Quinazolinone derivative, preparation method therefor, pharmaceutical composition, and applications

Abstract

Disclosed are a quinazolinone derivative, a preparation method therefor, a pharmaceutical composition, and applications. Provided are a compound represented by formula I, a pharmaceutically acceptable salt, a solvate, a crystal form, a eutectic crystal, a stereoisomer, an isotope compound, a metabolite, or a prodrug thereof. Generation or activity of a cell factor can be regulated, and accordingly, cancers and inflammatory diseases can be effectively treated. ##STR00001##

Claims

1. A method for treating a disease, disorder or condition comprising administering to a subject in need thereof a therapeutically effective amount of a compound of Formula I, or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic compound thereof; ##STR00069## wherein, X is selected from the group consisting of halogen, hydroxyl, cyano, substituted or unsubstituted C.sub.1-C.sub.6 alkyl, and C.sub.1-C.sub.6 alkoxy substituted with 6-10 membered aryl; wherein the 6-10 membered aryl in the C.sub.1-C.sub.6 alkoxy substituted with 6-10 membered aryl is optionally substituted with one or more selected from the group consisting of D, halogen, hydroxyl, cyano, substituted or unsubstituted C.sub.1-C.sub.6 alkyl, and substituted or unsubstituted C.sub.1-C.sub.6 alkoxy, wherein when more than one substituents are present, they are identical or different; Z is ##STR00070## wherein the carbon marked with * is asymmetric center; R.sup.1 is selected from the group consisting of hydroxyl, substituted or unsubstituted C.sub.1-C.sub.6 alkoxy and —NR.sup.1′R.sup.2′; wherein R.sup.1′ and R.sup.2′ are each independently selected from the group consisting of H, D, substituted or unsubstituted C.sub.1-C.sub.6 alkyl and —C(O)R.sup.3′; R.sup.3′ is substituted or unsubstituted C.sub.1-C.sub.6 alkyl; R.sup.2, R.sup.3, R.sup.5, R.sup.6, R.sup.7, R.sup.8, R.sup.9, and R.sup.10 are each independently H or D; R.sup.4 is CH.sub.3, CH.sub.2D, CHD.sub.2 or CD.sub.3; wherein the substituted in the above substituted or unsubstituted C.sub.1-C.sub.6 alkoxy and the above substituted or unsubstituted C.sub.1-C.sub.6 alkyl independently represents substitution with one or more selected from the group consisting of D, halogen, amino, hydroxyl, cyano, C.sub.1-C.sub.6 alkoxy, and 4-10 membered heterocycloalkyl, wherein when more than one substituents are present, they are identical or different; and wherein the disease, disorder or condition is a TNF-α associated disorder selected from the group consist of: myelodysplastic syndrome, multiple myeloma, mantle cell lymphoma, diffuse large B cell lymphoma, central nervous system lymphoma, non-Hodgkin's lymphoma; papillary and follicular thyroid carcinoma; breast cancer, chronic lymphocytic leukemia, chronic myelogenous leukemia, amyloidosis, type I complex regional pain syndrome, malignant melanoma, radiculopathy, myelofibrosis, glioblastoma, gliosarcoma, malignant glioma, refractory plasma cell tumor, chronic myelomonocytic leukemia, follicular lymphoma, ciliary body and chronic melanoma, iridic melanoma, recurrent interocular melanoma, extraocular extension melanoma, solid tumor, T-cell lymphoma, erythroid lymphoma, monoblastic and monocytic leukemia; myeloid leukemia, brain tumor, meningioma, spinal tumor, thyroid cancer, non-small cell lung cancer, ovarian cancer, renal cell carcinoma, Burkitt's lymphoma, Hodgkin's lymphoma, large cell lymphoma, astrocytoma, hepatocellular carcinoma, and primary macroglobulinemia.

2. The method of claim 1, wherein in the compound of Formula I, or the pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic compound thereof the halogen in X is fluorine, chlorine, bromine or iodine; and/or when the 6-10 membered aryl is optionally substituted with halogen, the halogen is fluorine, chlorine, bromine or iodine; and/or when the substituted in the substituted or unsubstituted C.sub.1-C.sub.6 alkoxy and the substituted or unsubstituted C.sub.1-C.sub.6 alkyl independently represents substitution with halogen, the halogen is fluorine, chlorine, bromine or iodine; and/or the C.sub.1-C.sub.6 alkyl in the substituted or unsubstituted C.sub.1-C.sub.6 alkyl is C.sub.1-C.sub.4 alkyl; and/or the C.sub.1-C.sub.6 alkoxy in the substituted or unsubstituted C.sub.1-C.sub.6 alkoxy is C.sub.1-C.sub.4 alkoxy; and/or the 4-10 membered heterocycloalkyl is selected from the group consisting of 5-6 membered heterocycloalkyl, wherein the heteroatom is one or more selected from the group consisting of N, O and S, and wherein the number of heteroatom is 1 or 2; and/or the C.sub.1-C.sub.6 alkoxy substituted with 6-10 membered aryl is selected from the group consisting of C.sub.1-C.sub.4 alkoxy substituted with phenyl; wherein the phenyl is optionally substituted with one or more of the following groups: D, halogen, hydroxyl, cyano, or C.sub.1-C.sub.4 alkyl substituted with pyrrolidinyl, piperidinyl, piperazinyl or morpholinyl, wherein when more than one substituents are present, they are identical or different.

3. The method of claim 1, wherein in the compound of Formula I, or the pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic compound thereof Z is selected from the group consisting of any of the following structures: ##STR00071## ##STR00072## ##STR00073##

4. The method of claim 1, wherein in the compound of Formula I, or the pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic compound thereof
R.sup.1 is —NR.sup.1′R.sup.2′; and/or R.sup.1′ and R.sup.2′ are each independently selected from the group consisting of H, D, substituted or unsubstituted C.sub.1-C.sub.4 alkyl, and —C(O)R.sup.3′; and/or R.sup.3′ is selected from the group consisting of substituted or unsubstituted C.sub.1-C.sub.4 alkyl.

5. The method of claim 1, wherein in the compound of Formula I, or the pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic compound thereof X is selected from the group consisting of halogen, hydroxyl, cyano, substituted or unsubstituted C.sub.1-C.sub.4 alkyl, and methoxy substituted with phenyl; wherein the phenyl is optionally substituted with one or more selected from the group consisting of D, halogen, hydroxyl, cyano, and C.sub.1-C.sub.4 alkyl substituted with morpholinyl, wherein when more than one substituents are present, they are identical or different.

6. The method of claim 5, wherein in the compound of Formula I, or the pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic compound thereof X is selected from the group consisting of fluorine, chlorine, bromine, hydroxyl, cyano, benzyloxy, 2-fluoro-4-(morpholinyl-1-methyl)benzyloxy, methyl, ethyl, CD.sub.3, C.sub.2D.sub.5 and CH.sub.2CD.sub.3.

7. The method of claim 1, wherein in the compound of Formula I, or the pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic compound thereof X is halogen, R.sup.1 is NH.sub.2, NHD or ND.sub.2; R.sup.2, R.sup.3, R.sup.5, R.sup.6, R.sup.7, R.sup.8, R.sup.9 and R.sup.10 are independently H or D; R.sup.4 is CH.sub.3, CH.sub.2D, CHD.sub.2 or CD.sub.3.

8. The method of claim 1, wherein the compound of Formula I is selected from any of the following structures: ##STR00074## ##STR00075## ##STR00076## ##STR00077## ##STR00078## ##STR00079## ##STR00080## ##STR00081## ##STR00082## ##STR00083## ##STR00084## ##STR00085## ##STR00086## ##STR00087## ##STR00088##

9. The method according to claim 1, further comprising administering to the subject a therapeutically effective amount of one or more other therapeutic agents selected from the group consisting of anti-angiogenic agent, immunoregulating agent, immunotherapeutic agent, chemotherapeutic agent or hormone compound.

10. The method of claim 4, wherein in the compound of Formula I, or the pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic compound thereof, R.sup.1′ and R.sup.2′ are each independently selected from the group consisting of H, D, methyl, ethyl, isopropyl, acetyl, propionyl and isobutyryl.

