, m), 0.81-0.83 (1H, m), 3.67 (1H, d, J = 11.6 Hz), 4.21



(1H, d, J = 7.8 Hz), 4.35 (1H, d, J = 7.8 Hz), 4.44 (1H, d, J = 11.6 Hz), 6.21 (2H, brs),



6.91-6.93 (1H, d, J = 8.5 Hz), 7.45 (1H, s), 7.55-7.58 (1H, m), 8.99 (2H, s), 9.12 (1H, s)


RP
ESI+: 323


101


RP
ESI+: 327


102
NMR-DMSO-d6: 0.42-0.48 (3H, m), 0.81-0.86 (1H, m), 3.71 (1H, d, J = 11.7 Hz), 4.19



(1H, d, J = 8.2 Hz), 4.35 (1H, d, J = 8.2 Hz), 4.49 (1H, dd, J = 1.6, 11.7 Hz), 6.20 (2H,



brs), 6.83 (1H, d, J = 12.0 Hz), 7.29 (1H, d, J = 9.0 Hz), 8.91 (2H, d, J = 1.4 Hz), 9.17 (1H,



s)


RP
ESI+: 326


103
NMR-DMSO-d6: 0.39-0.46 (3H, m), 0.83-0.85 (1H, m), 3.65 (1H, d, J = 10.9 Hz), 4.16



(1H, d, J = 8.0 Hz), 4.33 (1H, d, J = 8.0 Hz), 4.44 (1H, d, J = 10.9 Hz), 6.20 (2H, brs), 6.89



(1H, d, J = 8.4 Hz), 7.33-7.34 (1H, m), 7.37-7.40 (1H, m), 7.42-7.45 (1H, m), 7.97-8.02



(1H, m), 8.17-8.18 (1H, m)


RP
ESI+: 360, 362


104
NMR-DMSO-d6: 0.42-0.46 (3H, m), 0.81-0.83 (1H, m), 3.67 (1H, d, J = 11.7 Hz), 4.16



(1H, d, J = 8.2 Hz), 4.34 (1H, d, J = 8.2 Hz), 4.45 (1H, dd, J = 11.7, 1.6 Hz), 6.21 (2H,



brs), 6.89 (1H, d, J = 8.5 Hz), 7.37-7.38 (1H, m), 7.41-7.44 (1H, m), 8.16-8.19 (1H, m),



8.24-8.25 (1H, m)


RP
ESI+: 338


105
NMR-DMSO-d6: 0.41-0.45 (3H, m), 0.82-0.86 (1H, m), 3.65 (1H, d, J = 11.7 Hz), 3.89



(3H, s), 4.20 (1H, d, J = 8.1 Hz), 4.32 (1H, d, J = 8.1 Hz), 4.41-4.44 (1H, m), 6.21 (2H,



brs), 6.87 (1H, d, J = 8.4 Hz), 7.37-7.38 (1H, m), 7.44-7.45 (1H, m), 7.48-7.51 (1H, m),



8.24 (1H, d, J = 2.8 Hz), 8.34 (1H, d, J = 1.9 Hz)


RP
ESI+: 352


106
NMR-DMSO-d6: 0.42-0.44 (3H, m), 0.82-0.84 (1H, m), 1.37 (3H, t, J = 7.0 Hz), 3.65 (1H,



d, J = 11.7 Hz), 4.15-4.20 (3H, m), 4.32-4.34 (1H, m), 4.41-4.44 (1H, m), 6.21 (2H, brs),



6.87 (1H, d, J = 8.5 Hz), 7.38 (1H, s), 7.44 (1H, s), 7.48-7.50 (1H, m), 8.22 (1H, d, J = 2.7 Hz),



8.33 (1H, d, J = 1.2 Hz)


RP
ESI+: 342, 344


107
NMR-DMSO-d6: 0.38-0.48 (3H, m), 0.81-0.84 (1H, m), 3.67 (2H, d, J = 11.9 Hz),



4.20-4.25 (1H, m), 4.32-4.37 (1H, m), 4.45 (1H, d, J = 11.9 Hz), 6.21 (2H, brs), 6.89 (1H,



d, J = 8.2 Hz), 7.42-7.45 (1H, m), 7.55-7.59 (1H, m), 8.09 (1H, brs), 8.55-8.56 (1H, m),



8.74 (1H, brs)


RP
ESI+: 362


108


RP
ESI+: 402


109


RP
ESI+: 390


110


RP
ESI+: 346


111
NMR-DMSO-d6: 0.41-0.45 (3H, m), 0.81-0.86 (1H, m), 2.11 (3H, s), 3.65 (1H, d, J = 11.6 Hz),



4.21 (1H, d, J = 8.1 Hz), 4.33 (1H, d, J = 8.1 Hz), 4.43 (1H, d, J = 11.6 Hz), 6.21 (2H,



brs), 6.87 (1H, d, J = 8.4 Hz), 7.39 (1H, d, J = 2.4 Hz), 7.50-7.53 (1H, m), 7.91 (1H, t, J =



