NZ744387B2 - Quinazolinone derivative, preparation method therefor, pharmaceutical composition, and applications - Google Patents

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.

Description

OLINONE DERIVATIVE, PREPARATION METHOD THEREFOR, PHARMACEUTICAL COMPOSITION, AND APPLICATIONS The present application claims priorities of Chinese Patent Application CN201610023840.9 filed on January 14, 2016, the contents of which are incorporated herein by reference in their entireties.

FIELD OF THE INVENTION Provided are a quinazolinone tive, a ation s, a pharmaceutical composition and use thereof.

BACKGROUND OF THE INVENTION Tumor necrosis factor-a ) is a kind of proinflammatory cytokine, which plays an important role in immune homeostasis, inflammation, and host defense. TNF-a has been proved to be one of the major mediators of inflammation. TNF-a can also be produced by tumors. While being capable of promoting the fomiation of tumors, TNF-a can also cause the programmed death of tumor cells. In addition, TNF-a also affects the processes such as apoptosis, necrosis, angiogenesis, immune cell activation, difTerentiation and cell migration, all of which play important roles in genesis and tumor progression.

Uncontrolled activity of TNF-a or overproduction of TNF-a is associated with the pathology of various diseases, including but not limited to cancers, such as, colon, rectum, breast, brain and intestinal cancer; and inflammatory diseases, especially cancer-associated inflammation. The dysrcgulation of TNF-a can also lead to autoimmune es, toxic shock syndrome, cachexia, arthritis, psoriasis, HIV ion and AIDS, nervous system diseases and central nervous system diseases, sepsis, congestive heart failure, transplant rejection and virus infections. Thus, reducing the level of TNF—a, or regulating the activity of TNF—a is a ing strategy in ng many immunological, inflammatory and malignant diseases (e.g., cancers and inflammation). Such as, Sethi et a1. Front. Biosci. (2008) 13, 107 and Results Prob. Cell Differ. (2009) 49, 1-15.

Lenalidomide (3-(4—amino—l,3-dihydro—1~oxo-2H—isoindol—2-yl)-piperidine-2,6-dione) is a small molecule immune regulator. It has been proved that lenalidomide can inhibit the secretion of TNF-a and other proinflammatory cytokines, and increase the secretion of anti-inflammatory nes. Lenalidomide was approved for treating multiple myeloma (in 2006), myelodysplastic syndrome (in 2005) and mantle cell lymphoma (in 2013). In addition, in clinical trials, Lenalidomide alone or in combination with other therapeutic agents, can treat non-Hodgkin's lymphoma, papillary and follicular d carcinoma, chronic lymphocytic ia, chronic myelogenous leukemia, amyloidosis, type I complex regional pain syndrome, malignant ma, radiculopathy, myelofibrosis, glioblastoma, gliosarcoma, malignant glioma, myeloid leukemia, refractory plasma cell tumor, chronic myelomonocytic ia, follicular lymphoma, y body and chronic melanoma, iridic melanoma, recurrent interocular melanoma, extraocular spreading melanoma, solid tumor, T cell lymphoma, erythroid lymphoma, monoblastic and tic leukemia; myeloid leukemia and brain , meningioma, spinal tumor, thyroid cancer, mantle cell lymphoma, non-small cell lung cancer, ovarian cancer, renal cell carcinoma, Burkitt's lymphoma, Hodgkin's lymphoma, large cell lymphoma and lobulinemia (see ). r, Lenalidomide has many side effects. In fact, Lenalidomide’s prescription information clearly recites that the drug has a risk of myelosuppression, deep vein thrombosis, pulmonary embolism and teratogenesis. During the clinical trials, a majority of patients taking Lenalidomide need a reduction of dose due to the hematologic toxicity. Therefore, although Lenalidomide is of useful activity, its effectiveness is limited by the significant occurrence of side effects. Therefore, it is desirable in the field to have Lenalidomide derivatives with improved structures to optimize its performance.

DISCLOSURE OF THE INVENTION The present invention relates to quinazolinone tives, a preparation process, a pharmaceutical composition and use f. The olinone derivatives of the invention can te generation or activity of cytokines such as TNF-α, thereby effectively treating cancer and inflammatory disease.

In a first aspect the present invention es a compound of Formula I, or a pharmaceutically acceptable salt, solvate, co-crystal, stereoisomer or isotopic compound thereof: wherein, X is selected from the group consisting of halogen and cyano; Z is , wherein the carbon marked with * is asymmetric center; R1 is -NR1'R2'; wherein R1' and R2' are each independently selected from the group consisting of H, D, methyl and ethyl; R2, R3, R5, R6, R7, R8, R9, and R10 are each independently H or D; R4 is CH3, CH2D, CHD2 or CD3.

In another aspect the present invention provides a process for preparing the compound of Formula I according to the first aspect of the invention, or the pharmaceutically acceptable salt, solvate, co-crystal, stereoisomer or isotopic compound thereof, which is selected from the group ting of any one of the following: s A comprising the following step: reducing or deprotecting compound A1 to give the nd of Formula I wherein R1a is nitro, azide or ; R1b' and R1b'' are ndently H, D or amino protecting group, provided that R1b' and R1b'' are not aneously H or D; and the definitions of R1, R2, R3, R4, X and Z are as defined in the first aspect of the invention; Process B-1, comprising the following steps: deprotecting compound B3 to give compound B2; and then subjecting compound B2 to amidation to give the compound of formula I; Process B-2, comprising the ing steps: subjecting compound B3 to cyclization reaction to give the compound of formula I: wherein in s B-1 and Process B-2, one of Ra and Rb is and the other is , ,or ; one of Ra' and Rb' is , and the other is ; Ra'' and Rb'' are each independently H or D; and the definitions of R1, R2, R3, R4, R5, R6, R7, R8, R9, X and Z are as defined in the first aspect of the invention; s C-1, comprising the following steps: reacting compound C1 and compound Z-NH2 as shown below to give the compound of Formula I wherein the definitions of R1, R2, R3, R4, X and Z are as defined in the first aspect of the invention.

In another aspect the present invention provides an intermediate compound of Formula A1, A1-1, A1-2, B2, B3, C1, or C1-1: wherein R1a, Ra, Rb, Ra', Rb', R1, R2, R3, R4, R5, R6, R7, R8, R9, X and Z are as defined in any one of claims 1-10.

In another aspect the present invention provides a pharmaceutical composition comprising the compound of formula I according to the first aspect of the invention, or the pharmaceutically acceptable salt, solvate, co-crystal, stereoisomer or isotopic compound thereof, and one or more pharmaceutically acceptable excipients.

In another aspect the present invention provides the use of the compound of Formula I according to the first aspect of the invention, or the ceutically acceptable salt, solvate, cocrystal , stereoisomer, isotopic compound thereof in the manufacture of a regulator for the generation or ty of TNF-α; or use of the combination of the compound of Formula I according to the first aspect of the ion, or the pharmaceutically acceptable salt, solvate, stal, stereoisomer, isotopic compound thereof, and one or more other therapeutic agents selected from the group consisting of ngiogenic agent, immunoregulating agent, immunotherapeutic agent, chemotherapeutic agent or hormone compound, in the manufacture of a tor for the generation or activity of TNF-α.

Another aspect of the invention provides the use of the compound of Formula I according to the first aspect of the invention, or the pharmaceutically acceptable salt, solvate, co-crystal, stereoisomer, isotopic compound thereof in the manufacture of a medicament for treating or preventing a disease, disorder or condition; or use of the combination of the compound of Formula I ing to the first aspect of the invention, or the pharmaceutically able salt, solvate, co-crystal, stereoisomer, isotopic compound f, and one or more other therapeutic agents selected from the group consisting of anti-angiogenic agent, immunoregulating agent, immunotherapeutic agent, chemotherapeutic agent or hormone compound, in the manufacture of a medicament for treating or preventing a disease, disorder or ion; wherein the disease, disorder or condition is selected from the group consist of: TNF-α associated disorders, cancers, diseases and disorders ated with red angiogenesis, pains, macular degeneration syndrome, skin diseases, keratosis, atory system disease, immunodeficiency diseases, central nervous system diseases, mune diseases, atherosclerosis, heredity, allergy, viruses, sleep disorders and associated syndrome, inflammatory diseases, PDE-4 associated diseases or IL-2 associated diseases.

Other aspects of the present ion relate to a compound of Formula I, or a ceutically acceptable salt, solvate, polymorph, co-crystal, stereoisomer, isotopic compound, lite, or prodrug thereof MARKED-UP VERSION wherein, X is selected from the group consisting of halogen, hydroxyl, cyano, substituted or tituted C1-C6 alkyl, and C1-C6 alkoxy substituted with 6-10 membered aryl; wherein 2d (followed by page 3) "6-10 membered aryl" in the "C1-C6 alkoxy substituted with 6-10 membered aryl" is optionally substituted with one or more of the following groups: D, halogen, hydroxyl, cyano, substituted or unsubstituted C1-C6 alkyl, and substituted or unsubstituted C1-C6 alkoxy, wherein when more than one substituents are present, they are identical or different; Z is. R8 Rm wherein the carbon marked w1th * lS asymmetrlc center;. . . .

Rl is selected from the group consisting of hydroxyl, substituted or unsubstituted C1—C6 alkoxy and -NR"R2'; wherein R" and R2. are each independently selected from the group consisting of H, D, substituted or unsubstituted C1-C6 alkyl and —C(O)R3‘; Ry is substituted or unsubstituted C1-C6 alkyl; R2, R3, R5, R6, R7, R8, R9, and R'0 are each independently H or D; R4 is CH3, CHzD, CHDZ or CD3; the "substituted" in the above "substituted or unsubstituted C1—C6 " and "substituted or unsubstituted C1-C6 alkyl" independently represents substitution with one or more of the following groups: D, halogen, amino, hydroxyl, cyano, C1-C6 alkoxy, and 4-10 membered heterocycloalkyl, wherein when more than one substituents are present, they are identical or different.

In an embodiment of the invention, the asymmetric center is preferably S configured carbon atom, enriched S configured carbon atom, R configured carbon atom, enriched R configured carbon atom or racemic carbon atom.

In an embodiment of the invention, the en" in X is ably fluorine, chlorine, bromine or iodine, more preferably fluorine, chlorine or bromine.

In an embodiment of the invention, when the "6-10 membered aryl" is optionally substituted with halogen, the "halogen" is preferably fluorine, chlorine, bromine or iodine, more preferably fluorine, chlorine or e.

In an embodiment of the invention, when the term ituted" in "substituted or unsubstituted C1-C6 alkoxy" and ituted or unsubstituted C1—C6 alkyl" ndently refers to substitution with "halogen", the "halogen" is preferably fluorine, chlorine, bromine or iodine, more preferably fluorine, chlorine or e.

In an ment of the invention, the "Cl-C6 alkyl" in ituted or unsubstituted C1-C6 alkyl" is preferably "Cl-C4 alkyl". The "Cl-C4 alkyl" is preferably methyl, ethyl, isopropyl, n-propyl, n-butyl, isobutyl or tert-butyl, more preferably methyl or ethyl.