Description

DETAILED DESCRIPTION OF THE EMBODIMENTS

(1) The invention will be further illustrated by the following examples, but it should not be constructed that the invention is limited to the scope of the examples. The experimental methods that are not specified in details in the following examples are those according to conventional methods and conditions, or according to the product manuals.

Example 1 Synthesis of Compound K101

(2) Synthesis Scheme

(3) ##STR00038## ##STR00039##

(4) Step 1: Synthesis of Compound K101-b

(5) To a solvent of K101-a (60 g, 38.7 mmol) in con. H.sub.2SO.sub.4 (150 mL), a mixture of con. HNO.sub.3 (36 mL) and con. H.sub.2SO.sub.4 (200 mL) was added dropwise over 2 hours at 0° C. while the temperature of the reaction mixture was controlled at 0° C.-15° C. The reaction mixture was stirred for another 1 hour and quenched by pouring into crushed ice. Then the water layer was extracted with DCM (100 mL×2). The combined organic solution was dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to afford a crude product, which was purified by chromatography column on silica gel to afford the product K101-b (16 g, 21%).

(6) .sup.1H NMR (CDCl.sub.3, 300 MHz): δ 7.78 (d, J=6.9 Hz, 2H), 2.55 (s, 3H).

(7) Step 2: Synthesis of Compound K101-c

(8) To a solution of K101-b (9.0 g, 45.0 mmol) in EtOH (100 mL), a solution of Na.sub.2S (16.2 g, 67.5 mmol) in H.sub.2O (50 mL) was added dropwise over 30 min at 25° C. The mixture was stirred for 4 hours, then concentrated to afford a crude product. The crude product was diluted with H.sub.2O (200 mL) and extracted with EtOAc (100 mL×2), the combined organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated, the residue was purified by chromatography column (PE/EtOAc=10/1) to afford the product K101-c (5.0 g, 65%).

(9) .sup.1H NMR (DMSO-d.sub.6, 300 MHz): δ 6.84 (dd, J=6.3, 1.8 Hz, 1H), 6.68 (dd, J=8.4, 1.8 Hz, 1H), 5.90 (br s, 2H), 2.03 (s, 3H).

(10) Step 3: Synthesis of Compound K101-e

(11) To a solution of K101-c (1.60 g, 9.40 mmol) in THF (150 mL) was added (Boc).sub.2O (2.25, 10.0 mmol) and DMAP (1.15 g, 9.40 mmol). The mixture was stirred for 18 hours at 25° C. and concentrated to remove THF. The residue was diluted with EtOAc (200 mL), then washed with 1N/HCl (100 mL×2) and dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to afford a crude product (1.7 g). The crude product was added to a mixture of pyridine (30 mL) and H.sub.2O (15 mL). The mixture was heated to 80° C., then KMnO.sub.4 (3.2 g, 19.8 mmol) was added in 4 batches over 2 hours (one batch every 30 minutes). The resulting mixture was stirred overnight. The reaction solution was filtered and the cake was washed with hot water. The filtrate was extracted with DCM (150 mL×3). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered and concentrated to afford a crude product. The crude product was purified by chromatography column on silica gel (EtOAc/PE=1/5) to afford the product K101-e (1.0 g, 30% for 2 steps).

(12) .sup.1H NMR (CDCl.sub.3, 300 MHz): δ 10.53 (br s, 1H), 8.01 (dd, J=11.4, 2.4 Hz, 1H), 7.46 (dd, J=7.8, 2.4 Hz, 1H), 1.45 (s, 9H).

(13) Step 4: Synthesis of Compound K101-g

(14) To a solution of 4N/HCl in 1,4-dioxane (80 mL) was added K101-e (1.0 g, 3.3 mmol). The mixture was stirred for 2 hours at 25° C. and concentrated to afford a crude product (800 mg). A mixture of the crude product and Ac.sub.2O (10 mL) was heated to reflux and stirred for 4 hours. The reaction solution was concentrated and the residue was stirred with (EtOAc/Et.sub.2O=½, 30 mL) for 30 min. The solid impurities were remove by filtration. The filtrate was concentrated to afford the product K101-g (670 mg, 91% for 2 steps)

(15) .sup.1H NMR (DMSO-d.sub.6, 300 MHz): δ 8.14 (dd, J=8.1, 2.4 Hz, 1H), 7.74 (dd, J=9.0, 2.4 Hz, 1H), 2.42 (s, 3H).

(16) Step 5: Synthesis of Compound K101-h

(17) To a mixture of K101-g (500 mg, 2.23 mmol) in MeCN (25 mL) was added (S)-tert-butyl 4,5-diamino-5-oxopentanoate hydrochloride (640 mg, 2.68 mmol), imidazole (334 mg, 4.91 mmol) and triphenyl phosphite (832 mg, 2.68 mmol). The reaction solution was stirred at reflux for 16 hours. This mixture was concentrated and diluted with H.sub.2O (150 mL) and extracted with EtOAc (100 mL×2). The combined organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified with chromatography column on silica gel (EtOAc/PE=1/3) to afford the product K101-h (600 mg, 66%).

(18) .sup.1H NMR (DMSO-d.sub.6, 300 MHz): δ 7.98 (dd, J=8.7, 2.4 Hz, 1H), 7.68 (dd, J=9.9, 2.4 Hz, 1H), 7.42-7.49 (m, 1H), 7.13-7.21 (m, 1H), 4.68-4.92 (m, 1H), 2.54 (s, 3H), 2.05-2.43 (m, 4H), 1.28 (s, 9H).

(19) Step 6: Synthesis of Compound K101-j

(20) To a solution of K101-h (600 mg, 1.47 mmol) in EtOH (60 mL) was added saturated aq. NH.sub.4Cl solution (20 mL). The mixture was heated to 80° C. and Fe powder (600 mg, 10.7 mmol) was added. The reaction mixture was heated with stirring for another 3 hours, filtered and concentrated to remove the majority of EtOH. The remaining mixture was extracted with EtOAc (150 mL×2). The combined organic layer was dried and concentrated to afford the product K101-j (540 mg, 97%)

(21) .sup.1H NMR (DMSO-d.sub.6, 300 MHz): δ 6.97-7.50 (m, 4H), 6.30-6.33 (m, 2H), 4.56-4.73 (m, 1H), 2.44 (s, 3H), 2.06-2.32 (m, 4H), 1.32 (s, 9H).

(22) Step 7: Synthesis of Compound K101-k

(23) To a solution of 4N/HCl in 1,4-dioxane (20 mL) was added K101-j (540 mg, 1.43 mmol). This mixture was stirred at 25° C. for 2 hour, then concentrated to afford the product K101-k (492 mg).

(24) .sup.1H NMR (DMSO-d.sub.6, 300 MHz): δ 7.12-7.56 (m, 4H), 6.64 (d, J=6.0 Hz, 1H), 6.51 (d, J=6.0 Hz, 1H), 4.80 (br s, 1H), 2.76 (s, 3H), 1.98-2.38 (m, 4H).

(25) Step 8: Synthesis of Compound K101

(26) To a solution of K101-k (400 mg, 1.24 mmol) in MeCN was added CDI (400 mg, 2.48 mmol). The reaction solution was heated to 95° C. and stirred overnight, then concentrated to afford a crude product. The crude product was purified by HPLC to afford the product K101 (210 mg, 56%).

(27) .sup.1H NMR (DMSO-d.sub.6, 300 MHz): δ 10.99 (s, 1H), 7.32 (br s, 2H), 6.34 (d, J=10.8 Hz, 2H), 5.13-5.19 (m, 1H), 2.82-2.88 (m, 1H), 2.58-2.78 (m, 2H), 2.53 (s, 3H), 2.11-2.18 (m, 1H). LCMS: 305.1 ([M+1].sup.+).