2.1 Hz), 8.51 (1H, d, J = 2.1 Hz), 8.70 (1H, d, J = 2.1 Hz)


RP
ESI+: 364


112
NMR-DMSO-d6: 0.41-0.49 (3H, m), 0.81-0.88 (1H, m), 2.10 (3H, s), 3.69 (1H, d, J = 11.8 Hz),



4.18-4.41 (2H, m), 4.48 (1H, d, J = 11.8 Hz), 6.22 (2H, brs), 6.79 (1H, d, J = 11.8 Hz),



7.26 (1H, brs), 7.86 (1H, s), 8.56 (1H, d, J = 2.0 Hz), 8.59 (1H, s)


RP
ESI+: 360


113
NMR-DMSO-d6: 1.57-1.62 (2H, m), 1.68-1.76 (1H, m), 1.85-2.00 (2H, m), 2.11 (3H, s),



2.14-2.18 (1H, m), 3.97-4.10 (3H, m), 4.37-4.43 (1H, m), 6.26 (2H, brs), 6.83-6.86 (1H,



m), 7.42 (1H, s), 7.48-7.50 (1H, m), 7.91 (1H, s), 8.50-8.51 (1H, m), 8.69 (1H, s)


RP
ESI+: 339, 341


114


RP
ESI+: 389


115
NMR-DMSO-d6: 1.82-1.97 (2H, m), 2.15-2.34 (2H, m), 2.44-2.52 (1H, m), 2.94-3.03 (1H,



m), 4.10 (1H, d, J = 8.7 Hz), 4.20 (1H, d, J = 5.5 Hz), 4.26 (1H, d, J = 8.7 Hz), 4.48 (1H,



d, J = 6.8 Hz), 4.53 (1H, d, J = 6.8 Hz), 4.66 (1H, d, J = 5.5 Hz), 6.36 (2H, s), 6.99 (1H, d,



J = 8.5 Hz), 7.28 (1H, d, J = 2.3 Hz), 7.45 (1H, dd, J = 8.5, 2.3 Hz), 7.80 (1H, dd, J = 8.0,