In an embodiment of the invention, the "Cl-C6 " in "substituted or unsubstituted C1-C6 alkoxy" is preferably "Cl-C4 alkoxy". The "C1-C4 alkoxy" is preferably methoxy, ethoxy, isopropoxy, oxy, n-butoxy, isobutoxy, tert-butoxy, yloxy or loxy, more preferably methoxy.

In an embodiment of the ion, the "4-10 membered heterocycloalkyl" is preferably "5-6 membered heterocycloalkyl, wherein the heteroatom is one or more selected from the group ting of N, O and S, and wherein the number of heteroatom is 1 or 2" (e.g., pyrrolidinyl, piperidinyl, piperazinyl, or morpholinyl), most preferably [0] .

In an embodiment of the invention, the "C1-C6 alkoxy substituted with 6-10 membered aryl" is preferably a CI-C4 alkoxy substituted with phenyl; wherein the phenyl is optionally tuted with one or more of the following groups: D, halogen, hydroxyl, cyano, and 0-0; alkyl substituted with 4-10 membered heterocycloalkyl (e.g., C1—C4 alkyl substituted with pyrrolidinyl, dinyl, piperazinyl, or morpholinyl); more preferably selected from methoxy substituted with phenyl, wherein the phenyl is optionally substituted with one or more of the following groups: D, halogen, hydroxyl, cyano and C1-C4 alkyl substituted with morpholinyl, wherein when more than one substituents are present, they are identical or different.

In an embodiment of the invention, Z is selected from any of the ing structures: H, H, H , H , H, D, o H o D o p o ,H o H o D o D N D N D N D N D N D N D N ,\ o . 0‘5. o, - o, . o . o . o D I H I o D D I H I D D D o HH, DHH, HHH, HD, DH, DDH,HHD, o D o H o p o, op o H 0 D D N D N D N D N D N H N H N . o \D o . o . o . o . o . o I o D I H I D "\D D ’\H H I H DHH, DD, 000, DHD, DD, H, H o H o H o ,H o [H o H o o H N H N H N H N H N H N H N . - . o . o . o ,\o 0, o, o, o,\ H \H H \H o H D I H :5. H HH, HD, HD, DH, HH, DHH,HH0, o H o H 0 p o p O o H o p H N H N H N H N H N H N H N o, o . o - o . o, o . o D D I H I D I o D 3‘: H D D D DH HD, DDH, HHD DHH, 00,000, H0 0 ,H o ,H 4: mg H N D N - ,\. ° ’\' 0 I H H :Zis prcfcrablx H H H H or H H In an embodiment of the invention, Rl is -NR"R2..

In an embodiment of the ion, R" and R2. are each independently selected from the group consisting of H, D, substituted or unsubstituted C1-C4 alkyl, and -C(O)R3'.

In an embodiment of the invention, Ry is a substituted or unsubstituted C1-C4 alkyl.

In an embodiment of the invention, R3. is selected from the group consisting of methyl, ethyl and isopropyl.

In an ment of the invention, R" and R2 are each independently selected from the group consisting of H, D, methyl, ethyl, isopropyl, acetyl, propionyl and isobutyryl.

In an ment of the invention, X is selected from the group consisting of halogen, hydroxyl, cyano, substituted or unsubstituted C1-C4 alkyl, and methoxy substituted with phenyl; n the phenyl is optionally substituted with one or more of the ing groups: D, halogen, hydroxyl, cyano, and C1-C4 alkyl substituted with morpholinyl, wherein when more than one substituents are t, they are identical or different.

In an embodiment of the invention, X is selected from the group consisting of fluorine, chlorine, bromine, yl, cyano, benzyloxy, 2—fluoro(morpholinyl-l-methyl)benzyloxy, methyl, ethyl, CD3, C2D5 and CH2CD3.

In an embodiment of the invention, X is halogen, Rl is NHz, NHD or NDz; R2, R3, R5, R6, R7, R8, R9 and R'0 are independently H or D; R4 is CH3, CHzD, CHD2 or CD3.

Preferably, the compound of formula I is selected from any of the following structures: H H O N O 0 O N O NH2 0 $61"NHZO NHzo NH; 0 H D FL I AN N N F F x K101 K102 K103N K104 o H CENNNN}3%}: NH 0 0 2 HUG NHZO DU NH20 D, NH20 K113 KIN/N: K105 K116N K113-1 K116 1 K401 K402 O O NH20 NH2 0 giro NH2 0 EU0 NH20 DU Cl,CKNNJL iN)\ K403 K404 K405 K406N o N N \\‘ \\‘ C! N 003 CI Niimj c1 NAC03 K409 K4094 K410N K410 1 n o 0 Mkdb!"CCU Cl N’K A Km K431 1 K432N K432 1 NH2 NLU0:U 0 N o NH2 0 "U NH20 "If NH2 08 :NNJ\ Br’ BrN’)\W/Ei/K K501 K502 K503 K504 Br CD: Br K614 o 0 H NH 0 DU ok "W N/ NA K620 K620-1 o H o o H o 0 U \o o D D N N‘" NJ\/ F N¢K K628 K628—1 H o o OH 0 {121 21 HO D N/J A K704 K701 K70 I -‘I C(\ F N 0000° ~’* w: K,N K706-1 01:00 \,N K707 H H o N o o N o 0 NH 0 N N\" N NA NA NA K720 K720-1 K720 2 N 21 K729 K7294 K722 NC N CD3 NC N CD15 fli (It K726 and K727 Provided is also a process for preparing the nd of formula I, which can be synthesized by known processes using commercially available starting materials. The invention gives particular preference to any one of the following.

Process A comprises the following step: reducing or deprotecting compound A1 to give the compound of Formula I; Rm R1 0 o R2 N’2 ,2 x NAR‘Q X N/ R4 R3 R3 R1b' ‘é‘N—R1bI I ll wherein Rla is nitro, azide or ; Rlb and Rlb are independently H, D or amino protecting group, provided that R'b' and R'b" are not aneously H or D. The definitions of R', R2, R3, R4, X and Z are as defined above.

The amino protecting group may be an amino protecting group commonly used in the art, non-limiting examples being benzyloxycarbonyl (Cbz), utoxycarbonyl (Boc), fluorenylmethyloxycarbonyl (Fmoc), p-methoxybenzyl (PMB), benzyl (Bn), etc.

Process B-l comprises the following steps: deprotecting compound B3 to give compound 82; and then ting compound 82 to amidation to give the compound of formula I; O R8 1 O 9 R8 R O N _.

AR‘R6 R7 0 x NAR‘ 33 I Process B-2 comprises the following steps: subjecting compound B3 toOcyclRization reaction to give the compound of formula I; RR8 RQR R1 O R2 N’Z NRR: R70 x NAR" In Process B-1 and Process B-2, one of Ra1 and Rb is n" and the other is 'é'O'B", —§—osn,or é'OMe; one of Ra. and Rb. is uRbu, and the other is ‘éOH; Ra" and Rb" are each independently H or D. The definitions of R1, R2, R3, R4, R5, R6, R7, R8, R9, X and Z are as defined above.

Process C-l comprises the following steps: reacting compound C1 and compound Z-NHZ as shown below to give the compound of Formula I; R‘ o R1 o R2 R2 ,z i + ~ 1 x N R4 x N R4 wherein, the definitions of R', R2, R3, R4, X and Z are as defined above. ile, in Process C-l, Z group in compound Z-NHz may be replaced with o R3 o R R6 R7 ’7" R6 R7 0 or 0 and/0r RI , group in compound C1 may be replaced with Rla (ie., C1-1 ) and the corresponding reactions are performed to give intermediate compound B3 or B2 or A1. Likewise, in Process A, Z group in compound Al is 05 R" R18 0R R8 R9 R2 Rb o R3 N 0 R3' 6 7 8 9 R R a 9 R6 R7 R3 :3 R6 R7 replaced with 0 (ie., A" ) or 0 (ie., 0 R3.

R1a O R5 R8 R9 R2 Rb‘ N/1R6 0 x R4 A14 ) and the corresponding reaction are performed to give intermediate compound B3 or 82; wherein R", Ra, Rb, Ra', R", R2, R3, R4, R5, R6, R7, R8 and R9, X and Z are as defined above.

The conditions and steps applied in the chemical ons involved in the above processes can be performed by referring to the routine conditions and steps for this type of reaction in the art, and the compound ed by the above process can be r modified on the peripheral positions to obtain other target compounds of the invention.

Provided is also an intermediate compound of Formulae A1, Al-l, A1-2, BZ, BS, Cl, Cl-l: R1a R13 9 R1a X NE: R70 A1-1 A1-2 O Ra. O R8 R13 0 R1 O N N AWR6 R7 MAJ;6 R7 0 o NiRc: NiR‘z R3 R3 82 B3 C1 C1-1 , , , n R'", R", R", R"', R", R', R3, R3, R4, R5, R", R7, R8, R’, x and z are as defined above.

Provided is also a pharmaceutical ition comprising the compound of formula I, or the pharmaceutically acceptable salt, solvate, polymorph, co-crystal, stereoisomer, isotopic compound, metabolite or prodrug thereof, and one or more pharmaceutically acceptable excipients.

The pharmaceutically acceptable excipient may be those widely used in drug manufacture.

Excipients are mainly used to provide a safe, stable and fimctionalized pharmaceutical composition, and can also provide a method which makes the active ingredient dissolve at a d rate or facilitates effective absorption of the active ingredient after being administered to a subject. The excipient can be an inert filler, or one that provides some functions, such as izing the overall pH value of the composition or preventing the degradation of the active ingredient of the composition. The phamiaceutically acceptable excipient may comprise one or more excipients selected from the group consisting of binder, suspending agent, emulsifier, diluent, filler, granulating agent, adhesive, disintegrating agent, lubricant, anti-adhesive agent, glidant, wetting agent, gelling agent, absorption retarder, dissolution tor or rcing agent, adsorbent, buffer, chelating agent, preservative, colorant, ent and sweetening agent.

The pharmaceutical composition of the invention can be ed based on the contents disclosed herein ing to any method known by one skilled in the art. For example, the ceutical composition can be prepared by mixing the compound of fomtula l, or the pharmaceutically acceptable salt, solvate, polymorph, co-crystal, stereoisomer, isotopic compound, metabolite or prodrug thereof, with one or more pharmaceutically acceptable excipients, based on common preparation technology for medicaments. The technologies include but not limited to conventional mixing, dissolving, granulating, emulsifying, levigating, wrapping, ing or freeze-dry s.

The pharmaceutical ition according to the invention may be formulated for administration in any route, including injection (intravenous), mucosal, oral administration (solid and liquid preparation), inhalation, ocular administration, rectal administration, topical or parenteral (infusion, injection, implantation, subcutaneous, vein, artery, intramuscular) administration. The pharmaceutical composition of the invention can also be controlled e or delayed release dosage me'lS. Examples of solid oral preparation include but not limited to powder, capsule, caplet, soft capsule or tablet. Examples of liquid preparation for oral or mucosal administration e but not limited to sion, emulsion, elixir and solution.