Example 2 Synthesis of Compound K105

(28) ##STR00040##

(29) Step 1: Synthesis of Compound K105-b

(30) To a mixture of K101-g (800 mg, 3.57 mmol) in CH.sub.3CN (30 mL), K105-a (726 mg, 3.57 mmol), imidazole (533 mg, 7.85 mmol) and triphenyl phosphite (1.33 g, 4.28 mmol) were added. The reaction solution was stirred at reflux for 16 hour. This mixture was concentrated and diluted with EtOAc (500 mL), washed successively with water, saturated aq. NaHCO.sub.3 solution and brine. The organic layer was dried over Na.sub.2SO.sub.4, filtered and concentrated. The residue was purified with chromatography column on silica gel (EtOAc/PE=1/1) to afford the product K105-b (480 mg, yield: 33%).

(31) .sup.1H NMR (DMSO-d.sub.6, 300 MHz): δ 8.00 (dd, J=8.1, 2.4 Hz, 1H), 7.70 (dd, J=9.6, 2.1 Hz, 1H), 7.21-7.38 (m, 2H), 2.57 (s, 3H), 2.24-2.51 (m, 2H), 2.08-2.18 (m, 2H), 1.31 (s, 9H).

(32) Step 2: Synthesis of Compound K105-c

(33) To a solution of K105-b (480 mg, 1.17 mmol) in EtOH (60 mL) was added saturated aq. NH.sub.4Cl solution (20 mL). The mixture was heated to 80° C. and Fe powder (6570 mg, 11.72 mmol) was added to the reaction solution. The mixture was stirred for 3 hour at 80° C., then cooled to room temperature, filtered and concentrated under reduced pressure to remove the majority of EtOH. The remaining water layer was extracted with EtOAc (100 mL×3). The combined organic layer was dried, filtered and concentrated. The residue was purified with chromatography column on silica gel (EtOAc/PE=1/1) to afford the product K105-c (437 mg, yield: 98%)

(34) .sup.1H NMR (DMSO-d.sub.6, 300 MHz): δ 7.07-7.49 (m, 4H), 6.30-6.35 (m, 2H), 2.45 (s, 3H), 2.07-2.34 (m, 4H), 1.33 (s, 9H).

(35) Step 3: Synthesis of Compound K105

(36) To a solution of 6N/HCl in 1,4-dioxane (30 mL) was added K105-c (437 mg, 1.15 mmol). This mixture was stirred at 25° C. for 2 hours, and then concentrated. The residue was dissolved in DMF (3 mL) and DCM (30 mL). The mixture was cooled to −40° C., and SOCl.sub.2 (685 mg, 5.76 mmol) in DCM (2 mL) was added dropwise. Then the mixture was reacted at −40˜−30° C. for 1.5 hours. Pyridine (912 mg, 11.52 mmol) in DCM (2 mL) was added and the mixture was stirred at −40˜−30° C. for 1 hour. Et.sub.3N (237 mg, 2.7 mmol) in DCM (1 mL) was added and the mixture was stirred at −40˜−30° C. for 1 hour. H.sub.2O (10 mL) was then added to quench the reaction. The mixture was concentrated under reduced pressure to afford a crude product. The crude product was purified by Prep-HPLC to afford K105 (68 mg).

(37) .sup.1H NMR (DMSO-d.sub.6, 400 MHz): δ 10.94 (br s, 1H), 7.35 (br s, 2H), 6.36 (s, 1H), 6.33 (s, 1H), 5.14-5.19 (m, 0.21H), 2.87-2.77 (m, 1H), 2.63-2.54 (m, 2H), 2.51 (s, 3H), 2.11-2.16 (m, 1H). MS: 306.1 ([M+1].sup.+).

Example 3 Synthesis of Compound K102

(38) ##STR00041##

(39) Compound K102 was synthesized by a similar method as K105 described in Example 2 except the corresponding substrate was used instead of compound K105-a in step 1.

(40) .sup.1H NMR (DMSO-d.sub.6, 400 MHz): δ 11.01 (s, 1H), 7.32 (br s, 2H), 6.32-6.36 (m, 2H), 5.16 (dd, J=11.6, 5.6 Hz, 1H), 2.78-2.83 (m, 1H), 2.57-2.66 (m, 2H), 2.54 (s, 3H), 2.08-2.17 (m, 1H). MS: 305.1 ([M+1].sup.+).

Example 4 Synthesis of Compound K106

(41) ##STR00042##

(42) Compound K106 was synthesized by a similar method as compound K105 described in Example 2 except the corresponding substrate

(43) ##STR00043##
was used instead of K105-a in step 1.

(44) .sup.1H NMR (400 MHz, DMSO-d6) δ 11.03 (s, 1H), 7.34 (br s, 2H), 6.36 (s, 1H), 6.33 (s, 1H), 5.14-5.18 (m, 0.11H), 2.76-2.87 (m, 1H), 2.59-2.63 (m, 2H), 2.54 (s, 3H), 2.13-2.17 (m, 1H). LCMS: 306.0 ([M+1]+).

Example 5 Synthesis of Compound K103

(45) ##STR00044##

(46) Compound K103 was synthesized by a similar method as compound K105 described in Example 2 except the corresponding substrate

(47) ##STR00045##
was used instead of K105-a in step 1.

(48) .sup.1H NMR (400 MHz, DMSO-d6) δ 11.00 (s, 1H), 7.32 (br s, 2H), 6.36 (s, 1H), 6.33 (s, 1H), 5.14-5.18 (m, 1H), 2.78-2.87 (m, 1H), 2.59-2.67 (m, 2H), 2.54 (s, 3H), 2.08-2.17 (m, 1H). LCMS: 305.1 ([M+1]+).

Example 6 Synthesis of Compound K104

(49) ##STR00046##

(50) Compound K104 was synthesized by a similar method as compound K105 described in Example 2 except the corresponding substrate

(51) ##STR00047##
was used instead of K105-a in step 1.

(52) .sup.1H NMR (400 MHz, DMSO-d6) δ 11.01 (s, 1H), 7.33 (brs, 1H), 6.33-6.36 (m, 2H), 2.79-2.83 (m, 1H), 2.57-2.62 (m, 2H), 2.53 (s, 3H), 2.12-2.16 (m, 1H). LCMS: 306.1 ([M+1]+).

Example 7 Synthesis of Compound K501

(53) ##STR00048##

(54) Step 1: Synthesis of Compound K501-B

(55) To the solution of compound K501-A (14.0 g, 51.1 mmol) in THF (90 mL) was added LiOH (6.4 g, 153 mmol) and H.sub.2O (30 mL). The reaction mixture was stirred at 25° C. overnight and then concentrated. The remaining liquid was diluted with Et.sub.20 (60 mL) and water (100 mL). The organic layer was separated. The water layer was adjusted with 2N HCl to pH=2, extracted with EtOAc (150 mL). The organic layers were washed with sat. brine (200 mL), dried, filtered and concentrated to afford K501-B (12.9 g) as yellow solid.

(56) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ 8.30 (d, J=2.0 Hz, 1H), 8.14 (d, J=2.0 Hz, 1H), 2.45 (s, 3H)

(57) Step 2: Synthesis of Compound K501-C

(58) To the solution of K501-B (12.9 g, 49.61 mmol) in t-BuOH (200 mL) was added phosphorazidic acid diphenyl ester (20.5 g, 74.42 mmol) and Et.sub.3N (7.5 g, 74.4 mmol). The mixture was stirred at 80° C. overnight. The reaction solution was concentrated and the remaining liquid was diluted with EtOAc (300 mL) and water (200 mL), the organic layer was washed with sat. brine (200 mL), dried, filtered and concentrated. The solid residue was purified by column chromatography on silica gel PE:EA (10:1) to afford K501-C (15.3 g) as yellow solid.

(59) .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.34 (s, 1H), 7.64 (d, J=2.0 Hz, 1H), 6.46 (s, 1H), 2.29 (s, 3H), 1.53 (s, 9H).

