Pyrrolobenzodiazepine dimer prodrug and ligand-linker conjugate compound of the same

Abstract

The present disclosure relates to pyrrolobenzodiazepine dimer prodrugs and ligand-linker conjugates thereof. The present disclosure also relates to compositions and uses of said prodrugs and conjugates.

Claims

1. A conjugate of Chemical Formula IIIa, IIIb, or a pharmaceutically acceptable salt thereof: ##STR00091## wherein a dotted line represents an optional double bond; ligand is an protein; R.sub.1 is selected from H, OH, ═O, ═CH.sub.2, CN, R.sup.m, OR.sup.m, halo, and dihalo; R.sup.m′ is selected from R.sup.m, CO.sub.2R.sup.m, COR.sup.m, CHO, CO.sub.2H, and halo; R.sup.m is selected from optionally substituted C.sub.1-12 alkyl, optionally substituted C.sub.6 aryl, optionally substituted C.sub.6 heteroaryl, and optionally substituted C.sub.3-6 cycloalkyl, wherein the optional substituent is C.sub.1-12 alkyl; R.sub.2, R.sub.3, and R.sub.5 are each independently selected from H, R.sup.m, OH, OR.sup.m, NH.sub.2, NHR.sup.m, NR.sup.mR.sup.m′, NO.sub.2, and halo; R.sub.4 is selected from H, R.sup.m, OH, OR.sup.m, NH.sub.2, NHR.sup.m, NR.sup.mR.sup.m, NO.sub.2, halo, C.sub.1-6 alkyl, or C.sub.1-6 alkoxy; R.sup.n, R.sup.o, R.sup.p, and R.sup.q are each independently selected from H, C.sub.1-7 alkyl, C.sub.2-7 alkenyl, C.sub.2-7 alkynyl, C.sub.3-13 cycloalkyl, 3- to 7-membered heterocycloalkyl, C.sub.6-10 aryl, and 5- to 7-membered heteroaryl; X and X′ are each independently selected from —C(O)O—, —C(O)—, and —C(O)NR—; Xa and Xa′ are each independently a bond or C.sub.1-6 alkylene; Y and Y′ are each independently selected from O, S, and N(H); R.sup.6 is a substituted or unsubstituted saturated or unsaturated C.sub.3-12 hydrocarbon chain, wherein the chain may comprise one or more heteroatoms, NMe, or a substituted or unsubstituted aromatic ring, the chain or aromatic ring may be substituted with —NH, —NR.sup.m, —NHC(O)R.sup.m, —NHC(O)CH.sub.2—[OCH.sub.2CH.sub.2].sub.n—R, or —[CH.sub.2CH.sub.2O].sub.n—R, wherein n is 1 to 12; R.sup.7 is H or C.sub.1-6 alkyl; Z.sub.a and Z.sub.b are each independently O, N, or S; R.sup.12a, R.sup.13a, and R.sup.14a are each independently H, C.sub.1-6 alkyl: or wherein R.sup.13a and R.sup.14a combine with the carbon to which they are attached to form a - to 7-membered heterocyclyl; R.sub.1′, R.sub.2′, R.sub.3′, R.sub.4′, and R.sub.5′ are as defined for R.sub.1, R.sub.2, R.sub.3, R.sub.4, and R.sub.5, respectively; G and G′ denote a glucuronide group; each Z and Z′ is selected from H, C.sub.1-8 alkyl, halo, NO.sub.2, CN, ##STR00092## R.sub.9, R.sub.10, and R.sub.16 are each independently selected from H, C.sub.1-8 alkyl, C.sub.2-6 alkenyl, C.sub.1-6 alkoxy, and methyloxyalkyl; n is an integer from 1 to 3; W is —C(O)—, —C(O)NR″—, or —C(O)O—, R″ is H or C.sub.1-8 alkyl; L is one or more units selected from a branching unit, a connection unit, and a binding unit or a combination of these units: wherein the connection unit connects W with a binding unit, W with a branching unit, a branching unit with another branching unit, or a branching unit with a binding unit, and the branching unit connects a connection unit with W or the connection unit with another connection unit; the branching unit is a C.sub.2-100 alkenyl (wherein a carbon atom of the alkenyl may be substituted with one or more heteroatoms selected from the group consisting of N, O, and S and the alkenyl may be further substituted with one or more C.sub.1-20 alkyls), a hydrophilic amino acid, —C(O)—, —C(O)NR″″—, —C(O)O—, —(CH.sub.2).sub.s—NHC(O)—(CH.sub.2).sub.t—, —(CH.sub.2).sub.u—C(O)NH—(CH.sub.2).sub.v—, —(CH.sub.2).sub.s—NHC(O)—(CH.sub.2).sub.t—C(O)—, —(CH.sub.2).sub.u—C(O)NH—(CH.sub.2).sub.v—C(O)—, —S(O).sub.2NR″″—, —P(O)R′″″NR″″—, —S(O)NR″″—, or —PO.sub.2NR″″—; R″″ and R′″″ each independently denote H, C.sub.1-8 alkyl, C.sub.3-8 cycloalkyl, C.sub.1-8 alkoxy, C.sub.1-8 alkylthio, mono- or di-C.sub.1-8 alkylamino, C.sub.3-20 heteroaryl, or C.sub.5-20 aryl; s, t, u, and v are each independently an integer from 0 to 10; the connection unit is —(CH.sub.2).sub.r(V(CH.sub.2).sub.p).sub.q—; r is an integer from 0 to 10; p is an integer from 0 to 12; q is an integer from 1 to 20; V denotes a single bond, —O—, or S—; the binding unit is ##STR00093## and L.sub.1 is a single bond or C.sub.2-30 alkenyl; R.sub.11 is H or C.sub.1-10 alkyl; and L.sub.2 denotes C.sub.2-30 alkenyl.

2. The conjugate of claim 1, wherein a dotted line represents presence of a double bond between the carbons bearing R.sub.1 and R.sub.7 or R.sub.1′ and R.sub.7′.

3. The conjugate of claim 1, wherein: R.sub.1 is selected from substituted or unsubstituted C.sub.1-6 alkyl, substituted or unsubstituted C.sub.2-6 alkenyl, substituted or unsubstituted C.sub.5-7 aryl, and substituted or unsubstituted C.sub.3-6 heteroaryl; R.sub.2, R.sub.3, and R.sub.5 are each independently H or OH; and R.sub.4 is C.sub.1-6 alkoxy.

4. The conjugate of claim 1, wherein X and X′ are each independently selected from —C(O)O*, —C(O)*, and —C(O)NR*, wherein each R independently is H, OH, N.sub.3, CN, NO.sub.2, SH, NH.sub.2, ONH.sub.2, NNH.sub.2, halo, substituted or unsubstituted C.sub.1-8 alkyl, or substituted or unsubstituted C.sub.1-8 alkoxy, wherein C.sub.1-8 alkyl or C.sub.1-8 alkoxy may be substituted with OH, N.sub.3, CN, NO.sub.2, SH, NH.sub.2, ONH.sub.2, NNH.sub.2, or halo.

5. The conjugate of claim 1, wherein Y and Y′ are each O.

6. The conjugate of claim 1, wherein R.sub.6 is a substituted or unsubstituted saturated or unsaturated C.sub.3-8 hydrocarbon chain and the chain comprises one or more heteroatoms or a substituted or unsubstituted aromatic ring, wherein the heteroatom is O, S, or N(H) and the aromatic ring is benzene, pyridine, imidazole, or pyrazole, and a hydrogen in the chain or aromatic ring may be replaced with —NHC(O)CH.sub.2—[OCH.sub.2CH.sub.2].sub.n—R or —[CH.sub.2CH.sub.2O].sub.n—R; and n is 1 to 6.

7. The conjugate of claim 1, wherein W is —C(O)—, —C(O)NR′″—, or —C(O)O—, wherein R′″ is H or C.sub.1-8 alkyl, L is one or more units selected from a branching unit, a connection unit, and a binding unit or a combination of these units, wherein the connection unit connects W with a binding unit, W with a branching unit, a branching unit with another branching unit, or a branching unit with a binding unit, and the branching unit connects a connection unit with W or the connection unit with another connection unit; the branching unit is a C.sub.2-8 alkenyl wherein a carbon atom of the alkenyl may be replaced with one or more heteroatoms selected from N, O, and S and the alkenyl may be further substituted with one or more C.sub.1-6 alkyls, a hydrophilic amino acid, —C(O)—, —C(O)NR″″—, —C(O)O—, —(CH.sub.2).sub.s—NHC(O)—(CH.sub.2).sub.t—, —(CH.sub.2).sub.u—C(O)NH—(CH.sub.2).sub.v—, —(CH.sub.2).sub.s—NHC(O)—(CH.sub.2).sub.t—C(O)—, or —(CH.sub.2).sub.u—C(O)NH—(CH.sub.2).sub.v—C(O)— (wherein R″″ is H, C.sub.1-8 alkyl, C.sub.3-8 cycloalkyl, C.sub.1-8 alkoxy, C.sub.1-8 alkylthio, mono- or di-C.sub.1-8 alkylamino, C.sub.3-20 heteroaryl, or C.sub.5-20 aryl and s, t, u, and v are each independently 0 to 5; the connection unit is —(CH.sub.2).sub.r(V(CH.sub.2).sub.p).sub.q—, wherein r is 0 to 10, p is 0 to 12, q is 1 to 20, and V is a single bond or —O—, the binding unit is ##STR00094##  wherein L.sub.1 is a single bond or C.sub.2-8 alkenyl, R.sub.11 is H or C.sub.1-6 alkyl, and L.sub.2 is C.sub.2-8 alkenyl; and the connection unit is —(CH.sub.2).sub.r(V(CH.sub.2).sub.p).sub.q—, wherein r is 0 to 8, p is 1 to 12, q is 1 to 10, and V is a single bond or —O—.

8. The conjugate of claim 7, wherein the connection unit is —(CH.sub.2).sub.r(V(CH.sub.2).sub.p).sub.q—; r is 0, 1, or 2; p is 2, q is 2, 3, or 6; and V is O.

9. The conjugate of claim 1, wherein Xa and Xa′ each independently a bond or C.sub.1-3 alkyl.

10. The conjugate of claim 1, wherein Z and Z′ are each independently selected from H, ##STR00095## and —(CH.sub.2).sub.m—OCH.sub.3.

11. The conjugate of claim 1, wherein the ligand is a protein.

12. The conjugate of claim 11, wherein the protein is an antibody.

13. The conjugate of claim 1, wherein the protein has an amino acid motif represented by CYYX, XXCC, XCXC or CXX, wherein C is cysteine, Y is an aliphatic amino acid, and X is an amino acid that determines substrate specificity of an isoprenoid transferase.

14. The conjugate of claim 1, wherein D has a structure selected from: ##STR00096## ##STR00097## ##STR00098## ##STR00099## ##STR00100## and the bond overlaid with a dashed line represents a connection point to L.

15. The conjugate of claim 1, wherein: D has a structure selected from: ##STR00101## ##STR00102## ##STR00103## ##STR00104## the bond overlaid with a dashed line represents a connection point to L; L is one or more units selected from a branching unit, a connection unit, and a binding unit or a combination of these units, wherein the connection unit connects W with a binding unit, W with a branching unit, a branching unit with another branching unit, or a branching unit with a binding unit, and the branching unit connects a connection unit with W or the connection unit with another connection unit; the branching unit is a C.sub.2-8 alkenyl, wherein a carbon atom of the alkenyl may be replaced with one or more heteroatoms selected from N, O, and S and the alkenyl may be substituted with one or more C.sub.1-6 alkyls, a hydrophilic amino acid, —C(O)—, —C(O)NR″″—, —C(O)O—, —(CH.sub.2).sub.s—NHC(O)—(CH.sub.2).sub.t—, —(CH.sub.2).sub.u—C(O)NH—(CH.sub.2).sub.v—, —(CH.sub.2).sub.s—NHC(O)—(CH.sub.2).sub.t—C(O)—, or —(CH.sub.2).sub.u—C(O)NH—(CH.sub.2).sub.v—C(O)—, wherein R″″ is H, C.sub.1-8 alkyl, C.sub.3-8 cycloalkyl, C.sub.1-8 alkoxy, C.sub.1-8 alkylthio, mono- or di-C.sub.1-8 alkylamino, C.sub.3-20 heteroaryl, or C.sub.5-20 aryl and s, t, u, and v are each independently 0 to 5; the connection unit is —(CH.sub.2).sub.r(V(CH.sub.2).sub.p).sub.q—, wherein r is 0 to 10, p is 0 to 12, q is 1 to 20, and V is a single bond or —O—, the binding unit is ##STR00105##  wherein L.sub.1 is a single bond or C.sub.2-8 alkenyl, R.sub.11 is H or C.sub.1-6 alkyl, and L.sub.2 is C.sub.2-8 alkenyl; the connection unit is —(CH.sub.2).sub.r(V(CH.sub.2).sub.p).sub.q—, wherein r is 0 to 8, p is 1 to 12, q is 1 to 10, and V is a single bond or —O—; and the ligand is an antibody.

16. The conjugate of claim 15, wherein the protein has one or more amino acid motifs that can be recognized by an isoprenoid transferase represented by CYYX, XXCC, XCXC or CXX, wherein C is cysteine, Y is an aliphatic amino acid, and X is an amino acid that determines substrate specificity of isoprenoid transferase.

17. A pharmaceutical composition comprising the conjugate of claim 1 and a pharmaceutically acceptable carrier.

18. A method of treating a proliferative disease in a subject, comprising administering the conjugate of claim 1 to the subject.

Description

BRIEF DESCRIPTION OF DRAWINGS

(1) FIG. 1 illustrates an example of the synthesis process of Compound No. 28 according to the present invention.

DESCRIPTION OF EMBODIMENTS

(2) Hereinafter, the present invention will be described in more detail with reference to Examples. However, the following Examples are intended to assist in understanding the present invention and are not intended to limit the scope of the present invention thereto.

<Example 1> Preparation of Compound 4

(3) ##STR00020##

(4) Preparation of Compound 2

(5) Oxalyl chloride (3.1 mL, 36.2 mmol) was dissolved in dichloromethane (40 mL) and dimethylsulfoxide (4.7 mL, 66.4 mmol) was added thereto at −78° C. under a nitrogen atmosphere. After 10 minutes, a solution of Compound 1 (10 g, 30.2 mmol, Compound 1 was prepared by the method described in J. Org. Chem., 2003, 68, 3923-3931) in dichloromethane (140 mL) was gradually added to the mixture, the reaction solution was stirred for 1 hour, then triethylamine (16.7 mL, 120.6 mmol) was added thereto, and the reaction temperature was gradually raised to 0° C. over 2 hours. The reaction solution was diluted with dichloromethane (200 mL), and the organic layer was washed with a saturated aqueous ammonium chloride solution (200 mL) and brine (200 mL) and then dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated, and purified by column chromatography to obtain Compound 2 (9.5 g, 95%).

(6) .sup.1H-NMR (400 MHz, CDCl.sub.3) (rotamers) δ 4.39-4.26 (m, 1H), 4.03-3.80 (m, 2H), 3.69-3.64 (m, 1H), 3.63-3.51 (m, 1H), 2.70-2.60 (m, 1H), 2.43 (d, J=17.6 Hz), 1.61-1.41 (m, 10H), 0.98-0.67 (m, 6H), 0.08-0.05 (s, 6H).

(7) Preparation of Compound 3

(8) Methyltriphenylphosphonium bromide (7.6 g, 21.2 mmol) was diluted with tetrahydrofuran (80 mL), and then potassium t-butoxide (1 M in THF, 21.2 mL, 21.2 mmol) was added thereto at 0° C. under a nitrogen atmosphere. The mixture was stirred for 1 hour, and then a solution of Compound 2 (5.0 g, 15.2 mmol) in tetrahydrofuran (10 mL) was gradually added thereto. The mixture was stirred for 4 hours while gradually raising the reaction temperature to room temperature. A saturated aqueous ammonium chloride solution (200 mL) was added to the reaction solution and then the mixture was subjected to extraction using diethyl ether (2×200 mL). The combined organic layers were washed with brine (200 mL) and then dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated, and purified by column chromatography to obtain Compound 3 (4.27 g, 86%).

(9) 1H-NMR (400 MHz, CDCl.sub.3) (rotamers) δ 4.97-4.91 (m, 2H), 4.09-3.93 (m, 2H), 3.84-3.80 (m, 1H), 3.65-3.61 (m, 1H), 3.59-3.34 (m, 1H), 2.64-2.55 (m, 2H), 1.69 (s, 9H), 0.87 (s, 9H), 0.03 (s, 6H).

(10) Preparation of compound 4

(11) Compound 3 (15.5 g, 47.2 mmol) was dissolved in dichloromethane (120 mL), then hydrochloric acid (4 N 1,4-dioxane solution, 82.6 mL, 330.4 mmol) was added thereto at 0° C., and the mixture was stirred for 2 hours under a nitrogen atmosphere. The reaction solution was concentrated under reduced pressure to obtain Compound 4 (6.53 g, 92%) as a white solid.

(12) .sup.1H-NMR (400 MHz, CDCl.sub.3) (rotamers) δ 9.79 (br s, 1H), 9.17 (br s, 1H), 5.15 (d, J=8 Hz, 1H), 4.91 (br s, 1H), 4.10 (m, 5H), 2.76-2.70 (m, 1H), 2.60-2.54 (m, 1H).

<Example 2> Preparation of Compound 9

(13) ##STR00021##

(14) Preparation of Compound 6

(15) Compound 5 (10 g, 20.2 mmol, Compound 5 was prepared by the method described in J. Med. Chem., 2004, 47, 1161-1174) was dissolved in dichloromethane (100 mL) and then oxalyl chloride (6.1 mL, 70.8 mmol) and N,N-dimethylformamide (2 drops) were added thereto at 0° C. under a nitrogen atmosphere. The reaction solution was stirred for 4 hours, then the temperature was raised to room temperature, and the reaction solution was stirred for 10 hours, concentrated under reduced pressure, and vacuum dried. The compound obtained was dissolved in dichloromethane (120 mL), and then Compound 4 (6.2 g, 41.4 mmol) and triethylamine (9.9 mL, 70.8 mmol) were added thereto at 0° C. under a nitrogen atmosphere. The reaction temperature was raised to room temperature, and the mixture was stirred for 3 hours, then a saturated aqueous ammonium chloride solution (200 mL) was added to the reaction solution, and the mixture was subjected to extraction using dichloromethane (2×200 mL). The combined organic layers were washed with brine (200 mL) and then dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated, and purified by column chromatography to obtain Compound 6 (12 g, 87%).

(16) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 7.71 (s, 2H), 6.80 (s, 2H), 5.13 (s, 2H), 4.88 (s, 2H), 4.61 (m, 2H), 4.17-4.14 (t, J=6.2 Hz, 4H), 3.98 (s, 6H), 3.94-3.74 (m, 10H), 2.89-2.83 (m, 2H), 2.52-2.48 (m, 2H), 2.04-1.96 (m, 4H), 1.77-1.71 (m, 2H).

(17) Preparation of Compound 7

(18) Compound 6 (6.4 g, 9.36 mmol) was dissolved in dichloromethane (100 mL), and then imidazole (2.5 g, 37.4 mmol) and t-butyldimethylsilyl chloride (3.5 g, 23.4 mmol) were added thereto at 0° C. under a nitrogen atmosphere. The reaction solution was stirred for 2 hours, then a saturated aqueous ammonium chloride solution (100 mL) was added to the reaction solution, and the mixture was subjected to extraction using dichloromethane (2×100 mL). The combined organic layers were washed with brine (200 mL) and then dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated, and purified by column chromatography to obtain Compound 7 (6.88 g, 75%).

(19) .sup.1H-NMR (400 MHz, CDCl.sub.3) (rotamers) δ 7.70 (s, 1H), 6.76 (s, 2H), 4.99 (s, 2H), 4.83 (s, 2H), 4.59 (br s, 2H), 4.14, (t, 4H), 3.95 (s, 6H), 3.90 (d, 2H), 3.77-3.69 (m, 4H), 3.57 (q, J=6.2 Hz, 1H), 3.31-3.29 (m, 1H), 2.82-2.67 (m, 4H), 1.99 (t, J=7.2 Hz, 4H), 1.75-1.72 (m, 2H), 0.89 (s, 18H), 0.09 (s, 12H).