Examples of topical preparation include but not limited to emulsion, gel, ointment, cream, patch, paste, foam, lotion, drops or serum ation. Examples of preparation for parenteral administration include but not limited to injection on, dry preparation which can be dissolved or suspended in a pharmaceutically acceptable r, injectable suspension and able emulsion. Examples of other le preparations of the pharmaceutical composition include but not limited to eye drops and other ophthalmic preparations; aerosol, such as nasal spray or inhalation; liquid dosage forms le for parenteral administration; itory and pastille.

The phamiaceutical composition of the invention may further comprise one or more other therapeutic agents. More infomiation on the other eutic agents that may be comprised in the pharmaceutical composition of the invention is disclosed below. The amount and type of the other therapeutic agents depend on the e, disorder or condition to be treated or prevented, the severity of disease, disorder or condition, and the factors of the subject to be administered with the composition, such as age, weight, physical condition, etc; and administration route, etc.

The eutic or prophylactic amount of the compound of formula I, or the pharmaceutically acceptable salt, solvatc, rph, co-crystal, stereoisomer, isotopic compound, metabolite or prodrug thereof, any pharmaceutical composition or preparation thereof ctc., may be administrated to a subject over a period (drug delivery cycle), followed by a period free of the compound (non-drug delivery cycle). The drug delivery cycle and ug delivery cycle can be repeated for required times. The required length and time of the drug delivery cycle and non-drug ry cycle depend on the type and/or severity of the disease, disorder or condition being treated or prevented, and the gender, age, weight of the subject, and other parameters (e.g., the subject's ical, physical and physiological conditions, etc.).

One skilled in the an can sufficiently determine a suitable length and time for the drug delivery cycle and non-drug delivery cycle based on the contents disclosed herein.

In another aspect of the invention, provided is a method for regulating the generation or activity of TNF-a, which ses administering to a subject in need thereof a therapeutically effective amount of the compound of fomiula l, or the pharmaceutically acceptable salt, solvate, polymorph, co-crystal, stereoisomer, isotopic compound, metabolite or prodrug thereof, or the pharmaceutical ition thereof.

In another aspect of the invention, provided is use of the compound of formula I, or the pharmaceutically acceptable salt, solvatc, polymorph, co-crystal, stereoisomer, isotopic compound, metabolite or g thereof in the manufacture of a regulator for generation or activity of TNF—a.

In an embodiment, when the term "regulate" is used to describe the activity or generation of a c molecule, it refers to inhibiting the activity or generation of the molecule. In r ment, when the term "regulate" is used to describe the activity or generation of a specific le, it refers to increasing or enhancing the activity or generation of the molecule. However, in another embodiment, when the term ate" is used to describe the activity or generation of a specific molecule, it refers to decreasing or increasing the activity or generation of the le.

Provided is use of the compound of formula I, or the pharmaceutically acceptable salt, solvatc, stcrcoisomcr, isotopic compound, metabolite or prodrug thereof in the manufacture of a ment for treating or preventing a disease, er or condition. In another aspect, provided is a method for treating or preventing a disease, disorder or condition comprising administering to a subject a therapeutically or prophylactically effective amount of the compound of formula I, or the pharmaceutically acceptable salt, solvatc, stcrcoisomcr, isotopic compound, metabolite and prodrug thereof, or the pharmaceutical composition thereof.

Examples of the disease, disorder or condition to be treated or prevented include but not limited to TNF-a associated disorders, cancers, diseases and disorders associated with undesired angiogenesis, pains, macular degeneration (MD) syndrome, skin diseases, keratosis, respiratory system disease (such as pulmonary diseases), immunodeficiency es, central nervous system (CNS) diseases, mune diseases, atherosclerosis, heredity, allergy, viruses, sleep disorders and associated syndrome, matory diseases, PDE-4 associated diseases or lL-2 associated es. Well-known examples of the disease, disorder or condition in the field include but not d to those described in PCT patent publications W02012015986 and W0200601 8182 and US patent publication U820100204227.

Examples of TNF-a associated disease of the ion include but are not limited to, those diseases or disorders described in WO9803502. Specific examples include but are not limited to inflammation; cancers; endotoxemia or toxic shock me; cachexia; adult atory distress syndrome; bone resorption es such as arthritis; hypercalcemia; versus-host reaction; brain type disease; chronic pulmonary inflammatory disease; reperfusion injury; myocardial infarction; ; circulatory shock; rheumatoid arthritis; Crohn's disease; HIV infection and AIDS; other diseases such as rheumatoid spondylitis, osteoarthritis, psoriatic arthritis, septic shock, sepsis, g disease, ulcerative colitis, multiple sclerosis, systemic lupus erythematosus; asthma; autoimmune diseases; radiation damage; hyperoxic alveoli ; viral infections such as those caused by the herpes virus; viral conjunctivitis or atopic dermatitis.

In a preferred embodiment, the TNF—a ated disease, disorder or condition of the invention is selected from myelodysplastic syndrome, multiple myeloma, mantle cell lymphoma, diffuse large B cell lymphoma, central nervous system lymphoma, non-Hodgkin's lymphoma; papillary and follicular d carcinoma; breast cancer, chronic lymphocytic leukemia, chronic myelogenous leukemia, dosis, type 1 complex regional pain syndrome, malignant melanoma, radiculopathy, myelofibrosis, glioblastoma, gliosarcoma, malignant , refractory plasma cell tumor, chronic myelomonocytic leukemia, follicular lymphoma, ciliary body and chronic melanoma, iridic melanoma, recurrent interocular melanoma, extraocular ion 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, n cancer, renal cell carcinoma, Burkitt’s ma, Hodgkin's lymphoma, large cell lymphoma, ytoma, hepatocellular carcinoma, primary macroglobulinemia (Waldenstrom macroglobulinemia). In an embodiment, the cancer is metastatic. In another embodiment, the cancer is refractory or ineffective with the treatment of chemotherapy or radiation therapy.

The method for treating or preventing a disease, disorder or condition of the ion comprises administering the compound of formula I, or the pharmaceutically acceptable salt, solvate, isomer, isotopic nd, metabolite and prodrug thereof to a subject by any le means, such as ion, mucosa], oral, inhalation, ocular, rectal, long-acting implant, liposome, emulsion or sustained release method.

One skilled in the art understands that the therapeutically effective or prophylactically effective amount of the compound used in the invention may vary with factors for a ic subject, such as age, diet, health, etc., the severity, complication and type of the disease, er or condition to be treated or prevented, and the preparation used etc. Based on the disclosures of the invention, one d in the art can easily ine therapeutically effective or prophylactically effective amount of the compound to be administered to the subject, so as to induce the desired ical or medical response in the subject.

In any method or application described in the invention, the compound of formula I, or the pharmaceutically acceptable salt, solvate, polymorph, co-crystal, stereoisomer, ic nd, metabolite or prodrug thereof, can be used alone or in combination with radiation therapy or radioimmunotherapy and the like, and may be used in further combination with one or more therapeutic agents having pharmaceutical activity (hereinafter referred to as "other therapeutic agent(s)") In an embodiment of the invention, the other therapeutic agent(s) may be a natural, semisynthetic or tic compound. In another embodiment, the other therapeutic agent(s) may be a small molecule, such as a synthetic organic or inorganic molecule, or a larger molecule or biomolecule, such as proteins or nucleic acids with pharmacologically activity. In other embodiment, the other therapeutic s) may be anti-angiogenic agent, immunoregulating agent, immunotherapeutic agent, chemotherapeutic agent or hormone compound.

In an embodiment of the invention, a composition comprising the compound of fomiula I, or the pharmaceutically acceptable salt, solvate, polymorph, co-crystal, stereoisomer, isotopic compound, metabolite or prodrug thereof, and another therapeutic agent is administrated to a subject aneously. In another embodiment, the compound of formula I, or the phamiaceutically acceptable salt, solvate, polymorph, co-crystal, stereoisomer, ic compound, metabolite or prodrug thereof, and the other therapeutic agent are administrated sequentially. In another embodiment, the compound of formula I, or the pharmaceutically acceptable salt, solvate, stereoisomer, ic compound, metabolite or prodrug thereof, and the other therapeutic agent are strated separately. The other therapeutic agent can be administrated before, in succession to, or after the administration of the compound of formula I, or the ceutically acceptable salt, solvate, polymorph, co-crystal, stereoisomer, isotopic compound, metabolite or prodrug thereof.

According to the ion, one or more other therapeutic agents, which can bc administrated in combination with the compound of formula I, or the pharmaceutically acceptable salt, solvate, polymorph, stal, stereoisomer, isotopic compound, metabolite or prodrug thereof, depend on a variety of factors, such as the disease, er or condition to be treated or prevented, etc. One skilled in the art can easily determine suitable other therapeutic agent(s) to be administrated in combination with the compound of formula I, or the pharmaceutically acceptable salt, solvate, rph, stal, isomer, isotopic compound, metabolite or prodrug thereof, based on the contents disclosed herein.

The eutically effective amount of the other therapeutic agent used in the method of the invention is known by one skilled in the art, and administration guidance can be referred to the patents and published ations cited herein, and Wells et al, eds., Pharmacotherapy Handbook, 2nd Edition, Appleton and Lange, Stamford, Conn. (2000); PDR Pharmacopoeia, Tarascon Pocket copoeia 2000, Deluxe Edition, Tarascon Publishing, Loma Linda, Calif. (2000) and other medical literatures cited herein. However, one skilled in the art is capable of detemiining the optimal dose range of the other therapeutic agent.

According to an embodiment of the invention, when being administered in combination with other therapeutic s), the therapeutically ive amount of the compound of formula 1, or the pharmaceutically acceptable salt, solvate, polymorph, co-crystal, stereoisomer, isotopic compound, metabolite or prodrug thereof is less than the required therapeutically ive amount of the compound of formula I, or the ceutically acceptable salt, solvate, polymorph, co-crystal, stereoisomer, isotopic compound, metabolite or prodrug thereof not in combination with other therapeutic agent(s). In another embodiment, the therapeutically effective amount of the other therapeutic agent(s) is less than that when the administration is performed without the compound of formula I, or the pharmaceutically acceptable salt, solvate, polymorph, co-crystal, stereoisomer, isotopic compound, metabolite or prodrug f. By this means, the side effects ated with high dose of any of the drugs can be minimized. Other potential advantages, for example, improving the administration regimen and/or lowering the cost of the drugs, are obvious to one skilled in the an.

According to an ment of the invention, when the compound of formula I, or the phamiaceutically able salt, solvate, polymorph, co-crystal, stereoisomer, isotopic compound, metabolite or prodrug f, and the other therapeutic agent(s) are administered to a subject to treat or prevent a disease, disorder or condition, the compound of formula I, or the phamiaceutically acceptable salt, solvate, polymorph, co-crystal, stereoisomer, isotopic compound, metabolite or prodrug thereof, and the other therapeutic agent(s) can be administered in the same or different routes. The other therapeutic agent(s) can be administered in any ways described herein, including but not limited to, oral, inhalation, injection, ocular, mucosal, rectal, emulsion, liposome, long-acting implant or sustained release . The specific administration route of the other eutic agent(s) depends on the agent itself and the ation, and the e, disorder or condition to be prevented or treated. According to the disclosures herein, one skilled in the art is capable of determining the stration route of the other therapeutic agent(s).