(60) Step 3: Synthesis of Compound K501-D

(61) To a mixture of pyridine (300 mL) and H.sub.2O (150 mL) was added K501-C (15.3 g, 46.2 mmol). This mixture was heated to 80° C., KMnO.sub.4 (29.2 g, 184.8 mmol) was added in 6 batches over 3 hours (one batch every 30 minutes). The resulting mixture was stirred overnight and then the reaction solution was filtered. The filter cake was washed with EtOAc (800 mL) and hot water (200 mL). The combined filtrate was concentrated and adjusted with 1N HCl to pH=2, extracted with EtOAc (800 mL). The combined organic layer was dried over Na.sub.2SO.sub.4 then filtered and concentrated to afford the solid residue. The residue was purified by column chromatography on silica gel (PE:EA 30:1˜5:1) to afford K501-D (9.8 g) as yellow solid.

(62) .sup.1H NMR (400 MHz, DMSO-d6) δ 9.55 (s, 1H), 8.29 (d, J=2.0 Hz, 1H), 7.91 (d, J=2.0 Hz, 1H), 1.47 (s, 9H).

(63) Step 4: Synthesis of Compound K501-E

(64) To a solution of K501-D in DCM (100 mL) was added CF.sub.3COOH at 0° C. The mixture was reacted overnight at 25° C., then concentrated. HCl in 1,4-dioxane (30 ml) was added. The mixture was stirred for 20 min at 25° C., then concentrated to afford K501-E (6.7 g).

(65) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 7.19-7.24 (m, 1H), 7.11-7.13 (m, 1H).

(66) Step 5: Synthesis of Compound K501-F

(67) A solution of K501-E (2.8 g) in Ac.sub.2O (20 mL) and HOAc (60 mL) was heated to reflux and stirred for 3 hours. The reaction solution was concentrated and the residue was stirred and slurried in EtOAc:PE (2:1, 15 mL) for 1 hour, then filtered to afford K501-F (2.3 g) as yellow solid.

(68) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 8.36 (d, J=1.6 Hz, 1H), 8.08 (d, J=2.0 Hz, 1H), 2.42 (s, 3H).

(69) Step 6: Synthesis of Compound K501-G

(70) To a mixture of K501-F (500 mg, 1.75 mmol), 3-aminopiperidine-2,6-dione hydrochloride (433 mg, 2.63 mmol) in CH.sub.3CN (20 mL) was added imidazole (262 mg, 3.86 mmol), (PhO).sub.3P (816 mg, 2.63 mmol). The mixture was stirred for 16 hours at 85° C. After the reaction was completed, the solvent was removed in vacuum. To the residue was added 9 mL of EtOAc and 9 mL of H.sub.2O, the mixture was stirred and slurried for 1 hour and filtered to afford K501-G (382 mg, crude) as gray solid.

(71) Step 7: Synthesis of Compound K501

(72) A mixture of K501-G in HOAc (15 mL) was heated to 80° C., and then Fe powder (965 mg, 17.3 mmol) was added. The mixture was reacted for 2 hours, then filtered to remove Fe powder. HOAc was removed in vacuum to afford a crude product. The crude product was purified by column chromatography on silica gel (CH.sub.3CN:DCM 1:1) to give a product which was further purified by prep-HPLC to afford K501 (180 mg).

(73) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ 11.00 (s, 1H), 7.24 (s, 2H), 6.74 (dd, J=13.2, 1.6 Hz, 2H), 5.15-5.19 (m, 1H), 2.78-2.87 (m, 1H), 2.59-2.65 (m, 2H), 2.58 (s, 3H), 2.14-2.18 (m, 1H). LCMS: 367.0 ([M+2].sup.+).

Example 8 Synthesis of Compound K401

(74) Compound K401 was synthesized by a similar method as compound K501 described in Example 7 except the corresponding starting material 5-chloro-2-methyl-3-nitrobenzoic acid was used instead of compound K501-B.

(75) ##STR00049##

(76) .sup.1H NMR (400 MHz, DMSO-d.sub.6): δ 11.00 (s, 1H), 7.24 (br s, 2H), 6.76 (d, J=1.6 Hz, 1H), 6.73 (d, J=1.6 Hz, 1H), 5.15-5.19 (m, 1H), 2.82-2.83 (m, 1H), 2.58-2.62 (m, 2H), 2.54 (s, 3H), 2.07-2.16 (m, 1H). LCMS: 321.0 ([M+1].sup.+).

Example 9 Synthesis of Compound K633 and K635

(77) ##STR00050##

(78) Synthesis of Compound K633

(79) To a solution of K101 (300 mg, 0.99 mmol) in 10 mL of DMF was added Ac.sub.2O (1 mL). The mixture was heated to 50° C. in oil bath and reacted for 5 hours, cooled to 25° C. and concentrated to dryness under reduced pressure. The residue was recrystallized from CH.sub.3CN and then purified by prep-HPLC to afford K633 (250 mg).

(80) .sup.1H NMR (DMSO-d.sub.6, 400 MHz): δ 11.82 (s, 1H), 11.09 (s, 1H), 8.36 (dd, J=12.8, 2.4 Hz, 1H), 7.08 (dd, J=9.6, 2.4 Hz, 1H), 5.32-5.36 (m, 1H), 2.81-2.91 (m, 1H), 2.61-2.73 (m, 5H), 2.19-2.23 (m, 4H). LCMS: 347.1 [(M+1)]+.

(81) Synthesis of Compound K635

(82) Compound K635 was synthesized by a similar method as compound K633 except the corresponding substrate isobutyric anhydride was used instead of Ac.sub.2O.

(83) .sup.1H NMR (DMSO-d.sub.6, 400 MHz): δ 11.96 (s, 1H), 11.11 (s, 1H), 8.39 (dd, J=12.4, 2.4 Hz, 1H), 7.08 (dd, J=9.6, 2.8 Hz, 1H), 5.34 (dd, J=11.6, 5.6 Hz, 1H), 2.81-2.90 (m, 1H), 2.51-2.73 (m, 6H), 2.17-2.23 (m 1H), 1.16 (d, J=7.2 Hz, 6H). LCMS: 375.0 [(M+1)]+.

Example 10 Synthesis of Compound K627

(84) ##STR00051##

(85) Step 1: Synthesis of Compound K627-B

(86) K627-A (10.0 g, 37.0 mmol) was dissolved in HCl/dioxane (5 M, 100 mL) and stirred at 15° C. for 2 hours. The solvent was removed by rotary evaporation. The residue was slurried with PE (100 mL) at 15° C. for 1 hour to afford the product K627-B (7.1 g) as solid.

(87) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 8.26 (br s, 2H), 6.92 (dd, J=8.8, 2.8 Hz, 1H), 6.79 (dd, J=11.4, 2.8 Hz, 1H), 2.06 (s, 3H).

(88) Step 2: Synthesis of Compound K627-C

(89) To a mixed solvent of conc. H.sub.2SO.sub.4 (75 mL) and water (37 mL) was added K627-B (6.5 g) at 0° C. NaNO.sub.2 (2.86 g, 42 mmol) was added slowly and the reaction solution was stirred at 0° C. for another 2 hours. The mixture was heated to 115° C. and H.sub.2SO.sub.4 (50%, 110 mL) was added dropwise. Then the mixture was stirred at 115° C. for another 2 hours. After cooled to room temperature, the mixture was extracted with EtOAc (300 mL×2). The organic layer was washed with sat. brine (300 mL), dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to afford a crude product K627-C (5.4 g).

(90) .sup.1H NMR (400 MHz, DMSO) δ 10.91 (s, 1H), 7.26 (dd, J=8.4, 2.4 Hz, 1H), 6.93 (dd, J=10.0, 2.4 Hz, 1H), 2.18 (s, 3H).

(91) Step 3: Synthesis of Compound K627-D

(92) K627-C (5.4 g, 31.6 mmol) and K.sub.2CO.sub.3 (21.8 g, 158 mmol) were dissolved in DMF (100 mL). To the mixture was added CH.sub.3I (13.5 g, 94.7 mmol) at 0° C. The mixture was stirred at 20° C. overnight, then concentrated under reduced pressure to remove the solvent. The residue was dissolved with EtOAc (500 mL), washed with water (300 mL×2) and sat. brine (300 mL), dried and concentrated to afford K627-D (5.23 g) as brown solid.