(20) Preparation of Compound 8

(21) Compound 7 (3.0 g, 3.29 mmol) was dissolved in ethanol (44 mL), and then zinc dust (12.9 g, 197 mmol) and formic acid (5% ethanol solution, 128 mL) were added thereto. The reaction solution was stirred at room temperature for 15 minutes and then filtered through Celite, and ethyl acetate (500 mL) was added thereto. The organic layer was washed with distilled water (200 mL), a saturated aqueous sodium hydrogencarbonate solution (200 mL), and brine (200 mL) and then dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated, and purified by column chromatography to obtain Compound 8 (2.76 g, 98%).

(22) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 6.74 (s, 2H), 6.24 (s, 2H), 4.97 (s, 2H), 4.90 (s, 2H), 4.54 (br s, 2H), 4.33 (br s, 4H), 4.18 (br s, 1H), 4.14 (br s, 2H), 4.14-4.09 (m, 2H), 4.00 (t, J=8 Hz, 4H), 3.77 (s, 6H), 3.62 (br s. 2H), 2.68 (s, 4H), 1.95-1.88 (m, 4H), 1.66-1.64 (m, 2H), 0.87 (s, 18H), 0.02 (s, 12H).

(23) Preparation of Compound 9

(24) Compound 8 (5.0 g, 5.86 mmol) was dissolved in dichloromethane (300 mL) and then pyridine (0.94 mL, 11.7 mmol) and allyl chloroformate (0.62 mL, 5.86 mmol) were added thereto at −78° C. under a nitrogen atmosphere. The reaction solution was stirred for 1 hour, then the reaction temperature was raised to room temperature, and reaction solution was concentrated and then purified by column chromatography to obtain Compound 9 (2.23 g, 41%).

(25) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 7.84 (s, 1H), 6.81 (s, 1H), 6.74 (s, 1H), 6.24 (s, 1H), 5.98-5.92 (m, 1H), 5.37, (dd, J=17.6 Hz, J=1.2 Hz, 1H), 5.25 (dd, J=10.4 Hz, J=1.2 Hz, 1H), 4.97 (br s, 2H), 4.90 (br s, 2H), 4.63-4.62 (m, 4H), 4.34 (br s, 2H), 4.21-4.18 (m, 2H), 4.10 (t, J=6.4 Hz, 3H), 3.99 (t, J=6.4 Hz, 3H), 3.83 (s, 3H), 3.77 (s, 3H), 3.63 (bs, 1H), 2.68 (br s, 4H), 1.97-1.89 (m, 4H), 1.69-1.61 (m, 2H), 0.87 (s, 18H), 0.02 (br s, 12H).

<Example 3> Preparation of Compound 12

(26) ##STR00022##

(27) Preparation of Compound 11

(28) In acetonitrile (40 mL), 4-hydroxybenzaldehyde (475 mg, 3.89 mmol) and Compound 10 (1.7 g, 4.28 mmol, Compound 10 was prepared by the method described in Korean Patent No. 1,628,872) were dissolved, then 4 Å molecular sieves (4 g) and silver(I) oxide (3.6 g, 15.6 mmol) were added thereto, and the mixture was stirred at room temperature for 3 hours under a nitrogen atmosphere. The reaction solution was concentrated under reduced pressure, diluted with distilled water (40 mL), and the subjected to extraction using ethyl acetate (2×50 mL). The organic layer extracted was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and then purified by column chromatography to obtain Compound 11 (1.3 g, 69%).

(29) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 9.93 (s, 1H), 7.86 (d, J=8 Hz, 2H), 7.11 (d, J=8.4 Hz, 2H), 5.38-5.29 (m, 4H), 4.25-4.23 (m, 1H), 3.71 (s, 3H), 2.06 (s, 9H).

(30) Preparation of Compound 12

(31) Compound 11 (1.3 g, 2.96 mmol) was dissolved in chloroform/isopropanol (50 mL/10 mL), then silica gel (1.3 g) and sodium borohydride (134 mg, 3.55 mmol) were added thereto at 0° C. under a nitrogen atmosphere, and the mixture was then stirred for 2 hours. Distilled water (40 mL) was added to the reaction solution, and the mixture was subjected to extraction using ethyl acetate (2×50 mL). The organic layer extracted was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and then purified by column chromatography to obtain Compound 12 (600 mg, 45%).

(32) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 7.31 (d, J=8.4 Hz, 2H), 6.99 (d, J=8.4 Hz 2H), 5.35-5.26 (m, 3H), 5.13 (d, J=7.6 Hz, 1H), 4.64 (d, J=5.6 Hz, 2H), 4.18-4.16 (m, 1H), 3.73 (s, 3H), 2.06-2.04 (m, 9H), 1.61 (t, J=5.6 Hz, 1H).

<Example 4> Preparation of Compound 15

(33) ##STR00023##

(34) Preparation of Compound 13

(35) In methanol (50 mL), 5-formylsalicylic acid (5.0 g, 30.1 mmol) was dissolved, and concentrated sulfuric acid (2 mL) was added thereto. The reaction solution was heated under reflux for 24 hours, then concentrated under reduced pressure, and diluted with ethyl acetate (100 mL). The organic layer was washed with distilled water (100 mL), saturated aqueous sodium hydrogencarbonate solution (200 mL), and brine (200 mL), and then dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated, and vacuum dried to obtain Compound 13 (4.62 g, 85%) as a white solid.

(36) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 11.36 (s, 1H), 9.88 (s, 1H), 8.38 (d, J=2.4 Hz, 1H), 8.00 (dd, J=8.4 Hz, J=2 Hz, 1H), 7.11 (d, J=8.8 Hz, 1H), 4.01 (s, 3H).

(37) Preparation of Compound 14

(38) Compound 13 (1.7 g, 9.38 mmol) and Compound 10 (4.1 g, 10.3 mmol) were dissolved in acetonitrile (50 mL), then 4 Å molecular sieves (4 g) and silver(I) oxide (8.7 g, 37.5 mmol) were added thereto, and the mixture was stirred at room temperature for 3 hours under a nitrogen atmosphere. The reaction solution was concentrated under reduced pressure, diluted with distilled water (50 mL), and then subjected to extraction using ethyl acetate (2×50 mL). The organic layer extracted was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and then purified by column chromatography to obtain Compound 14 (2.85 g, 61%).

(39) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 9.95 (s, 1H), 8.29 (d, J=2 Hz, 1H), 8.01 (dd, J=8.4 Hz, J=2 Hz, 1H), 7.26 (d, J=8.8 Hz, 1H), 5.42-5.30 (m, 4H), 4.27 (d, J=9.2 Hz, 1H), 3.89 (s, 3H), 3.72 (s, 3H), 2.08 (s, 3H), 2.07 (s, 3H), 2.06 (s, 3H).

(40) Preparation of Compound 15

(41) Compound 14 (2.85 g, 5.74 mmol) was dissolved in chloroform: isopropanol (50 mL/10 mL), then silica gel (2.8 g) and sodium borohydride (434 mg, 11.5 mmol) were added thereto at 0° C. under a nitrogen atmosphere, and the mixture was stirred for 2 hours. Distilled water (40 mL) was added to the reaction solution, and the mixture was subjected to extraction using dichloromethane (2×50 mL). The organic layer extracted was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and then purified by column chromatography to obtain Compound 15 (1.42 g, 49%).

<Example 5> Preparation of Compound 20

(42) ##STR00024##

(43) Preparation of Compound 16 With tetrahydrofuran (30 mL), 5-formylsalicylic acid (10.0 g, 60.1 mmol) was diluted, and then N,N-diisopropylethylamine (29.8 mL, 180 mmol) and benzyl bromide (7.15 mL, 60.1 mmol) were added thereto at room temperature. The reaction solution was heated under reflux for 18 hours, then the temperature was lowered to room temperature, and a 2 N aqueous hydrochloric acid solution (100 mL) was added thereto. The mixture was subjected to extraction using ethyl acetate (2×100 mL), and the combined organic layers were dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated under reduced pressure, and purified by column chromatography to obtain Compound 16 (12.9 g, 83%).

(44) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 11.38 (s, 1H), 9.86 (s, 1H), 8.40 (s, 1H), 8.01 (d, J=8.8 Hz, 1H), 7.44 (m, 5H), 7.12 (d, J=8.0 Hz, 1H), 5.42 (s, 2H).

(45) Preparation of Compound 17

(46) Compound 16 (5.0 g, 19.5 mmol) and Compound 10 (8.5 g, 21.4 mmol) were dissolved in acetonitrile (100 mL), 4 Å molecular sieves (10 g) and silver(I) oxide (18.0 g, 78.0 mmol) were added thereto, and the mixture was stirred at room temperature for 12 hours under a nitrogen atmosphere. The reaction solution was concentrated under reduced pressure, diluted with distilled water (100 mL), and subjected to extraction using ethyl acetate (2×200 mL). The organic layer extracted was dried over anhydrous magnesium sulfate, filtered, concentrated under reduced pressure, and then purified by column chromatography to obtain Compound 17 (8.63 g, 77%).

(47) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 9.94 (s, 1H), 8.28 (s, 1H), 8.02 (d, J=8.8 Hz, 1H), 7.46-7.28 (m, 6H), 5.41-5.32 (m, 6H), 4.27 (d, J=9.2 Hz, 1H), 3.71 (s, 3H), 2.06-2.04 (m, 9H).

(48) Preparation of Compound 18

(49) Compound 17 (3.10 g, 5.41 mmol) was dissolved in chloroform/isopropanol (45 mL/9 mL), then silica gel (3 g) and sodium borohydride (0.41 g, 10.8 mmol) were added thereto at 0° C. under a nitrogen atmosphere, and the mixture was stirred for 2 hours. Distilled water (100 mL) was added to the reaction solution, and then the mixture was subjected to extraction using ethyl acetate (200 mL). The organic layer extracted was dried over anhydrous magnesium sulfate, filtered, concentrated under reduced pressure, and then purified by column chromatography to obtain Compound 18 (2.73 g, 87%) as a white solid.

(50) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 7.74 (s, 1H), 7.48-7.34 (m, 6H), 7.16 (d, J=8.8 Hz, 1H), 5.35-5.26 (m, 5H), 5.16-5.14 (m, 1H), 4.17-4.15 (m, 1H), 3.73 (s, 3H), 2.04 (s, 9H), 1.73 (t, J=7.2 Hz, 1H).

(51) Preparation of Compound 19

(52) Compound 18 (2.40 g, 4.17 mmol) was dissolved in ethanol (150 mL) and then Raney nickel (240 mg) was added thereto. The reaction solution was stirred at room temperature for 10 minutes under a hydrogen atmosphere. The reaction solution was filtered through Celite and concentrated to obtain Compound 19 (2.10 g) as a white solid.

(53) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 8.06 (s, 1H) 7.61 (d, J=8.8 Hz, 1H), 7.23 (d, J=8.0 Hz 1H), 5.43-5.29 (m, 5H), 4.17 (s, 2H), 4.32 (d, J=8.4 Hz, 1H) 3.69 (s, 3H), 2.11-2.08 (m, 9H), 1.24 (t, 1H).

(54) Preparation of Compound 20

(55) Compound 19 (7.0 g, 14.5 mmol) and 2-methoxyethylamine (1.38 mL, 1.59 mmol) were dissolved in N,N-dimethylformamide (14 mL) and then N,N,N′,N′-tetramethyl-O-(1H-benzotrazol-1-yl)uronium hexafluorophosphate (6.57 g, 17.3 mmol) and N,N-diisopropylethylamine (5 mL, 28.9 mmol) were added thereto at 0° C. under a nitrogen atmosphere. The reaction solution was stirred at room temperature for 2 hours, then a saturated aqueous ammonium chloride solution (100 mL) was added to the reaction solution, and the mixture was subjected to extraction using ethyl acetate (2×100 mL). The combined organic layers were washed with brine (200 mL) and then dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated, and purified by column chromatography to obtain Compound 20 (7.53 g, 96%).

(56) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 7.98 (d, J=2 Hz, 1H), 7.49 (br s, 1H), 7.46 (dd, J=8.4 Hz, J=2.4 Hz, 1H), 7.04 (d, J=8.4 Hz, 1H), 5.42-5.28 (m, 4H), 4.66 (s, 1H), 4.19 (d, J=9.2 Hz, 1H), 3.72 (s, 3H), 3.57 (s, 3H), 3.42 (s, 3H), 2.05 (s, 9H).

<Example 6> Preparation of Compound 22

(57) ##STR00025##

(58) Compound 19 (1.0 g, 2.06 mmol) and Compound 21 (1.49 g, 2.80 mmol, Compound 21 was prepared by the method described in PCT/US2016/063564) were dissolved in N,N-dimethylformamide (10 mL), and then N,N,N′,N′-tetramethyl-O-(1H-benzotrazol-1-yl)uronium hexafluorophosphate (1.56 g, 4.12 mmol) and N,N-diisopropylethylamine (1.07 mL, 6.18 mmol) were added thereto at 0° C. under a nitrogen atmosphere. The reaction solution was stirred at room temperature for 12 hours, then a saturated aqueous ammonium chloride solution (100 mL) was added to the reaction solution, and the mixture was subjected to extraction using ethyl acetate (2×100 mL). The combined organic layers were washed with brine (200 mL) and dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated, and purified by column chromatography to obtain Compound 22 (1.6 g, 80%).

(59) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 7.98 (s, 1H), 7.46 (dd, J=8.4 Hz, J=2.4 Hz, 1H), 7.41 (br s, 1H), 7.04 (d, J=8.4 Hz, 1H), 5.93-5.25 (m, 4H), 4.67 (d, J=5.2 Hz, 2H), 4.20 (d, J=9.6 Hz, 1H), 4.08 (t, J=4.8 Hz, 2H), 3.74 (s, 6H), 3.72-3.49 (m, 22H), 2.06 (s, 9H), 1.53 (s, 18H).

<Example 7> Preparation of Compound 25

(60) ##STR00026##

(61) Preparation of Compound 23

(62) Compound 9 (2.2 g, 2.34 mmol) was dissolved in toluene (65 mL), then triphosgene (250 mg, 0.84 mmol) and triethylamine (0.44 mL, 3.16 mmol) were added thereto at −10° C., and the mixture was stirred for 1 hour under a nitrogen atmosphere. Compound 20 (1.39 g, 2.58 mmol) was dissolved in dry tetrahydrofuran (65 mL), triethylamine (0.44 mL, 3.16 mmol) was added thereto, and then this solution was gradually added to the reaction solution. After 30 minutes, the reaction solution was heated under reflux and stirred for 4 hours. The reaction solution was concentrated, diluted with dichloromethane (100 mL), washed with brine (50 mL), and dried over anhydrous sodium sulfate. The resultant was filtered, concentrated under reduced pressure, and then purified by column chromatography to obtain Compound 23 (2.5 g, 72%).

(63) EI-MS m/z: [M+H].sup.+ 1504.7, 1/2[M+H].sup.+ 753.5.

(64) Preparation of Compound 24

(65) Compound 23 (2.0 g, 1.33 mmol) was dissolved in dichloromethane (15 mL), then pyrrolidine (0.13 mL, 1.59 mmol) and tetrakis(triphenylphosphine)palladium(O) (76 mg, 0.066 mmol) were added thereto, and the mixture was stirred at room temperature for 6 hours under a nitrogen atmosphere. The reaction solution was concentrated under reduced pressure and then purified by column chromatography to obtain Compound 24 (1.7 g, 90%).

(66) EI-MS m/z: [M+H].sup.+ 1420.6, 1/2[M+H].sup.+ 711.2.

(67) Preparation of Compound 25

(68) Compound 24 (1.2 g, 0.84 mmol) was dissolved in toluene (24 mL), triphosgene (90 mg, 0.30 mmol) and pyridine (0.33 mL, 4.22 mmol) were added thereto at −10° C., and the mixture was stirred for 1 hour under a nitrogen atmosphere. Compound 22 (974 mg, 1.01 mmol) was dissolved in dry tetrahydrofuran (24 mL), N,N-diisopropylethylamine (0.21 mL, 1.26 mmol) was added thereto, and then this solution was gradually added to the reaction solution. After 30 minutes, the reaction solution was heated under reflux and stirred for 4 hours. The reaction solution was concentrated, diluted with dichloromethane (50 mL), washed with brine (30 mL), and dried over anhydrous sodium sulfate. The resultant was filtered, concentrated under reduced pressure, and then purified by column chromatography to obtain Compound 25 (800 mg, 40%).

(69) EI-MS m/z: [M+H].sup.+ 2409.9, 1/2[M+Na].sup.+1214.3.

<Example 8> Preparation of Compound 28

(70) ##STR00027##

(71) Preparation of Compound 26

(72) Compound 25 (800 mg, 0.33 mmol) was dissolved in tetrahydrofuran/distilled water (4 mL/4 mL), acetic acid (8 mL) was added thereto, and then the mixture was stirred at room temperature for 16 hours under a nitrogen atmosphere. The reaction solution was concentrated under reduced pressure and then purified by column chromatography to obtain Compound 26 (660 mg, 90%).

(73) EI-MS m/z: [M+H].sup.+ 2181.6, 1/2[M-Boc+H].sup.+1041.5.

(74) Preparation of Compound 27

(75) Compound 26 (660 mg, 0.15 mmol) was dissolved in dichloromethane (15 mL), then Dess-Martin periodinane (141 mg, 0.33 mmol) was added thereto, and the mixture was stirred at room temperature for 3.5 hours under a nitrogen atmosphere. The reaction solution was concentrated under reduced pressure and then purified by column chromatography to obtain Compound 27 (477 mg, 70%).

(76) EI-MS m/z: [M+H].sup.+ 2177.6, 1/2[M+H].sup.+ 1089.5.

(77) Preparation of Compound 28

(78) Compound 27 (150 mg, 0.068 mmol) was dissolved in methanol/tetrahydrofuran (3 mL/3 mL), and then a solution of lithium hydroxide (26 mg, 0.62 mmol) in distilled water (3 mL) was gradually added thereto at −40° C. The mixture was stirred for 2 hours while gradually raising the reaction temperature to 0° C. The reaction solution was neutralized with acetic acid, then concentrated under reduced pressure, and vacuum dried. The solid obtained was diluted with dichloromethane (5 mL), then trifluoroacetic acid (1.2 mL) was added thereto at 0° C., and the mixture was stirred for 2 hours. The reaction solution was concentrated under reduced pressure, then purified by HPLC, and freeze-dried to obtain Compound 28 (20 mg, 16%) as a white solid.

(79) EI-MS m/z: [M+H].sup.+ 1697.5, 1/2[M+H].sup.+ 849.3.

<Example 9> Preparation of Compound 29

(80) ##STR00028##

(81) Compound 29 was prepared from Compound 9 and Compound 12 by a method similar to that for the synthesis of Compound 28. EI-MS m/z: [M+H].sup.+ 1596.9, 1/2[M+H].sup.+ 799.3.

<Example 10> Preparation of Compound 30

(82) ##STR00029##

(83) Compound 30 was prepared from Compound 9 and Compound 15 by a method similar to that for the synthesis of Compound 28.

<Example 11> Preparation of Compound 32

(84) ##STR00030##

(85) Compound 32 was prepared from Compound 19 and Compound 31 (Compound 31 was prepared by the method described in PCT/US2016/063564) by a method similar to that for the synthesis of Compound 22.

(86) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 7.97 (s, 1H), 7.46 (dd, J=8.4 Hz, J=2.4 Hz, 1H), 7.41 (bs, 1H), 7.04 (d, J=8.4 Hz, 1H), 5.72 (s, 1H), 5.42-5.27 (m, 4H), 4.66 (d, J=5.2 Hz, 2H), 4.25 (d, J=9.6 Hz, 1H), 3.97 (t, J=4.8 Hz, 2H), 3.78 (s, 3H), 3.74-3.64 (m, 10H), 2.04 (s, 9H), 1.53 (s, 9H). EI-MS m/z: [M+H].sup.+ 731.5.

<Example 12> Preparation of Compound 34

(87) ##STR00031##

(88) Compound 34 was prepared from Compound 24 and Compound 32 by a method similar to that for the synthesis of Compound 28.

(89) EI-MS m/z: [M+H].sup.+ 1565.5, 1/2[M+H].sup.+ 783.4.