4.7 Hz), 8.02 (1H, dd, J = 8.0, 1.6 Hz), 8.72 (1H, dd, J = 4.7, 1.6 Hz)


RP
ESI+: 396


116


RP
ESI+: 379


117


RP
ESI+: 414


118


RP
ESI+: 382


119


RP
ESI+: 389


120


RP
ESI+: 400


121


RP
ESI+: 412


122


RP
ESI+: 363


123


RP
ESI+: 381


124


RP
ESI+: 381


125


RP
ESI+: 393


126


RP
ESI+: 377


127


RP
ESI+: 388


128


RP
ESI+: 475


129


RP
ESI+: 399


130


RP
ESI+: 407


131


RP
ESI+: 414


132


RP
ESI+: 425


133


RP
ESI+: 406


134


RP
ESI+: 367


135


RP
ESI+: 411


136


RP
ESI+: 411


137


RP
ESI+: 399


138


RP
ESI+: 385, 387


139


RP
ESI+: 391


140


RP
ESI+: 419


141


RP
ESI+: 381


142


RP
ESI+: 449


143


RP
ESI+: 395


144


RP
ESI+: 411


145


RP
ESI+: 403, 405


146


RP
ESI+: 387


147


RP
ESI+: 383


148


RP
ESI+: 370


149


RP
ESI+: 387, 389


150


RP
ESI+: 400, 402


151


RP
ESI+: 420, 422


152


RP
ESI+: 404, 406


153


RP
ESI+: 404, 406


154


RP
ESI+: 416, 418


155


RP
ESI+: 386, 388


156


RP
ESI+: 384


157


RP
ESI+: 404, 406


158


RP
ESI+: 420, 422


159


RP
ESI+: 377


160


RP
ESI+: 400


161


RP
ESI+: 416, 418


162


RP
ESI+: 386, 388


163


RP
ESI+: 382


164


RP
ESI+: 369


165


RP
ESI+: 369


166


RP
ESI+: 343


167


RP
ESI+: 391


168


RP
ESI+: 385


169


RP
ESI+: 394


170


RP
ESI+: 426


171


RP
ESI+: 383


172


RP
ESI+: 386, 388


173


RP
ESI+: 390


174


RP
ESI+: 390


175


RP
ESI+: 391


176


RP
ESI+: 391


177


RP
ESI+: 393


178


RP
ESI+: 402


179


RP
ESI+: 402


180


RP
ESI+: 345


181


RP
ESI+: 395


182


RP
ESI+: 435


183


RP
ESI+: 435


184


RP
ESI+: 405


185


RP
ESI+: 390


186


RP
ESI+: 417


187


RP
ESI+: 359, 361


188


RP
ESI+: 430, 432


189
NMR-DMSO-d6: 0.32-0.46 (3H, m), 0.78-0.90 (1H, m), 3.58 (1H, d, J = 11.6 Hz), 4.09



(1H, d, J = 8.1 Hz), 4.31 (1H, d, J = 8.1 Hz), 4.36 (1H, dd, J = 1.3, 11.6 Hz), 6.18 (2H, bs),



6.71-6.77 (1H, m), 7.58-7.65 (2H, m), 9.19 (2H, s), 10.60 (1H, s)


190
ESI+: 474, 476



NMR-DMSO-d6: 1.18 (3H, s), 1.73 (3H, s), 4.09-4.22 (2H, m), 4.25-4.42 (3H, m), 4.90



(1H, d, J = 5.6 Hz), 6.33 (2H, bs), 6.63-6.71 (1H, m), 7.54-7.63 (2H, m), 9.19 (2H, s),



10.64 (1H, s)


RP
ESI+: 390


191
NMR-DMSO-d6: 0.34-0.45 (3H, m), 0.78-0.86 (1H, m), 2.52 (3H, s), 3.58 (1H, d, J = 11.6 Hz),



4.08 (1H, d, J = 8.1 Hz), 4.31 (1H, d, J = 8.1 Hz), 4.36 (1H, dd, J = 1.5, 11.6 Hz),



6.18 (2H, bs), 6.71-6.76 (1H, m), 7.53-7.59 (2H, m), 8.34-8.38 (1H, m), 8.92-8.97 (1H,



m), 10.49 (1H, s)


RP
ESI+: 410, 412


192
NMR-DMSO-d6: 0.35-0.45 (3H, m), 0.78-0.86 (1H, m), 3.58 (1H, d, J = 11.6 Hz), 4.09



(1H, d, J = 8.1 Hz), 4.31 (1H, d, J = 8.1 Hz), 4.37 (1H, dd, J = 1.4, 11.6 Hz), 6.20 (2H, bs),



6.76 (1H, d, J = 8.8 Hz), 7.45 (1H, d, J = 2.7 Hz), 7.55 (1H, dd, J = 2.7, 8.8 Hz), 8.77 (1H,



d, J = 1.8 Hz), 9.07 (1H, d, J = 1.8 Hz), 10.64 (1H, s)


RP
ESI+: 382


193
NMR-DMSO-d6: 0.35-0.44 (3H, m), 0.80-0.88 (1H, m), 3.58 (1H, d, J = 11.5 Hz), 4.01



(3H, s), 4.09 (1H, d, J = 8.1 Hz), 4.31 (1H, d, J = 8.1 Hz), 4.34 (1H, br.d, J = 11.6 Hz),



6.16 (2H, s), 6.72 (1H, d, J = 8.8 Hz), 7.56 (1H, dd, J = 2.7, 8.8 Hz), 7.71 (1H, d, J = 2.6 Hz),



8.38 (1H, d, J = 1.3 Hz), 8.86 (1H, d, J = 1.3 Hz), 10.30 (1H, s)


RP
ESI+: 399, 401


194
NMR-DMSO-d6: 0.37-0.43 (3H, m), 0.82-0.88 (1H, m), 2.54 (3H, s), 3.57 (1H, d, J = 11.7 Hz),



4.08 (1H, d, J = 8.1 Hz), 4.31 (1H, d, J = 8.1 Hz), 4.34-4.37 (1H, m), 6.18 (2H, s),



6.72 (1H, d, J = 8.7 Hz), 7.54-7.58 (2H, m), 7.98-7.99 (1H, m), 8.536-8.543 (1H, m), 10.34 (1H, s)


195
ESI+: 443, 445



NMR-DMSO-d6: 1.18 (3H, s), 1.73 (3H, s), 2.53 (3H, s), 4.13-4.18 (2H, m), 4.30 (2H, d, J =



6.1 Hz), 4.36 (1H, d, J = 6.8 Hz), 4.90 (1H, d, J = 5.6 Hz), 6.33 (2H, s), 6.64-6.66 (1H,



m), 7.52-7.55 (2H, m), 7.99 (1H, dd, J = 2.4, 0.7 Hz), 8.54-8.55 (1H, m), 10.38 (1H, s)