The present application cites or describes a y of publications, articles and patents, the e of citing or describing these references or incorporating these references by their tics or discussing these references is to illustrate the background of the invention rather than admission that the contents of these references contribute to a part of the prior art of the invention.

Unless ise defined, the technical and scientific temls used herein have the same gs as those commonly understood by one skilled in the art. Otherwise, certain terms used herein have the meanings specified in the present description. All the patents, published applications and publications cited herein are incorporated herein by reference, just like elaborating in detail herein. It should be noted that, unless otherwise indicated explicitly in the context, the singular fomi used herein and in the attached claims encompass the plural meaning.

As used herein, when the specific salt, composition, and excipient etc. are referred to as "pharmaceutically acceptable", it means that the salt, composition, or excipient etc. are generally non-toxic, safe, and suitable for administration to a subject, preferably mammalian, more ably human.

The temi "pharmaceutically acceptable salt" used herein refers to a pharmaceutically acceptable c or inorganic salt. Examples of the salt include but are not limited to, sulfate, citrate, e, e, chloride, bromide, iodide, nitrate, hydrosulfate, phosphate, acid phosphate, isonicotinate, lactate, salicylate, acid citrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, suecinate, maleate, gentisinate, fumarate, gluconate, glucuronate, saccharate, fomiate, benzoate, glutamate, methanesulfonate, ethanesulfonate, ulfonate, p—toluenesulfonatc, and embonate (i.e. 1—l-methylene-bis(2-hydroxylnaphthoate)). The compounds of the ion may be used to form ceutically acceptable salts with various amino acids. Suitable alkali salt includes but is not limited to, aluminum salt, calcium salt, lithium salt, magnesium salt, potassium salt, sodium salt, zinc salt, bismuth salt and diethanolamine salt. Review regarding pharmaceutically acceptable salts is referred to Handbook of Pharmaceutical Salts: ties, Selection, and Use (P. ch Stahl and Camille G. Wermuth, ed., Wiley-VCH, 2002).

As used herein, the term olite" refers to an active substance produced by a drug molecule which has gone through chemical structure changes in vivo, the active substance is generally a tive of the aforementioned drug molecule, and also can be chemically modified.

As used herein and unless ise specified, the term "polymorph" refers to one or more kinds of crystal structure formed by different arrangements of molecules in the lattice space when crystallizing.

As used herein, the term "co-crystal" refers to a multi—component system comprising one or more AP] (active ceutical ingredient) molecules and one or more object (or ligand) molecules. In the co-crystal, API molecules and object (or ligand) molecules exist as solids at room temperature when they are used in their pure form alone (in order to guish co-crystal from solvate or e). From this particular definition, salts in which significant or complete proton exchange occurs between APl molecules and guest molecules are excluded. In the co-crystal, API and ligands interact through hydrogen bonds and other possible non-covalent interactions. It is noted that the co-crystal itself may form solvates, including hydrates.

As used herein, the term "solvate" refers to a crystal form of the compound of formula I, or the phamiaceutically acceptable salt, rph, co-crystal, stereoisomer, isotopic compound, metabolite or prodrug thereof, which further has one or more solvent molecules incorporated into the crystal structure. The solvate may include a stoichiometric amount or a non-stoichiometric amount of solvent, and the solvent molecule in the solvent may exist in an ordered or non-ordered arrangement. The solvate containing a non-stoichiometric amount of solvent molecules may be formed by losing at least one solvent molecule (but not all) from the solvate. In a particular embodiment, a solvate refers to a hydrate, which means the crystal of the compound further includes water molecule, and water is used as a solvent.

As used herein and unless otherwise specified, the temi "prodrug" refers to a derivative of the compound comprising a ically reactive fiinctional group, the biological reactive functional group can be cleaved from the compound or react in other ways to give the compound under biological conditions (in vivo or in vitro). Usually, the prodrug is inactive, or at least has lower activity than the compound, which makes the compound exhibit its activity after it is cleaved from the biologically ve nal group. The biologically reactive onal group can be hydrolyzed or oxidized under ical conditions to give the compound. For instance, the prodrug may contain a biologically hydrolysable group. Examples of the biologically hydrolysable group include, but are not limited to, a ically hydrolysable phosphate, a biologically hydrolysable ester, a biologically hydrolysable amide, a biologically hydrolysablc carbonic ester, a biologically hydrolysablc atc and a biologically hydrolysablc ureide. Review regarding the prodrug refers to, such as J. Rautio et al., Nature Reviews Drug Discovery (2008) 7, 255-270 and Prodrugs: Challenges PFIJ Rewards (V. Stella et al. ed., Springer, 2007).

The compound of formula I in the ion, the pharmaceutically acceptable salt, solvate, rph, co-crystal, stereoisomer, ic compound, metabolite or prodrug thereof, can contain one or more asymmetric centers eoisomer"). As used herein, the term "stereoisomer" refers to all stereoisomers ing enantiomer, diastereoisomer, epimer, xo isomer, atropisomer, regioisomer, cis- and trans-isomer. The "stereoisomer" herein also includes "pure stereoisomer" and "enriched stereoisomer" or "racemic isomer" of the various aforementioned stereoisomers. These stereoisomers can be prepared according to an asymmetric synthesis process, or separated, purified and enriched by a chiral separation process (including but not limited to thin layer chromatography, rotating chromatography, column chromatography, gas chromatography, high pressure liquid chromatography, etc.), as well as obtained by chiral separation by means of bonding (chemical binding etc.) or salifying (physical binding etc.) with other chiral compound(s). The term "pure stereoisomer" herein refers to that the mass content of a stereoisomer of the compound is no less than 95% relative to other stereoisomers of the compound. The term "enriched stereoisomer" herein refers to that the mass content of a stereoisomer of the nd is no less than 50% relative to other stereoisomers of the compound. The term "racemic isomer" herein refers to that the mass t of a stereoisomer of the compound is equal to that of another isomer of the compound.

The term pic compound" used herein refers to that there is one or more atomic isotopes with natural or non-natural abundance contained in the nd of formula 1, or the ceutically acceptable salt, solvate, rph, co-crystal, stereoisomer, metabolite or g thereof. Atomic es with non-natural abundance include, but are not limited to, deuterium (3H or D), tritium (hi or T), iodine-125 ("51), phosphorus-32 (32p), carbon-l3 (BC) or carbon-l4 (MC). The aforementioned isotopic compound can also be used as a therapeutic or diagnostic agent (i.e., internal developing agent) or research tool. All the isotopic variants of the compound of the invention, whether or not radioactive, are included in the scope of the invention.

The term "isotope ed" used herein refers to that there is one or more atomic isotopes with non-natural abundance contained in the compound of formula I, or the pharmaceutically acceptable salt, solvate, polymorph, co-crystal, stereoisomer, isotopic compound, metabolite or prodrug thereof. The term "isotope enriched" also refers to that the compound of formula I, or the pharmaceutically acceptable salt, solvate, polymorph, co-crystal, stereoisomer, isotopic nd, metabolite or prodrug compound f, contains at least one isotopic atom with tural abundance.

As used herein, the term nt" or "subject" refers to any animal to be treated or have been treated with the compound or the composition according to an ment of the invention, mammalian is preferable, and human is the most preferable. The term "mammalian" used herein includes any s. Examples of mammal include but are not d to cattle, horse, sheep, pig, cat, dog, mice, rat, rabbit, guinea pig, monkey, human, etc., human is the most preferable. The terms "subject" and nt" are used interchangeably .

In an embodiment, the terms "treat" and "treating" refers to an improvement, tion or reversal of a disease or disorder or at least one of identifiable symptoms thereof, such as treating cancer by ng or stabilizing the symptoms of cancer or a e. In another embodiment, "treat" or "treating" refers to an improvement, prevention or reversal of at least one measurable body parameter of a e or disorder which is being treated, the disease or disorder may not be identified in mammal. However, in another embodiment, the term "treat" or "treating" refers to slow the progress of a disease or disorder, in physical, such as stabilizing identifiable symptoms, or in physiological, such as stabilizing physical parameters, or in both.

In another ment, the term "treat" or "treating" refers to delaying the onset of a disease or disorder.

In some embodiments, the compound is administered for a prevention purpose. As used herein, "prevent" or "preventing" refers to a reduction in a risk of given disease or symptom. In a preferred mode of embodiment, the designated compound is administered to a subject for a prevention purpose, such as the subject with family history or tendency of cancer or autoimmune disease.

As used herein, "therapeutically effective amount" refers to an amount of the compound or the composition that can cause a biological or medical response (which is sought by researchers, veterinarians, physicians, or other clinicians) for a tissue system, an animal or a person, where may include relieving symptoms of the disease or symptom which is being treated. In a preferred embodiment, the therapeutically ive amount is an amount which is enough to effectively treat, improvably treat or prevent cancer, m or disorder associated with undesirable angiogenesis or TNF—a.

The term "prophylactically effective amount" refers to an amount of an active compound or agent (sought by researchers, veterinarians, physicians or other ians), that can inhibit the onset of a disease in a subject. A prophylactically effective amount of a compound refers to an amount of a therapeutic agent used alone or in combination with other active compound, which can e a therapeutic benefit for treating or preventing the disease, er or condition.

Unless otherwise specified, the singular form of the term used herein, "a" or "an", also includes a plural meaning.

Unless otherwise specified, the term "or" or "and" used herein refers to "and/ or".

Unless otherwise specified, the "MN" or "— ,9 in the specific group herein refers to a connection position.

The temi "optional" or "optionally" means the event or circumstance described subsequent thereto may or may not happen. This term encompasses the cases that the event or circumstance may or may not happen. For example, "optional substitution" or "optionally substituted" encompasses the cases that being unsubstituted or substituted.

Deuterium (D or 2H) is a stable non-radioactive isotope of hydrogen, its atomic weight is 2.0144. Hydrogen exists in the form of an isotopic mixture ofH (hydrogen or protium), D (2H or deuterium) and T (3H or tritium) in natural, where the deuterium abundance is 0.0156%.

According to the common technical knowledge in the field, of all the nds whose structures contain natural hydrogen atoms, the en atom actually represents a mixture of H, D and T. Therefore, if a compound contains a deuterium whose nce greater than its natural abundance 0.0156% at any on, these compounds should be considered to be non-natural or deuterium enriched, and thus these compounds are novel ve to its non-enriched analogues.

In the ion, "deuterium enriched" compound refers to a compound of formula I, or the ceutically acceptable salt, solvate, polymorph, co-crystal, stereoisomer, isotopic compound, metabolite or prodrug thereof, where the deuterium abundance is greater than its l abundance at any relevant position. Therefore, in the rium enriched" compound, the ium abundance at any of the relevant positions is likely between more than 0.0156% and 100%. The deuterium ed position is represented by D, whereas the non-deuterium ed position is ented by H. According to the common technical knowledge in the field, the symbol H may be elided at the uterium enriched position. An e of a s for preparing a deuterium enriched compound is replacing the hydrogen with the deuterium, or employing deuterium-enriched starting material to synthesize the compound.