<Example 13> Preparation of Compound 39

(90) ##STR00032##

(91) Compound 35 and Compound 36 were prepared by the method described in PCT/US2016/063564.

(92) ##STR00033##

(93) Preparation of Compound 37

(94) Compound 24 (400 mg, 0.28 mmol) was dissolved in toluene (10 mL), then triphosgene (30 mg, 0.10 mmol) and triethylamine (0.053 mL, 0.38 mmol) were added thereto at −10° C., and the mixture was stirred for 1 hour under a nitrogen atmosphere. Compound 35 (177 mg, 0.33 mmol) was dissolved in dry tetrahydrofuran (10 mL), triethylamine (0.053 mL, 0.38 mmol) was added thereto, and then this solution was gradually added to the reaction solution. After 30 minutes, the reaction solution was heated under reflux and stirred for 4 hours. The reaction solution was concentrated, diluted with dichloromethane (50 mL), washed with brine (30 mL), and dried over anhydrous sodium sulfate. The resultant was filtered, concentrated under reduced pressure, and then purified by column chromatography to obtain Compound 37 (192 mg, 34%).

(95) EI-MS m/z: [M+H].sup.+ 1971.8, 1/2[M+H].sup.+ 986.6.

(96) Preparation of Compound 38

(97) Compound 37 (192 mg, 0.097 mmol) was dissolved in dichloromethane (5 mL), then pyrrolidine (0.012 mL, 0.14 mmol) and tetrakis(triphenylphosphine)palladium(O) (11.2 mg, 0.096 mmol) were added thereto, and the mixture was stirred at room temperature for 6 hours under a nitrogen atmosphere. The reaction solution was concentrated under reduced pressure and then purified by column chromatography to obtain Compound 38 (180 mg, 96%).

(98) EI-MS m/z: [M+H].sup.+ 1932.8, 1/2[M+H].sup.+ 966.5

(99) Preparation of Compound 39

(100) Compound 38 (180 mg, 0.093 mmol) and Compound 36 (112 mg, 0.116 mmol) were dissolved in N,N-dimethylformamide (2 mL), and then 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (HATU, 46 mg, 0.121 mmol) and N,N-diisopropylethylamine (0.032 mL, 0.186 mmol) were added thereto at 0° C. under a nitrogen atmosphere. The reaction solution was stirred at room temperature for 36 hours, then distilled water (20 mL) was added to the reaction solution, and the mixture was subjected to extraction using ethyl acetate (2×20 mL). The combined organic layers were washed with brine (20 mL) and then dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated, and purified by column chromatography to obtain Compound 39 (133 mg, 50%).

(101) EI-MS m/z: [M+H].sup.+ 2876.4, 1/2[M+H].sup.+ 1438.6.

<Example 14> Preparation of Compound 42

(102) ##STR00034##

(103) Preparation of Compound 40

(104) Compound 39 (133 mg, 0.046 mmol) was dissolved in tetrahydrofuran/distilled water (1 mL/1 mL), acetic acid (2 mL) was added thereto, and then the mixture was stirred at room temperature for 16 hours under a nitrogen atmosphere. The reaction solution was concentrated under reduced pressure and then purified by column chromatography to obtain Compound 40 (67.4 mg, 55%).

(105) EI-MS m/z: [M+H].sup.+ 2647.4, 1/2[M+H].sup.+ 1324.5.

(106) Preparation of Compound 41

(107) Compound 40 (67.4 mg, 0.025 mmol) was dissolved in dichloromethane (2 mL), then Dess-Martin periodinane (23.7 mg, 0.056 mmol) was added thereto, and the mixture was stirred at room temperature for 3.5 hours under a nitrogen atmosphere. The reaction solution was concentrated under reduced pressure and then purified by column chromatography to obtain Compound 41 (43 mg, 65%).

(108) EI-MS m/z: [M+H].sup.+ 2643.1, 1/2[M+H].sup.+ 1322.5.

(109) Preparation of Compound 42

(110) Compound 41 (43 mg, 0.016 mmol) was dissolved in methanol/tetrahydrofuran (0.5 mL/0.5 mL) and then a solution of lithium hydroxide (6.8 mg, 0.16 mmol) in distilled water (0.5 mL) was gradually added thereto at −40° C. The mixture was stirred for 2 hours while gradually raising the reaction temperature to −10° C. The reaction solution was neutralized with acetic acid, then concentrated under reduced pressure, and vacuum dried. The solid obtained was diluted with dichloromethane (1 mL), then trifluoroacetic acid (0.2 mL) was added thereto at 0° C., and the mixture was stirred for 2 hours. The reaction solution was concentrated under reduced pressure, then purified by HPLC, and freeze-dried to obtain Compound 42 as a white solid (7.0 mg).

(111) EI-MS m/z: [M+H].sup.+ 2263.4, 1/2[M+H].sup.+ 1132.3.

<Example 15> Preparation of Compound 48

(112) ##STR00035##

(113) Preparation of Compound 44

(114) Compound 43 (37 g, 40.2 mmol, Compound 43 was prepared by the method described in J. Med. Chem., 2004, 47, 1161-1174) was dissolved in dichloromethane (400 mL), then trichloroisocyanuric acid (14.9 g, 64.3 mmol) and 2,2,6,6-tetramethyl-1-piperidinyloxy (1.3 g, 8.0 mmol) were added thereto at 0° C., and the mixture was stirred for 1 hour under a nitrogen atmosphere. The reaction solution was diluted by addition of dichloromethane (400 mL), washed with a saturated aqueous sodium hydrogencarbonate solution (400 mL), sodium thiosulfate (0.2 M, 400 mL), and brine (200 mL) in this order, and then dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated under reduced pressure, and purified by column chromatography to obtain Compound 44 (35 g, 83%).

(115) .sup.1H-NMR (400 MHz, CDCl.sub.3) (rotamers) δ 7.72 (s, 2H), 6.73 (s, 2H), 4.97 (d, 2H), 4.31 (d, 2H), 4.12 (t, 4H), 3.95-3.96 (m, 6H), 3.71 (d, 2H), 3.64 (d, 2H), 3.45 (d, 2H), 2.82-2.75 (m, 2H), 2.55 (d, 2H), 1.99 (m, 4H), 1.72 (m, 2H), 0.85 (s, 18H), 0.08 (d, 12H).

(116) Preparation of Compound 45

(117) Compound 44 (5 g, 5.45 mmol) was dissolved in dichloromethane (90 mL), then 2,6-lutidine (5.1 ml, 43.8 mmol) and triflic anhydride (5.5 ml, 39.0 mmol) were added thereto at −40° C., and the mixture was stirred for 1 hour under a nitrogen atmosphere. The reaction solution was diluted by addition of dichloromethane (90 mL), washed with a saturated aqueous sodium hydrogencarbonate solution (90 mL), distilled water (90 mL), and brine (90 mL), and then dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated under reduced pressure, and purified by column chromatography to obtain Compound 45 (4.0 g, 62%).

(118) .sup.1H-NMR (400 MHz, CDCl.sub.3) (rotamers) δ 7.71 (s, 2H), 6.77 (s, 2H), 6.08 (s, 2H), 4.79-4.78 (m, 2H), 4.18-4.09 (m, 6H), 4.02-3.92 (m, 8H), 3.22-3.14 (m, 2H), 3.01-2.97 (m, 2H), 2.02-1.97 (m, 4H), 0.91 (s, 18H), 0.11 (s, 12H).

(119) Preparation of Compound 46

(120) Compound 45 (3.1 g, 2.6 mmol) was dissolved in toluene (45 mL), then methylboronic acid (1.1 g, 18.2 mmol), silver(I) oxide (4.8 g, 20.9 mmol), potassium phosphate (6.6 g, 31.5 mmol), triphenylarsine (642 mg, 2.1 mmol), and bis(triphenylphosphine)palladium(II) dichloride (184 mg, 0.3 mmol) were added thereto under an argon atmosphere, and the mixture was heated and stirred at 80° C. for 3 hours. The reaction solution was filtered through Celite, then concentrated, and purified by column chromatography to obtain Compound 46 (955 mg, 40%).

(121) .sup.1H-NMR (400 MHz, CDCl.sub.3) (rotamers) δ 7.68 (s, 2H), 6.77 (s, 2H), 5.52 (s, 2H), 4.66-4.64 (m, 2H), 4.14-4.07 (m, 6H), 3.94-3.92 (m, 8H), 2.75-2.73 (m, 2H), 2.55-2.51 (m, 2H), 1.99-1.93 (m, 4H), 1.72-1.68 (m, 2H), 1.60 (s, 6H), 0.88 (s, 18H), 0.09 (s, 12H).

(122) Preparation of Compound 47

(123) Compound 46 (2.9 g, 3.17 mmol) was dissolved in ethanol (44 mL), and then zinc dust (12.9 g, 197 mmol) and formic acid (5% ethanol solution, 128 mL) were added thereto. The reaction solution was stirred at room temperature for 15 minutes and then filtered through Celite, and ethyl acetate (500 mL) was added thereto. The organic layer was washed with distilled water (200 mL), a saturated aqueous sodium hydrogencarbonate solution (200 mL), and brine (200 mL) in this order and then dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated, and purified by column chromatography to obtain Compound 47 (3.0 g, 82%).

(124) .sup.1H-NMR (400 MHz, CDCl.sub.3) (rotamers) δ 6.74 (s, 2H), 6.23 (s, 2H), 6.18 (bs, 2H), 4.64 (bs, 2H), 4.34 (s, 3H), 4.07-3.93 (m, 6H), 3.80-3.76 (m, 7H), 2.74-2.68 (m, 2H), 2.53 (d, 2H), 1.91 (m, 4H), 1.67-1.62 (m, 8H), 0.88 (s, 18H), 0.05 (d, 12H).

(125) Preparation of Compound 48

(126) Compound 47 (3.0 g, 3.51 mmol) was dissolved in dichloromethane (175 mL), and then pyridine (0.57 mL, 7.03 mmol) and allyl chloroformate (0.34 mL, 3.16 mmol) were added thereto at −78° C. under a nitrogen atmosphere. The reaction solution was stirred for 1 hour, then the reaction temperature was raised to room temperature, and the reaction solution was concentrated and then purified by column chromatography to obtain Compound 48 (1.33 g, 44%).

(127) .sup.1H-NMR (400 MHz, CDCl.sub.3) (rotamers) δ 8.80 (br s, 1H), 7.82 (s, 1H), 6.78 (s, 1H), 6.74 (s, 1H), 6.23 (s, 1H), 6.19 (br s, 2H), 5.99-5.90 (m, 1H), 5.34 (d, 1H), 5.23 (d, 1H), 4.63 (m, 4H), 4.35 (br s, 2H), 4.10 (t, 2H), 3.99 (t, 3H), 3.99 (m, 2H), 3.80 (s, 5H), 3.76 (s, 4H), 2.73 (m, 2H), 2.55 (m, 2H), 1.95-1.90 (m, 4H), 1.68-1.63 (m, 8H), 0.88 (s, 18H), 0.05 (d, 12H).

<Example 16> Preparation of Compound 49

(128) ##STR00036##

(129) Compound 19 (3.7 g, 7.56 mmol) and propargylamine (0.43 mL, 7.07 mmol) were dissolved in N,N-dimethylformamide (50 mL), and then N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (2.32 g, 12.1 mmol) and 1-hydroxybenzotriazole (2.04 g, 15.1 mmol) were added thereto. The reaction solution was stirred at room temperature for 12 hours, then distilled water (100 mL) was added to the reaction solution, and the mixture was subjected to extraction using ethyl acetate (2×100 mL). The combined organic layers were washed with brine (200 mL) and then dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated, and purified by column chromatography to obtain Compound 49 (3.4 g, 86%).

(130) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 8.01 (d, 1H), 7.57 (t, 1H), 7.49 (dd, 1H), 7.02 (d, 1H), 5.42-5.38 (m, 1H), 5.36-5.28 (m, 2H), 4.67 (d, 2H), 4.31-4.13 (m, 3H), 2.23 (t, 1H), 2.07-2.06 (m, 9H), 1.88 (t, 1H).

<Example 17> Preparation of Compound 53

(131) ##STR00037##

(132) Preparation of Compound 51

(133) Compound 50 (4.5 g, 25.68 mmol) was dissolved in N,N-dimethylformamide (50 mL), then sodium hydride (1.23 g, 30.82 mmol) was added thereto at 0° C. under a nitrogen atmosphere, the mixture was stirred for 30 minutes, then propargyl bromide (up to 80% toluene solution, 4.96 mL, 33.4 mmol) was added thereto, and then this mixture was stirred at room temperature for 3 hours. Distilled water (40 mL) was added to the reaction solution, and then the mixture was subjected to extraction using ethyl acetate (2×50 mL). The combined organic layers were washed with brine (100 mL) and then dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated, and purified by column chromatography to obtain Compound 51 (4.35 g, 79%).

(134) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 4.21 (d, J=2.4 Hz, 2H), 3.70-3.38 (m, 10H), 3.39 (t, J=5.2 Hz, 2H), 2.43 (t, J=2.4 Hz, 1H).

(135) Preparation of Compound 52

(136) Compound 51 (1.55 g, 7.03 mmol) was dissolved in dry tetrahydrofuran (30 mL)/distilled water (2.53 mL), then triphenylphosphine (2.21 g, 8.44 mmol) was added thereto, and the mixture was stirred at room temperature for 24 hours. The resultant mixture was concentrated and purified by column chromatography to obtain Compound 52 (1.3 g, 99%).

(137) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 4.21 (s, 2H), 3.69-3.64 (m, 8H), 3.52-3.49 (m, 2H), 2.88-2.85 (m, 2H), 4.23 (s, 1H). EI-MS m/z: [M+H].sup.+ 188.2.

(138) Preparation of Compound 53

(139) Compound 52 (2.0 g, 10.68 mmol) and Compound 19 (4.7 g, 9.71 mmol) were dissolved in N,N-dimethylformamide (50 mL), and then N,N,N′,N′-tetramethyl-O-(1H-benzotrazol-1-yl)uronium hexafluorophosphate (3.71 g, 11.6 mmol) and N,N-diisopropylethylamine (3.38 mL, 19.4 mmol) were added thereto at 0° C. under a nitrogen atmosphere. The reaction solution was stirred at room temperature for 24 hours, then distilled water (100 mL) was added to the reaction solution, and the mixture was subjected to extraction using ethyl acetate (2×100 mL). The combined organic layers were washed with brine (200 mL) and then dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated, and purified by column chromatography to obtain Compound 53 (4.78 g, 75%).

(140) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 7.95 (s, 1H), 7.46 (d, J=8.4 Hz, 1H), 7.41-7.37 (m, 1H), 7.04 (d, J=8.8 Hz, 1H), 5.41-5.25 (m, 4H), 4.65 (d, J=4.4 Hz, 2H), 4.21 (d, J=9.2 Hz, 1H), 4.17 (s, 2H), 3.74 (s, 3H), 3.68 (s, 11H), 3.56-3.50 (m, 1H), 2.05 (s, 9H).

<Example 18> Preparation of Compound 55

(141) ##STR00038##

(142) Preparation of Compound 55

(143) Compound 19 (3.68 g, 7.60 mmol) and Compound 54 (1.46 g, 8.40 mmol, Compound 54 was prepared by the method described in PCT/US2016/063564) were dissolved in N,N-dimethylformamide (10 mL), then N,N,N′,N′-tetramethyl-O-(1H-benzotrazol-1-yl)uronium hexafluorophosphate (4.53 g, 11.40 mmol) and N,N-diisopropylethylamine (3.97 mL, 22.80 mmol) were added thereto at 0° C. under a nitrogen atmosphere, and then the mixture was stirred at room temperature for 12 hours. A saturated aqueous ammonium chloride solution (100 mL) was added to the reaction solution, the mixture was subjected to extraction using ethyl acetate (2×100 mL), and then the extract was dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated, and purified by column chromatography to obtain Compound 55 (3.31 g, 68%).

(144) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 7.99 (s, 1H), 7.47 (d, J=8.4 Hz, 1H), 7.41 (s, 1H), 5.42-5.25 (m, 4H), 4.68 (d, J=5.6 Hz, 2H), 4.20 (d, J=9.2 Hz, 1H), 3.78-3.68 (m, 11H), 3.58-3.52 (m, 1H), 3.39-3.36 (m, 2H), 2.06 (s, 9H), 1.89-1.86 (m, 1H).

<Example 19> Preparation of Compound 58

(145) ##STR00039##

(146) Preparation of Compound 56

(147) Compound 48 (1.33 g, 1.41 mmol) was dissolved in toluene (40 mL), then triphosgene (151 mg, 0.51 mmol) and triethylamine (0.26 mL, 1.91 mmol) were added thereto at −10° C., and the mixture was stirred for 1 hour under a nitrogen atmosphere. Compound 20 (845 mg, 1.56 mmol) was dissolved in dry tetrahydrofuran (40 mL), triethylamine (0.26 mL, 1.91 mmol) was added thereto, and then this solution was gradually added to the reaction solution. After 30 minutes, the reaction solution was heated under reflux and stirred for 4 hours. The reaction solution was concentrated, diluted with dichloromethane (30 mL), then washed with brine (20 mL), and dried over anhydrous sodium sulfate. The resultant was filtered, concentrated under reduced pressure, and then purified by column chromatography to obtain Compound 56 (1.15 mg, 54%).

(148) EI-MS m/z: [M+H].sup.+ 1504.7, 1/2[M+H].sup.+ 753.5.

(149) Preparation of Compound 57

(150) Compound 56 (1.15 g, 0.79 mmol) was dissolved in dichloromethane (10 mL), then pyrrolidine (0.08 mL, 1.35 mmol) and tetrakis(triphenylphosphine)palladium(O) (45 mg, 0.057 mmol) were added thereto, and the mixture was stirred at room temperature for 2 hours under a nitrogen atmosphere. The reaction solution was concentrated under reduced pressure and then purified by column chromatography to obtain Compound 57 (820 mg, 72%).

(151) EI-MS m/z: [M+H].sup.+ 1420.6, 1/2[M+H].sup.+ 711.2.

(152) Preparation of Compound 58

(153) Compound 57 (730 mg, 0.51 mmol) was dissolved in toluene (20 mL), then triphosgene (54 mg, 0.36 mmol) and pyridine (0.2 mL, 2.56 mmol) were added thereto at −10° C., and the mixture was stirred for 1 hour under a nitrogen atmosphere. Compound 49 (321 mg, 0.61 mmol) was dissolved in dry tetrahydrofuran (20 mL), N,N-diisopropylethylamine (0.14 mL, 0.77 mmol) was added thereto, and then this solution was gradually added to the reaction solution. After 30 minutes, the reaction solution was heated under reflux and stirred for 4 hours. The reaction solution was concentrated, diluted with dichloromethane (50 mL), then washed with brine (30 mL), and dried over anhydrous sodium sulfate. The resultant was filtered, concentrated under reduced pressure, and then purified by column chromatography to obtain Compound 58 (650 mg, 64%).

(154) EI-MS m/z: [M+H].sup.+ 1969.2, 1/2[M+H].sup.+ 985.2.

<Example 20> Preparation of Compound 61

(155) ##STR00040##

(156) Preparation of Compound 59

(157) Compound 58 (650 mg, 0.33 mmol) was dissolved in tetrahydrofuran/distilled water (3.5 mL/3.5 mL), acetic acid (7 mL) was added thereto, and the mixture was stirred at room temperature for 16 hours under a nitrogen atmosphere. The reaction solution was concentrated under reduced pressure and then purified by column chromatography to obtain Compound 59 (440 mg, 78%).

(158) EI-MS m/z: [M+H].sup.+ 1740.0, [M+Na].sup.+1762.0, 1/2[M+H].sup.+ 871.0.

(159) Preparation of Compound 60

(160) Compound 59 (440 mg, 0.25 mmol) was dissolved in dichloromethane (25 mL), then Dess-Martin periodinane (236 mg, 0.55 mmol) was added thereto, and the mixture was stirred at room temperature for 2.5 hours under a nitrogen atmosphere. The reaction solution was concentrated under reduced pressure and then purified by column chromatography to obtain Compound 60 (365 mg, 84%).

(161) EI-MS m/z: [M+H].sup.+ 1736.0, 1/2[M+H].sup.+ 869.5.