RP
ESI+: 376


196
NMR-DMSO-d6: 0.36-0.44 (3H, m), 0.82-0.85 (1H, m), 3.58 (1H, d, J = 11.6 Hz), 4.09



(1H, d, J = 8.1 Hz), 4.30-4.37 (2H, m), 6.18 (2H, s), 6.74 (1H, d, J = 8.8 Hz), 7.61 (1H, dd,



J = 8.8, 2.7 Hz), 7.74 (1H, d, J = 2.6 Hz), 8.26 (1H, dd, J = 8.2, 0.9 Hz), 8.56 (1H, dd, J =



8.2, 2.1 Hz), 9.16 (1H, dd, J = 2.1, 0.9 Hz), 10.64 (1H, s)


197
ESI+: 420



NMR-DMSO-d6: 1.18 (3H, s), 1.73 (3H, s), 4.13-4.19 (2H, m), 4.30-4.37 (3H, m), 4.90



(1H, d, J = 5.6 Hz), 6.33 (2H, s), 6.67 (1H, d, J = 8.8 Hz), 7.59 (1H, dd, J = 8.8, 2.6 Hz),



7.74 (1H, d, J = 2.5 Hz), 8.26 (1H, dd, J = 8.2, 0.9 Hz), 8.56 (1H, dd, J = 8.2, 2.1 Hz),



9.16-9.17 (1H, m), 10.69 (1H, s)


RP
ESI+: 375


198


RP
ESI+: 373


199
NMR-DMSO-d6: 0.18-0.22 (1H, m), 0.39-0.46 (2H, m), 0.61-0.63 (1H, m), 4.14 (1H, d, J =



11.5 Hz), 4.58 (1H, d, J = 11.5 Hz), 6.47 (2H, brs), 6.82 (1H, s), 7.06 (1H, d, J = 8.7 Hz),



7.21-7.27 (2H, m), 7.42 (1H, s), 7.58 (1H, d, J = 8.7 Hz), 8.81 (2H, s), 9.05 (1H, s)


RP
ESI+: 415


200
NMR-DMSO-d6: 2.89-2.95 (1H, m), 3.26-3.32 (1H, m), 4.30 (1H, d, J = 7.0 Hz), 4.45



(1H, d, J = 6.3 Hz), 4.60-4.72 (4H, m), 4.79-4.83 (1H, m), 6.28 (2H, brs), 6.82 (1H, d, J =



8.4 Hz), 7.23-7.29 (1H, m), 7.32-7.37 (4H, m), 7.51 (1H, d, J = 2.3 Hz), 7.56 (1H, dd, J =



2.3, 8.4 Hz), 9.02 (2H, s), 9.14 (1H, s)



a compound prepared from Reference Example 15a


RP
ESI+: 452


201
NMR-DMSO-d6: 2.11 (3H, s), 2.87-2.94 (1H, m), 3.24-3.34 (1H, m), 4.31 (1H, d, J = 7.0 Hz),



4.44 (1H, d, J = 6.3 Hz), 4.59-4.71 (4H, m), 4.79 (1H, dd, J = 1.9, 10.7 Hz), 6.29 (2H,



brs), 6.77 (1H, d, J = 8.5 Hz), 7.23-7.28 (1H, m), 7.32-7.37 (4H, m), 7.44 (1H, d, J = 2.3 Hz),



7.51 (1H, dd, J = 2.3, 8.5 Hz), 7.95-7.96 (1H, m), 8.52 (1H, d, J = 1.9 Hz), 8.71 (1H,



d, J = 2.3 Hz)



a compound prepared from Reference Example 15a


RP
ESI+: 415


202
NMR-DMSO-d6: 3.31-3.39 (1H, m), 3.57 (1H, brd, J = 13.5 Hz), 4.10-4.17 (2H, m),



4.38-4.43 (2H, m), 4.48 (1H, d, J = 6.6 Hz), 4.53 (1H, d, J = 6.6 Hz), 4.63 (1H, d, J = 5.7 Hz),



6.45 (2H, brs), 6.76 (1H, d, J = 8.4 Hz), 7.24-7.28 (1H, m), 7.33-7.38 (2H, m),



7.46-7.53 (4H, m), 8.98 (2H, s), 9.13 (1H, s)



a compound prepared from Reference Example 15b


RP
ESI+: 452


203
NMR-DMSO-d6: 2.10 (3H, s), 3.31-3.38 (1H, m), 3.56 (1H, brd, J = 13.8 Hz), 4.10-4.15



(2H, m), 4.36-4.40 (1H, m), 4.42 (1H, d, J = 5.6 Hz), 4.47 (1H, d, J = 6.6 Hz), 4.53 (1H, d,



J = 6.6 Hz), 4.63 (1H, d, J = 5.6 Hz), 6.46 (2H, brs), 6.71 (1H, d, J = 8.4 Hz), 7.23-7.28