In the invention, the percentage of the deuterium in the enriched deuterium or the deuterium abundance refers to molar percentage.

In the invention, uterium enriched refers to the hydrogen in natural, which is in the form of a mixture of isotopes H (hydrogen or protium), D (2H or deuterium) and T (3H or tritium).

Each preferred conditions aforementioned can be combined in any way without departing from the common knowledge in the art and thereby forming various preferred embodiments of the invention.

The reagents and starting materials used herein are all commercially available.

The positive effects achieved by the invention are that the compound of fomiula I can regulate the generation and/or ty of cytokines (e.g. TNF-a) so as to effectively treat cancer and inflammatory diseases.

Detailed Description of the Embodiments The invention will be further illustrated by the following examples, but it should not be constructed that the ion is limited to the scope of the examples. The mental methods that are not specified in s in the following examples are those according to conventional methods and conditions, or according to the product manuals.

Example 1 Synthesis of compound K101 Synthesis scheme | OVOH NNoz 02N HN03 \ "02 N323 OZNJVNH21)(BOC)ZO.THF 02N\/J\/NH 1 /\ . > _. U — ‘Boc HSO H\/ Ii 2 4 EtOH 2 KMnO) ,. \‘ I ‘ i F F \‘y K101-a K101-b K101-c K101-e o NH2 N020 HCI 1’ NO2 Oo§/NH2 1)4N HCI> /|\/H\ H2N\"‘l\/\ro‘t-Bu 1 u Fe : H 0 \’i\/\/° .> 2)A020 I1 :3 WNW "'3" [I MW F/\/\N/\ FNMA K101—g K101-h 0 NH2 0 NH2 W20 Y NHzfiD Y O NH OOVN /\/"\\ 0\ 4NHCI M OH cm I H 1 m N" w/Mw N/\ O F FMNA 0 FNMA K101-j K101-k K101 Step 1: Synthesis of compound K101-b To a solvent of K101-a (60 g, 38.7 mmol) in con. H2804 (150 mL), a mixture of con.

HN03 (36 mL) and con. H3804 (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 d for another 1 hour and quenched by pouring into crushed ice. Then the water layer was ted with DCM (100 mL x 2). The combined organic solution was dried over anhydrous NagsO4, filtered and concentrated to afford a crude t, which was purified by tography column on silica gel to afford the product K101-b (16 g, 21%).

IH NMR (CDCl3, 300 MHz): 5 7.78 (d, J= 6.9 Hz, 2H), 2.55 (s, 3H).

Step 2: Synthesis of compound K101-c To a solution of K101-b (9.0 g, 45.0 mmol) in EtOH (100 mL), a solution of Nags (16.2 g, 67.5 mmol) in H30 (50 mL) was added dropwise over 30min at 25°C. The mixture was stirred for 4 hours, then concentrated to afford a crude product. The crude product was diluted with Is) H30 (200 mL) and extracted with EtOAc (100 mL x 2), the combined organic layer was dried over ous Na3804, filtered and concentrated, the residue was purified by chromatography column (PE/EtOAc=10/1) to afford the product K101-c (5.0 g, 65 %). lH NMR (DMSO-dé, 300 MHz): 8 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).

Step 3: Synthesis of compound K101-e To a solution of KlOl-c (1.60 g, 9.40 mmol) in THF (150 mL) was added (Boc)20 (2.25, .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 IN/HC1 (100 mL x 2) and dried over anhydrous Na2$04, filtered and concentrated to afford a crude product (1.7 g). The crude product was added to a mixture of pyridine (30 mL) and H30 (15 mL). The mixture was heated to 80°C, then KMn04 (3.2 g, 19.8 mmol) was added in 4 batches over 2 hours (one batch every 30 s). The resulting mixture was stirred overnight.

The reaction solution was d and the cake was washed with hot water. The filtrate was extracted with DCM (150 mL x 3).The combined organic phase was dried over Na3$04, ed and concentrated to afford a crude product. The crude product was purified by chromatography column on silica gel (Et0Ac/PE=l/5) to afford the product KlOl-e (1.0 g, 30% for 2 steps).

'H NMR (CDC13, 300 MHz): 5 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).

Step 4: Synthesis of compound K101-g To a solution of 4N /HCl in 1,4-dioxane (80 mL) was added K101-e (1.0 g, 3.3 mmol).

The e was stirred for 2 hours at 25°C. and concentrated to afford a crude product (800 mg). A mixture of the crude product and Aeg0 (10 mL) was heated to reflux and stirred for 4 hours. The on solution was trated and the residue was stirred with (EtOAc/Et30 = 1/2, 30 mL) for 30 min. The solid impurities were remove by filtration. The filtrate was concentrated to afford the product KlOl-g (670 mg, 91% for 2 steps) 1H NMR (DMSO-d,, 300 MHz): 6 8.14 (dd, J= 8.1, 2.4 Hz, 1H), 7.74 (dd, J= 9.0, 2.4 Hz, 1H ), 2.42 (s, 3H).

Step 5: Synthesis of nd KlOl-h To a mixture of K101-g (500 mg, 2.23 mmol) in MeCN (25 mL) was added (S)-tert-buty1 4,5-diamino-S-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 H30 (150 mL) and extracted with EtOAc (100 mL x 2). The combined c layer was dried over anhydrous NaZSO4, filtered and concentrated. The residue was purified with tography column on silica gel (EtOAc/PE = 1/3) to afford the product K101-h (600 mg, 66%). lH NMR (DMSO-dé, 300 MHz): 6 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,lH), .21 (m, 1H), 4.68-4.92 , 2.54 (s, 3H), 2.05-2.43 (m, 4H), 1.28 (s, 9H).

Step 6: Synthesis of compound K101-j To a solution of K101-h (600 mg, 1.47 mmol) in EtOH (60 mL) was added saturated aq.

NH4C1 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 x 2). The combined organic layer was dried and trated to afford the product K101—j (540 mg, 97%) lH NMR (DMSO-db, 300 MHz): 5 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).

Step 7: Synthesis of compound K101-k To a solution of 4N/HC1 in 1,4-dioxanc (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).

'H NMR (DMSO-d6, 300 MHz): 5 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).

Step 8: Synthesis of compound K101 To a solution of K101-k (400 mg, 1.24 mmol) in MeCN was added CD] (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 t K101 (210 mg, 56%). lH NMR (DMSO-db, 300 MHz): 5 10.99 (s, 1H), 7.32 (br s, 2H), 6.34 (d, J = 10.8 Hz, 2H ), .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+l]+).

Example 2 Synthesis of compound K105 OVNVO 0 NH Na + szlx , »Q J' 61 MM} FMNA Fwy/K D |\ F N’\ (3021-87Evhf/KO \COZt-Bu C0213" K101-g K105-a K105-b K105 K10" Step 1: Synthesis of nd K105-b To a mixture of K101-g (800mg, 3.57 mmol) in CH3CN (30 mL), K105-a (726 mg, 3.57 mmol), imidazole (533 mg, 7.85 mmol) and triphenyl phosphitc (1.33g, 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. NaHC03 solution and brine. The organic layer was dried over NaZSO4, d and concentrated. The e was purified with chromatography column on silica gel (EtOAc/PE = l/ l) to afford the product K105-b (480 mg, yield: 33%).

IH NMR (DMSO—dé, 300 MHz): 6 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).

Step2: Synthesis of compound K105-c To a solution of K105-b (480 mg, 1.17 mmol) in EtOH (60 mL) was added saturated aq.

NH4C1 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 x 3). The combined organic layer was dried, filtered and concentrated. The residue was purified with chromatography column on silica gel (EtOAc /PE=1/ l) to afford the product K105-c (437 mg, yield: 98%) 1H NMR (DMSO-dé, 300 MHz): 5 7.07-7.49 (m, 4H), .35 (m, 2H), 2.45 (s, 3H), 2.07-2.34 (m, 4H), 1.33 (s, 9H).

Step3: Synthesis of compound K105 To a solution of 6N/HC1 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 trated. The residue was dissolved in DMF (3 mL) and DCM (30 mL). The mixture was cooled to -40°C, and SOCig (685 mg, .76 mmol) in DCM (2 mL) was added dropwise. Then the mixture was reacted at °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. Et3N (237 mg, 2.7 mmol) in DCM (1 mL) was added and the mixture was stirred at 0°C for 1 hour. H20 (10 mL) was then added to quench the reaction. The mixture was concentrated under reduced pressure to afford a crude t. The crude product was purified by Prep-HPLC to atTord K105 (68 mg). lH NMR (DMSO'dfi, 400 MHZ): 6 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), .54 (m, 2H), 2.51 (s, 3H), 2.11—2.16 (m, 1H).MS: 306.1 ([M+l]‘).

Example 3 Synthesis of compound K102 H20 HU K102 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.

'H NMR (DMSO-db, 400 MHz): 6 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), .83 (m, 1H), 2.57—2.66 (m, 2H), 2.54 (s, 3H), 2.08—2.17 (m, 1H). MS: 305.] ([M+1]‘).

Example 4 Synthesis of compound K106 o N o ZO BM K106 Compound K106 was synthesized by a similar method as compound K105 described in o NH2 WNW00/1 Example 2 except the corresponding substrate 0 was used instead of K105-a in step 1.

'H NMR (400 MHz, DMSO-d6) 6 11.03 (s, 1H), 7.34 (br s, 2H), 6.36 (s, 1H), 6.33 (s, 1H), .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 sis of nd K103 Lift/LireNH2 0 :1), K103 Compound K103 was synthesized by a similar method as compound K105 described in H2N’V\H/0\1/H Example 2 except the corresponding substrate . 0 was used instead of K105-a in step 1. lH NMR (400 MHz, DMSO-d6) 5 11.00 (s, 1H), 7.32 (br s, 2H), 6.36 (s, 1H), 6.33 (s, 1H), .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 +).

Example 6 Synthesis of nd K104 K104 Compound K104 was synthesized by a similar method as compound K105 described in 0 NH; . Hsztfi/YOJ/ Example 2 except the corresponding substrate I . . 0 was used Instead of K105-a m step 1.

'H NMR (400 MHz, DMSO-d6) 5 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), .16 (m, 1H). LCMS: 306.1 ([M+1]+).