(162) Preparation of Compound 61

(163) Compound 60 (365 mg, 0.21 mmol) was dissolved in methanol/tetrahydrofuran (9 mL/2 mL), and then a solution of lithium hydroxide (58 mg, 1.4 mmol) in distilled water (9 mL) was gradually added thereto at −40° C. The mixture was stirred for 2 hours while gradually raising the reaction temperature to 0° C. The reaction solution was neutralized with acetic acid, then concentrated under reduced pressure, then purified by HPLC, and freeze-dried to obtain Compound 61 (100 mg, 32%) as a white solid.

(164) EI-MS m/z: [M+H].sup.+ 1456.8, 1/2[M+H].sup.+ 729.5.

<Example 21> Preparation of Compound 62

(165) ##STR00041##

(166) Compound 62 was prepared from Compound 53 and Compound 57 by a method similar to that for the synthesis of Compound 61.

(167) EI-MS m/z: [M+H].sup.+ 1588.7, 1/2[M+H].sup.+ 795.3.

<Example 22> Preparation of Compound 63

(168) ##STR00042##

(169) Compound 63 was prepared from Compound 55 and Compound 57 by a method similar to that for the synthesis of Compound 61.

(170) EI-MS m/z: [M+H].sup.+ 1575.8, 1/2[M+H].sup.+ 788.7.

<Example 23> Preparation of Compound 65

(171) ##STR00043##

(172) Preparation of Compound 65

(173) Compound 61 (100 mg, 0.068 mmol) was dissolved in dimethylsulfoxide (1.6 mL), then Compound 64 (136 mg, 0.302 mmol, Compound 64 was prepared by the method described in PCT/US2016/063564) was added thereto under a nitrogen atmosphere, and then a solution of a copper(II)sulfate pentahydrate (7.4 mg, 0.03 mmol) and sodium ascorbate (28 mg, 0.15 mmol) in distilled water (0.4 ml) was added to the reaction solution. After being stirred at room temperature for 30 minutes, the reaction solution was concentrated under reduced pressure, then purified by HPLC, and freeze-dried to obtain Compound 65 (15.2 mg, 13%) as a white solid.

(174) EI-MS m/z: [M+H].sup.+ 1645.8, 1/2[M+H].sup.+ 823.9.

<Example 24> Preparation of Compound 70

(175) ##STR00044##

(176) Preparation of Compound 66 In dichloromethane (150 mL), 3-amino-1-propanol (3.0 g, 66.57 mmol) was dissolved, and then di-t-butyl dicarbonate (16 g, 73.2 mmol) was added thereto at 0° C. under a nitrogen atmosphere. After being stirred at room temperature for 12 hours, the reaction solution was concentrated under reduced pressure and purified by column chromatography to obtain Compound 66 (6.4 g, 92%).

(177) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 4.78 (s, 1H), 3.65 (m, 2H), 3.30 (m, 2H), 2.90 (s, 1H), 1.68 (m, 2H), 1.48 (s, 9H).

(178) Preparation of Compound 67

(179) Compound 66 (6.04 g, 34.47 mmol) and triethylamine (14.4 mL, 103.4 mmol) were dissolved in tetrahydrofuran (100 mL) and then methanesulfonic anhydride (7.21 g, 41.36 mmol) was gradually added thereto at 0° C. under a nitrogen atmosphere. The temperature was gradually raised to room temperature, and then the mixture was stirred for 12 hours. The reaction solution was concentrated under reduced pressure and purified by column chromatography to obtain Compound 67 (9.01 g, 98%).

(180) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 4.73 (s, 1H), 4.30 (t, J=5.9 Hz, 2H), 3.31-3.24 (m, 2H), 3.04 (s, 3H), 1.94 (t, J=6.1 Hz, 2H), 1.44 (s, 9H).

(181) Preparation of Compound 68

(182) Compound 67 (3.0 g, 11.84 mol) was dissolved in N,N-dimethylformamide (40 mL), then sodium azide (924 mg, 14.21 mmol) was added thereto at room temperature under a nitrogen atmosphere, and the reaction mixture was stirred at 60° C. for 12 hours. Distilled water (50 mL) and a 1 N aqueous hydrochloric acid solution (5 mL) were added to the reaction solution, the mixture was subjected to extraction using ethyl acetate (100 mL), and then the extract was dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated, and purified by column chromatography to obtain Compound 68 (2.3 g, 99%).

(183) .sup.1H-NMR (600 MHz, CDCl.sub.3) δ 4.63 (s, 1H), 3.36 (t, J=6.6 Hz, 2H), 3.24-3.18 (m, 2H), 1.80-1.75 (m, 2H), 1.45 (s, 9H).

(184) Preparation of Compound 69

(185) Compound 68 (3.8 g, 18.98 mmol) was dissolved in dichloromethane (10 mL), and hydrochloric acid (4 M 1,4-dioxane solution, 10 mL) was gradually added thereto at 0° C. under a nitrogen atmosphere. The reaction mixture was stirred for 12 hours and then concentrated under reduced pressure to obtain Compound 69 (2.5 g, 99%).

(186) .sup.1H-NMR (600 MHz, DMSO-d.sub.6) δ 8.06 (s, 3H), 3.47 (t, J=6.6 Hz, 2H), 2.82 (t, J=7.2 Hz, 2H), 1.84-1.79 (m, 2H).

(187) Preparation of Compound 70

(188) Compound 19 (4.1 g, 8.46 mmol) and Compound 69 (1.1 g, 11.0 mmol) were dissolved in N,N-dimethylformamide (20 mL), then N,N,N′,N′-tetramethyl-O-(1H-benzotriazol-1-yl)uronium hexafluorophosphate (4.39 g, 11.0 mmol) and N,N-diisopropylethylamine (2.96 mL, 16.92 mmol) were added thereto at 0° C. under a nitrogen atmosphere, and then the mixture was stirred at room temperature for 12 hours. A saturated aqueous ammonium chloride solution (100 mL) was added to the reaction solution, and the mixture was subjected to extraction using ethyl acetate (2×100 mL), and then the extract was dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated, and purified by column chromatography to obtain Compound 70 (5.48 g, 88%).

(189) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 8.07 (s, 1H), 7.50-7.46 (m, 2H), 7.01 (d, J=8.4 Hz, 1H), 5.45-5.30 (m, 4H), 4.69 (d, J=5.6 Hz, 2H), 4.21 (d, J=9.6 Hz, 1H), 3.74 (s, 3H), 3.67-3.60 (m, 1H), 3.47-3.41 (m, 3H), 2.80 (s, 2H), 2.07-2.05 (m, 9H), 1.98-1.91 (m, 2H), 1.80-1.77 (m, 1H).

<Example 25> Preparation of Compound 72

(190) ##STR00045##

(191) Preparation of Compound 72

(192) Compound 19 (3.6 g, 7.42 mmol) and Compound 71 (1.0 g, 8.16 mmol, Compound 71 was prepared by a method similar to that for the synthesis of Compound 69) were dissolved in N,N-dimethylformamide (15 mL) and then N,N,N′,N′-tetramethyl-O-(1H-benzotrazol-1-yl)uronium hexafluorophosphate (4.2 g, 11.2 mmol) and N,N-diisopropylethylamine (3.2 mL, 18.6 mmol) were added thereto at 0° C. under a nitrogen atmosphere. The reaction solution was stirred at room temperature for 14 hours, then a saturated aqueous ammonium chloride solution (50 mL) was added to the reaction solution, and the mixture was subjected to extraction using ethyl acetate (2×50 mL). The combined organic layers were washed with brine (50 mL) and then dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated under reduced pressure, and purified by column chromatography to obtain Compound 72 (3.9 g, 95%).

(193) EI-MS m/z: [M+H].sup.+ 553.3, [M+Na].sup.+575.4.

<Example 26> Preparation of Compound 73

(194) ##STR00046##

(195) Compound 73 was prepared from Compound 24 and Compound 55 by a method similar to that for the synthesis of Compound 63.

(196) EI-MS m/z: [M+H].sup.+ 1575.7, 1/2[M+H].sup.+ 788.8.

<Example 27> Preparation of Compound 74

(197) ##STR00047##

(198) Compound 74 was prepared from Compound 24 and Compound 70 by a method similar to that for the synthesis of Compound 63.

(199) EI-MS m/z: [M+H].sup.+ 1500.9, 1/2[M+H].sup.+ 751.2.

<Example 28> Preparation of Compound 75

(200) ##STR00048##

(201) Compound 75 was prepared from Compound 24 and Compound 72 by a method similar to that for the synthesis of Compound 63.

(202) EI-MS m/z: [M+H].sup.+ 1486.42, [M+Na].sup.+1509.31.

<Example 29> Preparation of Compound 80

(203) ##STR00049##

(204) Preparation of Compound 77

(205) Compound 76 (7.30 g, 28.5 mmol, Compound 76 was prepared by the method described in Angew. Chem. Int. Ed., 2016, 55, 12338-12342) and Compound 16 (14.0 g, 29.4 mmol) were dissolved in acetonitrile (145 mL), then 4 Δ molecular sieves (14.6 g) and silver(I) oxide (27.0 g, 116.4 mmol) were added thereto, and the mixture was stirred at room temperature for 12 hours under a nitrogen atmosphere. The reaction solution was filtered through Celite, concentrated, and then purified by column chromatography to obtain Compound 77 (15.3 g, 92%).

(206) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 9.94 (s, 1H), 8.27 (s, 1H), 7.99 (d, J=8.8 Hz, 1H), 7.46-7.29 (m, 6H), 5.64-5.59 (m, 1H), 5.49-5.48 (m, 1H), 5.36 (s, 2H), 5.18 (d, J=8.0 Hz, 1H), 5.19-5.11 (m, 1H), 4.27-4.10 (m, 3H), 2.19 (s, 3H), 2.08 (s, 3H), 2.04 (s, 3H), 2.03 (s, 3H).

(207) Preparation of Compound 78

(208) Compound 77 (15.30 g, 26.10 mmol) was dissolved in chloroform/isopropanol (200 mL/40 mL), then silica gel (16 g) and sodium borohydride (1.53 g, 40.50 mmol) were added thereto at 0° C. under a nitrogen atmosphere, and the mixture was stirred for 30 minutes. Distilled water (200 mL) was added to the reaction solution, and then the mixture was subjected to extraction using ethyl acetate (400 mL). The organic layer extracted was dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure to obtain Compound 78 (14.0 g, 91%).

(209) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 7.73 (s, 1H), 7.47-7.31 (m, 6H), 7.19 (d, J=8.4 Hz, 1H), 5.57 (t, J=9.2 Hz, 1H), 5.46 (d, J=3.2 Hz, 1H), 5.36-5.28 (m, 2H), 5.12-5.04 (m, 2H), 4.66 (d, J=6.0 Hz, 2H), 4.26-4.04 (m, 3H), 2.18 (s, 3H), 2.07 (s, 3H), 2.05 (s, 3H), 2.02 (s, 3H), 1.67 (t, J=5.6 Hz, 1H).

(210) Preparation of Compound 79

(211) Compound 78 (14.0 g, 23.8 mmol) was dissolved in ethanol (550 mL) and then Raney nickel (14.0 g) was added thereto. The reaction solution was stirred at room temperature for 12 hours under a hydrogen atmosphere. The reaction solution was filtered through Celite and concentrated to obtain Compound 79 (11.4 g, 96%).

(212) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 8.07 (s, 1H) 7.58 (d, J=8.8 Hz, 1H), 7.17 (d, J=8.8 Hz 1H), 5.57 (t, J=9.2 Hz, 1H), 5.49 (d, J=3.2 Hz, 1H), 5.22 (d, J=8.0 Hz, 1H), 5.17-5.14 (m, 1H), 4.71 (s, 2H), 4.25-4.10 (m, 3H), 2.20 (s, 3H), 2.11 (s, 3H), 2.07 (s, 3H), 2.00 (s, 3H).

(213) Preparation of Compound 80

(214) Compound 79 (3.00 g, 6.00 mmol) and 2-methoxyethylamine (0.57 mL, 6.6 mmol) were dissolved in N,N-dimethylformamide (15 mL), then N,N,N′,N′-tetramethyl-O-(1H-benzotrazol-1-yl)uronium hexafluorophosphate (2.86 g, 7.20 mmol) and N,N-diisopropylethylamine (2.10 mL, 12.0 mmol) were added thereto at 0° C. under a nitrogen atmosphere, and then the mixture was stirred at room temperature for 12 hours. A saturated aqueous ammonium chloride solution (100 mL) was added to the reaction solution, and the mixture was subjected to extraction using ethyl acetate (2×100 mL), and then the extract was dried over anhydrous magnesium sulfate. The resultant was filtered, then concentrated, and purified by column chromatography to obtain Compound 80 (2.3 g, 68%).

(215) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 8.07 (s, 1H), 7.48-7.44 (m, 2H), 7.06 (d, J=8.4 Hz, 1H), 5.55 (t, J=9.2 Hz, 1H), 5.49 (d, J=2.8 Hz, 1H), 5.20-5.14 (m, 2H), 4.69 (d, J=5.2 Hz, 2H), 4.25-4.09 (m, 3H), 3.78-3.74 (m, 1H), 3.62-3.51 (m, 3H), 3.40 (s, 3H), 2.21 (s, 3H), 2.07 (m, 6H), 2.02 (s, 3H), 1.71 (t, J=6.0 Hz, 1H).

<Example 30> Preparation of Compound 81

(216) ##STR00050##

(217) Preparation of Compound 81

(218) Compound 79 (2.19 g, 4.38 mmol) and Compound 31 (1.50 g, 5.70 mmol) were dissolved in N,N-dimethylformamide (10 mL), then N,N,N′,N′-tetramethyl-O-(1H-benzotrazol-1-yl)uronium hexafluorophosphate (2.26 g, 5.7 mmol) and N,N-diisopropylethylamine (1.53 mL, 8.76 mmol) were added thereto at 0° C. under a nitrogen atmosphere, and then the mixture was stirred at room temperature for 12 hours. A saturated aqueous ammonium chloride solution (100 mL) was added to the reaction solution, the mixture was subjected to extraction using ethyl acetate (2×100 mL), and then the extract was dried over anhydrous magnesium sulfate. The resultant was filtered, then concentrated, and purified by column chromatography to obtain Compound 81 (2.73 g, 84%).

(219) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 8.07 (s, 1H), 7.68 (s, 1H), 7.46-7.44 (m, 2H), 7.05 (d, J=8.0 Hz, 1H), 5.56-5.49 (m, 2H), 5.18-5.14 (m, 2H), 4.68 (d, J=4.8 Hz, 2H), 4.27-4.10 (m, 3H), 3.97-3.95 (m, 2H), 3.83-3.78 (m, 1H), 3.71-3.67 (m, 7H), 3.57-3.52 (m, 1H), 2.22 (s, 3H), 2.07 (m, 6H), 2.03 (s, 3H), 1.47 (s, 9H).

<Example 31> Preparation of Compound 82

(220) ##STR00051##

(221) Compound 82 was prepared from Compound 9, Compound 80, and Compound 81 by a method similar to that for the synthesis of Compound 28.

(222) EI-MS m/z: [M+H].sup.+ 1537.7, 1/2[M+H].sup.+ 769.7.

<Example 32> Preparation of Compound 83

(223) ##STR00052##

(224) Compound 83 was prepared from Compound 9, Compound 80, and Compound 32 by a method similar to that for the synthesis of Compound 28.

(225) EI-MS m/z: [M+H].sup.+ 1551.6, 1/2[M+H].sup.+ 776.7.

<Example 33> Preparation of Compound 85

(226) ##STR00053##

(227) Compound 85 was prepared from Compound 9, Compound 84 (Compound 84 was prepared by the method described in WO2011/130598 A1), and Compound 32 by a method similar to that for the synthesis of Compound 28.

(228) EI-MS m/z: [M+H].sup.+ 1587.8, 1/2[M+H].sup.+ 794.7.

<Comparative Example 1> Preparation of Compound 86, Compound 87, and Compound 88

(229) ##STR00054##

(230) Compound 86, Compound 87, and Compound 88 were prepared by the methods described in PCT/US2016/063564.

<Example 34> Preparation of Compound 94

(231) ##STR00055##

(232) Preparation of Compound 90

(233) Compound 89 (4.5 g, 4.88 mmol, Compound 89 was prepared by the method described in J. Med. Chem., 2004, 47, 1161-1174) was dissolved in dichloromethane (100 mL), and then 2,2,6,6-tetramethyl-1-piperidinyloxy (153 mg, 0.98 mmol) and (diacetoxyiodo)benzene (7.0 g, 21.7 mmol) were added thereto at room temperature under a nitrogen atmosphere. The reaction solution was stirred for 24 hours, then distilled water (200 mL) was added to the reaction solution, and the mixture was subjected to extraction using dichloromethane (2×200 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, then concentrated under reduced pressure, and then purified by column chromatography to obtain Compound 90 (4.25 g, 95%).

(234) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 7.74 (s, 2H), 6.73 (s, 2H), 4.97 (d, J=8.8 Hz, 1H), 4.39-4.27 (m, 8H), 3.96 (s, 6H), 3.80-3.70 (m, 2H), 3.58-3.52 (m, 2H), 3.42-2.79 (m, 2H), 2.74-2.56 (m, 2H), 2.52-2.44 (m, 2H), 2.08 (s, 2H), 0.85 (s, 18H), 0.97 (s, 12H).

(235) Preparation of Compound 91

(236) Compound 90 (10.0 g, 10.9 mmol) was dissolved in dichloromethane (450 mL) and then 2,6-lutidine (10.0 mL, 87.2 mmol) and triflic anhydride (11.0 mL, 65.4 mmol) were added thereto at −40° C. under a nitrogen atmosphere. The reaction solution was stirred for 2 hours, then a saturated aqueous sodium hydrogencarbonate solution (500 mL) was added to the reaction solution, and the mixture was subjected to extraction using dichloromethane (2×500 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, then concentrated under reduced pressure, and then purified by column chromatography to obtain Compound 91 (10.6 g, 47%).

(237) Preparation of Compound 92

(238) Compound 91 (1.7 g, 1.44 mmol) was dissolved in ethanol/toluene/distilled water (12 mL/24 mL/12 mL) and then 4-methylphenylboronic acid (568 mg, 3.74 mmol), sodium carbonate (793 mg, 7.48 mmol), and tetrakis(triphenylphosphine)palladium(O) (133 mg, 0.115 mmol) were added thereto at room temperature under a nitrogen atmosphere. The reaction solution was stirred for 2 hours and then diluted with ethyl acetate (100 mL), and the organic layer was washed with brine (100 mL) and distilled water (100 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, then concentrated under reduced pressure, and then purified by column chromatography to obtain Compound 92 (1.25 g, 79%).

(239) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 7.80 (s, 2H), 7.13 (d, J=8.8 Hz, 4H), 6.90 (s, 2H), 6.79 (d, J=8.0 Hz, 4H), 6.14 (s, 2H), 4.80-4.50 (m, 2H), 4.39-4.36 (m, 4H), 3.98 (s, 6H), 3.79 (s, 6H), 3.17 (bs, 2H), 3.02-2.98 (m, 2H), 2.50-2.47 (m, 2H), 0.88 (s, 18H), 0.11 (s, 12H). EI-MS m/z: [M+H].sup.+ 1069.8, 1/2[M+H].sup.+ 535.6.

(240) Preparation of Compound 93

(241) Compound 92 (8.0 g, 7.48 mmol) was dissolved in ethanol (300 mL) and then zinc dust (29 g, 28.1 mmol) and formic acid (5% in EtOH, 320 mL) were added thereto. The reaction solution was stirred at room temperature for 20 minutes and then filtered through Celite, and ethyl acetate (1 L) was added thereto. The organic layer was washed with distilled water (500 mL), a saturated aqueous sodium hydrogencarbonate solution (500 mL), and brine (500 mL) in this order and then dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated, and purified by column chromatography to obtain Compound 93 (4.85 g, 64%).

(242) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 7.16 (d, J=8.4 Hz, 4H), 6.78 (d, J=6.4 Hz, 8H), 6.30 (s, 2H), 4.71-4.41 (m, 2H), 4.25 (br s, 4H), 4.19-4.17 (m, 4H), 4.10-4.05 (m, 2H), 3.95-3.81 (m, 2H), 3.73 (s, 6H), 3.72 (s, 6H), 3.64-3.10 (m, 2H), 3.03-2.93 (m, 2H), 2.36-2.34 (m, 2H), 0.81 (s, 18H), 0.11 (s, 12H). EI-MS m/z: [M+H].sup.+ 1010.4, 1/2[M+H].sup.+ 505.7.