(1H, m), 7.33-7.51 (6H, m), 7.89-7.91 (1H, m), 8.51 (1H, d, J = 1.9 Hz), 8.67 (1H, d, J =



2.3 Hz)



a compound prepared from Reference Example 15b


RP
ESI+: 379


204
NMR-DMSO-d6: 0.11-0.24 (2H, m), 0.43-0.58 (2H, m), 1.00-1.08 (1H, m), 1.48-1.54 (1H,



m), 1.82-1.90 (1H, m), 4.19 (1H, d, J = 6.9 Hz), 4.34 (1H, d, J = 6.3 Hz), 4.40 (1H, d, J =



6.9 Hz), 4.54-4.70 (4H, m), 6.24 (2H, brs), 6.92 (1H, d, J = 8.5 Hz), 7.48 (1H, d, J = 2.3 Hz),



7.57 (1H, dd, J = 2.3, 8.5 Hz), 9.02 (2H, s), 9.14 (1H, s)



a compound prepared from Reference Example 16a


RP
ESI+: 416


205
NMR-DMSO-d6: 0.11-0.24 (2H, m), 0.43-0.58 (2H, m), 0.98-1.08 (1H, m), 1.46-1.52 (1H,



m), 1.80-1.87 (1H, m), 2.11 (3H, s), 4.19 (1H, d, J = 6.9 Hz), 4.33 (1H, d, J = 6.2 Hz), 4.39



(1H, d, J = 6.9 Hz), 4.51 (1H, d, J = 9.0 Hz), 4.58-4.63 (2H, m), 4.65-4.69 (1H, m), 6.25



(2H, brs), 6.87 (1H, d, J = 8.5 Hz), 7.41 (1H, d, J = 2.3 Hz), 7.51 (1H, dd, J = 2.3, 8.5 Hz),



7.95 (1H, dd, J = 1.9, 2.2 Hz), 8.52 (1H, d, J = 1.9 Hz), 8.71 (1H, d, J = 2.2 Hz)



a compound prepared from Reference Example 16a


RP
ESI+: 359


206


RP
ESI+: 379


207
NMR-DMSO-d6: 0.24-0.33 (2H, m), 0.44-0.50 (1H, m), 0.56-0.62 (1H, m), 1.09-1.19 (1H,



m), 1.69-1.75 (1H, m), 2.30-2.37 (1H, m), 4.05-4.12 (2H, m), 4.15-4.18 (1H, m), 4.25



(1H, d, J = 5.5 Hz), 4.30 (1H, d, J = 6.6 Hz), 4.39 (1H, d, J = 6.6 Hz), 4.55 (1H, d, J = 5.5 Hz),



6.43 (2H, brs), 6.91 (1H, d, J = 8.4 Hz), 7.46 (1H, d, J = 2.4 Hz), 7.57 (1H, dd, J =



2.4, 8.4 Hz), 8.99 (2H, s), 9.13 (1H, s)



a compound prepared from Reference Example 16b


RP
ESI+: 359


208


RP
ESI+: 364


209
NMR-DMSO-d6: 1.80-1.95 (2H, m), 2.13-2.24 (1H, m), 2.24-2.34 (1H, m), 2.37-2.48 (1H,



m), 2.94-3.04 (1H, m), 4.07-4.15 (1H, m), 4.15-4.23 (2H, m), 4.47-4.54 (2H, m), 4.66



(1H, d, J = 5.2 Hz), 6.40 (2H, brs), 6.92 (1H, d, J = 8.4 Hz), 7.28 (1H, s), 7.44-7.50 (2H,



m), 7.90 (1H, d, J = 7.6 Hz), 8.52 (1H, d, J = 4.8 Hz), 8.73 (1H, s)


RP
ESI+: 414


210


RP
ESI+: 414


211


RP
ESI+: 432


212


RP
ESI+: 361


213


RP
ESI+: 323, 325


214


RP
ESI+: 323


215
NMR-DMSO-d6: 0.20-0.24 (1H, m), 0.42-0.54 (2H, m), 0.95-0.98 (1H, m), 3.51 (1H, d, J =



11.6 Hz), 3.73 (1H, d, J = 11.4 Hz), 3.84 (1H, d, J = 11.4 Hz), 3.97 (1H, d, J = 15.6 Hz),



4.13 (1H, d, J = 15.6 Hz), 4.61 (1H, dd, J = 1.8, 11.6 Hz), 5.69 (2H, brs), 6.89 (1H, d, J =



8.4 Hz), 7.31 (1H, d, J = 2.4 Hz), 7.53 (1H, dd, J = 2.4, 8.4 Hz), 8.99 (2H, s), 9.12 (1H, s)


RP
ESI+: 359, 361


216


RP
ESI+: 337, 339


217


218
ESI+: 429, 431



NMR-DMSO-d6: 1.18 (3H, s), 1.73 (3H, s), 4.13-4.19 (2H, m), 4.30-4.32 (2H, m), 4.36