Example 7 Synthesis of compound K501 NR Ofi/OH NHBoc 9‘ NHBOC 1\ 20_ DPPA/t-BuOH KMnOt HM W CF3COOH w W —.Et3N N/1\/\Brpyridine/1120 DCM 02Mr O 2 O2N/\/\Br 2 K501 B K501": K501-D K501-A H H N02 0 A020 N020 (\1’NH2 "v0 NH 0W0 1\ 1 1 HCI 2 ,. |\ CANA0 N02 01 1 Fe/CH3000H 1 11 H O I 0nI/|\/LN/1\BfiNAA/ ' ’NW / / ‘ B NH2 B Imidazaole()3Ph0 BMWl K501-E CH3CN K501F K501-G K501 Step 1: Synthesis of compound K501-B To the solution of compound K501-A (14.0 g, 51.1 mmol) in THF (90 mL) was added LiOH (6.4g, 153 mmol) and H20 (30 mL). The reaction mixture was stirred at 25°C overnight and then concentrated. The remaining liquid was diluted with Eth (60 mL) and water (100 mL). The organic layer was separated. The water layer was adjusted with 2N HCl to pH = 2,extracted with EtOAe (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. lH NMR (400 MHz ,DMSO-db): 6 8.30 (d, J = 2.0 Hz, 1 H), 8.14 (d, J = 2.0 Hz, 1 H), 2.45 (s, 3H) Step 2: Synthesis of compound K501-C To the on 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 Eth (7.5 g, 74.4 mmol). The e 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 d by column chromatography on silica gel PEzEA (10:1) to afford K501-C (15.3 g) as yellow solid.

'H NMR (400 MHz, CDC13) 6 8.34 (s, 1H), 7.64 (d, J= 2.0 Hz, 1H), 6.46 (s, 1H), 2.29 (s, 3H), 1.53 (s, 9H).

Step 3: Synthesis of compound K501-D To a mixture of ne (300 mL) and H30 (150mL) was added K501-C (15.3 g, 46.2 mmol). This mixture was heated to 80°C, KMnO4 (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 Na3SO4 then filtered and concentrated to afford the solid residue. The residue was d by column chromatography on silica gel (PEzEA 30: l~ 5:1) to afford K501-D (9.8 g) as yellow solid. lH NMR (400 MHz, DMSO-dé) 5 9.55 (s, 1H), 8.29 (d, J = 2.0 Hz, 1H), 7.91 (d, J = 2.0 Hz, 1H), 1.47 (s, 9H).

Step 4: Synthesis of nd K501-E To a solution of K501-D in DCM (100 mL) was added CF3COOH at 0°C. The mixture was reacted overnight at 25°C, then concentrated. HCl in 1,4-dioxane (30ml) was added. The mixture was stirred for 20 min at 25°C, then concentrated to afford K501-E (6.7 g).

'H NMR (400 MHz, DMSO—dé) 5 7.19—7.24 (m, 1H), 7.11-7.13 (m, 1H).

Step 5: Synthesis of compound K501-F A solution of K501-E (2.8 g) in A020 (20 mL) and HOAc (60 mL) was heated to reflux and stirred for 3 hours. The reaction solution was trated and the residue was stirred and slurried in EtOAczPE (2:1, 15 mL) for 1 hour, then filtered to afford K501-F (2.3 g) as yellow solid.

'H NMR (400 MHz, DMSO—db) 6 8.36 (d, J= 1.6 Hz, 1H), 8.08 (d, J = 2.0 Hz, 1H), 2.42 (s, 3H).

Step 6: sis of compound K501-G To a mixture of K501-F (500 mg, 1.75 mmol), 3-aminopiperidine-2,6-dione hydrochloride (433 mg, 2.63 mmol) in CH3CN (20mL) was added imidazole (262 mg, 3.86 mmol), (PhO)3P (816 mg,2.63 mmol). The mixture was stirred for 16 hours at 85"C. After the reaction was completed, the solvent was d in vacuum. To the residue was added 9 mL of EtOAc and 9 mL of H30, the mixture was stirred and slurricd for 1 hour and filtered to afford K501-G (382 mg, crude) as gray solid.

Step 7: Synthesis of compound K501 A mixture of K501-G in HOAc (15 mL) was heated to 80°C, and then Fe powder (965mg, 17.3mmol) was added. The mixture was d 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 (CH3CN2DCM 1:1) to give a product which was further purified by prep-HPLC to afford K501 (180 mg).

IH NMR (400 MHz, DMSO-d6): 5 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), .18 (m, 1H).LCMS: 367.0 ‘).

Example 8 Synthesis of compound K401 Compound K401 was synthesized by a similar method as nd K501 described in Example 7 except the corresponding starting material 5-chloromethylnitrobenzoic acid was used instead of compound K501-B.

NH2 9 O§§/N\T90 CIA/\NA K401 '11 NMR (400 MHz dé): 6 11.00 (s, 1H), 7.24 (br s, 2H), 6.76 (d, .1 = 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), 207—2. 16 (m, 1H). LCMS: 321.0 ([M+l]+).

Example 9 Synthesis of compound K633 and K635 ANH1 Oo NVO A020 l /11 N o n 0 NHz? U K633 F N//l\ i i " \/\ o N o K101 WOW 1 "H? I) o o g/ N 1\ l N//l\ K635 Synthesis of compound K633 To a solution of K101 (300 mg, 0.99 mmol) in 10 mL of DMF was added AC3O (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 CH3CN and then purified by prep-HPLC to afford K633 (250 mg). lH NMR (DMSO-dg, 400 MHz): 5 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), .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)]+.

Synthesis of compound K635 Compound K635 was synthesized by a similar method as compound K633 except the corresponding substrate isobutyric anhydride was used d ofA030.

'H NMR (DMSO-do, 400 MHz): 5 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 BOC‘NH . NH2 OH ‘0 1;):2:%4/:6302 , W o o \ ioxane. K2003. Mel ‘ ‘K‘ KMnQ4 / 2 4 2 (\i/ NOH F N02 F N02 FMNOZ DMF /oNaOH/HZON110; K627-A K627-B K627—C K627—D K627-E ‘0 o NVO Pd/C'HZ. A%Q’A°,9H b1\O HCI H [Eli/O NOH 2 I O MeOH FMNHZ FMN,/1\P(0Ph)3ll1m1dazole 1\ FKi: K627-F K627 K6276 Step 1: Synthesis of compound K627-B K627-A (10.0 g, 37.0 mmol) was dissolved in HCl/dioxane (5 M, 100 mL) and stirred at °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. 1H NMR (400 MHz, DMSO-dé) 5 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).

Step 2: Synthesis of nd K627—C To a mixed solvent of cone. H3804 (75 mL) and water (37 mL) was added K627-B (6.5 g) at 0°C. NaNOz (2.86 g, 42 mmol) was added slowly and the on solution was stirred at 0°C for another 2 hours. The mixture was heated to 115°C and H2804 (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 x 2). The organic layer was washed with sat. brine (300 mL), dried over anhydrous N33804, filtered and trated to afford a crude product K627-C (5.4 g). 1H NMR (400 MHz, DMSO) 6 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).

Step 3: sis of compound K627-D K627-C (5.4 g, 31.6 mmol) and K3CO3 (21.8 g, 158 mmol) were dissolved in DMF (100 mL). To the mixture was added CH3] (13.5 g, 94.7 mmol) at 0°C. The mixture was stirred at °C overnight, then concentrated under reduced pressure to remove the solvent. The residue was dissolved with EtOAc (500 mL), washed with water (300 mL x 2) and sat. brine (300 mL), dried and concentrated to afford K627-D (5.23 g) as brown solid.

'H NMR (400 MHz, DMSO-dé) 6 7.39-7.41 (m, 1H), 7.28-7.31 (m, 1H), 3.90 (s, 3H), 2.19 (s, 3H).

Step 4: Synthesis of compound K627-E To a mixed solution of KMnO4 (13.6 g, 86 mmol) and H20 (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 on was filtered and the filter cake was washed with hot water (100 mL x 2), the filtrate was ed with 2 N HCl to pH = 2, extracted with EtOAc (500 mL x 3). The combined organic layer was dried over anhydrous NaZSO4, filtered and concentrated to afford K627-E (2.5 g) as yellow solid.

'I-I NMR (400 MHz, DMSO-db) 6 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).

Step 5: Synthesis of nd K627-F 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 H2 atmosphere (50 psi).

The e was filtered. The filtrate was concentrated by rotary evaporation to obtain K627-F (1.9 g) as white solid. 1H NMR (400 MHz, DMSO-dé) 5 6.14 (dd, J= 11.6, 2.4 Hz, 1H), 6.07 (dd, J= 11.6, 2.4 Hz, 1H), 3.75 (s, 3H).

Step 6: Synthesis of compound K627-G Compound K627-F (1.9 g, 10.3 mmol) was dissolved in AegO (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), d for 0.5 hour at 20°C, filtered to obtain K627-G (1.96 g) as yellow solid. 1H NMR (400 MHz, DMSO'dfi) 6 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).

Step 7: Synthesis of compound K627 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 CH3CN (20 mL) was heated at reflux overnight under N2. 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 d by prep-HPLC to afford K627 (168 mg). 1H NMR (300 MHz, DMSO-db) 5 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 Example 11 Synthesis of compound K631 o NH2 0 O HZNINO do OX ‘0 oo NH2 N’ OH 00 F BBr3/DCM FflLNIHj P(OPh)3,imidazoleF °050 J\ K627-G K631—G K631 Step 1: Synthesis of compound K631-G Compound K627-G (1.0 g, 4.78 mmol), utyl 4,5-diaminooxopentanoate (1.26 g, 6.21 mmol), imidazole (0.98 g, 14.34 mmol) and nyl phosphate (4.45 g, 14.34 mmol) was dissolved in CH3CN (100 mL), then the mixture was d overnight under N2. The mixture was cooled to 25°C and concentrated to dryness by rotary evaporation. The residue was purified by silica gel chromatography (PEzEtOAc 1:1) to afford K631-G (1.18 g) as off-white solid. lH NMR (400 MHz, DMSO-dé) 5 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).

Step 2: Synthesis of compound K631 K631-G (600 mg, 1.53 mmol) was dissolved in DCM (10 mL). BBr3 (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 e. 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.

'H NMR (300 MHz, DMSO'dfi) 6 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 02N O 1 02N\ Q NH2 HN O NH2 \1%ng :N4\io (PhO)3P N12 Fe/NH4CI Z/K/Q sf H2 OoVNt-i2 ‘ \ N’\ mda oleBI z : 11 . 1 Pddf—CICHCOONa N )\ / NA K(O\< L101! BrMNA EO<1A€dengn§ BMNL: klo\<‘ K501-F K700-A K700-B K700-C H202THF H2NQ05’ 2 NvaOfi’NH2 NHQOYNVO OHCI/dioxane M _CQ|_, NW NH4CIH20HWWO l CHacN NL: o HMNéK K700-D K700 K700-E Step 1: Synthesis of compound K700-A To a solution of K501-F (500 g,l.754mmol), tert-butyl 4,5-diaminooxopentanoate (433 mg, 2.631 mmol) in CH3CN (40 mL) was added imidazole (525 17 mmol), (PhO)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 . To the residue was added EtOAc (100 mL) and H20 (50 mL). The organic phase was separated and washed with sat.

NaHC03 aqueous solution (50 mL), dried and concentrated to afford a crude product. The crude product was purified by column chromatography on silica gel (PEzEtOAc 3:l~1:1) to afford K700-A (1.29 g) as yellow solid.

'H NMR (400 MHz, é) 5 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).