(243) Preparation of Compound 94

(244) Compound 93 (4.6 g, 4.56 mmol) was dissolved in dichloromethane (300 mL) and then pyridine (0.74 mL, 9.11 mmol) and allyl chloroformate (0.48 mL, 4.56 mmol) were added thereto at −78° C. under a nitrogen atmosphere. The reaction solution was stirred for 1 hour, then the reaction temperature was raised to room temperature, and the reaction solution was concentrated and then purified by column chromatography to obtain Compound 94 (1.46 g, 29%).

(245) EI-MS m/z: [M+H].sup.+ 1093.6.

<Example 35> Preparation of Compound 97

(246) ##STR00056##

(247) Preparation of Compound 95

(248) Compound 94 (200 mg, 0.18 mmol) was dissolved in toluene (7.5 mL), then triphosgene (19 mg, 0.067 mmol) and triethylamine (0.035 mL, 0.25 mmol) were added thereto at −10° C., and the mixture was stirred for 1 hour under a nitrogen atmosphere. Compound 20 was dissolved in dry tetrahydrofuran (7.5 mL), triethylamine (0.035 mL, 0.25 mmol) was added thereto, and then this solution was gradually added to the reaction solution. After 30 minutes, the reaction solution was heated under reflux and stirred for 4 hours. The reaction solution was concentrated, diluted with dichloromethane (30 mL), then washed with brine (20 mL), and dried over anhydrous sodium sulfate. The resultant was filtered, concentrated under reduced pressure, and then purified by column chromatography to obtain Compound 95 (130 mg, 43%).

(249) EI-MS m/z: [M+H].sup.+ 1661.6, 1/2[M+H].sup.+ 831.4.

(250) Preparation of Compound 96

(251) Compound 95 (380 mg, 0.23 mmol) was dissolved in dichloromethane (10 mL), then pyrrolidine (0.023 mL, 0.27 mmol), tetrakis(triphenylphosphine)palladium(O) (13 mg, 0.011 mmol), and triphenylphosphine (15 mg, 0.057 mmol) were added thereto in this order, and the mixture was stirred at room temperature for 6 hours under a nitrogen atmosphere. The reaction solution was concentrated under reduced pressure and then purified by column chromatography to obtain Compound 96 (260 mg, 72%).

(252) EI-MS m/z: [M+H].sup.+ 1577.6, 1/2[M+H].sup.+ 789.4.

(253) Preparation of Compound 97

(254) Compound 96 (260 mg, 0.16 mmol) was dissolved in toluene (5 mL), then triphosgene (17.6 mg, 0.06 mmol) and diisopropylethylamine (0.053 mL, 0.30 mmol) were added thereto at −10° C., and the mixture was stirred for 1 hour under a nitrogen atmosphere. Compound 22 was dissolved in dry tetrahydrofuran (5 mL), pyridine (0.066 mL, 0.80 mmol) was added thereto, and then this solution was gradually added to the reaction solution. After 30 minutes, the reaction solution was heated under reflux and stirred for 4 hours. The reaction solution was concentrated, diluted with dichloromethane (50 mL), then washed with brine (30 mL), and dried over anhydrous sodium sulfate. The resultant was filtered, concentrated under reduced pressure, and then purified by column chromatography to obtain Compound 97 (168 mg, 41%).

(255) EI-MS m/z: [M+H]+2567.1, 1/2[M+H].sup.+ 1283.8.

<Example 36> Preparation of Compound 100

(256) ##STR00057##

(257) Preparation of Compound 98

(258) Compound 97 (168 mg, 0.065 mmol) was dissolved in tetrahydrofuran/distilled water (1 mL/1 mL), acetic acid (2 mL) was added thereto, and then the mixture was stirred at room temperature for 16 hours under a nitrogen atmosphere. The reaction solution was concentrated under reduced pressure and then purified by column chromatography to obtain Compound 98 (130 mg, 85%).

(259) EI-MS m/z: [M+H].sup.+ 2337.8, 1/2[M+H].sup.+ 1169.5.

(260) Preparation of Compound 99

(261) Compound 98 (130 mg, 0.055 mmol) was dissolved in dichloromethane (5 mL), then Dess-Martin periodinane (57 mg, 0.13 mmol) was added thereto, and the mixture was stirred at room temperature for 3.5 hours under a nitrogen atmosphere. The reaction solution was concentrated under reduced pressure and then purified by column chromatography to obtain Compound 99 (96 mg, 82%).

(262) EI-MS m/z: [M+H].sup.+ 2333.7, 1/2[M+H].sup.+ 1167.5.

(263) Preparation of Compound 100

(264) Compound 99 (96 mg, 0.041 mmol) was dissolved in methanol/tetrahydrofuran (1 mL/1 mL) and then a solution of lithium hydroxide (16 mg, 0.41 mmol) in distilled water (1 mL) was gradually added thereto at −40° C. The mixture was stirred for 2 hours while gradually raising the reaction temperature to 0° C. The reaction solution was neutralized with acetic acid, then concentrated under reduced pressure, and vacuum dried. The solid obtained was diluted with dichloromethane (2 mL), then trifluoroacetic acid (0.5 mL) was added thereto at 0° C., and the mixture was stirred for 2 hours. The reaction solution was concentrated under reduced pressure, then purified by HPLC, and freeze-dried to obtain Compound 100 (2.4 mg) as a pale yellow solid.

(265) EI-MS m/z: [M+H].sup.+ 1853.8, 1/2[M+H].sup.+ 927.4.

<Example 37> Preparation of Compound 102

(266) ##STR00058##

(267) Preparation of Compound 101

(268) In dichloromethane (30 mL), 1,3-diaminopropane (0.93 mL, 11.1 mmol) was dissolved, and di-t-butyl dicarbonate (0.84 mL, 3.7 mmol) was added thereto at 0° C. under a nitrogen atmosphere. After the reaction solution was stirred at room temperature for 3 hours, brine (50 mL) was added to the reaction solution, the mixture was subjected to extraction using ethyl acetate (2×100 mL), and then the extract was dried over anhydrous sodium sulfate. After filtration, the reaction solution was filtered, then concentrated under reduced pressure, and purified by column chromatography to obtain Compound 101 (658 mg, 100% based on Boc.sub.2O).

(269) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 4.88 (br s, 1H), 3.26-3.14 (m, 2H), 2.77 (t, J=6.8 Hz, 2H), 1.66-1.57 (m, 2H), 1.44 (s, 9H), 1.32 (br, 2H).

(270) Preparation of Compound 102

(271) Compound 19 (1.50 g, 3.10 mmol) and Compound 101 (0.65 g, 3.73 mmol) were dissolved in N,N-dimethylformamide (10 mL), then N,N,N′,N′-tetramethyl-O-(1H-benzotrazol-1-yl)uronium hexafluorophosphate (1.60 g, 4.03 mmol) and N,N-diisopropylethylamine (1.08 mL, 6.20 mmol) were added thereto at 0° C. under a nitrogen atmosphere, and then the mixture was stirred at room temperature for 12 hours. A saturated aqueous ammonium chloride solution (100 mL) was added to the reaction solution, the mixture was subjected to extraction using ethyl acetate (2×100 mL), and then the extract was dried over anhydrous magnesium sulfate. The resultant was filtered, then concentrated, and purified by column chromatography to obtain Compound 102 (1.67 g, 84%).

(272) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 8.05 (s, 1H), 7.49-7.47 (m, 2H), 7.02 (d, J=8.4 Hz, 1H), 5.42-5.30 (m, 4H), 4.69 (d, J=6.0 Hz, 2H), 4.21 (d, J=9.2 Hz, 1H), 3.74 (s, 3H), 3.63-3.58 (m, 1H), 3.44-3.39 (m, 1H), 322-3.13 (m, 2H), 2.06-2.05 (m, 9H), 1.79-1.74 (m, 2H), 1.45 (s, 9H).

<Example 38> Preparation of Compound 103

(273) ##STR00059##

(274) Compound 103 was prepared from Compound 24 and Compound 102 by a method similar to that for the synthesis of Compound 28.

(275) EI-MS m/z: [M+H].sup.+ 1475.8, 1/2[M+H].sup.+ 738.3.

<Example 39> Preparation of Compound 104

(276) ##STR00060##

(277) Compound 103 (35 mg, 0.024 mmol) and maleimidoacetic acid N-hydroxysuccinimide ester (9 mg, 0.035 mmol) were dissolved in N,N-dimethylformamide (1.5 mL), and then N,N-diisopropylethylamine (0.021 mL, 0.23 mmol) was added thereto at 0° C. under a nitrogen atmosphere. The reaction temperature was gradually raised to room temperature and then the mixture was stirred for 3 hours. The reaction solution was concentrated under reduced pressure, then purified by HPLC, and freeze-dried to obtain Compound 104 (15.1 mg, 37%) as a white solid.

(278) EI-MS m/z: [M+H].sup.+ 1612.6, 1/2[M+H].sup.+ 807.2.

<Example 40> Preparation of Compound 110

(279) ##STR00061##

(280) Preparation of Compound 105

(281) L-asparagine (3.0 g, 22.7 mmol) was dissolved in 1 N aqueous sodium carbonate solution (30 mL), then benzyl chloroformate (6.3 mL, 45.4 mmol) was added thereto at 0° C., and the mixture was stirred for 12 hours under a nitrogen atmosphere. Distilled water (50 ml) was added to the reaction solution, and then the reaction solution was acidified (pH 2) with a 1 N aqueous hydrochloric acid solution. This mixture was subjected to extraction using ethyl acetate (3×50 mL), and the combined organic layers were dried over anhydrous sodium sulfate. The resultant was filtered, and then concentrated under reduced pressure to obtain Compound 105 (3.5 g, 58%).

(282) .sup.1H-NMR (400 MHz, DMSO-d.sub.6) δ 7.51-7.40 (d, J=8.0 Hz, 1H), 7.35 (s, 6H), 6.92 (s, 1H), 5.02 (s, 2H), 2.61-2.35 (m, 2H).

(283) Preparation of Compound 106

(284) Compound 105 (3.5 g, 13.1 mmol) was dissolved in ethyl acetate/acetonitrile/distilled water (30 mL/30 mL/15 mL), then (diacetoxyiodo)benzene (5.1 g, 15.7 mmol) was added thereto, and the mixture was stirred for 10 hours under a nitrogen atmosphere. The solid formed was filtered and concentrated under reduced pressure to obtain Compound 106 (2.8 g, 89%).

(285) .sup.1H-NMR (400 MHz, DMSO-d.sub.6) δ 9.94 (s, 1H), 7.96 (s, 2H), 7.72 (d, J=8.8 Hz, 1H), 7.37 (s, 5H), 5.07 (s, 2H), 4.29 (s, 1H), 3.23 (br, 1H), 3.02 (br, 1H)).

(286) Preparation of Compound 107

(287) Compound 106 (2.8 g, 11.7 mmol) was dissolved in 1,4-dioxane/distilled water (25 mL 46 mL), then sodium hydroxide (0.5 g, 11.7 mmol) and di-t-butyl dicarbonate (3.0 mL, 12.9 mmol) were added thereto, and the mixture was stirred at room temperature for 8 hours under a nitrogen atmosphere. Distilled water (50 ml) was added to the reaction solution, and then the mixture was washed with ethyl acetate (2×50 ml). The aqueous layer was acidified by addition of citric acid and subjected to extraction using ethyl acetate (3×50 mL), and then the extract was dried over anhydrous sodium sulfate. The resultant was filtered and concentrated under reduced pressure to obtain Compound 107 (2.7 g, 68%).

(288) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 7.31 (s, 5H), 5.15-5.01 (m, 2H), 4.82-4.02 (m, 1H), 3.68-3.43 (m, 2H), 1.39 (s, 9H).

(289) Preparation of Compound 108

(290) Compound 107 (2.7 g, 7.9 mmol) was dissolved in methanol (40 mL) and then palladium/charcoal (10%) (Pd/C, 0.5 g) was added thereto. The reaction solution was stirred at room temperature for 4 hours under a hydrogen atmosphere. The reaction solution was filtered through Celite and concentrated to obtain Compound 108 (1.2 g, 75%).

(291) .sup.1H-NMR (400 MHz, D.sub.2O) δ 3.70-3.65 (m, 1H), 3.55-3.25 (m, 2H), 1.28 (s, 9H).

(292) Preparation of Compound 109

(293) Compound 108 (0.40 g, 1.96 mmol) and maleic anhydride (192 mg, 1.96 mmol) were dissolved in acetic acid (1.6 mL) and then the solution was stirred at room temperature for 3 hours. The reaction solution was concentrated under reduced pressure, and dichloromethane (10 mL) was added thereto, and the solid produced was filtered and then vacuum dried. This solid dried was diluted with toluene (15 mL), then triethylamine (1.2 mL, 8.6 mmol) and N,N-dimethylacetamide (0.75 mL) were added thereto, and the mixture was heated under reflux. After 16 hours of the reaction, the reaction solution was concentrated under reduced pressure, purified by HPLC, and then freeze-dried to obtain Compound 109 (287 mg, 52%).

(294) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 6.66 (s, 2H), 5.17 (br, 1H), 4.61 (br, 1H), 3.68 (br, 1H), 1.35 (s, 9H).

(295) Preparation of Compound 110

(296) Compound 109 (0.15 g, 0.52 mmol) was dissolved in N,N-diisopropylethylamine (3 mL) and then N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (0.15 g, 0.79 mmol) and N-hydroxysuccinimide (0.09 g, 0.79 mmol) were added thereto. The reaction solution was stirred at room temperature for 12 hours. Distilled water (30 mL) was added to the reaction solution, and then the mixture was subjected to extraction using ethyl acetate (30 mL). The organic layer extracted was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and then purified by column chromatography to obtain Compound 110 (0.08 g, 40%).

(297) EI-MS m/z: [M+Na].sup.+404.3.

<Example 41> Preparation of Compound 112

(298) ##STR00062##

(299) Preparation of Compound 111

(300) Compound 103 (57 mg, 0.04 mmol) and Compound 110 (0.016 g, 0.04 mmol) were dissolved in N,N-dimethylformamide (3 mL), then N,N-diisopropylethylamine (0.02 mL, 0.12 mmol) was added thereto, and the mixture was stirred at room temperature for 6 hours under a nitrogen atmosphere. The reaction solution was concentrated under reduced pressure, then purified by HPLC, and freeze-dried to obtain Compound 111 (37 mg, 58%).

(301) EI-MS m/z: [M+H].sup.+ 1741.7, 1/2[M+H].sup.+ 871.7.

(302) Preparation of Compound 112

(303) Compound 111 (0.035 g, 0.02 mmol) was diluted with dichloromethane (3 mL), then trifluoroacetic acid (0.3 mL) was added thereto at 0° C., and the mixture was stirred for 3 hours. The reaction solution was concentrated under reduced pressure, then purified by HPLC, and freeze-dried to obtain Compound 112 (6.5 mg, 20%) as a white solid.

(304) EI-MS m/z: [M+H].sup.+ 1641.9, 1/2[M+H].sup.+ 821.8.

<Example 42> Preparation of Compound 115

(305) ##STR00063##

(306) Preparation of Compound 114

(307) In dichloromethane (10 mL), 2-[2-[2-(2-azidoethoxy)ethoxy]ethoxy]acetic acid (1.1 g, 4.71 mmol) was dissolved, then 1-hydroxybenzotriazole (0.75 g, 5.60 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.15 g, 6.03 mmol) were added thereto in this order at 0° C. under a nitrogen atmosphere, and the mixture was stirred for 30 minutes. A solution of Compound 113 (1.5 g, 4.31 mmol, Compound 113 was prepared by the method described in WO2017/160569 A1) and triethylamine (1.08 mL, 7.76 mmol) in dichloromethane (10 mL) was added to the mixture under a nitrogen atmosphere. The reaction temperature was raised to room temperature, and this mixture was stirred for 12 hours, then diluted with dichloromethane (100 mL), washed with a saturated aqueous sodium hydrogencarbonate solution (100 mL), and then dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated, and purified by column chromatography to obtain Compound 114 (2.1 g, 87%).

(308) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 6.83 (d, 1H), 4.15-4.11 (m, 1H), 3.96 (s, 2H), 3.72-3.62 (m, 14H), 3.39-3.37 (m, 2H) 1.81-1.60 (m, 4H), 0.88 (s, 18H) 0.42 (s, 12H).

(309) Preparation of Compound 115

(310) Compound 114 (2.1 g, 3.73 mmol) was dissolved in methanol (30 mL), then concentrated hydrochloric acid (0.5 mL) was added thereto at 0° C. under a nitrogen atmosphere, and then the mixture was stirred at room temperature for 2 hours. The reaction solution was neutralized with triethylamine, then concentrated, and purified by column chromatography to obtain Compound 115 (1.2 mg, 98%).

(311) 1H-NMR (400 MHz, CDCl.sub.3) δ7.54 (br s, 1H), 4.31-4.28 (m, 1H), 4.02 (s, 2H), 3.68-3.65 (m, 14H), 3.43-3.40 (m, 2H), 3.21 (br s, 2H), 1.93-1.85 (m, 2H), 1.64-1.57 (m, 2H).

<Example 43> Preparation of Compound 118

(312) ##STR00064##

(313) Preparation of Compound 117

(314) Compound 116 (1.32 g, 4.73 mmol, Compound 116 was prepared by the method described in PCT/US2016/063564) was dissolved in dichloromethane (20 mL), and then 1-hydroxybenzotriazole (0.86 g, 5.59 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride were added thereto at 0° C. under a nitrogen atmosphere. A solution of Compound 113 (1.5 g, 4.31 mmol) and triethylamine (1.08 mL, 7.74 mmol) in dichloromethane (5 mL) was added to the mixture at 0° C. under a nitrogen atmosphere. The reaction temperature was raised to room temperature, and this mixture was stirred for 12 hours, then diluted with dichloromethane (100 mL), washed with a saturated aqueous sodium hydrogencarbonate solution (100 mL), and then dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated, and purified by column chromatography to obtain Compound 117 (2.05 g, 79%).

(315) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 7.60 (s, 1H), 6.92 (d, J=9.2 Hz, 1H), 4.15-4.10 (m, 1H), 4.05-4.03 (m, 2H), 3.97 (s, 1H), 3.73-3.66 (m, 10H), 1.84-1.72 (m, 4H), 1.48 (s, 9H), 0.89 (m, 18H), 0.05 (s, 12H).

(316) Preparation of Compound 118

(317) Compound 117 (2.05 g, 3.37 mmol) was dissolved in methanol (10 mL), then camphorsulfonic acid (158 mg, 0.68 mmol) was added thereto at 0° C. under a nitrogen atmosphere, and then the mixture was stirred at 0° C. for 4 hours. The reaction solution was neutralized with triethylamine (1 mL), then concentrated, and purified by column chromatography to obtain Compound 118 (1.28 g, 99%).

(318) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 7.76 (d, J=7.2 Hz, 1H), 7.64 (s, 1H), 4.32-4.29 (m, 1H), 4.05-4.03 (m, 4H), 3.75-3.69 (m, 10H), 3.48 (br s, 2H), 1.94-1.85 (m, 2H), 1.68-1.61 (m, 4H), 1.48 (s, 9H).

<Example 44> Preparation of Compound 124

(319) ##STR00065##

(320) Preparation of Compound 119 In methanol (50 mL), 3,5-pyrazoldicarboxylic acid hydrate (5 g, 28.71 mmol) was dissolved, then thionyl chloride (6.28 mL, 86.15 mmol) was added thereto at 0° C. under a nitrogen atmosphere, and then the mixture was heated to 80° C. The reaction solution was stirred for 4 hours and then concentrated to obtain Compound 119 (7.1 g, 99%).

(321) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 7.34 (s, 1H), 3.96 (s, 6H).