(1H, d, J = 6.8 Hz), 4.90 (1H, d, J = 5.6 Hz), 6.33 (2H, s), 6.66 (1H, d, J = 8.8 Hz), 7.57



(1H, dd, J = 8.8, 2.6 Hz), 7.70 (1H, d, J = 2.6 Hz), 8.13 (1H, dd, J = 8.5, 0.8 Hz), 8.18 (1H,



dd, J = 8.5, 2.3 Hz), 8.75 (1H, dd, J = 2.4, 0.7 Hz), 10.52 (1H, s)



Melting point: 165° C. (differential scanning calorimetry onset temperature, Heating rate:



10° C./minute, under N2 flow of 50 mL/minute)



Crystals having characteristic peaks of powder X-ray diffraction shown at angles 2θ (°) of



about 5.7, 9.6, 11.4, 12.3, 13.7, 15.7, 15.9 and 25.0.



This is the same compound as Ex. 228b.


219
ESI+: 473, 475



NMR-DMSO-d6: 1.18 (3H, s), 1.73 (3H, s), 4.13-4.19 (2H, m), 4.30-4.32 (2H, m), 4.36



(1H, d, J = 6.7 Hz), 4.90 (1H, d, J = 5.6 Hz), 6.33 (2H, s), 6.66 (1H, d, J = 8.8 Hz), 7.57



(1H, dd, J = 8.8, 2.6 Hz), 7.70 (1H, d, J = 2.5 Hz), 8.05 (1H, dd, J = 8.4, 0.3 Hz), 8.29-8.32



(1H, m), 8.82-8.85 (1H, m), 10.52 (1H, s)


220
ESI+: 426



NMR-DMSO-d6: 1.18 (3H, s), 1.73 (3H, s), 4.01 (3H, s), 4.14 (1H, d, J = 8.8 Hz), 4.17



(1H, d, J = 8.8 Hz), 4.28-4.33 (2H, m), 4.35 (1H, d, J = 6.8 Hz), 4.90 (1H, d, J = 5.6 Hz),



6.32 (2H, s), 6.65 (1H, d, J = 8.8 Hz), 7.53 (1H, dd, J = 2.6, 8.8 Hz), 7.72 (1H, d, J = 2.5 Hz),



8.38 (1H, d, J = 1.3 Hz), 8.87 (1H, d, J = 1.4 Hz), 10.35 (1H, s)



This is the same compound as Ex. 229b.


221
ESI+: 413



NMR-CDCl3: 0.42-0.55 (2H, m), 0.74-0.82 (1H, m), 1.18 (3H, s), 1.23-1.30 (1H, m), 1.33



(3H, s), 4.45 (br s), 4.46 (1H, d, J = 8.0 Hz), 4.57 (1H, d, J = 8.0 Hz), 6.81 (1H, d, J = 8.8 Hz),



7.45 (1H, d, J = 2.6 Hz), 7.60 (1H, dd, J = 2.6, 8.8 Hz), 7.84 (1H, dd, J = 2.4, 8.4 Hz), 8.20-8.24



(1H, m), 8.50-8.54 (1H, m), 9.69 (1H, bs)


222
ESI+: 397



NMR-CDCl3: 0.41-0.55 (2H, m), 0.73-0.82 (1H, m), 1.18 (3H, s), 1.23-1.30 (1H, m), 1.33



(3H, s), 4.45 (br s), 4.46 (1H, d, J = 8.0 Hz), 4.57 (1H, d, J = 8.0 Hz), 6.81 (1H, d, J =



8.8 Hz), 7.44-7.47 (1H, m), 7.52-7.63 (2H, m), 8.28-8.33 (1H, m), 8.41 (1H, d, J = 2.8 Hz),



9.67 (1H, bs)


223
ESI+: 410



NMR-DMSO-d6: 0.23-0.32 (1H, m), 0.49-0.57 (1H, m), 0.59-0.68 (1H, m), 1.07-1.16 (1H,



m), 1.11 (3H, s), 1.27 (3H, s), 4.01 (3H, s), 4.26 (1H, d, J = 8.0 Hz), 4.37 (1H, d, J = 8.0 Hz),



6.09 (2H, bs), 6.66 (1H, d, J = 8.7 Hz), 7.53 (1H, dd, J = 2.6, 8.7 Hz), 7.61 (1H, d, J = 2.6 Hz),



8.38 (1H, d, J = 1.3 Hz), 8.86 (1H, d, J = 1.3 Hz), 10.29 (1H, s)



This is the same compound as Ex. 230.