Step 2: Synthesis of compound K700-B To a solution of K700-A (1.29 g, 2.76 mmol) in EtOH (180 mL), was added sat. aq.

NH4Cl on (60ml). 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 l:1~l :5) to afford the K700-B (994 mg) as yellow solid. lH NMR (300 MHz, DMSO‘dfi) 6 7.37-7.42 (m, 1H), 7.22-7.33 (m, 2H), .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).

Step 3: Synthesis of compound K700-C 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), CH3CO3K (399 mg, 4.07 mmol) and Pd(dppf)Cl3 (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 t, which was purified by column chromatography on silica gel DCMzMeOH (20:1) to afford the K700-C (1.26 g).

Step 4: Synthesis of compound K700-D To a on of K700-C (1.45 g, 2.98 mmol) in THF (30 mL), was added NH4Cl (159 mg, 2.98 mmol) in H30 (15 mL), and H303 (22.5 mL) was added dropwise at 25°C. The mixture was stirred overnight. The mixture was washed by aq. Na3SO3 solution and extracted with EtOAc (150 mL x 3). The combined organic layer was dried, concentrated and purified by prep-HPLC to afford K700-D (437mg) as yellow solid. ll-l NMR (300 MHz, DMSO-dé) 6 9.86 (s, 1H), 7.31—7.36 (m, 1H), 6.97-7.03 (m, 3H), .99 (s, 2H), 4.56 (br s, 1H), 2.39 (s, 3H), .27 (m, 4H), 1.34 (s, 9H).

Step 5: Synthesis of compound K700-E 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).

Step 6: Synthesis of compound K700 To a mixture of K700-E (307 mg, 0.96 mmol) in CH3CN (20 mL) was added CD] (466 mg, 2.88 mmol) at 25°C. The mixture was heated to 85°C and reacted overnight. To the reaction solution was added H30 (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 d and slurried in CH3CN (5 mL) for 1 hour to afford K700 (119 mg) as yellow solid. 1H NMR (400 MHz, DMSO-a’b) 6 10.91 (s, 1H), 9.92 (s, 1H), 6.93 (s, 2H), 5.99-6.01 (m, 2H), 5.04-5.08 (m, 1H), .81 (m, 1H), .61 (m, 2H), 2.48 (s, 3H), 2.009-2.l3 (m, 1H). LCMS: 303.0 ([M+l]+).

Example 13 Synthesis of compound K613 N02 0 N020 NHZO NH2 FA/LNHOBoc—>\i i ' ‘ Pd/C F/CrLO/Li0H(aq)> mCOOH CH3I 0/ ACZO DMF /' 052003 NBoc /\/\NBDc FMNBoc K101--e K613-B K613—c K613-D 0 H o \ HC|/|\/1\NBoc0:j/\/H\fvoV v H "fsm’fi . \NH (ROY V0N M H N TFA ,! .

FMNA/ P(OPh)3 /\/\ #:V M//'\ imidazole F N F N K613—E K613-F K613 Step 1: Synthesis of compound K6l3-B K101-e (3.4 g, 11.32 mmol) was dissolved in 30 mL of DMF at 25°C, and CszC03 (9.23 g, 28.31 mmol) was added. The mixture was stirred for 30 min. CH3] (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 ous Na3SO4, concentrated to dryness to afford K613-B (3.5 g). lH NMR (DMSO-dé, 300 MHz): 6 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).

Step 2: Synthesis of K613-C 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 H3 atmosphere at 50 psi. Pd/C was removed by filtration, the filtrate was concentrated to dryness to afford product K6l3-C (3.0 g). lH NMR (DMSO-dé, 300 MHz): 5 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).

Step 3: sis of compound K613~D To a solution of K613-C (3.0 g, 10 mmol) in 60 mL of MeOH and 20 mL of H20 was added LiOH.H30 (2.11 g, 50.2 mmol). The e was heated to 70°C and reacted for 5 hours, then cooled to 25°C, and 50 mL of H20 was added. The mixture was trated to remove MeOH, and then cooled with ice-water, ed 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 K6l3-D (2.8 g). lH NMR (DMSO-dé, 300 MHz): 6 6.40-6.45 (m, 1H), 6.27 (dd, J = 9.6, 2.4 Hz, 1H), 3.03 (s, 3H), 1.26 (s, 9H).

Step 4: Synthesis of compound K613-E K613-D (2.8 g, 9.85 mmol) was dissolved in 50 mL of AegO. The mixture was heated to 50°C and d 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.

Step 5: Synthesis of compound K613—F K613-E (3.0 g, 9.85 mmol) was dissolved in 50 mL of CH3CN. To the mixture was immediately added opiperidine-2,6—dione hloride (2.43 g, 14.78 mmol), triphenyl phosphite (6.72 g, 21.67 mmol) and imidazole (2.01 g, 29.55 mmol). The e 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 sively with icy water and petroleum etherzEtOAc (1:1), and dried to afford product K613-F (2.8 g).

Step 6: Synthesis of compound K613 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. NaHCO3. A solid precipitation was formed. Filtration was conducted. The solid was washed with icy water and petroleum ether, dried and ed by prep-HPLC to afford K613 (719 mg) as white solid. lH NMR (400 MHz, DMSO-dé) 5 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 ‘ H "NH QOYNVo FMMA K617 Compound K617 was synthesized by a similar method as compound K613 described in Example 13 except (CH3)3CH1 was used instead of CH3] in step 1.

'H NMR (400 MHz, b) 5 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), .20 (m, 6H). LCMS:[(M+1)]+ = 347.0 Example 15 Synthesis of compound K704 0 NH NHzO V 2 # H I ll 1 <\ >3 NHz? H2 HO/"::/\NLO>< 1) HCI/dioxane NH20 OWNVO K2003DMF1:? ‘1\; OX2) OAoWMN K70?-D K704-A K704 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-diaminooxopentanoate hydrochloride was used instead of tert-butyl 4,5-diamino—5—oxopentanoate in step]. 1H NMR (300 MHz, DMSO'dfi) 6 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).

Step 1: Synthesis of compound K704-A To a mixture of K702-D (800 mg, 2.12 mmol) in DMF (15 mL), was added K3CO3 (352 mg, 2.55 mmol) and benzyl bromide (436 mg, 2.55 mmol) at 25°C. The mixture was d at °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).

'H NMR (400 MHz, 6) 6 7.32-7.44 (m, 6H), 7.06 (br s, 3H), 6.19-6.22 (m, 2H), .14 (s, 2H), 4.60 (br s, 1H), 2.05-2.41 (m, 7H), 1.33 (s, 9H).

Step 2: Synthesis of compound K704 To a on of C1 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 CH3CN 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). CD] (675 mg, 4.17 mmol) was added. The mixture was heated to 85°C and d overnight. Icy water (100 mL) was added and the mixture was extracted with EtOAc (70 mL x 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).

'H NMR (400 MHz, DMSO-db) 6 10.96 (s, 1H), 7.35-7.43 (m, 5H), 7.02 (br s, 2H), .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]+).

Example 16 Synthesis of compound K706 o N 0 NW? Y F NNL aux; K706 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. lH NMR (400 MHz, DMSO‘dfi) 6 10.96 (s, 1H), 7.48 (t, J = 8.0 Hz, 1H), .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]').

Example 17 Synthesis of compound K720 0 NH WHztot Y 2 ,B.

"H2 0 OYNHz 1) HCI/dioxane Pdd fC|.KCO " > ""1200?"ka fir}! W . 1| .

\‘ N‘ii \‘ 2) CN WNW Né\ \io/O _(BP)—22_3" Br dioxane/HZO M t \(O7L MW K720-A O K720-B K720 Compound K720-A was synthesized by a similar method as compound K700-B described in Example 12 except (S)-tert-butyl 4,5-diamino-S-oxopcntanoatc hydrochloride was used instead of tert-butyl 4,5-diaminooxopentanoate.

Step 1: Synthesis of compound K720-B K720-A (1.5 g, 3.4 mmol), methylboronic acid (1.23 g, 3.4 mmol), Pd(dppf)Clg (0.5 g, 0.68 mmol), K3C03 (0.94 g, 6.8 mmol) was dissolved in the mixture solution of e (20 mL) and water (5 mL). The mixture was heated at 100°C under N3 overnight. Water (100 mL) was added. The mixture was ted with EtOAc (100 mL x 2), dried, concentrated and purified by column chromatography on silica gel (PE:EtOAc 1:2 )to afford compound K720-B (0.8 g) . lH NMR (300 MHz, DMSO—do) 5 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).

Step 2: Synthesis of compound K720 nd K720-B (0.8 g, 2.13 mmol) was added to HCl/dioxanc solution (4.5 N, 100 mL). The mixture was stirred at 20°C for 3 hours and concentrated to remove the solvent. The e was ved in CH3CN (60 mL). CDI (0.69 g, 4.26 mmol) was added to the solution.

The reaction on 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. 1H NMR (400 MHz, DMSO-dé) 8 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: +] = 301.0 Example 18 Synthesis of compound K722 NH2 0 OVNVO H \l N/\/ / N//\ K722 Compound K722 was sized by a similar method as compound K720 described in Example 17 except ethylboronic acid was used d of methylboronic acid in stepl. lH NMR (400 MHz, DMSO-d6) 5 10.97 (s, 1H), 6.94 (s, 2H), 6.47 (s, 1H), 6.42 (s, 1H), .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 OVNHZ OVNHZ H NHZ NHZO ‘ NHZO O NH OOWN NH 0 0Y 1 w TFA MNm 2 Y /\2/\ CuCN .» CDI/MeCN > \ N" ‘\ Nmo DMFNC/‘k/LNAH kKO N/K/ Br/k/LNAH NCU AHN 7:014 NC«(l/\A OX c") N 0 K724'A K720-A K724-B K724 Step 1: Synthesis of compound K724-A To a solvent of K720-A (1.0 g, 2.28 mmol) and CuCN (1.02 g,ll.4 mmol) in DMF (30 mL) was reacted at 140°C under N2 overnight. H30 (200 mL) was added. The mixture was extracted with EtOAc (100 mL x 2), washed with sat. brine (100 mL x 2), dried, concentrated and purified by column chromatography on silica gel OAc 1:3) to afford the product K724-A (0.24 g) as yellow solid. 1H NMR (400 MHz, DMSO-dé) 5 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).

Step 2: Synthesis of compound K724-B To a on of nd 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 (CH3CNzH20 10:90) to afford the product K724-B (0.18 g) as yellow solid.