(322) Preparation of Compound 120

(323) Compound 119 (3.8 g, 20.63 mmol) was dissolved in tetrahydrofuran (200 mL), then lithium aluminum hydride (1 M tetrahydrofuran solution, 41.2 mL, 41.26 mmol) was added thereto at 0° C. under a nitrogen atmosphere, and then the mixture was heated under reflux and stirred for 12 hours. The reaction mixture was cooled to 0° C., distilled water (50 mL) was gradually added thereto, and then the mixture was concentrated, diluted with methanol (200 mL), and then heated to 80° C. again. The hot reaction product was filtered, and the filtrate was concentrated. The filtrate was diluted with ethanol (10 mL), then hydrochloric acid (4 N 1,4-dioxane solution, 82.6 mL, 22.7 mmol) was added thereto, and the mixture was stirred for 20 minutes. Diethyl ether (200 mL) was added to the reaction solution, and the solid produced was filtered and dried to obtain Compound 120 (2.6 g, 78%).

(324) .sup.1H-NMR (400 MHz, DMSO-d.sub.6) δ 6.32 (s, 1H), 4.52 (s, 4H).

(325) Preparation of Compound 121

(326) Compound 120 (2.59 g, 15.73 mmol) was dissolved in N,N-dimethylformamide (75 mL), then imidazole (5.35 g, 78.68 mmol) and t-butyldimethylsilyl chloride (5.69 g, 37.8 mmol) were added thereto at 0° C. under a nitrogen atmosphere. The reaction solution was stirred for 4 hours, then diluted with ethyl acetate (100 mL), washed with a saturated aqueous ammonium chloride solution (100 mL) and brine (100 mL) in this order, and dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated, and purified by column chromatography to obtain Compound 121 (4.56 g, 81%).

(327) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 6.09 (s, 1H), 4.74 (s, 4H), 0.88 (s, 18H), 0.09 (s, 12H).

(328) Preparation of Compound 122

(329) Compound 121 (1.6 g, 4.48 mmol) was dissolved in N,N-dimethylformamide (25 mL), then cesium carbonate (3.2 g, 9.8 mmol) and triethylene glycol ditosylate (4.05 g, 8.97 mmol) were added thereto at 0° C. under a nitrogen atmosphere, and then the mixture was heated to 50° C. The reaction solution was stirred for 4 hours, then diluted with ethyl acetate (100 mL), then washed with brine (100 mL), and dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated, and purified by column chromatography to obtain Compound 122 (1.69 g, 60%).

(330) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 7.78 (d, J=8 Hz, 2H), 7.33 (d, J=8 Hz, 2H), 6.10 (s, 1H), 4.67 (d, J=5.2 Hz, 4H), 4.25-4.22 (m, 2H), 4.13-4.11 (m, 2H), 3.79-3.76 (m, 2H), 3.62-3.60 (m, 2H), 3.49-3.48 (m, 2H), 3.46-3.45 (m, 2H), 0.89 (d, J=18 Hz, 18H), 0.06 (d, J=5.6 Hz, 12H).

(331) Preparation of Compound 123

(332) Compound 122 (1.69 g, 2.62 mmol) was dissolved in acetonitrile (25 mL), then t-butyl N-hydroxycarbamate (1.35 g, 5.51 mmol) and 1,8-diazabicyclo[5.4.0]-7-undecene (0.8 mL, 5.38 mmol) were added thereto at 0° C. under a nitrogen atmosphere, and then the mixture was heated to 50° C. The reaction solution was stirred for 12 hours, then diluted with ethyl acetate (100 mL), then washed with brine (100 mL), and dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated, and purified by column chromatography to obtain Compound 123 (1.5 g, 60%).

(333) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 6.14 (s, 1H), 4.70 (d, J=8 Hz, 4H), 4.29-4.26 (m, 2H), 3.83-3.74 (m, 4H), 3.63-3.62 (m, 2H), 3.55-3.53 (m, 2H), 1.49 (s, 9H), 0.92 (d, J=18 Hz, 18H), 0.09 (d, J=0.8 Hz, 12H).

(334) Preparation of Compound 124

(335) Compound 123 (1.5 g, 2.13 mmol) was dissolved in tetrahydrofuran (20 mL), and then tetrabutylammonium fluoride (1 M tetrahydrofuran solution, 8.18 mL, 8.18 mmol) was added thereto at 0° C. under a nitrogen atmosphere. The reaction solution was stirred for 12 hours, then diluted with ethyl acetate (50 mL), washed with a saturated aqueous ammonium chloride solution (50 mL), and then dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated, and purified by column chromatography to obtain Compound 124 (660 mg, 66%).

(336) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 6.22 (s, 1H), 4.65 (d, J=4.8 Hz, 2H), 4.57 (d, J=6 Hz, 2H), 4.34-4.31 (m, 2H), 3.38-3.38 (m, 2H), 3.66-3.54 (m, 8H), 1.52 (s, 9H).

<Example 45> Preparation of Compound 131

(337) ##STR00066## ##STR00067##

(338) Preparation of Compound 126

(339) Compound 125 (1.12 g, 5.67 mmol, Compound 125 was prepared by the method described in WO2016/148674 A1) was dissolved in dichloromethane (30 mL), then pyridine (0.67 mL, 8.51 mmol) and allyl chloroformate (0.66 mL, 6.24 mmol) were added thereto at 0° C. under a nitrogen atmosphere, and then the mixture was stirred for 1 hour. The reaction solution was concentrated and then purified by column chromatography to obtain Compound 126 (1.17 g, 73%).

(340) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 10.52 (s, 1H), 8.06 (s, 1H), 7.44 (s, 1H), 6.07 (s, 1H), 6.02-5.92 (m, 1H), 5.36 (d, J=17.2 Hz, 1H), 5.24 (d, J=10.4 Hz, 1H), 4.66 (d, J=5.2, 2H), 3.89 (s, 6H).

(341) Preparation of Compound 127

(342) Compound 115 (920 mg, 2.75 mmol), Compound 126 (1.7 g 6.05 mmol), and triphenylphosphine (2.52 g, 9.35 mmol) were dissolved in dry tetrahydrofuran, then diisopropyl azodicarboxylate (1.66 mL, 8.52 mmol) was added thereto at 0° C. under a nitrogen atmosphere, and then the mixture was stirred at room temperature for 2 hours. The resultant was concentrated and purified by column chromatography to obtain Compound 127 (1.54 g, 65%).

(343) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 10.54 (s, 2H), 8.07 (s, 2H), 7.41 (s, 2H), 6.00-5.93 (m, 2H), 5.36 (d, J=17.2 Hz, 2H), 5.25 (d, J=10.0 Hz, 2H), 4.64-4.63 (m, 4H), 4.44 (bs, 1H), 4.23-4.20 (m, 4H), 3.99 (s, 2H), 3.89 (s, 3H), 3.83 (s, 3H), 3.66-3.60 (m, 11H), 3.35-3.34 (m, 2H), 2.25-2.13 (m, 4H).

(344) Preparation of Compound 128

(345) Compound 127 (1.54 g, 1.78 mmol) was dissolved in methanol/tetrahydrofuran/distilled water (15 mL/15 mL/15 mL), then sodium hydroxide (0.28 g, 7.15 mmol) was added thereto, and then the mixture was stirred at 40° C. for 5 hours. The reaction solution was diluted with ethyl acetate (100 mL) and subjected to extraction using distilled water (100 mL). The combined aqueous layers were acidified with 1 N aqueous hydrochloric acid solution and then subjected to extraction using ethyl acetate (100 mL), and the extract was dried over anhydrous sodium sulfate. The resultant was filtered and then concentrated to obtain Compound 128 (1.48 g).

(346) .sup.1H-NMR (400 MHz, DMSO-d.sub.6) δ 10.84 (s, 2H), 7.94 (s, 2H), 7.40 (s, 2H), 6.00-5.95 (m, 2H), 5.34 (d, J=17.2 Hz, 2H), 5.24 (d, J=10.0 Hz, 2H), 4.64-4.63 (m, 4H), 4.18 (br s, 1H), 4.04-4.01 (m, 4H), 3.88 (s, 2H), 3.74 (s, 6H), 3.55-3.51 (m, 12H), 2.05-1.98 (m, 4H).

(347) Preparation of Compound 129

(348) Compound 128 (1.63 g, 1.95 mmol) was dissolved in N,N-dimethylformamide (5 mL), then N,N,N′,N′-tetramethyl-O-(1H-benzotrazol-1-yl)uronium hexafluorophosphate (2.22 g, 5.87 mmol) was added thereto at 0° C. under a nitrogen atmosphere, and the mixture was stirred for 30 minutes. A solution of Compound 4 (0.64 g, 4.3 mmol) and N,N-diisopropylethylamine (1.7 mL, 9.78 mmol) in N,N-dimethylformamide (5 mL) was added to the mixture under a nitrogen atmosphere. The reaction temperature was raised to room temperature, and this mixture was stirred for 12 hours, then diluted with ethyl acetate (100 mL), washed with a saturated aqueous sodium hydrogencarbonate solution (200 mL), and then dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated, and purified by column chromatography to obtain Compound 129 (1.2 g).

(349) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 8.65 (br s, 2H), 7.34 (br s, 2H), 7.21 (d, J=8.8 Hz, 1H), 6.75 (s, 2H), 6.00-5.90 (m, 2H), 5.35 (d, J=16.8 Hz, 2H), 5.23 (d, J=10.4 Hz, 2H), 5.00-4.92 (m, 4H), 4.68-4.57 (m, 6H), 4.48-4.40 (m, 1H), 4.20-4.08 (m, 8H), 3.97 (s, 2H), 3.79 (s, 6H), 3.67-3.63 (m, 14H), 3.39-3.37 (m, 2H), 2.80-2.72 (m, 2H), 2.48-2.44 (m, 2H), 2.22-2.17 (m, 2H), 2.10-2.04 (m, 2H).

(350) Preparation of Compound 130

(351) Compound 129 (1.2 g, 1.17 mmol) was dissolved in dichloromethane (10 mL), and then imidazole (0.4 g, 5.86 mmol) and t-butyldimethylsilyl chloride (0.53 g, 3.5 mmol) were added thereto at 0° C. under a nitrogen atmosphere. The reaction solution was stirred for 12 hours, then brine (50 mL) was added to the reaction solution, and the mixture was subjected to extraction using dichloromethane (2×100 mL), and then the extract was dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated, and purified by column chromatography to obtain Compound 130 (0.98 g, 3 steps 40%).

(352) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 8.81 (br s, 2H), 7.77 (s, 2H), 7.12 (d, J=8 Hz, 1H), 6.80 (s, 2H), 5.99-5.89 (m, 2H), 5.34 (d, J=17.2 Hz, 2H) 5.23 (d, J=10.4 Hz, 2H), 4.98-4.91 (m, 4H), 4.65-4.56 (m, 6H), 4.54-4.44 (m, 1H), 4.19-4.14 (m, 8H) 4.01 (s, 2H), 3.80 (s, 6H), 3.66-3.61 (m, 14H), 3.39-3.36 (m, 2H), 2.69 (s, 4H), 2.28-2.19 (m, 2H), 2.15-2.05 (m, 2H), 0.87 (s, 18H), 0.03 (s, 12H).

(353) Preparation of Compound 131

(354) Compound 130 (0.98 g, 0.78 mmol) was dissolved in dichloromethane (5 mL), then pyrrolidine (0.16 mL, 1.95 mmol) and tetrakis(triphenylphosphine)palladium(O) (18 mg, 0.015 mmol) were added thereto, and the mixture was stirred at room temperature for 6 hours under a nitrogen atmosphere. Distilled water (50 mL) was added to the reaction solution, the mixture was subjected to extraction using dichloromethane (50 mL), and then the extract was dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated, and purified by column chromatography to obtain Compound 131 (0.59 g, 70%).

(355) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 7.12. (d, J=9.2 Hz, 1H), 6.73 (s, 2H), 6.26 (s, 2H), 4.96-4.90 (m, 4H), 4.52 (bs, 1H), 4.38-4.35 (m, 4H), 4.21-4.17 (m, 2H), 4.11-4.03 (m, 6H), 4.00 (s, 2H), 3.75 (s, 6H), 3.66-3.61 m, 12H), 3.37-3.34 (m, 2H), 2.7-2.68 (m, 4H) 2.21-2.18 (m, 2H), 2.12-2.05 (m, 2H), 0.87 (s, 18H), 0.02 (s, 12H).

<Example 46> Preparation of Compound 135

(356) ##STR00068## ##STR00069##

(357) Preparation of Compound 132

(358) Compound 131 (590 mg, 0.54 mmol) was dissolved in dry tetrahydrofuran (5 mL), then triphosgene (116 mg, 0.39 mmol) and triethylamine (0.2 mL, 1.47 mmol) were added thereto at −10° C., and the mixture was stirred under a nitrogen atmosphere for 1 hour. Compound 20 (707 mg, 1.30 mmol) was dissolved in dry tetrahydrofuran (5 mL), triethylamine (0.2 mL, 01.47 mmol) was added thereto, and this solution was gradually added to the reaction solution. After 1 hour, the reaction solution was heated under reflux and stirred for 12 hours. The reaction solution was diluted with ethyl acetate (30 mL), then washed with brine (20 mL), and dried over anhydrous sodium sulfate. The resultant was filtered, concentrated under reduced pressure, and then purified by column chromatography to obtain Compound 132 (1.0 g, 83%).

(359) EI-MS m/z: [M+H].sup.+ 2219.10, 1/2[M+H].sup.+ 1110.30

(360) Preparation of Compound 133

(361) Compound 132 (1 g, 0.45 mmol) was dissolved in tetrahydrofuran/distilled water (5 mL/5 mL), acetic acid (15 mL) was added thereto, and then the mixture was stirred at room temperature for 16 hours under a nitrogen atmosphere. The reaction solution was diluted with ethyl acetate (50 mL), then washed with distilled water (50 mL), and then dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated, and purified by column chromatography to obtain Compound 133 (720 mg, 80%).

(362) EI-MS m/z: [M+H].sup.+ 1990.95, 1/2[M+H].sup.+ 996.06.

(363) Preparation of Compound 134

(364) Compound 133 (370 mg, 0.18 mmol) was dissolved in dichloromethane (10 mL), then Dess-Martin periodinane (181 mg, 0.42 mmol) was added thereto, and the mixture was stirred at room temperature for 16 hours under a nitrogen atmosphere. The reaction solution was diluted with dichloromethane (20 mL), then washed with a saturated aqueous sodium hydrogencarbonate solution (20 mL), and dried over anhydrous sodium sulfate. The resultant was filtered, concentrated under reduced pressure, and then purified by column chromatography to obtain Compound 134 (350 mg, 90%).

(365) EI-MS m/z: [M+H].sup.+ 1986.61, 1/2[M+H].sup.+ 994.11.

(366) Preparation of Compound 135

(367) Compound 134 (350 mg, 0.17 mmol) was dissolved in methanol/tetrahydrofuran (7.5 mL/7.5 mL), and then lithium hydroxide (66 mg, 1.58 mmol) dissolved in distilled water (7.5 mL) was gradually added thereto at −40° C. The reaction temperature was gradually raised to 0° C., and the mixture was stirred for 2 hours. The reaction solution was neutralized with acetic acid, then concentrated under reduced pressure, then purified by HPLC, and freeze-dried to obtain Compound 135 (150 mg, 50%) as a white solid.

(368) EI-MS m/z: [M+H].sup.+ 1706.20, 1/2[M+H].sup.+ 854.00.

<Example 47> Preparation of Compound 137

(369) ##STR00070##

(370) Preparation of Compound 136

(371) Compound 136 was prepared from Compound 126 and Compound 118 by a method similar to that for the synthesis of Compound 134.

(372) EI-MS m/z: [M+H].sup.+ 2032.98, 1/2[M+H].sup.+ 1017.03.

(373) Preparation of Compound 137

(374) Compound 136 (205 mg, 0.10 mmol) was dissolved in methanol/tetrahydrofuran (4 mL/6 mL) and then a solution of lithium hydroxide (38 mg, 0.91 mmol) in distilled water (4 mL) was gradually added thereto at −40° C. The mixture was stirred for 4 hours while gradually raising the reaction temperature to 0° C. The reaction solution was neutralized with acetic acid, then concentrated under reduced pressure, and freeze-dried. The solid obtained was diluted with dichloromethane (5 mL), then trifluoroacetic acid (1.5 mL) was added thereto at 0° C., and the mixture was stirred for 4 hours. The reaction solution was concentrated under reduced pressure, then purified by HPLC, and freeze-dried to obtain Compound 137 (29 mg, 17%) as a white solid.

(375) EI-MS m/z: [M+H].sup.+ 1653.01, 1/2[M+H].sup.+ 826.89.

<Example 48> Preparation of Compound 138

(376) ##STR00071##

(377) Compound 138 was prepared from Compound 126 and Compound 124 by a method similar to that for the synthesis of Compound 137.

(378) EI-MS m/z: [M+H].sup.+ 1647.60, 1/2[M+H].sup.+ 824.31.

<Example 49> Preparation of Compound 142

(379) ##STR00072##

(380) Preparation of Compound 139

(381) Dimethyl sulfoxide (3.53 mL, 3.88 mmol) was dissolved in dichloromethane (30 mL), and then oxalyl chloride (2.0 M dichloromethane solution, 13 mL, 23.9 mmol) was added thereto at −78° C. under a nitrogen atmosphere. Compound 6 (6.8 g, 9.94 mmol) was dissolved in dichloromethane (20 mL) and then this solution was gradually added to the mixture at −78° C. under a nitrogen atmosphere. The reaction solution was stirred for 10 minutes, then the temperature was raised to 0° C., triethylamine (13.85 mL, 4.41 mmol) was gradually added thereto, and then the mixture was stirred at room temperature for 2 hours. A saturated aqueous ammonium chloride solution (200 mL) was added to the reaction solution, and the mixture was subjected to extraction using dichloromethane (3×100 mL). The combined organic layers were washed with brine (2×100 mL) and then dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated, and purified by column chromatography to obtain Compound 139 (5.76 g, 85%).

(382) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 9.80 (s, 2H), 7.72 (s, 2H), 6.85 (s, 2H), 5.07 (s, 2H), 4.93 (s, 2H), 4.22-4.10 (m, 6H), 4.00 (s, 6H), 3.94-3.80 (m, 4H), 3.15-2.70 (m, 4H), 2.30-2.10 (m, 2H), 2.12-1.90 (m, 4H), 1.75-1.68 (m, 6H)). EI-MS m/z [M+H].sup.+ 681.6.

(383) Preparation of Compound 140

(384) Compound 139 (1.84 g, 2.71 mmol) was dissolved in benzene and N,N-dimethylformamide (v/v=10:1, 30 mL), and then ethylene glycol (1.5 mL, 27.11 mmol) and camphorsulfonic acid (251 mg, 0.81 mmol) were sequentially added thereto at room temperature under a nitrogen atmosphere. The reaction solution was stirred for 5 minutes, then heated under reflux using a Dean-Stark apparatus, and stirred for 2 hours. The reaction solution was concentrated, diluted with ethyl acetate (100 mL), then a saturated aqueous sodium hydrogencarbonate solution (100 mL) was added to the reaction solution, and the mixture was subjected to extraction using ethyl acetate (3×100 mL). The combined organic layers were washed with brine (2×100 mL) and then dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated, and purified by column chromatography to obtain Compound 140 (1.53 g, 72%).

(385) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 7.70 (s, 2H), 6.78 (s, 2H), 5.07 (s, 2H), 4.81 (s, 2H), 4.69 (s, 2H), 4.20-4.02 (m, 6H), 4.00-3.90 (m, 8H), 3.87-3.80 (m, 2H), 3.78-3.70 (m, 4H), 3.60-3.68 (m, 4H), 2.72-2.60 (m, 4H), 2.12-1.92 (m, 4H), 1.65-1.60 (m, 2H). EI-MS m/z: [M+H].sup.+769.8.

(386) Preparation of Compound 141

(387) Compound 140 (1.11 g, 1.45 mmol) was dissolved in ethanol (22 mL), and then zinc dust (2.84 g, 43.39 mmol) and formic acid (5% ethanol solution, 1.96 mL) were added thereto. The reaction solution was stirred at room temperature for 3 hours and then filtered through Celite, and ethyl acetate (300 mL) was added thereto. The organic layer was washed with distilled water (2×100 mL), a saturated aqueous sodium hydrogencarbonate solution (2×200 mL), and brine (200 mL) in this order and then dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated, and purified by column chromatography to obtain Compound 141 (800 mg, 78%).