(93) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 7.39-7.41 (m, 1H), 7.28-7.31 (m, 1H), 3.90 (s, 3H), 2.19 (s, 3H).

(94) Step 4: Synthesis of Compound K627-E

(95) To a mixed solution of KMnO.sub.4 (13.6 g, 86 mmol) and H.sub.2O (550 mL) was added K627-D (5.2 g, 28.1 mmol) and 5% NaOH aqueous solution (55 mL). This mixture was heated at refluxed for 3 hours. The reaction solution was filtered and the filter cake was washed with hot water (100 mL×2), the filtrate was adjusted with 2 N HCl to pH=2, extracted with EtOAc (500 mL×3). The combined organic layer was dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated to afford K627-E (2.5 g) as yellow solid.

(96) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 13.70 (br s, 1H), 7.64 (dd, J=8.8, 2.4 Hz, 1H), 7.56 (dd, J=10.8, 2.4 Hz, 1H), 3.91 (s, 3H).

(97) Step 5: Synthesis of Compound K627-F

(98) K627-E (2.5 g, 11.6 mmol) was dissolved in MeOH (30 mL), and 10% Pd/C (0.5 g, 50% water) was added. The mixture was stirred overnight at 25° C. under H.sub.2 atmosphere (50 psi). The mixture was filtered. The filtrate was concentrated by rotary evaporation to obtain K627-F (1.9 g) as white solid.

(99) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 6.14 (dd, J=11.6, 2.4 Hz, 1H), 6.07 (dd, J=11.6, 2.4 Hz, 1H), 3.75 (s, 3H).

(100) Step 6: Synthesis of Compound K627-G

(101) Compound K627-F (1.9 g, 10.3 mmol) was dissolved in Ac.sub.2O (20 mL) and AcOH (60 mL). The mixture was heated to 100° C. and reacted for 6 hours. The mixture was concentrated by rotary evaporation to give a solid. The solid was dispersed in EtOAc (5 mL) and PE (5 mL), stirred for 0.5 hour at 20° C., filtered to obtain K627-G (1.96 g) as yellow solid.

(102) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 7.09 (dd, J=12.0, 2.4 Hz, 1H), 6.90 (dd, J=9.6, 2.4 Hz, 1H), 3.93 (s, 3H), 2.34 (s, 3H).

(103) Step 7: Synthesis of Compound K627

(104) K627-G (250 mg, 1.2 mmol), 3-aminopiperidine-2,6-dione hydrochloride (257 mg, 1.56 mmol), imidazole (245 mg, 3.6 mmol) and triphenyl phosphate (1.12 g, 3.6 mmol) in CH.sub.3CN (20 mL) was heated at reflux overnight under N.sub.2. The mixture was cooled to 25° C. and concentrated to dryness by rotary evaporation. The residue was purified by silica gel chromatography (EtOAc) to afford a crude product. The crude product was further purified by prep-HPLC to afford K627 (168 mg).

(105) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 10.98 (s, 1H), 6.88-6.94 (m, 2H), 5.12-5.18 (m, 1H), 3.85 (s, 3H), 2.77-2.87 (m, 1H), 2.57-2.64 (m, 5H), 2.08-2.15 (m, 1H). LCMS: 320.1 [(M+1)]+.

Example 11 Synthesis of Compound K631

(106) ##STR00052##

(107) Step 1: Synthesis of Compound K631-G

(108) Compound K627-G (1.0 g, 4.78 mmol), tert-butyl 4,5-diamino-5-oxopentanoate (1.26 g, 6.21 mmol), imidazole (0.98 g, 14.34 mmol) and triphenyl phosphate (4.45 g, 14.34 mmol) was dissolved in CH.sub.3CN (100 mL), then the mixture was refluxed overnight under N.sub.2. The mixture was cooled to 25° C. and concentrated to dryness by rotary evaporation. The residue was purified by silica gel chromatography (PE:EtOAc 1:1) to afford K631-G (1.18 g) as off-white solid.

(109) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 6.99-7.36 (m, 2H), 6.83-6.91 (m, 2H), 4.67 (br s, 1H), 3.86 (s, 3H), 2.43-2.45 (m, 3H), 2.07-2.31 (m, 4H), 1.32 (s, 9H).

(110) Step 2: Synthesis of Compound K631

(111) K631-G (600 mg, 1.53 mmol) was dissolved in DCM (10 mL). BBr.sub.3 (1.15 g, 4.6 mmol) was added at 0° C. The mixture was stirred at 50° C. overnight and then poured into ice (10 g). The solvent was removed by rotary evaporation. Water (20 mL) was added to the residue. The mixture was stirred at 25° C. for 3 hours, filtered and the solid was purified by prep-HPLC to afford K631 (80 mg) as off-white solid.

(112) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 11.48 (br s, 1H), 11.17 (br s, 1H), 6.90 (dd, J=10.2, 2.4 Hz, 1H), 6.78 (dd, J=11.1, 2.4 Hz, 1H), 5.33-5.39 (m, 1H), 2.85-2.86 (m, 1H), 2.58-2.80 (m, 5H), 2.19-2.26 (m, 1H). LCMS: 306.1 [(M+1)]+

Example 12 Synthesis of Compound K700

(113) ##STR00053##

(114) Step 1: Synthesis of Compound K700-A

(115) To a solution of K501-F (500 g, 1.754 mmol), tert-butyl 4,5-diamino-5-oxopentanoate (433 mg, 2.631 mmol) in CH.sub.3CN (40 mL) was added imidazole (525 mg, 7.717 mmol), (PhO).sub.3P (1.3 g, 4.209 mmol). The reaction mixture was heated to 85° C. and reacted for 16 hours. When the reaction was completed, the solvent was removed via vacuum. To the residue was added EtOAc (100 mL) and H.sub.2O (50 mL). The organic phase was separated and washed with sat. NaHCO.sub.3 aqueous solution (50 mL), dried and concentrated to afford a crude product. The crude product was purified by column chromatography on silica gel (PE:EtOAc 3:1˜1:1) to afford K700-A (1.29 g) as yellow solid.

(116) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 8.19 (d, J=2.0 Hz, 1H), 8.07 (d, J=2.0 Hz, 1H), 7.47 (br s, 1H), 7.19 (br s, 1H), 4.83 (br s, 1H), 2.56 (s, 3H), 2.27-2.47 (m, 2H), 2.20-2.23 (m, 1H), 2.07-2.09 (m, 1H), 1.23 (s, 9H).

(117) Step 2: Synthesis of Compound K700-B

(118) To a solution of K700-A (1.29 g, 2.76 mmol) in EtOH (180 mL), was added sat. aq. NH.sub.4Cl solution (60 ml). The mixture was heated to 80° C. and Fe powder (1.54 g, 27.6 mmol) was added. The mixture was reacted for 3 hours. Then the reaction solution was filtered to remove Fe powder. EtOH was removed via vacuum. The residue was extracted with EtOAc (150 mL), partitioned, dried, concentrated, and purified by column chromatography on silica gel (PE:EtOAc 1:1˜1:5) to afford the K700-B (994 mg) as yellow solid.

(119) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 7.37-7.42 (m, 1H), 7.22-7.33 (m, 2H), 7.02-7.06 (m, 1H), 6.69-6.73 (m, 2H), 4.70 (br s, 1H), 2.44 (s, 3H), 2.02-2.37 (m, 4H), 1.32 (s, 9H).

(120) Step 3: Synthesis of Compound K700-C

(121) To a solution of K700-B (894 mg, 2.04 mmol) in dioxane (50 mL), was added bis(pinacolato)diboron (1.03 g, 4.07 mmol), CH.sub.3CO.sub.2K (399 mg, 4.07 mmol) and Pd(dppf)Cl.sub.2 (156 mg, 0.20 mmol). The mixture was heated to 100° C. under Ar and reacted for 3 hours. The reaction solution was filtered and concentrated to afford a crude product, which was purified by column chromatography on silica gel DCM:MeOH (20:1) to afford the K700-C (1.26 g).