224
ESI+: 430



NMR-DMSO-d6: 0.24-0.32 (1H, m), 0.50-0.58 (1H, m), 0.60-0.68 (1H, m), 1.08-1.16 (1H,



m), 1.12 (3H, s), 1.27 (3H, s), 4.27 (1H, d, J = 8.0 Hz), 4.38 (1H, d, J = 8.0 Hz), 6.11(2H, bs),



6.69 (1H, d, J = 8.7 Hz), 7.24 (1H, t, J = 53.9 Hz), 7.59 (1H, dd, J = 2.6, 8.7 Hz), 7.65 (1H, d,



J = 2.6 Hz), 9.06 (1H, s), 9.34-9.37 (1H, m), 10.71(1H, s)



This is the free form of Ex. 231.


RP
ESI+:337, 339


225a
HPLC retention time: 4.5 minutes (CHIRALCEL OD-RH, MeCN:20 mM aqueous



KH2PO4 = 80:20; flow rate 1.0 ml/minute, detected by 254 nm UV absorption, 2nd peak of



the enantiomer pair)


RP
ESI+:337, 339


225b
HPLC retention time: 2.4 minutes (CHIRALCEL OD-RH, MeCN:20 mM aqueous



KH2PO4 = 80:20; flow rate 1.0 ml/minute, detected by 254 nm UV absorption, 1st peak of



the enantiomer pair)


RP
ESI+: 353, 355


226


227
ESI+: 426


228a
ESI+: 429, 431



Supercritical fluid chromatography retention time: 5.23 minutes: (eluent: supercritical



CO2/(EtOH with 0.1% diethylamine) = 60:40; Column: CHIRALCEL OD-H column (10 ×



250 mm); flow rate 10 mL/minute; column temperature: 40° C., 1st peak of the



enantiomer pair)


228b
ESI+: 429, 431



Supercritical fluid chromatography retention time: 8.16 minutes (eluent: supercritical CO2/



(EtOH with 0.1% diethylamine) = 60:40; Column: CHIRALCEL OD-H column (10 × 250 mm);



flow rate 10 mL/minute; column temperature: 40° C., 2nd peak of the enantiomer



pair)



This is the same compound as Ex. 218.


229a
ESI+: 426



Supercritical fluid chromatography retention time: 5.94 minutes (eluent: supercritical



CO2/EtOH = 60:40; Column: CHIRALCEL OD-H column (4.6 × 250 mm); flow rate 3 mL/minute;



column temperature: 40° C., 2nd peak of the enantiomer pair)


229b
ESI+: 426



Supercritical fluid chromatography retention time: 3.58 minutes (eluent: supercritical CO2/



EtOH = 60:40; Column: CHIRALCEL OD-H column (4.6 × 250 mm); flow rate 3 mL/minute;



column temperature: 40° C., 1st peak of the enantiomer pair)



This is the same compound as Ex. 220.


230
ESI+: 410



NMR-DMSO-d6: 0.23-0.32 (1H, m), 0.49-0.57 (1H, m), 0.59-0.68 (1H, m), 1.07-1.16 (1H,



m), 1.11 (3H, s), 1.27 (3H, s), 4.01 (3H, s), 4.26 (1H, d, J = 8.0 Hz), 4.37 (1H, d, J = 8.0



Hz), 6.10 (2H, bs), 6.66 (1H, d, J = 8.7 Hz), 7.53 (1H, dd, J = 2.6, 8.7 Hz), 7.61 (1H, d, J =



2.6 Hz), 8.38 (1H, d, J = 1.3 Hz), 8.86 (1H, d, J = 1.3 Hz), 10.29 (1H, s)



Melting point: 191° C. (differential scanning calorimetry onset temperature, Heating rate:



10° C./minute, under N2 flow of 50 mL/minute)



Crystals having characteristic peaks of powder X-ray diffraction shown at angles 2θ (°) of



about 5.0, 7.9, 8.0, 8.8, 12.6, 15.2, 16.3, 17.7 and 20.2.



This is the same compound as Ex. 223.


231
ESI+: 430



NMR-DMSO-d6: 0.54-0.66 (1H, m), 0.76-0.92 (2H, m), 0.94-1.05 (1H, m), 1.18 (3H, s),



1.30 (3H, s), 5.07 (2H, s), 6.87 (1H, d, J = 8.9 Hz), 7.26 (1H, t, J = 53.9 Hz), 7.90 (1H, dd,



J = 2.4, 8.9 Hz), 7.98 (1H, d, J = 2.4 Hz), 9.02-9.21 (2H, m), 9.37-9.41 (1H, m), 9.57 (1H, bs),



10.55 (1H, s), 10.98 (1H, s)



Melting point: 254° C. (differential scanning calorimetry onset temperature, Heating rate:



10° C./minute, under N2 flow of 50 mL/minute)



Crystals having characteristic peaks of powder X-ray diffraction shown at angles 2θ (°) of



about 4.8, 6.5, 8.4, 12.8, 16.0, 17.4, 23.4, 26.6 and 27.6.