(388) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 6.78 (s, 2H), 6.23 (s, 2H), 5.12 (s, 2H), 4.97 (s, 2H), 4.91 (s, 2H), 4.78 (br s, 2H), 4.54 (br s, 2H), 4.33-4.21 (m, 2H), 4.10-3.91 (m, 12H), 3.90-3.82 (m, 4H), 3.78 (s, 6H), 2.72-2.58 (m, 4H), 1.98-1.84 (m, 4H), 1.74-1.58 (m, 2H), 0.87 (s, 18H), 0.02 (s, 12H). EI-MS m/z: [M+H].sup.+ 709.8.

(389) Preparation of Compound 142

(390) Compound 141 (640 mg, 0.90 mmol) was dissolved in dichloromethane (45 mL), and then pyridine (0.15 mL, 1.80 mmol) and allyl chloroformate (86 L, 0.81 mmol) were added thereto at −78° C. under a nitrogen atmosphere. After the reaction solution was stirred for 1 hour, the reaction temperature was raised to room temperature, and the reaction solution was concentrated and then purified by column chromatography to obtain Compound 142 (320 mg, 43%).

(391) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 8.75 (br, 1H), 7.83 (s, 1H), 6.83 (s, 1H), 6.77 (s, 1H), 6.23 (s, 1H), 5.97-5.91 (m, 1H), 5.34, (d, J=17.2 Hz, 1H), 5.23 (d, J=10.0 Hz, 1H), 5.08 (br s, 1H), 5.02-4.88 (m, 6H), 4.80 (br s, 1H), 4.68-4.56 (m, 2H), 4.44 (br s, 2H), 4.30-4.18 (m, 2H), 4.16-4.06 (m, 3H), 3.92-3.84 (m, 3H), 3.83 (s, 3H), 3.78 (s, 3H), 2.74-2.56 (m, 4H), 1.99-1.86 (m, 4H), 1.72-1.60 (m, 2H).

<Example 50> Preparation of Compound 146

(392) ##STR00073## ##STR00074##

(393) Preparation of Compound 143

(394) Compound 142 (260 mg, 0.35 mmol) was dissolved in tetrahydrofuran (4 mL), then triphosgene (40 mg, 0.13 mmol) and triethylamine (0.078 mL, 0.56 mmol) were added thereto at −10° C., and the mixture was stirred for 1 hour under a nitrogen atmosphere. Compound 20 (208 mg, 0.39 mmol) and triethylamine (0.087 mL, 0.62 mmol) were dissolved in dry tetrahydrofuran (3 mL), and this solution was gradually added to the reaction solution. After 30 minutes, the reaction solution was heated under reflux and stirred for 3 hours. The reaction solution was concentrated, diluted with dichloromethane (50 mL), then washed with brine (2×20 mL), and dried over anhydrous sodium sulfate. The resultant was filtered, concentrated under reduced pressure, and then purified by column chromatography to obtain Compound 143 (340 mg, 71%)

(395) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 8.78 (br s, 1H), 7.95 (d, J=12.2 Hz, 1H), 7.83 (br s, 1H), 7.52-7.41 (m, 2H), 7.27 (s, 1H), 7.04 (d, J=8.4 Hz, 1H), 6.83 (s, 1H), 6.00-5.88 (m, 1H), 5.42-5.23, (m, 10H), 5.20-5.08 (m, 4H), 5.06-4.82 (m, 8H), 4.67 (s, 1H), 4.28-4.18 (m, 6H), 4.16-4.06 (m, 8H), 4.05-3.86 (m, 6H), 3.86 (s, 3H), 3.76 (s, 3H), 3.52-3.62 (m, 3H), 3.41 (s, 3H), 2.78-2.58 (m, 2H), 2.12-2.06 (m, 2H), 2.05 (s, 9H), 1.86-2.01 (m, 4H), 1.72-1.60 (m, 2H), 1.27 (t, J=7.2 Hz, 2H).

(396) EI-MS m/z: [M+H].sup.+ 1361.5, 1/2[M+H].sup.+ 681.6.

(397) Preparation of Compound 144

(398) Compound 143 (330 mg, 0.24 mmol) was dissolved in dichloromethane (5 mL), then pyrrolidine (0.026 mL, 0.365 mmol) and tetrakis(triphenylphosphine)palladium(O) (14 mg, 0.012 mmol) were added thereto, and the mixture was stirred at room temperature for 5 hours under a nitrogen atmosphere. The reaction solution was concentrated under reduced pressure and then purified by column chromatography to obtain Compound 144 (290 mg, 90%).

(399) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 8.82 (br s, 1H), 8.05 (s, 1H), 7.52-7.42 (m, 2H), 7.04 (d, J=8.2 Hz, 1H), 6.82 (s, 1H), 6.78 (s, 1H), 6.24 (s, 1H), 5.44-5.26, (m, 4H), 5.16-5.04 (m, 4H), 5.02-4.86 (m, 5H), 4.52-4.38 (m, 2H), 4.30-4.10 (m, 6H), 4.16-4.07 (m, 5H), 4.04-3.92 (m, 6H), 3.91-3.84 (m, 6H), 3.83 (s, 3H), 3.78 (s, 3H), 3.72 (s, 3H), 3.60-3.52 (m, 3H), 3.41 (s, 3H), 2.71-2.58 (m, 4H), 2.05 (s, 9H), 1.97-1.85 (m, 4H), 1.72-1.58 (m, 4H), 1.25 (t, J=7.2 Hz, 2H). EI-MS m/z: [M+H].sup.+ 1277.2, 1/2[M+H].sup.+ 639.4.

(400) Preparation of Compound 145

(401) Compound 144 (340 mg, 0.29 mmol) was dissolved in dry tetrahydrofuran (3 mL), then triphosgene (25 mg, 0.09 mmol) and triethylamine (0.060 mL, 0.43 mmol) were added thereto at −10° C., and the mixture was stirred for 1 hour under a nitrogen atmosphere. Compound 32 (229 mg, 0.31 mmol) was dissolved in dry tetrahydrofuran (3 mL), triethylamine (0.060 mL, 0.43 mmol) was added thereto, and then this solution was gradually added to the reaction solution. After 30 minutes, the reaction solution was heated under reflux and stirred for 4 hours. The reaction solution was concentrated, diluted with dichloromethane (100 mL), then washed with brine (2×50 mL), and dried over anhydrous sodium sulfate. The resultant was filtered, concentrated under reduced pressure, and then purified by column chromatography to obtain Compound 145 (250 mg, 43%).

(402) EI-MS m/z: [M+Na].sup.+2056.4, 1/2[M+H].sup.+ 967.7.

(403) Preparation of Compound 146

(404) Compound 145 (230 mg, 0.113 mmol) was dissolved in methanol/tetrahydrofuran (3 mL/3 mL), and then a solution of lithium hydroxide (48 mg, 1.13 mmol) in distilled water (6 mL) was gradually added thereto at −40° C. The mixture was stirred for 2 hours while gradually raising the reaction temperature to 0° C. The reaction solution was neutralized with acetic acid, then concentrated under reduced pressure, and vacuum dried. The solid obtained was diluted with dichloromethane (10 mL), then trifluoroacetic acid (2 mL) was added thereto at 0° C., and the mixture was stirred for 2 hours. The reaction solution was concentrated under reduced pressure, then purified by HPLC, and freeze-dried to obtain Compound 146 (15.6 mg) as a white solid.

(405) EI-MS m/z: [M+H].sup.+ 1653.7, 1/2[M+H].sup.+ 827.6.

<Example 51> Preparation of Compound 148

(406) ##STR00075##

(407) Preparation of Compound 147

(408) Compound 144 (266 mg, 0.21 mmol) was dissolved in dry tetrahydrofuran (3 mL), then triphosgene (16 mg, 0.06 mmol) and triethylamine (0.044 mL, 0.31 mmol) were added thereto at −10° C., and the mixture was stirred for 1 hour under a nitrogen atmosphere. Compound 55 (147 mg, 0.23 mmol) was dissolved in dry tetrahydrofuran (3 mL), triethylamine (0.044 mL, 0.31 mmol) was added thereto, and then this solution was gradually added to the reaction solution. After 30 minutes, the reaction solution was heated under reflux and stirred for 4 hours. The reaction solution was concentrated, diluted with dichloromethane (100 mL), then washed with brine (2×50 mL), and dried over anhydrous sodium sulfate. The resultant was filtered, concentrated under reduced pressure, and then purified by column chromatography to obtain Compound 147 (170 mg, 42%).

(409) EI-MS m/z: [M+Na].sup.+1944.6, 1/2[M+H].sup.+ 972.8.

(410) Preparation of Compound 148

(411) Compound 147 (120 mg, 0.087 mmol) was dissolved in methanol/tetrahydrofuran (3 mL/3 mL), and then a solution of lithium hydroxide (37 mg, 0.87 mmol) in distilled water (6 mL) was gradually added thereto at −40° C. The mixture was stirred for 2 hours while gradually raising the reaction temperature to 0° C. The reaction solution was neutralized with acetic acid, then concentrated under reduced pressure, and vacuum dried. The solid obtained was diluted with dichloromethane (8 mL), then trifluoroacetic acid (2 mL) was added thereto at 0° C., and the mixture was stirred for 2 hours. The reaction solution was concentrated under reduced pressure, then purified by HPLC, and freeze-dried to obtain Compound 148 (31 mg) as a white solid.

(412) EI-MS m/z: [M+H].sup.+ 1663.4, 1/2[M+H].sup.+ 832.7.

<Example 52> Preparation of Compound 155

(413) ##STR00076##

(414) Preparation of Compound 149

(415) Compound 4 (13.8 g, 92.5 mmol) was dissolved in dichloromethane (400 mL), and then imidazole (18.8 g, 277.5 mmol) and t-butyldimethylsilyl chloride (15.3 g, 101.7 mmol) dissolved in dichloromethane (100 mL) were added thereto at 0° C. under a nitrogen atmosphere. The reaction solution was stirred at room temperature for 2 hours, then brine (30 mL) was added to the reaction solution, the mixture was subjected to extraction using dichloromethane (2×300 mL), and the extract was dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated, and purified by column chromatography to obtain Compound 149 (17 g, 80%).

(416) .sup.1H-NMR (400 MHz, CDCl.sub.3) (rotamers) δ 4.91, (d, J=14.4 Hz, 2H), 3.66-3.47 (m, 4H), 3.27-3.24 (m, 1H), 2.47-2.42 (m, 1H), 2.24-2.18 (m, 1H), 0.91 (s, 9H), 0.05 (s, 6H).

(417) Preparation of Compound 151

(418) Compound 150 (17.3 g, 46.8 mmol, Compound 150 was prepared by the method described in ACS Med. Chem. Lett. 2016, 7, 983) was dissolved in N,N-dimethylformamide (100 mL), then 1-hydroxybenzotriazole (6.8 g, 50.7 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (10.4 g, 54.6 mmol) were added thereto in this order at 0° C. under a nitrogen atmosphere, and then the mixture was stirred for 30 minutes. A solution of Compound 149 (8.8 g, 39.0 mmol) and triethylamine (9.78 mL, 70.2 mmol) in dichloromethane (50 mL) was added to the mixture under a nitrogen atmosphere. The reaction temperature was raised to room temperature, and this mixture was stirred for 12 hours, then diluted with dichloromethane (100 mL), washed with a saturated aqueous sodium hydrogencarbonate solution (100 mL), and then dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated, and purified by column chromatography to obtain Compound 151 (19.9 g, 89%).

(419) .sup.1H-NMR (400 MHz, CDCl.sub.3) (rotamers) δ 7.69 (s, 1H), 6.72 (s, 1H), 4.97 (s, 1H), 4.82 (s, 1H), 4.57-4.54 (m, 1H) 3.89 (s, 4H), 3.74-3.71 (m, 2H) 3.30-3.27 (m, 1H), 2.76-2.52 (m, 2H), 1.31-1.23 (m, 3H), 1.08 (s, 18H), 0.89 (s, 9H), 0.08 (s, 3H).

(420) Preparation of Compound 152

(421) Compound 151 (29.5 g, 50.9 mmol) was dissolved in ethanol (720 mL), and then zinc dust (66.6 g, 1019.1 mmol) and formic acid (38 mL, 1019.1 mmol) were added thereto. The reaction solution was stirred at room temperature for 40 minutes and then filtered through Celite, and ethyl acetate (500 mL) was added thereto. The organic layer was washed with distilled water (500 mL), a saturated aqueous sodium hydrogencarbonate solution (500 mL), and brine (500 mL) in this order and then dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated, and purified by column chromatography to obtain Compound 152 (27.9 g, 99%).

(422) .sup.1H-NMR (400 MHz, CDCl.sub.3) (rotamers) δ 6.71 (s, 1H), 6.25 (s, 1H), 4.96-4.89 (m, 2H), 4.53 (br s, 1H), 4.21-4.09 (m, 4H), 3.74 (br s, 1H), 3.71 (s, 3H), 3.62 (br s, 1H), 2.73-2.63 (m, 2H), 1.29-1.21 (m, 3H), 1.05 (s, 18H), 0.87 (s, 9H), 0.02 (s, 6H).

(423) Preparation of Compound 153

(424) Compound 152 (27.9 g, 50.8 mmol) was dissolved in dichloromethane (300 mL), and then pyridine (9 mL, 111.8 mmol) and allyl chloroformate (5.9 mL, 55.9 mmol) were added thereto at −78° C. under a nitrogen atmosphere. After the reaction solution was stirred for 1 hour, the reaction temperature was raised to room temperature, and the reaction solution was concentrated and then purified by column chromatography to obtain Compound 153 (31.8 g, 99%).

(425) .sup.1H-NMR (400 MHz, CDCl.sub.3) (rotamers) δ 8.67 (br s, 1H), 7.75 (s, 1H), 6.78 (s, 1H), 5.99-5.89 (m, 1H), 5.33, (d, J=17.2 Hz, 1H), 5.21 (d, J=10.4 Hz, 1H), 4.98-4.90 (m, 2H), 4.66-4.57 (m, 3H), 4.19-4.11 (m, 1H), 4.01 (br s, 1H), 3.86 (br s, 1H), 3.76 (s, 3H), 3.65 (br s, 1H), 2.68 (s, 2H), 1.33-1.24 (m, 3H), 1.05 (s, 18), 0.87 (s, 9H), 0.03 (s, 6H).

(426) Preparation of Compound 154

(427) Compound 153 (31.8 g, 50.2 mmol) was dissolved in N,N-dimethylformamide (300 mL) and distilled water (6 mL), then sodium acetic acid (5 g, 60.2 mmol) was added thereto at 0° C. under a nitrogen atmosphere, and the mixture was stirred at room temperature for 2 hours. The reaction solution was diluted with ethyl acetate (300 mL), then washed with distilled water (300 mL), and dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated, and purified by column chromatography to obtain Compound 154 (17.7 g, 74%).

(428) .sup.1H-NMR (400 MHz, CDCl.sub.3) (rotamers) δ 8.75 (br s, 1H), 7.75 (s, 1H), 6.78 (s, 1H), 6.14 (s, 1H), 5.94-5.90 (m, 1H), 5.32, (d, J=17.2 Hz, 1H), 5.21 (d, J=10.4 Hz, 1H), 4.97-4.90 (m, 2H), 4.65-4.56 (m, 3H), 4.18-4.15 (m, 1H), 4.01 (br s, 1H), 3.85 (s, 4H), 3.65 (br s, 1H), 2.68 (s, 2H), 0.87 (s, 9H), 0.02 (s, 6H).

(429) Preparation of Compound 155

(430) Compound 154 (18.6 g, 39.0 mmol) was dissolved in acetone (200 mL), then 1,5-diiodopentane (11.6 mL, 156 mmol) and potassium carbonate (5.9 g, 42.9 mmol) were added thereto in this order under a nitrogen atmosphere, and then the mixture was stirred at 60° C. for 12 hours. The reaction solution was concentrated and purified by column chromatography to obtain Compound 155 (23 g, 87%).

(431) .sup.1H-NMR (400 MHz, CDCl.sub.3) (rotamers) δ 8.88 (br s, 1H), 7.83 (s, 1H), 6.81 (s, 1H), 5.98-5.90 (m, 1H), 5.34, (d, J=17.2 Hz, 1H), 5.24 (d, J=10.4 Hz, 1H), 4.98-4.90 (m, 2H), 4.67-4.58 (m, 3H), 4.21-4.12 (m, 1H), 4.10-4.06 (m, 3H) 3.82 (s, 4H), 3.64 (br s, 1H), 3.23-3.19 (m, 2H), 2.69 (s, 2H), 1.94-1.84 (m, 4H), 1.62-1.55 (m, 2H), 0.87 (s, 9H), 0.03 (s, 6H).

<Example 53> Preparation of Compound 162

(432) ##STR00077## ##STR00078##

(433) Preparation of Compound 156

(434) Compound 151 (9.3 g, 16.0 mmol) was dissolved in N,N-dimethylformamide (100 mL) and distilled water (2 mL), then sodium acetate (1.6 g, 19.2 mmol) was added thereto at 0° C. under a nitrogen atmosphere, and the mixture was stirred at room temperature for 30 minutes. The reaction solution was diluted with ethyl acetate (100 mL), then washed with distilled water (100 mL), and dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated, and purified by column chromatography to obtain Compound 156 (5.4 g, 80%).

(435) .sup.1H-NMR (400 MHz, CDCl.sub.3) (rotamers) δ 7.75 (s, 1H), 6.76 (s, 1H), 6.07 (s, 1H), 4.98 (s, 1H), 4.83 (s, 1H), 4.58-4.54 (m, 1H), 3.99 (s, 3H), 3.89-3.87 (m, 1H), 3.77-3.70 (m, 2H), 3.33-3.29 (m, 1H), 2.81-2.53 (m, 2H), 0.89 (s, 9H), 0.09 (s, 6H).

(436) Preparation of Compound 157

(437) Compound 156 (3.0 g, 7.1 mmol) was dissolved in N,N-dimethylformamide (30 mL), and then potassium carbonate (1.1 g, 7.8 mmol) and benzyl bromide (0.9 ml, 7.8 mmol) were added thereto at 0° C. under a nitrogen atmosphere. The reaction solution was stirred for 3 hours, then a saturated aqueous ammonium chloride solution (50 mL) was added to the reaction solution, and the mixture was subjected to extraction using ethyl acetate (2×50 mL). The combined organic layers were washed with distilled water (2×100 mL) and brine (100 mL) and then dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated, and purified by column chromatography to obtain Compound 157 (3.6 g, 97%).

(438) .sup.1H-NMR (400 MHz, CDCl.sub.3) (rotamers) δ 7.77 (d, J=4.8 Hz, 1H), 7.46-7.33 (m, 5H), 6.79 (d, J=18.8 Hz, 1H), 5.22 (d, J=5.2 Hz, 2H), 5.09 (d, J=7.6 Hz, 1H), 4.98 (s, 1H), 4.83 (s, 1H), 4.58 (br s, 1H), 3.96 (s, 3H), 3.87 (br s, 1H), 3.77-3.69 (m, 2H), 3.30-3.28 (m, 1H), 2.81-2.53 (m, 2H), 0.89 (s, 9H), 0.09 (s, 6H).

(439) Preparation of Compound 158

(440) Compound 157 (3.6 mg, 6.9 mmol) was dissolved in tetrahydrofuran/distilled water (15 mL/15 mL), acetic acid (30 mL) was added thereto, and then the mixture was stirred at room temperature for 16 hours under a nitrogen atmosphere. The reaction solution was concentrated under reduced pressure and then purified by column chromatography to obtain Compound 158 (2.8 g, 99%).

(441) .sup.1H-NMR (400 MHz, CDCl.sub.3) (rotamers) δ7.77 (s, 1H), 7.47-7.35 (m, 5H), 6.81 (s, H), 5.22 (s, 2H), 5.02 (s, 1H), 4.87 (s, 1H), 4.60 (br s, 1H), 3.99 (s, 3H), 3.88 (br s, 1H), 3.83-3.72 (m, 3H), 3.54 (br s, 1H), 2.88-2.82 (m, 1H), 2.52-2.48 (m, 1H).