(122) Step 4: Synthesis of Compound K700-D

(123) To a solution of K700-C (1.45 g, 2.98 mmol) in THF (30 mL), was added NH.sub.4Cl (159 mg, 2.98 mmol) in H.sub.2O (15 mL), and H.sub.2O.sub.2 (22.5 mL) was added dropwise at 25° C. The mixture was stirred overnight. The mixture was washed by aq. Na.sub.2SO.sub.3 solution and extracted with EtOAc (150 mL×3). The combined organic layer was dried, concentrated and purified by prep-HPLC to afford K700-D (437 mg) as yellow solid.

(124) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 9.86 (s, 1H), 7.31-7.36 (m, 1H), 6.97-7.03 (m, 3H), 5.99 (s, 2H), 4.56 (br s, 1H), 2.39 (s, 3H), 2.05-2.27 (m, 4H), 1.34 (s, 9H).

(125) Step 5: Synthesis of Compound K700-E

(126) To a solution of 8N HCl in 1,4-dioxane (20 mL) was added K700-D (300 mg, 0.80 mmol). The mixture was stirred for 2 hours at 40° C., and then concentrated to afford crude product K700-E (307 mg).

(127) Step 6: Synthesis of Compound K700

(128) To a mixture of K700-E (307 mg, 0.96 mmol) in CH.sub.3CN (20 mL) was added CDI (466 mg, 2.88 mmol) at 25° C. The mixture was heated to 85° C. and reacted overnight. To the reaction solution was added H.sub.2O (20 mL). The mixture was heated to 60° C. and reacted for 3 hours, concentrated and purified by prep-HPLC to afford a crude product, which was then stirred and slurried in CH.sub.3CN (5 mL) for 1 hour to afford K700 (119 mg) as yellow solid.

(129) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 10.91 (s, 1H), 9.92 (s, 1H), 6.93 (s, 2H), 5.99-6.01 (m, 2H), 5.04-5.08 (m, 1H), 2.76-2.81 (m, 1H), 2.55-2.61 (m, 2H), 2.48 (s, 3H), 2.009-2.13 (m, 1H). LCMS: 303.0 ([M+1].sup.+).

Example 13 Synthesis of Compound K613

(130) ##STR00054##

(131) Step 1: Synthesis of Compound K613-B

(132) K101-e (3.4 g, 11.32 mmol) was dissolved in 30 mL of DMF at 25° C., and Cs.sub.2CO.sub.3 (9.23 g, 28.31 mmol) was added. The mixture was stirred for 30 min. CH.sub.3I (2.1 mL, 34.0 mmol) was added. The mixture was stirred at 25° C. overnight, diluted with 200 mL of EtOAc, washed successively with water and sat. brine, dried over anhydrous Na.sub.2SO.sub.4, concentrated to dryness to afford K613-B (3.5 g).

(133) .sup.1H NMR (DMSO-d.sub.6, 300 MHz): δ 8.09 (d, J=9.3 Hz, 1H), 7.92 (d, J=9.6 Hz, 1H), 3.80 (s, 3H), 3.10 (s, 3H), 1.28 (s, 9H).

(134) Step 2: Synthesis of K613-C

(135) To a solution of K613-B (3.5 g, 10.66 mmol) in 100 mL of MeOH was added 10% Pd/C (700 mg, 50% water). The mixture was stirred overnight under H.sub.2 atmosphere at 50 psi. Pd/C was removed by filtration, the filtrate was concentrated to dryness to afford product K613-C (3.0 g).

(136) .sup.1H NMR (DMSO-d.sub.6, 300 MHz): δ 6.43-6.48 (m, 1H), 6.38 (s, 2H), 6.29-6.33 (m, 1H), 3.72 (s, 3H), 3.03 (s, 3H), 1.24 (s, 9H).

(137) Step 3: Synthesis of Compound K613-D

(138) To a solution of K613-C (3.0 g, 10 mmol) in 60 mL of MeOH and 20 mL of H.sub.2O was added LiOH.Math.H.sub.2O (2.11 g, 50.2 mmol). The mixture was heated to 70° C. and reacted for 5 hours, then cooled to 25° C., and 50 mL of H.sub.2O was added. The mixture was concentrated to remove MeOH, and then cooled with ice-water, adjusted with 2N HCl to pH=2. A solid precipitation was formed. Filtration was conducted. The solid was washed with cold water and petroleum ether, then dried to afford product K613-D (2.8 g).

(139) .sup.1H NMR (DMSO-d.sub.6, 300 MHz): δ 6.40-6.45 (m, 1H), 6.27 (dd, J=9.6, 2.4 Hz, 1H), 3.03 (s, 3H), 1.26 (s, 9H).

(140) Step 4: Synthesis of Compound K613-E

(141) K613-D (2.8 g, 9.85 mmol) was dissolved in 50 mL of Ac.sub.2O. The mixture was heated to 50° C. and reacted for 5 hours, then cooled to 25° C. and concentrated to dryness to afford crude product K613-E (3.0 g), which was used directly in the next step.

(142) Step 5: Synthesis of Compound K613-F

(143) K613-E (3.0 g, 9.85 mmol) was dissolved in 50 mL of CH.sub.3CN. To the mixture was immediately added 3-aminopiperidine-2,6-dione hydrochloride (2.43 g, 14.78 mmol), triphenyl phosphite (6.72 g, 21.67 mmol) and imidazole (2.01 g, 29.55 mmol). The mixture was refluxed overnight, then cooled to 25° C. and concentrated to dryness. 50 mL of icy water and 30 mL of petroleum ether/EtOAc (1:1) was added. The mixture was stirred for 30 min, filtered, washed successively with icy water and petroleum ether:EtOAc (1:1), and dried to afford product K613-F (2.8 g).

(144) Step 6: Synthesis of Compound K613

(145) K613-F (2.8 g) was dissolved in 50 mL DCM, cooled with ice-water, and 50 mL of TFA was added dropwise. Then the reaction solution was stirred at 25° C. for 2 hours and concentrated to dryness. Then 20 mL icy water was added and the mixture was basified with sat. NaHCO.sub.3. A solid precipitation was formed. Filtration was conducted. The solid was washed with icy water and petroleum ether, dried and purified by prep-HPLC to afford K613 (719 mg) as white solid.

(146) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.01 (s, 1H), 8.47 (s, 1H), 6.38 (dd, J=10.0, 2.4 Hz, 1H), 6.26 (dd, J=12.8, 2.4 Hz, 1H), 5.19 (dd, J=11.2, 6.0 Hz, 1H), 2.80-2.84 (m, 4H), 2.56-2.65 (m, 5H), 2.13-2.19 (m, 1H). LCMS: ([M+1]+)=319.2

Example 14 Synthesis of Compound K617

(147) ##STR00055##

(148) Compound K617 was synthesized by a similar method as compound K613 described in Example 13 except (CH.sub.3).sub.2CHI was used instead of CH.sub.3I in step 1.

(149) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.02 (s, 1H), 8.52 (d, J=6.4 Hz, 1H), 6.35-6.38 (m, 2H), 5.17-5.21 (m, 1H), 3.67-3.72 (m, 1H), 2.79-2.83 (m, 1H), 2.55-2.63 (m, 5H), 2.11-2.17 (m, 1H), 1.18-1.20 (m, 6H). LCMS: [(M+1)]+=347.0

Example 15 Synthesis of Compound K704

(150) ##STR00056##

(151) The starting material K702-D was synthesized by a similar method as compound K700-D described in Example 12 except (S)-tert-butyl 4,5-diamino-5-oxopentanoate hydrochloride was used instead of tert-butyl 4,5-diamino-5-oxopentanoate in step 1.

(152) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 9.85 (s, 1H), 7.35 (br s, 1H), 6.98-6.99 (m, 3H), 5.98 (s, 2H), 4.54 (br s, 1H), 2.10-2.38 (m, 7H), 1.33 (s, 9H).

(153) Step 1: Synthesis of Compound K704-A

(154) To a mixture of K702-D (800 mg, 2.12 mmol) in DMF (15 mL), was added K.sub.2CO.sub.3 (352 mg, 2.55 mmol) and benzyl bromide (436 mg, 2.55 mmol) at 25° C. The mixture was reacted at 25° C. for 16 hours. The reaction solution was quenched with ice-water (100 mL) and then extracted with EtOAc (100 mL), dried, concentrated and purified by column chromatography on C18 to afford K704-A (567 mg).