This is the hydrochloride of Ex. 224.









The compounds shown in Tables below can be prepared using tert-butyl (6′-aminodispiro[cyclopropane-1,3′-chromene-4′,4′′-[1,3]oxazol]-2′′-yl)carbamate or tert-butyl (6′-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-2-yl)carbamate as a starting material in the same manner as the methods of Preparation Examples 70 and Reference Examples 19 or Examples 27 above. The structures, and the preparation methods, for the compounds are shown in [Table. 6] below.










TABLE 6





No.
Structure
1st_step
2nd_step







P1




R70
E27


 


P2




R70
E27


 


P3




R70
E27


 


P4




R70
E27


 


P5




R70
E27


 


P6




R70
E27


 


P7




R70
E27


 


P8




R70
E27


 


P9




R70
E27


 


P10




R70
E27


 


P11




R70
E27


 


P12




R70
E27


 


P13




R70
E27


 


P14




R70
E27


 


P15




R70
E27


 


P16




R70
E27


 


P17




R70
E27


 


P18




R70
E27


 


P19




R70
E27


 


P20




R70
E27


 


P21




R70
E27


 


P22




R70
E27


 


P23




R70
E27


 


P24




R70
E27


 


P25




R70
E27


 


P26




R70
E27


 


P27




R70
E27


 


P28




R70
E27


 


P29




R70
E27


 


P30




R70
E27


 


P31




R70
E27


 


P32




R70
E27


 


P33




R70
E27


 


P34




R70
E27


 


P35




R70
E27


 


P36




R70
E27


 


P37




R70
E27


 


P38




R70
E27


 


P39




R70
E27


 


P40




R70
E27


 


P41




R70
E27


 


P42




R70
E27


 


P43




R70
E27


 


P44




R70
E27


 


P45




R70
E27


 


P46




R70
E27


 


P47




R70
E27


 


P48




R70
E27


 


P49




R70
E27


 


P50




R70
E27


 


P51




R70
E27


 


P52




R70
E27


 


P53




R70
E27


 


P54




R70
E27


 


P55




R70
E27


 


P56




R70
E27


 


P57




R70
E27


 


P58




R70
E27


 


P59




R70
E27


 


P60




R70
E27


 


P61




R70
E27


 


P62




R70
E27


 


P63




R70
E27


 


P64




R70
E27


 


P65




R70
E27


 


P66




R70
E27


 


P67




R70
E27


 


P68




R70
E27


 


P69




R70
E27


 


P70




R70
E27


 


P71




R70
E27


 


P72




R70
E27


 


P73




R70
E27


 


P74




R70
E27


 


P75




R70
E27


 


P76




R70
E27


 


P77




R70
E27


 


P78




R70
E27


 


P79




R70
E27


 


P80




R70
E27


 


P81




R70
E27


 


P82




R70
E27


 


P83




R70
E27


 


P84




R70
E27


 


P85




R70
E27


 


P86




R70
E27


 


P87




R70
E27


 


P88




R70
E27


 


P89




R70
E27


 


P90




R70
E27


 


P91




R70
E27


 


P92




R70
E27


 


P93




R70
E27


 


P94




R70
E27


 


P95




R70
E27


 


P96




R70
E27


 


P97




R70
E27


 


P98




R70
E27


 


P99




R70
E27


 


P100




R70
E27









The teachings of all references cited in this specification are incorporated herein in their entirety.
INDUSTRIAL APPLICABILITY
The compounds of the formula (I) or a salt thereof have a beta-secretase inhibitory activity, and therefore can be used as an agent for preventing or treating diseases or conditions associated with and/or mediated by β-secretase activity, hydrolysis of a β-secretase site of a β-amyloid precursor protein, and/or β-amyloid protein accumulation, such as Glaucoma, MCI (Mild cognitive impairment) or Alzheimer's disease, especially, Alzheimer's disease, or the like.

## Claims

1. A hydrate of N-[(4S)-2-amino-2′,2′-dimethyldispiro[1,3-oxazole-4,4′-chromene-3′,3′′-oxetan]-6′-yl]-5-chloropyridine-2-carboxamide, wherein a powder X-ray diffraction pattern for the hydrate comprises peaks at angles 2 theta of about 5.7 °, 9.6 °, 11.4 °, 12.3°, 13.7°, 15.7°, 15.9°, and 25.0 °.

2. A pharmaceutical composition comprising the hydrate of claim 1 and a pharmaceutically acceptable carrier.