(442) Preparation of Compound 159

(443) Oxalyl chloride (2.1 mL, 14.1 mmol) was dissolved in dichloromethane (20 mL), and then dimethyl sulfoxide (1.5 mL, 21.1 mmol) was added thereto at −78° C. under a nitrogen atmosphere. After 1 hour, a solution of Compound 158 (2.7 g, 6.9 mmol) in dichloromethane (50 mL) was gradually added to the mixture. The reaction solution was stirred for 2 hours, and then triethylamine (3.4 mL, 42.3 mmol) was diluted with dichloromethane (30 mL) and gradually added thereto. The reaction temperature was gradually raised to 0° C. over 2 hours. The reaction solution was diluted with dichloromethane (100 mL), and the organic layer was washed with a saturated aqueous ammonium chloride solution (200 mL) and brine (200 mL) and then dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated, and purified by column chromatography to obtain Compound 159 (2.7 g, 96%).

(444) .sup.1H-NMR (400 MHz, CDCl.sub.3) (rotamers) δ 9.79 (s, 1H), 7.79 (s, 1H), 7.46-7.26 (m, 5H), 6.87 (s, 1H), 5.22 (s, 2H), 5.06-4.96 (m, 1H), 4.93-4.90 (m, 1H), 4.78 (br s, 1H), 4.62-4.56 (m, 1H), 3.99 (s, 3H), 3.93 (s, 1H), 3.85 (s, 1H), 2.91-2.62 (m, 2H).

(445) Preparation of Compound 160

(446) Compound 159 (2.7 g, 6.8 mmol) was dissolved in tetrahydrofuran/distilled water (60 mL/40 mL) and then sodium dithionite (Na.sub.2S.sub.2O.sub.4, 11.2 g, 64.4 mmol) was added thereto. The mixture was stirred for 20 hours under a nitrogen atmosphere.

(447) The reaction solution was diluted by addition of methanol (60 ml), acidified (pH 2) by addition of a 6 N aqueous hydrochloric acid solution, and stirred for 1 hour. The reaction solution was concentrated under reduced pressure to remove methanol. The reaction solution was acidified (pH 2) by addition of a 6 N aqueous hydrochloric acid solution and subjected to extraction using ethyl acetate (5×100 mL). The combined organic layers were dried over anhydrous sodium sulfate. The resultant was filtered, then concentrated under reduced pressure, and purified by column chromatography to obtain Compound 160 (1.8 g, 77%).

(448) EI-MS m/z: [M+H].sup.+ 349.3, [M+H.sub.2O].sup.+367.3.

(449) Preparation of Compound 161

(450) Compound 160 (1.8 g, 5.3 mmol) was dissolved in dichloromethane/N,N-dimethylformamide (20 mL/8 mL), then sodium triacetoxyborohydride (1.2 g, 5.8 mmol) was added thereto at 0° C. under a nitrogen atmosphere, and then the mixture was stirred for 2 hours. Distilled water (40 mL) was added to the reaction solution, and then the mixture was subjected to extraction using dichloromethane (2×50 mL). The organic layer extracted was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and then purified by column chromatography to obtain Compound 161 (1.2 g, 64%).

(451) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 7.61 (s, 1H), 7.41-7.30 (m, 5H), 6.05 (s, 1H), 5.12 (s, 2H), 5.06 (s, 1H), 5.02 (s, 1H), 4.38 (d, J=18 Hz, 1H), 4.27 (d, J=16.4 Hz, 1H), 4.04-3.96 (m, 1H), 3.86 (s, 3H), 3.49 (d, J=11 Hz, 1H), 3.29 (dd, J=9.2 Hz, 1H), 2.91-2.85 (m, 1H), 2.40 (dd, J=10 Hz, 1H).

(452) Preparation of Compound 162

(453) Compound 161 (1.3 g, 3.7 mmol) was dissolved in dichloromethane (70 mL), then methanesulfonic acid (25 mL) was added thereto, and the mixture was stirred for 2 hours under a nitrogen atmosphere. Distilled water (20 mL) was added to the reaction solution, and then the reaction solution was neutralized by addition of sodium carbonate. The reaction solution was diluted by addition of water (200 mL) and subjected to extraction using dichloromethane (3×50 mL). The organic layer extracted was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and then purified by column chromatography to obtain Compound 162 (620 mg, 64%).

(454) .sup.1H-NMR (400 MHz, CDCl.sub.3) δ 7.60 (s, 1H), 6.17 (s, 1H), 5.88 (br s, 1H), 5.09 (s, 1H), 5.06 (s, 1H), 4.41 (d, J=16.4 Hz, 1H), 4.31 (d, J=16.4 Hz, 1H), 4.08-3.99 (m, 1H), 3.88 (s, 3H), 3.54 (d, J=12.4 Hz, 1H), 3.49 (d, J=11 Hz, 1H), 3.34 (dd, J=9.2 Hz, 1H), 2.95-2.89 (m, 1H), 2.43 (dd, J=6.4 Hz, 1H).

<Example 54> Preparation of Compound 164

(455) ##STR00079##

(456) Preparation of Compound 163

(457) Compound 162 (374 mg, 1.4 mmol) and Compound 155 (1.0 g, 1.5 mmol) were dissolved in acetone/N,N-dimethylformamide (20 mL/20 mL), then potassium carbonate (258 mg, 1.8 mmol) was added thereto under a nitrogen atmosphere, and the mixture was heated and stirred at 80° C. for 12 hours. The reaction solution was filtered and then concentrated under reduced pressure, and distilled water (20 mL) was added to the reaction solution, and then the mixture was subjected to extraction using ethyl acetate (3×30 mL). The organic layer extracted was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and then purified by column chromatography to obtain Compound 163 (620 mg, 53%).

(458) .sup.1H-NMR (400 MHz, CDCl.sub.3) (rotamers) δ 8.86 (br s, 1H), 7.85 (s, 1H), 7.60 (s, 1H), 6.82 (s, 1H), 6.06 (s, 1H), 6.01-5.91 (m, 1H), 5.36 (d, J=17.2 Hz, 1H), 5.25 (d, J=10.4 Hz, 1H), 5.08 (s, 1H), 5.05 (s, 1H), 5.00 (s, 1H), 4.92 (br s, 1H), 4.63 (d, J=4.8 Hz, 2H), 4.41 (d, J=16.4 Hz, 1H), 4.30 (d, J=16.4 Hz, 1H), 4.20 (d J=14 Hz, 1H), 4.13-4.10 (m, 3H), 4.05-3.98 (m, 3H), 3.85 (s, 3H), 3.82 (s, 3H), 3.66 (bs, 1H), 3.55 (d, J=12.8 Hz, 1H), 3.32 (dd, J=9.2 Hz, 1H), 2.91 (dd, J=8.8 Hz, 1H), 2.70 (br s, 2H), 2.43 (dd, J=7.2 Hz, 1H), 1.97-1.91 (m, 4H), 1.69-1.64 (m, 2H), 0.89 (s, 9H), 0.04 (br s, 6H).

(459) Preparation of Compound 164

(460) Compound 164 was prepared from Compound 163 by a method similar to that for the synthesis of Compound 28.

(461) EI-MS m/z: [M+H].sup.+ 1548, 1/2[M+H].sup.+ 775.

<Example 55> Preparation of Compound 167

(462) ##STR00080##

(463) Preparation of Compound 165

(464) L-Histidine (5.0 g, 32.22 mmol) was dissolved in dichloromethane (45 mL), then dichlorodimethylsilane (3.9 mL, 32.22 mmol) and triethylamine (9.0 mL, 64.44 mmol) were added thereto at room temperature, and the reaction solution was heated under reflux for 4 hours under a nitrogen atmosphere. Trityl chloride (8.9 g, 32.22 mmol) and triethylamine (4.5 mL, 32.22 mmol) were added thereto, and the mixture was stirred for 2 hours under a nitrogen atmosphere. Methanol (50 mL) was added to the reaction solution, then the mixture was concentrated under reduced pressure, distilled water (50 mL) and triethylamine were added thereto to adjust the pH to about from 8 to 8.5, the insoluble slurry was filtered off therefrom, and the filtered product was washed with chloroform (50 mL), diethyl ether (50 mL), and distilled water (50 mL) in this order. The white solid compound formed was dried to obtain Compound 165 (triethylamine salt, 12.4 g, 95%).

(465) .sup.1H-NMR (400 MHz, CD.sub.3OD) δ 7.45-7.32 (m, 10H), 7.21-7.15 (m, 5H), 3.75-3.77 (m, 1H), 3.20 (q, 2H), 3.00-2.97 (m, 1H), 1.32 (t, 3H).

(466) Preparation of Compound 166

(467) Compound 165 (1.0 g, 2.52 mmol) and N-methoxycarbonylmaleimide (429 mg, 2.77 mmol) were dissolved in 1,4-dioxane/distilled water (5 mL/2.5 mL), then sodium carbonate (267 mg, 2.52 mmol) was added thereto, and the mixture was heated under reflux for 12 hours under a nitrogen atmosphere. The reaction solution was concentrated under reduced pressure and dissolved in N,N-dimethylformamide (3 mL), then triethylamine (0.16 mL, 1.12 mmol) was added to the reaction solution, and the mixture was stirred for 10 hours under a nitrogen atmosphere. Distilled water (5 ml) was added to the reaction solution, then the reaction solution was acidified (pH 4) by addition of a 0.5 N aqueous hydrochloric acid solution and subjected to extraction using dichloromethane (3×10 mL), and then the extract was dried over anhydrous sodium sulfate. The resultant was filtered and concentrated under reduced pressure to obtain Compound 166 (504 mg, 32%).

(468) EI-MS m/z: [M+H].sup.+ 478.4, [M+Na].sup.+500.4.

(469) Preparation of Compound 167

(470) Compound 166 (252 mg, 0.53 mmol) was dissolved in N,N-diisopropylethylamine (4 mL), and then N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (132 mg, 0.69 mmol) and N-hydroxysuccinimide (85 mg, 0.74 mmol) were added thereto. The reaction solution was stirred at room temperature for 12 hours. Distilled water (30 mL) was added to the reaction solution, and then the mixture was subjected to extraction using ethyl acetate (2×30 mL). The organic layer extracted was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and then purified by column chromatography to obtain Compound 167 (274 mg, 90%).

(471) EI-MS m/z: [M+H].sup.+ 575.3.

<Example 56> Preparation of Compound 169

(472) ##STR00081##

(473) Preparation of Compound 168

(474) Compound 103 (50 mg, 0.03 mmol) and Compound 167 (27.4 mg, 0.05 mmol) were dissolved in N,N-dimethylformamide (1 mL), then N,N-diisopropylethylamine (0.01 mL, 0.05 mmol) was added thereto, and the mixture was stirred at room temperature for 12 hours under a nitrogen atmosphere. The reaction solution was concentrated under reduced pressure, then purified by HPLC, and freeze-dried to obtain Compound 168 (11 mg, 18%).

(475) EI-MS m/z: [M+H].sup.+ 1934.8, 1/2[M+H].sup.+ 968.0.

(476) Preparation of Compound 169

(477) Compound 168 (11 mg, 6 mol) and anisole (6 μL, 60 μmol) were diluted with dichloromethane (0.75 mL), then trifluoroacetic acid (0.25 mL) was added thereto at 0° C., and the mixture was stirred for 3 hours. The reaction solution was concentrated under reduced pressure, then purified by HPLC, and freeze-dried to obtain Compound 169 (2 mg, 21%) as a white solid.

(478) EI-MS m/z: [M+H].sup.+ 1692.7, 1/2[M+H].sup.+ 846.9.

<Example 57> Preparation of Compound 171

(479) ##STR00082##

(480) Preparation of Compound 170

(481) DBCO-PEG4-acid (50 mg, 91 μmol) was dissolved in dichloromethane (1 mL), then N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (19 mg, 99 mol) and N-hydroxysuccinimide (11 mg, 99 μmol) were added thereto, and the mixture was stirred at room temperature for 3 hours under a nitrogen atmosphere. The reaction solution was concentrated under reduced pressure to obtain Compound 170 (59 mg).

(482) EI-MS m/z: [M+H].sup.+ 650.7.

(483) Preparation of Compound 171

(484) Compound 103 (47 mg, 32 μmol) and Compound 170 (24 mg, 38 μmol) were dissolved in N,N-dimethylformamide/dichloromethane (1 mL/0.25 mL), then N,N-diisopropylethylamine (51 μL, 38 μmol) was added thereto, and the mixture was stirred at room temperature for 3 hours under a nitrogen atmosphere. The reaction solution was concentrated under reduced pressure, then purified by HPLC, and freeze-dried to obtain Compound 171 (8.1 mg, 14%).

(485) EI-MS m/z: [M+H].sup.+ 2010.1, 1/2[M+H].sup.+ 1005.6.

(486) Preparation of Compounds 172 to 178

(487) Pyrrolobenzodiazepine dimer Compounds 172 to 178 having the structures presented in the following Table 1 were prepared referring to the references.

(488) TABLE-US-00001 TABLE 1 Structure of pyrrolobenzodiazepine dimer No. compound Reference 172 U.S. Pat. No. 8,697,688 173 U.S. Pat. No. 9,713,647 174 U.S. Pat. No. Application Laid- Open No. 2015- 0283258 175 U.S. Pat. Application Laid- Open No. 2015- 0283258 176 U.S. Pat. Application Laid- Open No. 2015- 0283258 177 U.S. Pat. Application Laid- Open No. 2015- 0283258 178 U.S. Pat. Application Laid- Open No. 2015- 0283258

<Example 58>

(489) Preparation of ADC

(490) ADCs were prepared through two steps, and LCB14-0511, LCB14-0512, and LCB14-0606 which were commonly used were prepared by the method described in Korean Patent Application Laid-Open No. 10-2014-0035393. The structural formulas of LCB14-0606, LCB14-0511, and LCB14-0512 are as follows:

(491) ##STR00090##

(492) Step 1: Preparation of Prenylated Antibody

(493) The prenylation reaction mixture of antibody was prepared and reacted at 30° C. for 16 hours. The reaction mixture was composed of 24 μM antibody, 200 nM FTase (Calbiochem #344145), and a buffer solution (50 mM Tris-HCl (pH 7.4), 5 mM MgCl.sub.2, 10 μM ZnCl.sub.2, 0.5 mM DTT) containing 0.144 mM LCB14-0511 or LCB14-0512 or LCB14-0606. After completion of the reaction, the prenylated antibody was desalted with a G25 Sepharose column (AKTA purifier, GE healthcare) equilibrated with PBS buffer solution.

(494) Step 2: Drug-Conjugation Method

(495) <Conjugation by Oxime Bond Formation>

(496) The oxime bond-forming reaction mixture between the prenylated antibody and the linker-drug was prepared by mixing 100 mM Na-acetate buffer solution pH 4.5, 10% DMSO, 24 M antibody, and 240 μM linker-drug (in house, compound in Table 1 as final product of Example 8 to 10 and Comparative Example 1) together and mildly stirred at 30° C. After 24 hours of reaction, the excess amount of low-molecular compounds used was removed through FPLC (AKTA purifier, GE healthcare), and the protein fractions were collected and concentrated.

(497) <Conjugation by Click Reaction>

(498) The click reaction mixture between the prenylated antibody and the linker-drug was prepared by mixing 10% DMSO, 24 μM antibody, 240 μM linker-drug (in house, compound in Table 1 as final product of Examples 20, 21, 22, 26, 27, 28, and 46), 1 mM copper(II) sulfate pentahydrate, 2 mM (BimC.sub.4A).sub.3 (Sigma-Aldrich 696854), 10 mM sodium ascorbate, and 10 mM aminoguanidine hydrochloride, reacted at 25° C. for 3 hours, then treated with 2.0 mM EDTA, and reacted for 30 minutes. After completion of the reaction, the excess amount of the low-molecular compounds used was removed through FPLC (AKTA purifier, GE healthcare), and the protein fractions were collected and concentrated.

(499) TABLE-US-00002 TABLE 2 List of ADC prepared # Compound ADC# Example 8 28 ADC1 Example 9 29 ADC2 Example 10 30 ADC3 Example 12 34 ADC4 Example 14 42 ADC5 Example 20 61 ADC6 Example 21 62 ADC7 Example 22 63 ADC8 Example 23 65 ADC9 Example 26 73 ADC10 Example 27 74 ADC11 Example 28 75 ADC12 Example 31 82 ADC13 Example 32 83 ADC14 Example 33 85 ADC15 Example 36 100 ADC16 Example 39 104 ADC17 Example 41 112 ADC18 Example 46 135 ADC19 Example 47 137 ADC20 Example 57 171 ADC21 Comparative 86 ADC22 Example 1 87 ADC23 88 ADC24

<Experimental Example 1> Evaluation on Cytotoxicity in Vitro

(500) The cell proliferation inhibitory activity of the drug and ADCs presented in the following Table 2 on cancer cell lines was measured. As the cancer cell lines, commercially available human breast cancer cell lines MCF-7 (HER2 negative to normal), SK-BR3 (HER2 positive), and JIMT-1 (HER2 positive) were used. MMAF-OMe was used as the drug and the ADCs in Table 1 were used as the ADC. Each of the cancer cell lines was seeded in a 96-well plate by from 5,000 to 13,000 per well for the 72 hour treatment group and from 1,500 to 3,000 per well for the 168 hour treatment group, incubated for 24 hours, and then treated with antibodies and ADCs at a concentration of from 0.0051 to 33.33 nM or from 0.0015 to 10.0 nM (three times serial dilutions) and drugs at a concentration of from 0.023 to 50 nM (three times serial dilutions). After 72/168 hours, the number of living cells was quantified using SRB (Sulforhodamine B) dye.

(501) TABLE-US-00003 TABLE 3 Comparison of cytotoxicity of ADC samples CC50 (nM) SK-BR3 JIMT-1 MCF7 Test Samples 72 hr.sup.a 168 hr 72 hr 168 hr 72 hr 168 hr MMAF-OMe 0.14 0.06 0.14 0.06 0.84 0.19 ADC1 0.05 0.004 >10 0.11 >10 >10 ADC2 0.06 0.01 >10 0.19 >10 >10 ADC3 0.11 0.01 >10 0.19 >10 >10 ADC4 NT.sup.b 0.003 NT 0.10 NT >1 ADC5 NT 0.005 NT 0.10 NT >1 ADC8 NT 0.01 NT 0.19 NT >1 ADC9 NT 0.007 NT 0.70 NT >10 ADC10 0.04 0.004 >100 0.09 >100 0.82 ADC12 NT 0.004 NT 0.15 NT >10 ADC13 NT 0.02 NT 0.12 NT 0.76 ADC14 NT 0.007 NT 0.11 NT >1 ADC15 NT 0.02 NT 2.39 NT >10 ADC16 0.06 0.01 >10 0.33 >10 0.48 ADC20 NT 18.5.sup.c NT >100.sup.c NT >100.sup.c ADC21 NT 0.006.sup.c NT 0.16.sup.c NT 27.1.sup.c ADC22 15.3 NT >33.3 NT >33.3 NT ADC23 0.40 0.09 >33.3 >10 >33.3 >10 ADC24 1.08 NT >33.3 NT >33.3 NT

(502) It has been found that ADC1, 2, and 3 samples into which prodrug linker-drugs 28, 29, and 30 having a carbamate structure at both N10 positions of pyrrolobenzodiazepine are introduced among antibody-drug conjugates are superior to ADC22, 23, and 24 samples in cytotoxicity in the SK-BR3 and JIMT-1 breast cancer cell lines.

(503) Since the compound is required to be converted into an effective drug by an additional reaction at the time of exposure to blood in the case of being administered in a form of the prodrug according to the present invention, and it is thus advantageous as compared to conventional PBD drugs in that the occurrence of side effects which may occur at the time of unexpected decomposition of linker can be prevented in advance, toxicity to normal cells diminishes, and the drug is more stable.

(504) In addition, in the preparation of antibody-drug conjugate, the content of impurities is high and there is a possibility that the exposed imine group is attacked by nucleophiles and a drug having an unwanted structure is thus formed in the case of an antibody-drug conjugate prepared by the conventional method. However, the antibody-drug conjugate prepared by the method according to the present invention has an advantage of being protected from the attack by nucleophiles since the imine group of PBD dimer is in a form of the prodrug and easily separated since the purity thereof is high and exhibits improved physical properties as compared to the conventional PBD or PBD dimer.

INDUSTRIAL APPLICABILITY

(505) The pyrrolobenzodiazepine dimer prodrug, pyrrolobenzodiazepine dimer prodrug-linker, or pyrrolobenzodiazepine dimer prodrug-linker-ligand conjugate according to the present invention can be used in the targeting and specific treatment of proliferative diseases such as cancer.