(155) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 7.32-7.44 (m, 6H), 7.06 (br s, 3H), 6.19-6.22 (m, 2H), 5.14 (s, 2H), 4.60 (br s, 1H), 2.05-2.41 (m, 7H), 1.33 (s, 9H).

(156) Step 2: Synthesis of Compound K704

(157) To a solution of 4.5N/HCl in 1,4-dioxane (20 mL) was added K704-A (567 mg, 1.2 mmol). The mixture was stirred for 4 hours at 25° C. After concentration, 15 mL of CH.sub.3CN was added and then concentrated (repeated for three times) to afford a crude product (570 mg). The crude product was dissolved in MeCN (15 mL). CDI (675 mg, 4.17 mmol) was added. The mixture was heated to 85° C. and reacted overnight. Icy water (100 mL) was added and the mixture was extracted with EtOAc (70 mL×2), dried and concentrated to afford a crude product which was purified by column chromatography on C18 to afford the crude product. The crude product was purified by Prep-HPLC to afford K704 (71 mg).

(158) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 10.96 (s, 1H), 7.35-7.43 (m, 5H), 7.02 (br s, 2H), 6.21-6.24 (m, 2H), 5.04-5.18 (m, 3H), 2.77-2.81 (m, 1H), 2.51-2.62 (m, 5H), 2.08-2.11 (m, 1H). LC-MS: 393.0 ([M+1].sup.+).

Example 16 Synthesis of Compound K706

(159) ##STR00057##

(160) Compound K706 was synthesized by a similar method as compound K704 described in Example 15 except the corresponding substrate was used instead of benzyl bromide in step 1.

(161) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 10.96 (s, 1H), 7.48 (t, J=8.0 Hz, 1H), 7.16-7.20 (m, 2H), 7.02 (s, 2H), 6.27 (d, J=2.4 Hz, 1H), 6.19 (d, J=2.4 Hz, 1H), 5.09-5.13 (m, 3H), 3.56-3.59 (m, 4H), 3.49 (s, 2H), 2.76-2.87 (m, 1H), 2.55-2.61 (m, 2H), 2.51 (s, 3H), 2.36 (s, 4H), 2.08-2.17 (m, 1H). LCMS: 510.0 ([M+1].sup.+).

Example 17 Synthesis of Compound K720

(162) ##STR00058##

(163) Compound K720-A was synthesized by a similar method as compound K700-B described in Example 12 except (S)-tert-butyl 4,5-diamino-5-oxopentanoate hydrochloride was used instead of tert-butyl 4,5-diamino-5-oxopentanoate.

(164) Step 1: Synthesis of Compound K720-B

(165) K720-A (1.5 g, 3.4 mmol), methylboronic acid (1.23 g, 3.4 mmol), Pd(dppf)Cl.sub.2 (0.5 g, 0.68 mmol), K.sub.2CO.sub.3 (0.94 g, 6.8 mmol) was dissolved in the mixture solution of dioxane (20 mL) and water (5 mL). The mixture was heated at 100° C. under N.sub.2 overnight. Water (100 mL) was added. The mixture was extracted with EtOAc (100 mL×2), dried, concentrated and purified by column chromatography on silica gel (PE:EtOAc 1:2) to afford compound K720-B (0.8 g).

(166) .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 6.96-7.39 (m, 4H), 6.40 (s, 1H), 6.36 (s, 1H), 4.61 (br s, 1H), 2.09-2.50 (m, 10H), 1.33 (s, 9H).

(167) Step 2: Synthesis of Compound K720

(168) Compound K720-B (0.8 g, 2.13 mmol) was added to HCl/dioxane solution (4.5 N, 100 mL). The mixture was stirred at 20° C. for 3 hours and concentrated to remove the solvent. The residue was dissolved in CH.sub.3CN (60 mL). CDI (0.69 g, 4.26 mmol) was added to the solution. The reaction solution was heated to 80° C. and stirred overnight. The mixture was concentrated and then purified by Prep-HPLC to afford the product K720 (85 mg) as off-white solid.

(169) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 10.97 (s, 1H), 6.94 (br s, 2H), 6.44 (s, 1H), 6.39 (s, 1H), 5.12 (dd, J=11.2, 6.0 Hz, 1H), 2.78-2.87 (m, 1H), 2.58-2.62 (m, 2H), 2.52 (s, 3H), 2.24 (s, 3H), 2.10-2.16 (m, 1H). LCMS: [(M+1)+]=301.0

Example 18 Synthesis of Compound K722

(170) ##STR00059##

(171) Compound K722 was synthesized by a similar method as compound K720 described in Example 17 except ethylboronic acid was used instead of methylboronic acid in step 1.

(172) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 10.97 (s, 1H), 6.94 (s, 2H), 6.47 (s, 1H), 6.42 (s, 1H), 5.12 (dd, J=11.2, 6.0 Hz, 1H), 2.78-2.82 (m, 1H), 2.51-2.63 (m, 7H), 2.11-2.14 (m, 1H), 1.16 (t, J=7.6 Hz, 3H). LCMS: [(M+1)+]=315.0.

Example 19 Synthesis of Compound K724

(173) ##STR00060##

(174) Step 1: Synthesis of Compound K724-A

(175) To a solvent of K720-A (1.0 g, 2.28 mmol) and CuCN (1.02 g, 11.4 mmol) in DMF (30 mL) was reacted at 140° C. under N.sub.2 overnight. H.sub.2O (200 mL) was added. The mixture was extracted with EtOAc (100 mL×2), washed with sat. brine (100 mL×2), dried, concentrated and purified by column chromatography on silica gel (PE:EtOAc 1:3) to afford the product K724-A (0.24 g) as yellow solid.

(176) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 7.05-7.44 (m, 4H), 6.90 (d, J=1.6 Hz, 1H), 6.83 (d, J=1.6 Hz, 1H), 4.71 (br s, 1H), 2.16-2.48 (m, 7H), 1.32 (s, 9H).

(177) Step 2: Synthesis of Compound K724-B

(178) To a solution of compound K724-A (0.22 g, 0.57 mmol) in DCM (10 mL) was added TFA (10 mL). The mixture was stirred at 20° C. for 4 hours, then concentrated and purified by C18 column (CH.sub.3CN:H.sub.2O 10:90) to afford the product K724-B (0.18 g) as yellow solid.

(179) Step 3: Synthesis of Compound K724

(180) To a solution of K724-B (180 mg, 0.547 mmol) in CH.sub.3CN (40 mL) was added CDI (133 mg, 0.82 mmol). The reaction solution was heated to 80° C. and stirred overnight, then concentrated and purified by prep-HPLC to afford K724 (45 mg).

(181) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 11.05 (s, 1H), 7.42 (s, 2H), 6.93 (d, J=1.6 Hz, 1H), 6.85 (d, J=2.0 Hz, 1H), 5.22 (dd, J=11.2, 5.6 Hz, 1H), 2.79-2.88 (m, 1H), 2.57-2.65 (m, 5H), 2.14-2.20 (m, 1H). LCMS: ([M+1]+)=312.0

Example 20 Synthesis of Compounds K402-K406 and Compounds K502-K506

(182) ##STR00061## ##STR00062##

(183) These compounds were synthesized by a similar method as compound K105 described in Example 2 except the starting compounds

(184) ##STR00063##
were used instead of compound K101-g in step 1, and corresponding substrates were used to replace K105-a. K401-F was synthesized by a similar method as compound K501-F described in Example 7.