Step 3: Synthesis of compound K724 To a solution of K724-B (180 mg, 0.547 mmol) in CH3CN (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). lH NMR (400 MHz, DMSO-db) 8 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 sis of compounds K402- K406 and compounds K502-K506 NHzONHU NHZONHI} 1}: Wzoijo K402 K40: K404 K405 C/ngE/w: NH20 HU o N o NH2 0 H, NHZO K406 K502 K503 K504 o N o f:r NH2 0 ON 8 Br’ :1}N)\ K505 K506 These nds were synthesized by a similar method as compound K105 described in T02 0 N02 0 ll o ’1 i 0 Br N CI N Example 2 except the starting compounds K501": or K4014: were used instead of nd 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—a activity inhibition assay Reagents for the Assay DPBS (10X): lnvitrogen, Cat # 14190 RPM] 1640: RPM] Medium 1640 (1X), liquid, GIBCO, Cat # 22400-105 Heat lnactivated FBS: lnvitrogen, Cat # 10100147 DMSO: Dimethyl sulphoxidc, Sigma, Cat # D8418 LPS: Sigma, Cat # L6529 Pen/Strep (lOOX): Gibco, Cat # 15140 hPBMC: CTL, Cat # CTL-UPl CTL-Anti-Aggregate Wash 20X: CTL, Cat # CTL-AA-005 Human TNF ELISA Set: BD, Cat # 555212 PBMC ry and Cell Plating Steps 1. Cell recovery 1) Agitation was performed continuously in a 37 °C water bath to rapidly thaw cells. 2) The cells were gently added to a 15 m1 centrifuge tube, to which was then added 10 ml of fresh, prewamied recovery medium gently and then fugation was performed at 1000 rpm for 10 min. 3) The atant medium was ded and resuspension was performed with 10 m1 of fresh, prewarmed RPM] 1640 complete medium. 2. 96-well plate plating l) The total number of cells needed for the experiment was calculated and adjusted to the appropriate cell concentration per m1. 100 L11 and 105 cells per well. 2) The cell suspension was d with appropriate volume of cell culture medium. 3) The cell suspension was added to a disposable e sample well. 4) 100 ul of cell suspension was added to each well of a 96-well plate.

) The plate was incubated in a 37 °C, 5% C03 incubator for 2 hours.

Compound Preparation Step 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 RPM] 1640 medium. 2. Test compound mM stock solution was ved in DMSO and the nd was checked for solubility, aliquoted, and stored at -80°C. 8X compound gradient preparation: 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 RPM] 1640 medium to the final 8X. The final concentration of DMSO in cell culture was 0.1%.

Compound Processing Experimental Procedures and Collection of Supernatants 1. Cell Plating: Fresh cells were plated in 96-well cell culture plates according to the procedure above, 100 ul and 105 cells per well, and then incubated in a 37 °C, 5% C03 incubator for 2 hours. 2. Compound Preparation: Before test, compounds were added to the plates according to the above description. A dose of compound in 8X concentration was prepared with serum-free RPM] 1640 medium and all gradients of on were added to the compound plate. 3. Compound on: 16.7 ul of compound solution in working tration was added to each well of the cell culture plate. The plate was incubated in a 37 °C, 5% C03 incubator for 1 hour. 4. 16.7 ul of 8X 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% C03 incubator. . 80 ul of supernatant per well was collected and then subjected to TNF-a 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-a standard curve, depending on the amount of TNF-a released in different .

Typically, 20-100ul of supernatant was diluted to 200ul and then used for ELISA experiments.

TNF-a ELISA Step The TNF-a ELISA test ure were referred to the BD human TNF-a ELISA kit experimental procedure.

Experimental design Four nds per plate. 5-fold dilution was med, starting from 10 uM, by 8 gradients, and parallel wells were made. A total of 16 compounds were tested.

The TNF-a standard was added to each plate. (15‘ well, starting from 500 pg/ml, 2-fold dilution, 7 nts) ZPE (0% inhibition) used 15 pg/ml LPS + 0.1% DMSO, while HPE (100% inhibition) used only 0.1% DMSO.

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 l and Table 1-1.

In the present efficacy example and efficacy example 2, the ures 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.

Table N Table l IC50 value for TNF-a tion (nM) Table 1-1 nd ICso value for TNF-u Inhibition (nM) K001 32.8 300] K10] 10.2 K501 cy Example 2: CTG Cell Proliferation mental Method MM.lS cells (myeloma cells) (ATCC, catalog number CRL-2974), DOHHZ cells (mantle cell lymphoma cells) (DSMZ, g 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) x 103 per well in white wall, transparent bottom 96—well plate containing specific media (Corning, catalog number CLS3903 ), which was ed in 37°C, 5% C02 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% C02 incubator for 72-120 hours. Afterwards, 100 pl 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 l 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 sofiware to obtain ICso values. The specific experimental data for various batches are shown in Table 2, Table 3, and Table 4.

Table 2 leo value ICso value leo value ICso value leo value for for for for for Namalwa.

MM. 1 S CLZ DOHHZ NCI-H929 CSN/70 Compound Inhibition Inhibition Inhibition Inhibition Inhibition (uM) (11M) (11M) (11M) (11M) K101 0.0133 0.1793 0.1254 0.0304 0.0068 BOO] 0.3618 1.0021 >300 K001 0.0498 0.4691 0.4589 0.0861 0.0658 Table 3 Compound leo value for MM. 1 S Inhibition (uM) K001 0.0375 Reference 1 2.4035 nce 2 >300 Reference 3 1.170 K627 5.929 K633 0.3144 K635 2.81 18 K700 0.5068 K401 0.022 K501 0.0267 K631 0.9565 Table 4 K104 0.192 K106 0.504 K103 0.465

Claims (15)

1. A compound of Formula I, or a pharmaceutically acceptable salt, solvate, co-crystal, stereoisomer or isotopic compound thereof: wherein, X is selected from the group consisting of halogen and cyano; Z is , wherein the carbon marked with * is asymmetric center; R1 is -NR1'R2'; wherein R1' and R2' are each independently selected from the group ting of H, D, methyl and ethyl; R2, R3, R5, R6, R7, R8, R9, and R10 are each independently H or D; R4 is CH3, CH2D, CHD2 or CD3.

2. The compound of Formula I according to claim 1, or the pharmaceutically acceptable salt, solvate, stal, stereoisomer or isotopic compound thereof, wherein the "halogen" in X is ne, chlorine, bromine or .

3. The compound of Formula I ing to claim 1, or the pharmaceutically acceptable salt, solvate, co-crystal, stereoisomer or isotopic compound thereof, wherein Z is selected from the group consisting of any of the following structures:

4. The compound of Formula I according to claim 1, or the pharmaceutically acceptable salt, solvate, co-crystal, stereoisomer or isotopic compound thereof, n Z is .

5. The compound of Formula I according to claim 1, or the pharmaceutically able salt, solvate, co-crystal, stereoisomer or isotopic compound thereof, wherein X is halogen.

6. The compound of Formula I according to claim 5, or the pharmaceutically acceptable salt, solvate, co-crystal, stereoisomer or isotopic compound thereof, n X is selected from the group ting of fluorine, chlorine and bromine.

7. The compound of Formula I according to claim 1, or the pharmaceutically acceptable salt, solvate, co-crystal, stereoisomer or isotopic compound thereof, wherein X is halogen, R1 is NH2, NHD or ND2; R2, R3, R5, R6, R7, R8, R9 and R10 are independently H or D; R4 is CH3, CH2D, CHD2 or CD3.

8. The compound of Formula I according to claim 1, or the pharmaceutically acceptable salt, e, stal, stereoisomer or isotopic compound thereof, wherein the carbon marked with * is asymmetric center, and the asymmetric center is S configured carbon atom or racemic carbon atom.

9. The compound of Formula I according to claim 1, or the pharmaceutically acceptable salt, solvate, co-crystal, stereoisomer or isotopic compound thereof, wherein the compound of formula I is ed from any of the following structures: K5091 50 50 and .

10. A process for preparing the compound of Formula I ing to any one of claims 1-9, or the pharmaceutically acceptable salt, solvate, co-crystal, stereoisomer or isotopic compound thereof, which is selected from the group consisting of any one of the following: Process A comprising the following step: reducing or deprotecting compound A1 to give the nd of Formula I n R1a is nitro, azide or ; R1b' and R1b'' are independently H, D or amino protecting group, provided that R1b' and R1b'' are not simultaneously H or D; and the definitions of R1, R2, R3, R4, X and Z are as defined in any one of claims 1-9; Process B-1, comprising the following steps: deprotecting nd B3 to give compound B2; and then subjecting compound B2 to amidation to give the compound of formula I; Process B-2, comprising the following steps: subjecting compound B3 to cyclization reaction to give the compound of formula I: wherein in Process B-1 and Process B-2, one of Ra and Rb is and the other is , ,or ; one of Ra' and Rb' is , and the other is ; Ra'' and Rb'' are each independently H or D; and the definitions of R1, R2, R3, R4, R5, R6, R7, R8, R9, X and Z are as d in any one of claims 1-9; Process C-1, comprising the following steps: reacting compound C1 and compound Z-NH2 as shown below to give the compound of Formula I wherein the definitions of R1, R2, R3, R4, X and Z are as defined in any one of claims 1-9.

11. An intermediate nd of Formula A1, A1-1, A1-2, B2, B3, C1, or C1-1: wherein R1a, Ra, Rb, Ra', Rb', R1, R2, R3, R4, R5, R6, R7, R8, R9, X and Z are as defined in any one of claims 1-10.

12. A pharmaceutical composition sing the compound of formula I according to any one of claims 1-9, or the pharmaceutically acceptable salt, solvate, co-crystal, stereoisomer or isotopic compound thereof, and one or more pharmaceutically acceptable excipients.

13. Use of the compound of Formula I according to any one of claims 1-9, or the pharmaceutically acceptable salt, solvate, co-crystal, stereoisomer, isotopic nd thereof in the manufacture of a tor for the generation or activity of TNF-α; or use of the combination of the nd of Formula I according to any one of claims 1-9, or the pharmaceutically acceptable salt, solvate, co-crystal, stereoisomer, isotopic nd f, and one or more other therapeutic agents selected from the group consisting of antiangiogenic agent, immunoregulating agent, immunotherapeutic agent, chemotherapeutic agent or hormone compound, in the manufacture of a regulator for the generation or activity of TNF-α.

14. Use of the compound of Formula I according to any one of claims 1-9, or the pharmaceutically acceptable salt, solvate, co-crystal, stereoisomer, isotopic compound thereof in the cture of a medicament for treating or preventing a disease, disorder or condition; or use of the combination of the compound of Formula I according to any one of claims 1-9, or the pharmaceutically acceptable salt, solvate, co-crystal, stereoisomer, isotopic compound thereof, and one or more other eutic agents selected from the group ting of antiangiogenic agent, immunoregulating agent, immunotherapeutic agent, chemotherapeutic agent or hormone compound, in the cture of a medicament for treating or preventing a disease, disorder or condition; wherein the disease, disorder or condition is selected from the group consist of: TNF-α associated disorders, cancers, diseases and ers associated with undesired angiogenesis, pains, macular degeneration syndrome, skin diseases, sis, respiratory system disease, deficiency diseases, central nervous system es, autoimmune diseases, atherosclerosis, heredity, allergy, viruses, sleep disorders and associated syndrome, matory es, PDE-4 associated es or IL-2 associated diseases.

15. Use of claim 14, wherein the disease, disorder or condition is 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; ary and follicular thyroid carcinoma; breast cancer, chronic lymphocytic leukemia, c myelogenous leukemia, dosis, 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, or primary macroglobulinemia.