Efficacy Example 1: TNF-α Activity Inhibition Assay

(185) Reagents for the Assay

(186) DPBS (10×): Invitrogen, Cat #14190

(187) RPMI 1640: RPMI Medium 1640 (1×), liquid, GIBCO, Cat #22400-105

(188) Heat Inactivated FBS: Invitrogen, Cat #10100147

(189) DMSO: Dimethyl sulphoxide, Sigma, Cat #D8418

(190) LPS: Sigma, Cat #L6529

(191) Pen/Strep (100×): Gibco, Cat #15140

(192) hPBMC: CTL, Cat #CTL-UP1

(193) CTL-Anti-Aggregate Wash 20×: CTL, Cat #CTL-AA-005

(194) Human TNF ELISA Set: BD, Cat #555212

(195) PBMC Recovery and Cell Plating Steps

(196) 1. Cell recovery

(197) 1) Agitation was performed continuously in a 37° C. water bath to rapidly thaw cells.

(198) 2) The cells were gently added to a 15 ml centrifuge tube, to which was then added 10 ml of fresh, prewarmed recovery medium gently and then centrifugation was performed at 1000 rpm for 10 min.

(199) 3) The supernatant medium was discarded and resuspension was performed with 10 ml of fresh, prewarmed RPMI 1640 complete medium.

(200) 2. 96-well plate plating

(201) 1) The total number of cells needed for the experiment was calculated and adjusted to the appropriate cell concentration per ml. 100 ul and 10.sup.5 cells per well.

(202) 2) The cell suspension was diluted with appropriate volume of cell culture medium.

(203) 3) The cell suspension was added to a disposable sterile sample well.

(204) 4) 100 ul of cell suspension was added to each well of a 96-well plate.

(205) 5) The plate was incubated in a 37° C., 5% CO.sub.2 incubator for 2 hours.

(206) Compound Preparation Step

(207) 1. LPS: The 1 mg/mL stock solution was diluted with water, aliquoted, and stored at −80° C. Prior to each test, the working solution of LPS was diluted from the stock solution with serum-free RPMI 1640 medium.

(208) 2. Test compound

(209) 20 mM stock solution was dissolved in DMSO and the compound was checked for solubility, aliquoted, and stored at −80° C.

(210) 8× compound gradient preparation:

(211) A series of compound concentration gradient was diluted with DMSO: 10 mM, 2 mM, 0.4 mM, 80 uM, 16 uM, 3.2 uM, 0.64 uM, 0.128 uM were obtained and then the compounds were diluted 125-fold with serum-free RPMI 1640 medium to the final 8×. The final concentration of DMSO in cell culture was 0.1%.

(212) Compound Processing Experimental Procedures and Collection of Supernatants

(213) 1. Cell Plating: Fresh cells were plated in 96-well cell culture plates according to the procedure above, 100 ul and 10.sup.5 cells per well, and then incubated in a 37° C., 5% CO.sub.2 incubator for 2 hours.

(214) 2. Compound Preparation: Before test, compounds were added to the plates according to the above description. A dose of compound in 8× concentration was prepared with serum-free RPMI 1640 medium and all gradients of solution were added to the compound plate.

(215) 3. Compound addition: 16.7 ul of compound solution in working concentration was added to each well of the cell culture plate. The plate was incubated in a 37° C., 5% CO.sub.2 incubator for 1 hour.

(216) 4. 16.7 ul of 8×LPS per well (final concentration of LPS is EC80, the amount of each PBMC needed to be determined) was added. The plate was incubated for 18 hours in 37° C., 5% CO.sub.2 incubator.

(217) 5. 80 ul of supernatant per well was collected and then subjected to TNF-α ELISA assay. The collected supernatant can be stored at −80° C. The supernatant needed to be diluted in various ratios to ensure that the experimental dose would not exceed the linear range of the TNF-α standard curve, depending on the amount of TNF-α released in different donors. Typically, 20-100 ul of supernatant was diluted to 200 ul and then used for ELISA experiments.

(218) TNF-α ELISA Step

(219) The TNF-α ELISA test procedure were referred to the BD human TNF-α ELISA kit experimental procedure.

(220) Experimental Design

(221) Four compounds per plate. 5-fold dilution was performed, starting from 10 uM, by 8 gradients, and parallel wells were made. A total of 16 compounds were tested.

(222) The TNF-α standard was added to each plate. (1.sup.st well, starting from 500 pg/ml, 2-fold dilution, 7 gradients)

(223) ZPE (0% inhibition) used 15 pg/ml LPS+0.1% DMSO, while HPE (100% inhibition) used only 0.1% DMSO.

(224) The inhibition rate statistics were calculated. The inhibition rate (%)=[1−(Max−Min)/(Test cpd−Min)]*100%. IC50 was used to evaluate the concentration of the test compound (nM) at 50% inhibition. The two experimental results are shown in Table 1 and Table 1-1.

(225) In the present efficacy example and efficacy example 2, the structures corresponding to the codes for the compound of the invention are all as described above. The codes and structures of the reference compounds are summarized in Table N.

(226) TABLE-US-00001 TABLE N Code Structure K001 B001 Reference 1 Reference 2 Reference 3

(227) TABLE-US-00002 TABLE 1 IC.sub.50 value for Compound TNF-α Inhibition (nM) K101 27.77 K001 32.54 B001 192.5

(228) TABLE-US-00003 TABLE 1-1 IC.sub.50 value for Compound TNF-α Inhibition (nM) K001 32.8 B001 99.1 K101 10.2 K102 7.3 K105 6.4 K103 282.7 K106 781.4 K501 9.6 K401 10.2 K633 392.5 Reference 1 2803 K720 137.8 K613 9.4 K704 465.5 K724 5.9 K706 319.2 K617 152.7

Efficacy Example 2: CTG Cell Proliferation Experimental Method

(229) MM.1S cells (myeloma cells) (ATCC, catalog number CRL-2974), DOHH2 cells (mantle cell lymphoma cells) (DSMZ, catalog number ACC-47), NCI-H929 cells (myeloma cells) (ATCC, catalog number CRL-9068), or WSU-DLCL-2 cells (diffuse large B cell lymphoma cells) (DSMZ, catalog number ACC-575), Namalwa.CSN/70 cells (non-Hodgkin's lymphoma cells (DSMZ, catalog number ACC-70) was inoculated as (1.8-15)×10.sup.3 per well in white wall, transparent bottom 96-well plate containing specific media (Corning, catalog number CLS3903), which was cultured in 37° C., 5% CO.sub.2 incubator for 24 hours. Compounds were formulated in DMSO (Sigma, Cat. No. 276855) as 150 mM stock, which was diluted to the required concentration (DMSO final concentration is 0.2%) in culture medium and added to each well, 2 wells/concentration. The plate was incubated in 37° C., 5% CO.sub.2 incubator for 72-120 hours. Afterwards, 100 μl of CellTiter-Glo® cell activity assay reagent (Promega, Cat. No. G7570) was added to each well, which was mixed on a plate shaker for 10 minutes to induce cell lysis. The 96-well plate was allowed to stand at room temperature for 10 minutes to stabilize the luminescence signal. A white base film was pasted on the bottom of the plate and the EnSpire was used to read the plate. Data processing was performed with XLfit software to obtain IC.sub.50 values. The specific experimental data for various batches are shown in Table 2, Table 3, and Table 4.

(230) TABLE-US-00004 TABLE 2 IC.sub.50 value IC.sub.50 value for IC.sub.50 value for IC.sub.50 value for IC.sub.50 value for for MM.1S WSU-DLCL2 DOHH2 NCI-H929 Namalwa. CSN/70 Compound Inhibition (μM) Inhibition (μM) Inhibition (μM) Inhibition (μM) Inhibition (μM) K101 0.0133 0.1793 0.1254 0.0304 0.0068 B001 0.3618 >300 >300 1.0021 >300 K001 0.0498 0.4691 0.4589 0.0861 0.0658

(231) TABLE-US-00005 TABLE 3 IC.sub.50 value for Compound MM.1S Inhibition (μM) K001 0.0375 Reference 1 2.4035 Reference 2 >300 Reference 3 1.170 K627 5.929 K633 0.3144 K635 2.8118 K700 0.5068 K401 0.022 K501 0.0267 K631 0.9565

(232) TABLE-US-00006 TABLE 4 IC.sub.50 value for Compound MM.1S Inhibition (μM) K001 0.275 K102 0.104 K105 0.033 K104 0.192 K106 0.504 K103 0.465